Secondly, oil-in-water emulsions improve vaccine responses agains

Secondly, oil-in-water emulsions improve vaccine responses against seasonal influenza in elderly populations, immunocompromised

patients and children [6]. They can also broaden the immunogenicity of pandemic vaccines as shown by the MF59-induced epitope spreading from HA2 to neuraminidase and HA1, thus providing cross-clade neutralization and potentially improving in vivo protection [7]. Thirdly, the safety profile of oil-in-water emulsions is well documented: MF59 and AS03 have been used successfully in over 100 million people including children. Novartis’ seasonal influenza vaccine containing MF59 is routinely and extensively used in the elderly [8], and both GSK’s AS03-adjuvanted pandemic (H1N1) 2009 influenza vaccine and Novartis’ MF59-adjuvanted pandemic (H1N1) I-BET151 datasheet 2009 influenza vaccine were used worldwide in 2009 and 2010. It is worth noting that the technology transfer of an emulsion containing metabolizable oil and surfactant in the absence of block-copolymer from a European centre to DCVMs does not infringe any intellectual property. Access by DCVMs to this adjuvant technology would therefore be highly advantageous not only

for pandemic influenza vaccines, but could trigger benefits for further applications since oil-in-water emulsions have been widely investigated in numerous clinical trials with several subunit antigens, such as HIV, hepatitis B virus and hepatitis C virus antigens see more [9]. In addition, the capital investment needed to produce oil-in-water

adjuvants is relatively modest and the cost of materials adds only marginally unless to the cost of antigen production. The manufacturing process for oil-in-water emulsions has been described in detail [10]. The Vaccine Formulation Laboratory has established the production processes and prepared oil-in-water emulsions that meet all expected physical, chemical and biological (adjuvant activity) parameters. We are currently screening a range of raw material sources and evaluating the acceptability of the products for use in clinical-grade emulsions. This is particularly important for materials of biological origin such as squalene (prepared from shark liver) and heterogeneous surfactants such as Tween80 and Span85. In order to develop all standard operating procedures and relevant documentation for Good Manufacturing Practice (GMP) production, a collaboration has been developed with The Netherlands Vaccine Institute (NVI) in Bilthoven, The Netherlands. Bio Farma, Indonesia, a grantee of the WHO initiative to transfer the capacity to produce influenza vaccines to DCVMs, is the first technology transfer partner of the Vaccine Formulation Laboratory. The first phase of the project comprises the installation of equipment required for production and characterization of oil-in-water emulsions, the establishment of relevant standard operating procedures, training of laboratory staff, and on-site validation of the transferred processes.

In a previous

publication we described the study design e

In a previous

publication we described the study design extensively.13 The effects of the physical activity stimulation program on social participation, quality of PI3K inhibitor life and self-perception will be reported in a separate paper. Participants were randomised 1:1 to the experimental or control intervention, with stratification by Gross Motor Function Classification System (GMFCS) level I versus level II/III. The GMFCS level I is walking without limitations, level II is walking with limitations and level III is walking with a hand-held mobility device.14 Sealed envelopes were used to conceal group allocation. Participants were informed of group allocation following the baseline assessments. The intervention group followed a 6-month physical activity stimulation program, involving a lifestyle intervention and 4 months of fitness training. The control group continued their usual paediatric physiotherapy.

Outcomes were assessed in the hospital: at baseline; at 4 months (ie, at the end of fitness training, when only walking capacity, functional strength and fitness were assessed); at 6 months (that is, at the end of the intervention); and at 12 months. The assessor (AB) was blinded to group allocation throughout the study. The parents’ attitudes towards sport were only assessed at baseline and 12 months. Children with spastic cerebral palsy, aged 7–13 years who could walk were recruited via paediatric physiotherapy practices and special schools for children with disabilities. Inclusion criteria were: Forskolin mouse classification in GMFCS level I–III, understanding of the Dutch language and fulfilling at least one of the following criteria as determined

in a telephone interview: less active than the international physical activity norm of less than 1 hour daily at >5 metabolic equivalents (METs), which is moderate or vigorous intensity;15 no regular participation in sports or (physiotherapeutic) fitness program (ie, less than three times a week for at least 20 minutes); and experience of problems related and to mobility in daily life or sports. Exclusion criteria were: surgery in the previous 6 months, botulinum toxin treatment or serial casting in the previous 3 months (or planned), unstable seizures, contra-indications for physical training, severe behavioural problems, severe intellectual disability and a predominantly dyskinetic or ataxic movement disorder. The intervention group followed the physical activity stimulation program, which involved a lifestyle intervention and fitness training followed by usual physiotherapy. The control group undertook only usual physiotherapy. The components of the interventions are presented in Figure 1 and described in more detail elsewhere.

Free radical generation during treatment with 5-FU, leading to li

Free radical generation during treatment with 5-FU, leading to lipid peroxidation and cell

membrane damage, could be one mechanism behind the toxic effects of 5-FU.4 BP is a well known ancient folk medicine, an intricate resinous hive product, and a blend of waxes, sugars and plant exudates collected by bees from plants. Flavonoids, aromatic acids, diterpenic acids and phenolic compounds appear to be the principal components responsible for its biological activities. It is alleged to exhibit a broad spectrum of activities including antibacterial, antifungal, antiviral, anti-inflammatory, local-anesthetic, anti-oxidant, immune stimulating, cytostatic and free radical scavenging activities.9 Recently, it is also being PARP inhibitor used in food and beverages to improve health and prevent diseases such as inflammation, heart disease, diabetes and cancer.10 To the best of our knowledge such an extensive study on renal toxicity by 5-FU has been reported GSI-IX in vivo for the first time. Glutathione reductase, oxidized (GSSG) and reduced glutathione, 1,2-dithio-bis-nitrobenzoic acid (DTNB), 1-chloro-2, 4-dinitrobenzene, bovine serum albumin (BSA), oxidized and reduced nicotinamide adenine dinucleotide phosphate (NADP), (NADPH), flavine adenine dinucleotide, 2,6-dichlorophenolindophenol,

thiobarbituric acid (TBA), 5-FU etc: were obtained from Sigma–Aldrich, USA. Sodium hydroxide, ferric nitrate, trichloroacetic acid (TCA) and perchloric acid (PCA) etc were purchased from CDH, India. Plant extract was purchased from Saiba Industries, Mumbai. Male Wistar rats (150–200 g), 6–8 weeks old, were obtained from the Central Thalidomide Animal House Facility of Hamdard University. Animals received humane

care in accordance with the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA),Government of India, and prior permission was sought from the Institutional Animal Ethics Committee (IAEC No: 173/CPCSEA, 28 January 2000). Rats were randomly divided into five groups of six rats each. Group I served as control and received water for 28 days and 0.9% saline intraperitoneally (i.p.) on day 25th, 26th. Group II received i.p. injections of 5-FU (75 mg/kg b.wt.) on 25th and 26th day. Groups III and IV were treated with an oral dose of BP 80 mg/kg b.wt. (D1) and 160 mg/kg b.wt. (D2), respectively, for 28 days and i.p. injections of 5-FU (75 mg/kg b.wt.) were administered on 25th and 26th day. Group V received only D2 (160 mg/kg b.wt.) of BP for 28 days. On the 28th day, the rats were sacrificed by cervical dislocation, blood was drawn for serum parameters and kidneys were taken after perfusion for examination of various biochemical, immunohistochemical and histopathological parameters.

The authors declare that there are no conflicts of interests The

The authors declare that there are no conflicts of interests. The Commuting and Health in Cambridge study was developed by David Ogilvie, Simon Griffin, Andy Jones and Roger Mackett and initially funded under the auspices of the Centre for Diet and Activity Research (CEDAR), a UKCRC Public Health Research Centre of Excellence. Funding from the British Heart Foundation, Economic and Social Research Council, Medical Research Council, National Institute for Health Research and the Wellcome Trust, under the auspices of the UK Clinical Research Collaboration, is gratefully acknowledged.

The study is now funded by the National Institute for Health Research Public Health Research programme (project number 09/3001/06: David Humphreys contributed to this study while funded by a CEDAR Career Development Fellowship. Anna Goodman’s contribution to this study was funded by an NIHR postdoctoral fellowship. CHIR-99021 cost David Ogilvie is supported by the Medical Research Council [Unit Programme number MC_UP_1001/1]. The views and opinions expressed herein are those of the authors and do not necessarily reflect those of the NIHR, the Department of Health or the NHS. The funding bodies had no part in the study design; in the collection, analysis or interpretation of data; in the writing Capmatinib concentration of the manuscript; or in the decision to submit the manuscript for publication. The study was

approved by the Hertfordshire Research Ethics Committee (reference number 08/H0311/208). We thank the study participants for their cooperation and the staff of the MRC Epidemiology Unit Functional Group Team, in particular for study coordination and data collection (led by Cheryl Chapman) and data management. “
“Many young people do not meet current UK physical activity guidelines (Craig et al., whatever 2009). Preventing the well-established decline in physical activity that occurs as children enter adolescence may reduce future risk of cardiovascular disease and obesity (Department of Health, 2004). Previous childhood physical

activity interventions have had little success (Van Sluijs et al., 2007), which could be due to a limited understanding of the complex factors which influence children’s physical activity. Time spent outdoors is a consistent predictors of children’s physical activity (Sallis et al., 2000), and physical activity levels are greater out of school than during school (Gidlow et al., 2008). Weekday evenings and weekend days are leisure time. Young people have more freedom of choice for physical activity in leisure periods than during the structured school day, when organised physical activity may be more easily promoted (Cardon et al., 2009 and Loucaides et al., 2009). Unstructured outdoor physical activity in children’s free time, (“active play”) could be a major contributor to total physical activity levels (Veitch et al., 2008).

Il incombe donc aux médecins de prendre en compte cette dimension

Il incombe donc aux médecins de prendre en compte cette dimension particulière. En fonction de la pathologie et

de l’état cardiaque des patients, une évaluation du risque lié à la pratique de l’activité sexuelle doit parfois être réalisée au cas par cas afin d’apporter un conseil personnalisé en ce domaine. L’activité sexuelle met en jeu une interaction complexe entre facteurs psychologiques, hormonaux, vasculaires et neurologiques. Les adaptations végétatives impliquent, chez la femme, essentiellement la mise en jeu du système nerveux sympathique alors que chez les hommes les interactions sont plus complexes avec, selon les phases, augmentation du tonus parasympathique et réduction de l’activité sympathique. Mais l’un des éléments clés chez l’homme est lié à la sécrétion de monoxyde d’azote (NO) au niveau de l’endothélium des corps caverneux, l’érection étant en effet un phénomène JQ1 essentiellement vasculaire (vasodilatation).

C’est chez l’homme que la contrainte cardiovasculaire lors de l’acte sexuel est la plus importante. En GDC-0199 solubility dmso général, la durée de l’acte sexuel pour parvenir à l’orgasme est d’environ cinq à six minutes, l’orgasme lui-même étant bien plus bref, environ 10 à 15 secondes. Les paramètres cardiovasculaires, fréquence cardiaque et pression artérielle, reviennent habituellement à un niveau basal dans les 2 à 3 minutes qui suivent l’orgasme. Compte tenu de ce caractère relativement bref et discontinu de l’activité sexuelle, l’évaluation détaillée des paramètres cardiovasculaires et respiratoires ne peut se concevoir qu’avec des enregistrements continus, notamment de la pression artérielle, la mesure discontinue par technique de mesure ambulatoire de pression artérielle (MAPA) ne permettant pas en effet d’avoir une évaluation précise au moment des pics d’activité [1]. De tels enregistrements invasifs (cathéters intra-artériels dans la mesure où les capteurs digitaux de pression

artérielle sont peu adaptés) n’ont été réalisés que dans de petites séries, qui plus est très anciennes [2] and [3]. La figure 1 montre schématiquement les adaptations cardiovasculaires et respiratoires chez also l’homme au cours d’une relation sexuelle avec un orgasme (adapté de Fox et al. [2] et Littler et al. [3]). L’augmentation de la fréquence cardiaque et de la pression artérielle est modérée en dehors du moment de l’orgasme et de l’éjaculation avec ensuite un retour aux valeurs basales en 2 à 3 minutes habituellement. La respiration est souvent hachée à l’approche de l’orgasme avec fréquemment de brèves apnées avant une augmentation nette de la ventilation après l’éjaculation. Les données chez les femme sont encore plus rares (une seulement dans chacune des deux études citées [2] and [3]).

47 (95% CI 0 20 to 0 73) (Figure 4, see also Figure 5 on the eAdd

47 (95% CI 0.20 to 0.73) (Figure 4, see also Figure 5 on the eAddenda for a detailed forest plot.) The effect of exercise training on the ‘sleep latency’ subscale of the Pittsburgh Sleep Quality Index was examined by pooling data from 239 participants across five trials. Participation in exercise training reduced (ie, improved) sleep latency, with an SMD of

0.58 (95% Cl 0.08 to 1.08) (Figure 6, see also Figure 7 on the eAddenda for a detailed forest plot.) Exercise training also reduced the use of medication to assist sleeping, with an SMD of 0.44 (95% Cl 0.14 to GW-572016 cost 0.74) on the ‘use of sleep medication’ subscale of the Pittsburgh Sleep Quality Index. This was based on pooled data from 196 participants across four trials (Figure 8, see also Figure 9 on the eAddenda for a detailed forest plot.) Exercise training did not cause significant improvement in other domains of the Pittsburgh Sleep Quality Index, including sleep duration, sleep efficiency, sleep disturbance, and daytime functioning Selisistat research buy (see Figures 10 to 13 on the eAddenda.) Objective sleep quality: Only one trial measured sleep quality objectively ( King et al 2008). Polysomnography indicated that the subjects who had participated in exercise training spent a significantly lower percentage of time in Stage 1 sleep (between-group difference 2.3%, 95% Cl 0.7 to 4.0,

effect size = 0.66) and a greater percentage in Stage 2 sleep (between-group difference 3.2%, 95% Cl 0.6 to 5.7, effect size = 0.41) relative to the control subjects. However, the study identified no other significant group differences regarding other polysomnographic parameters,

such as sleep latency and efficiency after participation in the 12-month exercise training program. This meta-analysis provides a comprehensive review of randomised trials examining the effects of an exercise training program on sleep quality in middle-aged and older adults with sleep complaints including insomnia, depression, and poor sleep quality. Pooled analyses of the results indicate that exercise training has a moderate beneficial effect on sleep quality, as indicated Oxygenase by decreases in the global Pittsburgh Sleep Quality Index score, as well as its subdomains of subjective sleep quality, sleep latency, and sleep medication usage. Other sleep time parameters, including sleep duration, efficiency, and disturbance, were not found to improve significantly. These findings demonstrate that the participants did not sleep for a longer duration after participation in exercise training but they nevertheless perceived better sleep quality. Since poor sleep quality and total sleep time each predict adverse health outcomes in the elderly (Pollack et al 1990, Manabe et al 2000), optimal insomnia treatment should not only aim to improve quantity but also self-reported quality of sleep.

Outcome measures: For standing up, weight distribution between th

Outcome measures: For standing up, weight distribution between the lower limbs was measured (2 trials). For standing, the measures used were directional control during reaching in standing (3 trials), Berg Balance Scale (3 trials),

Rivermead Mobility Index (1 trial), gross function subscale of the Rivermead Motor Assessment (1 trial), and the balance component of the Fugl-Meyer-Lindmark (1 trial). For walking, all trials measured gait parameters such as step/stride length or width of base of support or speed (11 trials). Outcomes were measured after intervention (20 trials) and from 1 to 5 months after cessation of intervention (11 trials). The short-term effect of biofeedback on activity limitations was examined by pooling data after intervention from 17 MEK inhibitor trials comprising 411 participants using a fixed-effect model. Biofeedback improved lower limb activities compared with usual therapy/placebo (SMD = 0.41, 95% CI 0.21 to 0.62) (see Figure 2 on the eAddenda for the detailed forest plot). There was, however, substantial statistical heterogeneity (I2 = 65%), indicating that the variation between the results of the trials is above that expected by chance. The results of a sensitivity analysis

MS-275 chemical structure revealed that the heterogeneity was best explained by the quality of the trials. When low quality trials (ie, seven trials with PEDro score 3 and 4) were excluded from the analysis, the magnitude of the effect PD184352 (CI-1040) was similar (SMD = 0.49,

95% CI 0.22 to 0.75) but with less heterogeneity (I2 = 43%) (Figure 3, see Figure 4 on eAddenda for the detailed forest plot). The long-term effect of biofeedback on activity limitations was examined by pooling data after the cessation of intervention from 5 high quality trials comprising 138 participants using a fixed-effect model. Biofeedback improved activity compared with usual therapy/placebo (SMD = 0.41, 95% CI 0.06 to 0.75, I2 = 42%) (Figure 5, see Figure 6 on the eAddenda for the detailed forest plot). Subgroup analysis by activity found that the short-term effect of biofeedback on standing up could only be examined in one high quality trial comprising 40 participants. Biofeedback tended to increase standing up compared with usual therapy (SMD = 0.54, 95% CI –0.09 to 1.17). The short-term effect of biofeedback on standing could be examined by pooling data after intervention from five high quality trials comprising 125 participants, using a fixed-effect model. Biofeedback increased standing compared with usual therapy/placebo (SMD = 0.42, 95% CI 0.05 to 0.78, I2 = 69%, see Figure 7 on the eAddenda for the detailed forest plot) and the magnitude of the effect was the same using a random-effects model (SMD = 0.42, 95% CI –0.08 to 0.93).

L L is an employee at Merck Sharp & Dome Corp , a subsidiary of

L.L. is an employee at Merck Sharp & Dome Corp., a subsidiary of Merck & Co., Inc., Whitehouse Station, New Jersey, and may own stock or stock options in Merck. L.T.T. has received a travel grant from Sanofi Pasteur MSD. K.E.J. has received a travel grant from Merck. C.M. received lecture fees and support for conference participation from Merck and Sanofi Pasteur MSD. M.N. has received research grants from /MSD/Merck through the affiliating institute. We wish to thank Jessica Pege, Lissa Churchward and Cecilia Olofsson

for organizing data collection, Pouran Almstedt and Suzanne Campbell for database administration, Miriam Elfström for help with dropout analyses, and Kirsten Frederiksen, Linda Vos and Tor Å. Myklebust for statistical advice. “
“Yellow fever is an acute arboviral disease with clinical presentations that include mild forms with a sudden onset of febrile symptoms buy Volasertib and severe forms with over 30% lethality, and also asymptomatic infections [1]. Yellow fever is one of the diseases requiring immediate report to the World Health Organization (WHO) CRM1 inhibitor under International Health Regulations [2]. In Brazil, most cases of yellow fever occur among adult males conducting occupational, tourism, or leisure activities in forested areas, where they become exposed to infected mosquitoes, mainly the wild species Haemagogus janthinomys. Although disease transmission in urban

areas have not been reported in

Brazil since 1942, sporadic outbreaks of yellow fever transmitted by jungle vectors in the southern and southeastern regions of the country, close to urban zones where Aedes aegypti is abundant, poses a threat of re-urbanisation of the disease [3]. There is no specific treatment for yellow fever. Disease prevention relies on current commercially available vaccines, which are highly immunogenic and safe. Immunisation is recommended to unvaccinated Electron transport chain residents and travellers to and from at-risk areas, aged ≥9 months [3] and [4]. Despite the lack of efficacy studies on yellow fever vaccines, vaccine effectiveness is evidenced by the dramatic reduction of disease incidence following mass vaccination. The duration of vaccine-induced immunity in primo-vaccinated adults appears to last for decades [5]. Previous recommendations [6] of revaccination have been revised by WHO experts in 2013 [5] and a systematic review of scientific evidence available until June 2012 [7]. The International Health Regulations have been ammended in May 2014 to stipulate that a single dose of the yellow fever vaccine is valid for the duration of the vaccinee’s life [2]. Data on the long-term immunity induced by yellow fever vaccine, which should guide vaccination policy are still scarce. Therefore, this study aimed to assess the level of neutralising antibodies persisting after years of primovaccination against yellow fever in adults.

HPLC data acquisition and processing

HPLC data acquisition and processing Paclitaxel research buy was performed by Shimadzu LC Solutions Ver 1.23 SP 1 software. PZA belongs to the basic class of drugs due to its amide functional group. Therefore adjusting the pH of mobile phase to the acidic side ionizes the PZA present in plasma thereby leading to poor recovery. In order to extract the un-ionized form of the drug, it is imperative to adjust the pH to the alkaline side, however, alkaline mobile phase characteristics causes deterioration of the bonded phase in the column due to alkaline hydrolysis of end-capped silica. Compared to acid catalyzed hydrolysis, the hydrolysis of end-capped

silica in alkaline conditions is usually very rapid. Therefore experiments were performed using potassium dihydrogen phosphate in a limited range of pH 7.0–8.0. The response was checked at the detector using a connector (without the column). A pH value of 7.4 ± 0.1 gave maximum FK228 response for the analyte at 268 nm. The run time of analysis was higher when a longer reverse phase column (250 × 4.6 mm i.d,) was used. The resolution

between the peaks was decreased and peaks were not of acceptable peak shape when the experiment was performed using a shorter column (50 × 4.6 mm i.d,). However better resolution, less tailing and high theoretical plates were obtained with Phenomenex column C18 150 × 4.6 cm 5 μm column. The mobile consists of 15:85 v/v methanol and 10 mM potassium dihydrogen phosphate (pH 7.4). The flow rate of the method was 1.0 ml/min. The column temperature was maintained at 25 °C. At the reported flow rate peak shape was excellent,

however, increasing or decreasing the Endonuclease flow rate increased the tailing factor and resulted in poor peak shape and in decreased resolution between the drug and internal standard. There was no interference in the drug and the internal standard, from the extracted blank. The peak shape and symmetry were found to be good when the mobile phase composition of 15:85 v/v was used with better resolution of the drug and internal standard. Increasing the organic portion of the mobile phase caused PZA to elute with high tailing and also merging of the peaks for PZA and MTZ. A mobile phase containing aqueous portion greater than 85% led to very late elution and very poor peak shape for MTZ. The peaks were also broad and had unacceptable asymmetry factor. Extraction methods were initially attempted using protein precipitation technique. Organic solvents such as acetonitrile and/or methanol were used as reagents for protein precipitation.13 Initial experiments of protein precipitation were done using 1:3 ratio of plasma:organic solvents. The recovery of the PZA was poor while that of the internal standard was relatively unchanged as compared with liquid–liquid extraction. Since the noise effects in solid phase extraction (SPE) method are similar to that of liquid–liquid extraction, the final analysis was carried out using liquid–liquid extraction (LLE).

We also explored the association between maternal serum and breas

We also explored the association between maternal serum and breast milk anti-rotavirus antibody concentrations

with the immune response in infants after two doses of this vaccine. The trial was conducted in typical urban resettlement neighborhoods of South Delhi, India. Infants aged less than 7 weeks were identified through a household survey. Families of infants aged 6–7 weeks were invited to the study clinic for screening and enrollment. Informed written consent was obtained from all parents and also specifically from the mothers. All enrolled infants received two Alectinib datasheet doses of Rotarix® at 6–7 weeks and at 10–14 weeks of age along with other childhood vaccines (Diphtheria, Pertussis, Tetanus, Haemophilus influenzae B, Hepatitis B and oral Polio). click here At the study clinic after consent was obtained, a physician examined the infant. Mother–infant pairs were enrolled if the parents gave consent, infants were aged 6–7 weeks, the weight for age was >−3SD of the WHO child growth standards, and the family had no plans to move out of the study area for the next 4 months. Infants were excluded if they were not breastfed,

had already received a rotavirus vaccine, had immunodeficiency disease, chronic enteric disease, and/or any other condition as warranting exclusion by the investigator. Infants were temporarily excluded if they had diarrhea or any illness requiring hospital referral on the day of enrollment. Eligible infants were either allocated to the group where mothers were requested to

withhold breastfeeding for 30 min before and after vaccine administration or to the group where MTMR9 mothers were encouraged to breastfeed their infants around the time of vaccination. There were two separate locations in the study clinic for the two groups to ensure that instructions for breastfeeding were followed by mothers. Clinical coordinators supervised each area. Activities were conducted in the following order: 30 min of withholding or encouraging breastfeeding; administration of Rotarix®; 30 min of withholding or encouraging breastfeeding; administration of other childhood vaccines; observation for 30 min to assess for immediate adverse events. The study team documented the time breastfeeding started and ended as well as the time when the other vaccines were administered. Infants were observed for immediate adverse events in the study clinic and referred to the hospital, if required. Families of infants were contacted weekly after each dose of the Rotarix® to ascertain presence of signs and symptoms of any illness requiring hospital referral including intussusception, or other serious adverse events. Minor illnesses not requiring hospital referral were managed by the study physician. Serious adverse events were reported to the relevant Ethics Committees. The randomization list was generated by a statistician independent of the study team in Stata 11 (StataCorp LP, TX, USA).