24203874 ( Fig. 3). The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) is shown next to the branches. 25 Overall average mean distance is 0.524. There were a total of 667 positions in the final dataset. Phylogenetic trees created by maximum parsimony and maximum-likelihood and UPGMA methods Alectinib research buy ( Fig. 4, Fig. 5 and Fig. 6) resulted in similar topologies of the strain to the tree

obtained by neighbour-joining method. In order to understand the significance in predicting the stability of chemical or biological molecules or entities of B. agaradhaerens strain nandiniphanse5; RNA secondary structure prediction has been performed. The 16S RNA gene sequence obtained was used to deduce the secondary structure of RNA using GeneBee ( Fig. 7A) and UNAFOLD ( Fig. 7C). The secondary structure showed helical regions which bind with proteins S1–S27, hairpin loops, bulge loops, interior loops and multi-branched loops that

may bind to 23S rRNA in the larger subunit of the ribosome. The free energy of the secondary structure of rRNA was −171.7 kcal/mol elucidated VX809 using GeneBee ( Fig. 7B). UNAFOLD results were obtained from .ct file and .reg file. Folding bases 1 to 770 of B. agaradhaerens strain nandiniphanse5 at 37 °C shows the Gibb’s free energy, ΔG = −265.13 kcal/mol. The thermodynamics result from the each base wise of the Bay 11-7085 dataset shows the average of External closing pair

Helix ΔG – 5.70, Stack ΔG – 3.40, Multi-loop ΔG – 2.50, Bulge loop ΔG – 1.70, Hairpin loop ΔG – 0.80, Closing pair and Interior loop of ΔG – 3.20 kcal/mol respectively. All rRNAs appear to be identical in function, because all are involved in the production of proteins. The overall three-dimensional rRNA structure that corresponds to this function shows only minor-but in highly significant-variation. However, within this nearly constant overall structure, molecular sequences in most regions of the molecule are continually evolving and undergoing change at the level of its primary structure while maintaining homologous secondary and tertiary structure, which never alters molecular function. The described results of phylogenetic distinctiveness and phenotypic disparities indicate that strain 2b represents a novel strain within B. agaradhaerens species, for which the name B. agaradhaerens strain nandiniphanse5 is proposed. All authors have none to declare. We extend our sincere thanks to Dr. Yogesh Shouche of National Center for Cell Sciences (NCCS), Pune, India; for performing 16S rRNA gene sequencing of our culture. Special thanks to Mr. Amit Yadav (NCCS) for his efforts. “
“Transdermal systems (TDS) are aimed to achieve the objective of delivering systemic medication through topical application to the intact skin surface.

2 These are analogous to primary colors, namely red, blue, and yellow, which are observed in case of vision. A drug substance is described by organoleptic properties, in terms of taste, color, and odor. These are important for pharmaceutical formulations, though these have applications in the areas of foods, beverages, pharmaceuticals, etc.3 The mechanisms leading to the sensation of taste are very complex and little is understood. Taste buds are responsible for sensing the taste.2 The up- and down-movements of the taste stimulant in the taste

bud may be termed as oscillation. There is STI571 price a need for evaluating the taste objectively. Electronic tongue has been proposed to handle the analysis. 4, 5 and 6 The electronic tongue utilizes the specially designed non-specific potentiometric chemical sensors

with enhanced PLX3397 cell line cross-sensitivity to as many components in solution as possible. Such analysis has practical applications, though lacked the support of principles of physical sciences. Any modeling based on the understanding and knowledge of physical and chemical principles would be ideal. 1 Yoshikawa et al recognized the non-linear dynamic character of the salt-water oscillations and were able to demonstrate that this is a simple system. 7 and 8 The rhythmic oscillations of water flow (up- and down-flows) were generated, when a sodium chloride solution filled in a capillary and was partially submerged in a beaker containing pure water. The hydrodynamic oscillations were considered analogs to the oscillations of taste generator potentials. The objective of the present write-up is to establish the evidence of instrument output of hydrodynamic oscillations.

Furthermore, each phase of an oscillation is enlarged for identifying MycoClean Mycoplasma Removal Kit the characteristic signals. These objectives are achieved using sour taste stimulants realizing the modeling of the sour taste in vitro. The experimental setup is the same as reported earlier, but improvements are made in terms of data acquisition card (DAQ) of NI-9234 as against the earlier DAQ card of NI-PCI 6024. 9 LabVIEW (version 8.6) was used for developing of software afresh independently, as against the earlier report of LabVIEW (version 5.1) and G programming. The present tools permit the analysis of oscillations even for a fraction of a second. The sour taste stimulants chosen are citric acid, hydrochloric acid, tartaric acid and lactic acid. These acids support the general understanding of sour taste as well as density oscillations. Citric acid, hydrochloric acid, lactic acid, and tartaric acid were AR grade (SD Fine Chem, Mumbai, India). The data acquisition card (DAQ, National Instruments, USA) No. NI-9234, Hi-speed USB carrier, NI USB-9162 (high speed processor), and LabVIEW (National Instruments, USA) version 8.6 were used. The Faraday cage was fabricated locally with aluminum.

After 30 min, the absorbance was measured

at 765 nm, and the results were expressed as mg/L catechin equivalent. High-performance liquid chromatography (HPLC) analysis was used to quantify the presence of individual phenolic compounds. Prior to selleck chemical the HPLC analysis, 1.5 mL of each sample was filtered through a cellulose membrane (diameter 0.2 μm). The equipment used in the analysis consisted of an LC-DAD Series 1100 liquid chromatographic system (Hewlett–Packard, Palo Alto, CA) with a diode array detector system. The chromatographic analyzes were a modification of the methods described by Lamuela-Raventós and Waterhouse (1994). A Zorbax SB C18 (250 × 4.6 mm), 5 m particle size, with a flow of 0.5 mL/min, was used for the stationary phase. After filtration on a 0.2 m Millipore membrane, five microliters of grape juice was injected into the HPLC system. The solvents used for the separation were as follows: solvent A (50 mM dihydrogen

ammonium phosphate adjusted to pH 2.6 with orthophosphoric acid), solvent B (20% of solvent A with 80% acetonitrile) and solvent C (0.2 M orthophosphoric acid adjusted with ammonia to pH 1.5). The gradient conditions were as follows: solvent A 100% (0–5 min), solvents A 96% and B 4% (5–15 min), solvents A 92% and B 8% (15–25 min), solvents B 8% and C 92% (25–45 min), solvents B 30% and C 70% (45–50 min), solvents B 40% and C 60% (50–55 min), solvents B 80% and C 20% (55–60 min) and solvent A 100% (60–65 min). Chromatograms were monitored at 204 nm, and identification was based on the retention time relative to authentic standards ((+)-catechin, (−)-epicatechin, HKI-272 price procyanidin B1, B2 and gallic acid). Quantification was performed PAK6 using the standards by establishing

calibration curves for each identified compound. Results are shown in mg/L. To determine cyanidin-3-glucoside, delphinidin-3-glucoside, peonidin-3-glucoside, malvidin-3-diglucoside and malvidin-3-glucoside, we used a mobile phase with solvents A (ultrapure water, formic acid, and acetonitrile) and B (ultrapure water, formic acid, and acetonitrile) in a constant flow of 0.8 mL/minute with a controlled temperature of 40 °C. The gradient conditions were as follows: solvents A 94% and B 6% (0 min), solvents A 70% and B 30% (0–15 min), solvents A 50% and B 50% (15–30 min), solvents A 40% and B 60% (30–35 min), solvents A 94% and B 6% (35–41 min). The peak was detected at 518 nm, and the amount of sample injected was 50 μL (OENO, 2003). To quantify the resveratrol compound, we used a mobile phase of ultrapure water and acetonitrile (75:25 vol/vol) (pH 3.0) with a constant flow of 1.0 mL/min for 20 min with a controlled temperature of 25 °C. The gradient conditions were as follows: solvents A 10% and B 90% (0 min), solvents A 85% and B 15% (0–23 min), solvents A 95% and B 5% (23–30 min), solvents A 10% and B 90% (30–35 min). The peak was detected at 385 nm, and the amount of sample injected was 20 μL (McMurtrey et al., 1994).

Studying the impact of social status on health

in a laboratory environment affords tighter controls over confounding factors such as status differences in physical environments, food quality and accessibility, ethnicity, and health care allowing for a focused evaluation of the biological impact of social status differentials. In the wild, cynomolgus monkeys (Macaca fascicularis) live in groups comprised of one or more adult males, multiple adult females, and their dependent offspring. Males are usually not related and emigrate between groups one to several times during their lifetime. Adult females are related through one or more matrilines and typically remain in their natal group for life. Female offspring have the same social status as their mother; maternal social status determines number of pregnancies, infant survival, and lifetime selleckchem reproductive success ( v. S. and van Noordwijk, 1999). Thus, this species C59 wnt experiences suppression of reproductive function

by social status relationships. We have studied the effects of social status on the health of adult female cynomolgus monkeys (M. fascicularis) in the laboratory for nearly 30 years. These monkeys were wild-caught as adults, and in recent years came from a purpose-bred free-ranging colony in Indonesia. The monkeys were housed in small social groups of 3–5 females in rooms approximately 8–10 m3 and enriched with perches, barrels, and manipulanda such as mirrors and toys. The monkeys were fed a diet containing moderate amounts of fat and cholesterol to mimic key dietary constituents consumed in Western societies. When placed in these groups, the monkeys quickly organize themselves into linear social status hierarchies which are usually stable over time ( Shively and Kaplan, 1991). Social status is evaluated by recording the outcomes of agonistic interactions. The animal to which all others in the

group direct submissive behaviors is considered dominant. The monkey that all but the most dominant submits to is considered second-ranking, and so on. Compared to dominant females subordinates receive more aggression else (Fig. 2A), are groomed less (Fig. 2B), and spend more time alone out of arm’s reach of another monkey (Fig. 2C). Thus, subordinates appear to be subject to more hostility and have less social support than their dominant counterparts. Vigilant scanning (Fig. 2D) of the social environment, a behavior which consists of head swiveling to visually scan the home pen while in a crouched posture, is also a characteristic of subordinate female cynomolgus monkeys in these small groups. These monkeys appear fearful and anxious when engaged in vigilant scanning, as it is often accompanied by lip smacking and grimacing (fear and appeasement behaviors in macaques) (Shively et al., Apr 15 1997) (Shively, Nov 1 1998). We have used telemetered heart rate as an indicator of autonomic function.

5 [31] was used to determine the

best-fit model that resulted in the selection of an uncorrelated exponential relaxed molecular clock. The tree was obtained using the Tree Annotator program in BEAST and the evolutionary trees were viewed in FigTree signaling pathway program 1.3.1. The relationship between predicted protection (r1-value ≥0.3) and changes in aa was analysed using a general linear model (GLM) with binomial error distribution. For this, a binomial variable ‘protected/not protected’ was created based on the estimated r1-values ≥0.3 (protected), which was used as the response variable. Summaries of the aa count differences between the query sequence of the vaccine strain and those of the field viruses were used as independent variables using either entire P1 aa sequence and each of the different viral proteins (VP1-4), alone or in combination. Both variables were analysed independently in a univariate analysis and together in a multivariate analysis. The GLM modelling and analysis of the data was carried out using R [32]. In FMD endemic settings, implementation of the progressive disease control pathway [13] requires vaccines that can protect against both circulating and emerging variants, regular vaccination campaigns, post-vaccination sero-monitoring and biosecurity measures in the form of livestock movement

control. Therefore, selection of appropriate vaccine strains is an important element in implementing vaccination policies for the control learn more of FMD. FMD is enzootic in East Africa, with outbreaks reported regularly [15], [33], [34] and [35]. Although the region has two vaccine

producing plants, there is little information available on the protective value of the supplied vaccines. The only report on vaccine strain selection in East Africa [21] was limited to a small selection of Ethiopian vaccines (two) and viruses (five). In addition, Kenya uses historic viruses such as A-KEN-05-1980 (A/K/5/80) and A-KEN-35-1980 (A/K/35/80) for vaccine production [22] and the vaccine matching tests are seldom carried out [15]. In these settings, where emergence of new variants is unpredictable, especially for serotype A FMDV, continuous serological and genetic characterisations of field viruses is needed to understand the cross-reactivity Mephenoxalone of existing vaccines and to trace patterns of viral spread. In this study, the ability of the three existing vaccine strains (A-ERI-1998, A-ETH-06-2000 and A-KEN-05-1980) and four putative candidate vaccine strains (A-EA-2007, A-EA-1984, A-EA-2005 and A-EA-1981) of serotype A FMDV to cross-protect (in-vitro) against the circulating viruses was measured by 2D VNT. The three existing vaccine strains were found to be least cross-reactive (r1-values ≥0.3 observed for only 5.4–46.4% of the sampled viruses) suggesting a poor suitability in the field, unless the low antigenic match can be compensated for by highly potent vaccine formulations [36].

In particular, the reference set of colonisation states should exclude all serotypes included in either of the two vaccines. The target set of serotypes can be chosen in different ways, depending on the question and purpose of the study: (a) The vaccines are compared with regard to serotypes common to both vaccines: the target set includes the common serotypes only. In the non-inferiority settings, the statistical power is defined as the probability for the lower bound of the confidence interval for the relative efficacy

(investigational vs. active control) to be larger than a pre-chosen non-inferiority margin. Equivalently, the margin defines an upper bound selleck compound for the rate of overall target-type acquisition for the investigational vaccine (see Appendix B). In general, there are several aspects to be considered when specifying non-inferiority margins [14]. For vaccine licensure, a natural argument follows from the requirement to show vaccine efficacy against colonisation as high as to induce herd immunity if the vaccine

were used in large scale. If the active control vaccine DNA Synthesis inhibitor is hypothesised to have at least 50% efficacy (VEacq) against overall target-type acquisition, the investigational vaccine can be allowed to have ρ100% smaller efficacy. A margin of ρ = 0.2 may be reasonable still to induce herd immunity. For example, if VEacq of 50% is considered for the active control vaccine, the power is calculated with 40% efficacy

for the investigational vaccine. The margin for the efficacies does not uniquely determine Adenosine the margin for the relative efficacy. However, it can be shown that in the range in which VEacq ≥ 0.5 for the active control vaccine, the margin of the hazard ratio is approximated by −ρ. If the efficacy of the active control is clearly >50%, a wider margin can be allowed (see Appendix B for more details). Fig. 3 presents the power of a non-inferiority study for different values of the sample size (number of individuals per study group) and the vaccine efficacy of the investigational vaccine, assuming 50% efficacy for the active pneumococcal control vaccine and a margin ρ = 0.2. The analysis is based on alternative (a), i.e. on comparing the rates of acquisition for the target set of serotypes common to both vaccines. For instance, to obtain 80% power requires a group size of 500 or more if the efficacy of the investigational vaccine is as high as 60% under scenario of the moderate overall rate of acquisition. If the investigational vaccine has only about 50% efficacy, the sample size needs to be very large for a high power. Smaller sample sizes or less strict requirements on the efficacy of the investigational vaccine are needed if comparisons are made against the union set of target serotypes (alternative (c)).

First, numerous studies have shown that despite a lack of sarcolemma depolarisation or crossbridge cycling, a stretched muscle cell can not be considered metabolically dormant. In 1932, Feng (1932) showed that a passively stretched in vitro muscle was metabolically active. He found that passively stretched muscles exhibited increased heat production and oxygen consumption. Later research corroborated these findings; Clinch (1968) reported increased heat production, while Whalen and colleagues (1962) and Barnes (1987) added reports of increased oxygen consumption. In other related studies, passive stretch increased carbon dioxide production

( Eddy and Downs, 1921), increased glycogen breakdown Ribociclib ( Barnes and Worrell, 1985), increased lactic acid production ( Barnes, 1987), and decreased phosphocreatine concentrations ( Barnes, 1987). Since increased metabolic activity is related to increased activation of the adenosine monophosphate kinase (AMPK) facilitated glucose transporter (GLUT 4) activation pathway ( Dohm, 2002), it is possible that the increased metabolic activity accompanying passive muscle stretching could have activated the incorporation of GLUT 4 into the stretched muscles. Other research also points to the possibility

of stretching increasing GLUT4 incorporation. For instance, protein kinase B activity Palbociclib cell line partially controls GLUT 4 incorporation and activation, and Sakamoto and colleagues (2003) found that protein kinase B was stimulated by passively stretching isolated muscles for ten minutes. Second, mitogenactivated protein kinase activity stimulates muscle cell glucose uptake (Ho et al 2004), and the activity of mitogenactivated protein kinases directly reflects the magnitude of the mechanical stress (ie, actively or passively generated

Rutecarpine tension) applied to the muscle (Martineau and Gardiner, 2001). Third, exercise-induced increases in nitric oxide result in increased glucose transport (Roberts et al 1997), and nitric oxide released from excised soleus muscles can be increased 20% by a single two-minute passive stretch (Tidball et al 1998). Finally, ischaemia can increase GLUT 4 translocation to the sarcolemma as well as increasing glucose uptake (Sun et al 1994, Young et al 1997), and passive stretching has the potential to cause ischaemia (Poole et al 1997, Wines and Kirkebo 1976). Wisnes and Kirkebo (1976) found an increased resistance to blood flow during passive stretching. In addition, Poole and colleagues (1997) reported that muscle stretching reduces bulk blood delivery, alters capillary flow dynamics, and impairs blood tissue oxygen exchange. Regardless of the responsible mechanisms, it is clear that passive static stretching had a significant positive effect on blood glucose levels.

5%) of the daily vial quantity taken out for the day came from unused vials from the day(s) before. A summary of the number of vials kept for multiple days across all scenarios can be found in Fig. 1. We observed that on the first day of the campaign, after the CTC training, health centre staff took out more vials than were necessary to reach the days’ target population in an attempt to prevent vaccination teams running out of vaccines, which had occurred in previous campaigns. Following supervisory visits by district staff, the health centre staff removed only the estimated quantity of vaccine needed,

plus a small buffer. Vaccination coverage in the district was high, with MAPK inhibitor 155,596 people vaccinated at the end of the campaign, equivalent to a coverage rate of 105.7%. This proportion is comparable to the results seen in the other zones of Benin: the overall coverage in the country was 104.7%. In Banikoara, the average time for a health care worker to reach their vaccination site was 36 min and 85% of the teams used motorbikes

for transport. Each team vaccinated on average 318 persons a day (range 249–433). Over the course of the campaign, 15,570 vials of MenAfriVac were used. Nine vials were discarded due to surpassing the 4 day CTC limit, five vials at day 4 and four vials on learn more the last day. No VVMs reached their endpoint. One vial was reported as broken. No indicators reached 40 °C and no vial was discarded

because of exposure to a temperature higher than Resveratrol 40 °C. A total of 21 supervisors and 77 vaccinators were surveyed, 92.2% of which had conducted outreach vaccination activities as part of the campaign. Overall confidence and perceived usefulness of the CTC approach were very high among both groups (Table 1). Most of the participants felt that the CTC practice was more useful for outreach sessions (Table 2). Health staff identified the top benefits as allowing them to vaccinate more people per day, reduced weight of the vaccine carrier, not needing to return to the health centre every night and not needing to freeze ice packs. More than half of the interviewees (52.4% of supervisors and 54.1% of vaccinators) felt that there was no risk associated with CTC. Those that spoke of risks often raised what can more accurately be termed as concerns, usually about the ability to respect the CTC limits; very few were about efficacy, adverse events or wastage (Table 3). The main difficulties in implementing CTC were identified as reading the indicator and managing the quantity of vaccine that should be taken out of the fridge. A small proportion of staff indicated that avoiding exposing the vaccine to the sun was a challenge (Table 4). 98.

Bicistronic vectors (pXsheLL, where X is the Papillomavirus type from which the codon optimized L1 and L2 genes were derived) representing the HPV types

16, 18, 31, 45, 52, 58 and Bovine Papillomavirus (BPV) were obtained from J.T. Schiller, National Cancer Institute, Bethesda, MD, USA, while p33sheLL and p68sheLL were obtained from H. Faust and J. Dillner, Malmö University Hospital, Malmö, Sweden. Constructs representing HPV35 (p35sheLL), HPV39 (p39sheLL) and HPV59 (p59sheLL) were generated by the insertion of codon optimized genes (Blue Heron, Inc., Bothell, WA, USA) based upon consensus L1 and L2 amino acid sequences into p5shell (http://home.ccr.cancer.gov/lco/default.asp). The consensus sequences were derived from NCBI database sequences (HPV35: M74117, X74477; HPV39: M62849; HPV59: X77858, EU918767) and contemporary sequences from anonymous, HPV-infected cytology samples (HPV35 L1: JN104062–64; HPV35 L2: JN104065–67; HPV39 L1: JN104068–70;

PD0332991 HPV39 L2: JN104071–72; HPV59 L1: JN104073–74; HPV59 L2: JN104075–77). The production of L1L2 pseudovirus stocks was performed as described elsewhere [24] using the alternative protocol (http://home.ccr.cancer.gov/lco/ripcord.htm), developed to reduce the inclusion of excess non-reporter-containing ‘cold capsids’, and by using luciferase (pGL4.51 [luc2/CMV/Neo]; Promega, Madison, WI) as the encapsidated reporter. Briefly, 293TT cells were transfected with equal amounts of pXsheLL and pGL4.51 [luc2/CMV/Neo] plasmids (Lipofectamine 2000; Invitrogen, Carlsbad, CA) and the encoded PFI-2 mw proteins expressed for 48 h before the cells were lysed, the capsids matured overnight in the presence of ribonucleases (RNase Cocktail; Applied Biosystems/Ambion, Austin, TX) and the double-clarified supernatant subjected to iodixanol gradient fractionation. Purified pseudovirus stocks were titrated on 293TT cells in quadruplicate, five-fold serial dilutions and the equivalent of a Tissue Culture Infectious Dose 50% Ergoloid (TCID50) was estimated using the Spearman–Karber equation. The average of three such estimations was made for each pseudovirus

stock used in this study. Pseudovirus-mediated reporter gene transduction of target cells in both the infectivity and neutralization assays was measured using the Steady-Glo Luciferase Assay Reagent (Promega) and the Glomax Multi Detection System (Promega) according to manufacturer’s instructions. The HPV pseudovirus neutralization assay was performed as originally described [25] with some modification. For the present study, heat-inactivated (56 °C, 30 min) serum samples were initially screened against all pseudoviruses (at a final serum dilution of 1/20 with pseudovirus) and any serum that demonstrated ≥80% reduction in the luciferase signal (RLU) relative to the pseudovirus and cell only controls was subsequently titrated and an 80% reciprocal neutralization titer estimated by interpolation.

Intervention: The experimental intervention was mechanically assisted walking training, such as treadmill or gait trainer without body weight support because the participants were able to walk a priori. The control intervention was defined as no intervention or an intervention that did not involve walking

training, ie, non-walking p38 MAPK apoptosis intervention. The experimental intervention was also compared with overground training. Session duration, session frequency, and program duration were recorded in order to assess the similarity of the studies. Outcome measures: Two walking outcomes were of interest speed (typically measured using 10-m Walk Test) and distance (typically measured using 6-min Walk Test). The timing of the measurements of outcomes and the procedure used to measure walking speed and distance were recorded in order to assess the similarity of the studies. Data were extracted from the included studies by a reviewer and cross checked by another reviewer. Information about the method (ie, design, participants, intervention, outcome measures) and outcome data (ie, mean (SD) walking speed and walking distance) were extracted. Authors were contacted where there was difficulty with data. The post-intervention scores were used to obtain the pooled estimate selleck inhibitor of the effect of intervention immediately (ie, post intervention) and beyond the intervention period (ie,

after a period of no intervention). A fixed effects model was used. In the case of significant SB-3CT statistical heterogeneity (I2 > 50%), a random effects model was applied to check the robustness of the results. The analyses were performed using The MIX–Meta-Analysis Made Easy programa (Bax et al 2006, Bax et al 2009). The pooled data for each outcome were reported as the weighted mean difference (MD) (95% CI). The search returned 5305 studies. After screening the titles, abstracts and reference lists, 65 papers

were retrieved for evaluation of full text. Fifty-six papers failed to meet the inclusion criteria and therefore nine papers (Pohl et al 2002, Ada et al 2003, Eich et al 2004, Weng et al 2006, Langhammer and Stanghelle 2010, Ivey et al 2011, Kuys et al 2011, Olawale et al 2011, Ada et al 2013) were included in the review. See Appendix 2 on the eAddenda for a summary of the excluded papers. Figure 1 outlines the flow of studies through the review. Six randomised trials investigated the effect of mechanically assisted walking training on walking speed and walking distance, two on walking speed, and one on walking distance. The quality of the included studies is outlined in Table 1 and a summary of the studies is presented in Table 2. Quality: The mean PEDro score of the included studies was 6.7. Randomisation was carried out in 100% of the studies, concealed allocation in 67%, assessor blinding in 67%, and intention-to-treat analysis in 44%.