Respondents were asked their views about a decent interval to wai

Respondents were asked their views about a decent interval to wait after death prior to organ recovery surgery. Focus group participants identified a tension between preserving organ viability and allowing families time with the deceased. Of the survey respondents, 45.2% selected a timeframe compatible with potentially viable donations; 34.1% selected a timeframe incompatible with viable donations; and 20.8% gave an indeterminate answer. These findings provide information about public perceptions of DCD, can be used to inform educational campaigns PFTα about

DCD and serve as a baseline for evaluating such campaigns, and identify a number of areas for further investigation.”
“Artesunate (AS) is a clinically versatile artemisinin derivative utilized for the treatment of mild to severe malaria infection. Given the therapeutic significance of PF-04929113 AS and the necessity of appropriate AS dosing, substantial research has been performed investigating the pharmacokinetics of AS and its active metabolite dihydroartemisinin (DHA). In this article, a comprehensive review is presented of AS clinical pharmacokinetics following administration of AS by the intravenous (IV), intramuscular (IM), oral or rectal routes. Intravenous AS is associated with high initial AS concentrations

which subsequently decline rapidly, with typical AS half-life estimates of less than 15 minutes. AS clearance and volume estimates average 2 – 3 L/kg/hr and 0.1 – 0.3 L/kg, respectively. DHA concentrations peak within 25 minutes post-dose, MEK inhibitor and DHA is eliminated with a half-life of 30 – 60 minutes. DHA clearance and volume average between 0.5 – 1.5 L/kg/hr and 0.5 – 1.0 L/kg, respectively. Compared to IV administration, IM administration produces lower peaks, longer half-life values, and higher volumes of distribution for AS,

as well as delayed peaks for DHA; other parameters are generally similar due to the high bioavailability, assessed by exposure to DHA, associated with IM AS administration (> 86%). Similarly high bioavailability of DHA (> 80%) is associated with oral administration. Following oral AS, peak AS concentrations (Cmax) are achieved within one hour, and AS is eliminated with a half-life of 20 – 45 minutes. DHA Cmax values are observed within two hours post-dose; DHA half-life values average 0.5 – 1.5 hours. AUC values reported for AS are often substantially lower than those reported for DHA following oral AS administration. Rectal AS administration yields pharmacokinetic results similar to those obtained from oral administration, with the exceptions of delayed AS Cmax and longer AS half-life.

Coat color, mane and tail color, age and gender were recorded wit

Coat color, mane and tail color, age and gender were recorded with results of ophthalmic examination.

Results Ocular abnormalities detected most frequently consisted of cysts arising from the posterior iris, ciliary body, and peripheral retina, curvilinear streaks of retinal pigmented epithelium extending from the peripheral temporal retina, and retinal dysplasia (folds). One Miniature Horse also had multiple ocular abnormalities (cornea globosa, goniosynechiae, decreased or absent and pupillary light reflexes, miotic and dyscoric pupils, iris hypoplasia, and epinuclear cataract). Ocular abnormalities were observed in horses that had

a flaxen or white mane and tail color and a chocolate coat color.

Conclusions Abnormal ocular phenotype detected in eyes of Miniature Horses was similar to multiple congenital and BX-795 ic50 JNK-IN-8 manufacturer hereditary ocular abnormalities reported in the Rocky Mountain Horse. Phenotypic ocular developmental abnormalities and coat, mane and tail color

were most compatible with a heterozygous or homozygous effect of the Silver Dapple locus.”
“The objective of this study is to perform a cross-country comparison of cancer treatment costs in the Nordic countries, and to demonstrate the added value of decomposing documented costs in interpreting national differences.

The study is based on individual-level data from national patient and prescription drug registers, and data on cancer prevalence from the NORDCAN database. Hospital costs were estimated on the basis of information on diagnosis-related groups (DRG) cost weights and national unit costs. Differences in per capita costs were decomposed into two stages: stage one separated the price and volume components, and stage two decomposed the volume component, relating the level of activity to service needs

and availability.

Differences in the per capita costs of cancer treatment between the Nordic countries may be as much as 30 per cent. National differences in the costs of treatment mirror observed differences in total health care costs. Differences in health care costs between countries may relate to different sources of variation with different policy implications. Comparisons of per capita spending alone can be misleading if the purpose is to evaluate, for example, differences 3-MA mw in service provision and utilisation. The decomposition analysis helps to identify the relative influence of differences in the prevalence of cancer, service utilisation and productivity. (C) 2014 Elsevier Ireland Ltd. All rights reserved.”
“In the present study, an enzyme-linked immunosorbent assay (ELISA) standardized with vesicular fluid of Taenia solium cysticerci was used to screen for IgG (total and subclasses) and IgE antibodies in cerebrospinal fluid (CSF) samples from patients with neurocysticercosis showing intrathecal production of specific IgG antibodies and patients with other neurological disorders.

In participants <= 60 y of age, a 6% (95% CI: 0 88, 0 97; P =

In participants <= 60 y of age, a 6% (95% CI: 0.88, 0.97; P = 0.002) reduction in risk was observed for the flavan-3-ol catechin when the

highest and the lowest quintiles were compared.

Conclusions: Anthocyanins and some flavone and flavan-3-ol compounds PF-04929113 manufacturer may contribute to the prevention of hypertension. These vasodilatory properties may result from specific structural similarities (including the B-ring hydroxylation and methyoxylation pattern). Am J Clin Nutr 2011;93:338-47.”
“Interindividual variability in pain sensitivity and the response to analgesic manipulations remains a considerable clinical challenge as well as all area of intense scientific investigation. Techniques in this field have matured rapidly so that much relevant data have emerged only in the past few years. Our increasing understanding of the genetic mediation of these biological phenomena have nonetheless revealed their surprising complexity. This review provides a comprehensive picture and critical analysis of the field and its prospects.”
“Background: GlaxoSmithKline Biologicals and the PATH Malaria Vaccine Initiative are working in partnership to develop a malaria vaccine to protect infants and children living in malaria endemic regions of sub-Saharan Africa, which can be delivered through MEK inhibitor the Expanded Programme on

Immunization. The RTS, S/AS candidate vaccine has been evaluated in multiple phase I/II studies and shown to have a favourable safety profile and to be well-tolerated in both adults and children. This paper details the design of the phase III multicentre efficacy trial of the RTS, S/AS01 malaria vaccine candidate, which is pivotal for licensure and Proteasome inhibitor policy decision-making.

Methods: The phase III trial is a randomized, controlled, multicentre, participant-and observer-blind study ongoing in 11 centres associated with different malaria transmission settings in seven

countries in sub-Saharan Africa. A minimum of 6,000 children in each of two age categories (6-12 weeks, 5-17 months) have been enrolled. Children were randomized 1: 1: 1 to one of three study groups: (1) primary vaccination with RTS, S/AS01 and booster dose of RTS, S/AS01; (2) primary vaccination with RTS, S/AS01 and a control vaccine at time of booster; (3) primary vaccination with control vaccine and a control vaccine at time of booster. Primary vaccination comprises three doses at monthly intervals; the booster dose is administered at 18 months post-primary course. Subjects will be followed to study month 32. The co-primary objectives are the evaluation of efficacy over one year post-dose 3 against clinical malaria when primary immunization is delivered at: (1) 6-12 weeks of age, with co-administration of DTPwHepB/Hib antigens and OPV; (2) 5-17 months of age.

Before and after intravenous administration of fluorescein, centr

Before and after intravenous administration of fluorescein, central airways and alveolar structures were evaluated. Results: Fluorescein administration did not permit imaging of epithelial cells in the central airways. In the lung periphery, alveolar walls and partially macrophages could be seen in native imaging, as expected. After administration of fluorescein, alveoli were almost filled with foam in areas with normal lung tissue.

The origin of this foam was shown to be artificial. Furthermore, in patients with pathologies of the lung parenchyma, dark neoplastic and inflammatory cells adjacent to the alveolar walls were identified. No relevant side effects of fluorescein administration could NCT-501 inhibitor be observed. Conclusions: Fluorescein-aided CLE of the lung Sonidegib in vivo appeared to be safe and well tolerated. While the lack of staining of cells in the central airways was a major limitation, it permitted analysis of the lung interstitium and alveolar space and thus emerges as a new

approach for the in vivo analysis of interstitial lung diseases. Copyright (C) 2010 S. Karger AG, Basel”
“Percutaneous spine procedures may occasionally be difficult and subject to complications. Navigation using a dynamic reference base (DRB) may ease the procedure. Yet, besides other shortcomings, its fixation demands additional incisions and thereby defies the percutaneous character of the procedure.

A new concept of atraumatic referencing was invented including a special epiDRB. The accuracy of navigated needle placement in soft tissue and bone was experimentally scrutinised. Axial and pin-point deviations

from the buy LCL161 planned trajectory were investigated with a CT-based 3D computer system. Clinical evaluation in a series of ten patients was also done.

The new epiDRB proved convenient and reliable. Its fixation to the skin with adhesive foil provided a stable reference for navigation that improves the workflow of percutaneous interventions, reduces radiation exposure and helps avoid complications.

Percutaneous spine interventions can be safely and accurately navigated using epiDRB with minimal trauma or radiation exposure and without additional skin incisions.”
“Background: Recently, nicotine administration has been shown to be a potent inhibitor of a variety of innate immune responses, including endotoxin-induced sepsis. Objective: It was the aim of this study to evaluate the effect of nicotine on attenuating lung injury and improving the survival in mice with lipopolysaccharide (LPS)-induced acute lung injury (ALI). Methods: ALI was induced in mice by intratracheal instillation of LPS (3 mg/ml). The mice received intratracheal instillation of nicotine (50, 250 and 500 mu g/kg) before or after LPS administration. Pulmonary histological changes were evaluated by hematoxylin-eosin stain, and lung wet/dry weight ratios were observed.

We

hypothesized that starvation survival is a key microbi

We

hypothesized that starvation survival is a key microbial property in species selection. This study analyzed starvation-survival behavior over 60 days of species representative of the untreated root canal infection: Fusobacterium nucleatum, Peptostreptococcus anaerobius, Prevotella intermedia and Pseudoramibacter alactolyticus. All species did not survive 1 day in water. In 1% serum, the 4 species could not survive beyond 2-3 weeks. They required a high initial cell density and >= 10% serum to survive the observation period. The results highlight a poor starvation-survival capacity of these 4 species compared with species prevalent in post-treatment infection, which are well equipped to endure starvation Vorinostat and survive in low numbers on minimal serum. These findings point to starvation-survival capacity as a selection factor for microbial participation in post-treatment disease. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2009; 108: 129-134)”
“The this website poly2-vinylpyridine and cationic polymer-mixed metal complexes (P2VPMC) have been prepared and characterized by analytical measurement such as: molecular weight, elemental analysis, IR-spectroscopy, NMR-spectroscopy, XRD, and UV. The degree of N-alkylation of poly vinyl pyridine (P-2VP) was 10% determined by chlorine content according to elemental

microanalysis. Through ICP instrument the complexation between P-2VPC and the mixed metal chlorides was proven. The structure of the samples was characterized using X-ray diffraction. It was found that the complexes have amorphous properties. The antitumor activity of this compound was examined in vitro on cell lines. The results reflect pronounced cytotoxicity www.selleckchem.com/products/lcl161.html of P-2VPMC against animal experimental EAC cells line, and human, HeLa, HCT116 and Hep-G2, cell lines. Antitumor activity of the novel compound as applied on mice bearing Ehrlich solid carcinoma, revealed delays

tumor growth compared with untreated animals and decreases tumor volume. The antitumor activity of the P2VPMC might be due to its ability to pass through the membrane and interact with, DNA causing cross-linking action which might be the key factor for great antitumor activity of the P2VPMC compound. (C) 2010 Wiley Periodicals, Inc. J Appl Polym Sci 117: 501-508, 2010″
“In this paper, a transient magnetohydrodynamic (MHD) model of an anode melting pool (AMP) flow (AMPF) is established. Mass equation, momentum equations along axial, radial and azimuthal directions, energy equation, and current continuity equations are considered in the model. In the momentum equations, the influence of electromagnetic force, viscosity force and Marangoni force (anode surface shear stress) are included. Joule heating is also included in the energy equations.

The yield of fVL testing in women with previous adverse pregnancy

The yield of fVL testing in women with previous adverse pregnancy outcomes is up to six times higher than in the general population. Calculated post-test probabilities illustrate that the combined effect of fVL and poor pregnancy history places these women at a high-risk of recurrent events. The results to date of low molecular weight heparin (LMWH) treatment trials

cannot buy MK-0518 be extrapolated to all women with thrombophilia; however, the results provide a rationale for randomized prophylactic anticoagulant treatment trials in thrombophilic women with severe adverse pregnancy outcomes. While we await the results of well-designed, adequately powered treatment trials, we propose that post-test probabilities, in addition to the preliminary treatment data in high-risk women, justify consideration of screening for fVL in women with a strong past history of poor pregnancy outcome.”
“Purpose of review

To update the reader on immunogenetic advances in idiopathic

inflammatory myopathy (IIM) over the past 18 months.

Recent findings

In Caucasian IIM, despite a shared association with the human leukocyte antigen (HILA) 8.1 ancestral haplotype (HLA-DRB1*03-DQA1*05-DQB1*02), anti-Jo-1 and anti-PM-Scl anti body-positive cases have differing IN clinical phenotypes. A study of the HLA-DPB1 region has shown that DPB1*0101 is associated with anti-Jo-1 positivity but not with anti-PM-Scl. IN single nucleotide polymorphism studies have demonstrated HM781-36B nmr associations in the protein tyrosine phosphatase, nonreceptor type 22, tumour necrosis factor alpha and interleukin-1 genes. The GM 13 allotype has been confirmed as a risk factor in Caucasian IIM. In inclusion body myositis, the HLA 8.1 ancestral haplotype may find more not only influence disease susceptibility but also disease expression. A follow-up study including a meta-analysis of the apolipoprotein E gene in inclusion body myositis suggests

that this gene does not confer risk of disease.

Summary

Although a substantial part of the genetic risk for developing adult and juvenile IN lies within the major histocompatibility complex, recent research suggests that genetic regions outside of the major histocompatibility complex are also potentially involved in conferring IN disease susceptibility, although with more modest effect sizes. An ongoing and internationally coordinated IN genome-wide association scan may provide further insights into IN immunogenetics.”
“Motion occurring during magnetic resonance imaging acquisition is a major factor of image quality degradation. Self-navigation can help reduce artefacts by estimating motion from the acquired data to enable motion correction.

Further research is needed based on individual patient data from

Further research is needed based on individual patient data from ongoing randomized trials.”
“Objective: Impairment

of baroreceptor sensitivity (BRS) has been shown to be associated with blood pressure instability after carotid endarterectomy (CEA). The aim of this study was to determine whether there is a difference in postoperative BRS changes following eversion CEA (E-CEA) and conventional CEA (C-CEA).

Methods: Sixty-four patients undergoing E-CEA (n = 37) and C-CEA (n = 27) were prospectively studied. Non-invasive measurements of mean arterial pressure (MAP), cardiac output (CO) and total peripheral resistance (TPR) were perioperatively obtained over three 10-min periods. Baroreflex gain was calculated as the sequential cross-correlation between heart rate P505-15 and beat-to-beat selleck chemicals llc systolic blood pressure.

Results: Compared with changes observed after C-CEA, E-CEA was associated with an increase in systolic pressure (SP) (P = 0.01), diastolic pressure (DP) (P = 0.008), MAP (P = 0.002) and heart rate (HR) (P = 0.03) on postoperative day 1 (POD-1). BRS decreased after E-CEA from 6.33 to 4.71 ms mmHg(-1) on POD-1 (P = 0.001) and to 5.26 ms mmHg(-1) on POD-3 (P = 0.0004).

By contrast, BRS increased after CCEA from 4.59 to 6.13 ms mmHg(-1) on POD-1 (P = 0.002) and to 6.27 ms mmHg(-1) on POD-3 (P < 0.0001).

Conclusion: E-CEA and C-CEA have different effects on BRS. This is associated with an altered haemodynamic behaviour after E-CEA and C-CEA, respectively. These findings are likely the result of carotid sinus nerve interruption during E-CEA and preservation with C-CEA. (C) 2012 European Society for Vascular Surgery. Published by Elsevier Ltd. All rights reserved.”
“A high throughput culture methodology of unicellular eukaryote Tetrahymena pyriformis, strain GL were used for the determination of catecholamines toxicity and their metabolism. Metabolism inhibitor Catecholamines exhibited acute toxicity to Tetrahymena cells where dopamine and L-DOPA showed higher toxic potential

of EC(10) (0.39 and 0.63 mg/L, respectively) and EC(20) (1.1 and 1.0 mg/L, respectively). All the testing catecholamines were highly degradable in the PPY-medium due to the oxidizing environment during incubation. They were also naturally synthesized and released by Tetrahymena cells into the culture medium and increasingly accumulated with time where as noradrenalin demonstrated significant results. Cells were exposed with physiological concentration (0.12 mg/L) and one higher concentration (8.0 mg/L) of catecholamines, resulting noradrenalin depletion and in vivo generation of a metabolite in response to dopamine with higher concentration treatment. This dopamine metabolite was relatively nonpolar compared with the catecholamines and was eluted later from the reverse phase C-18 column. (C) 2008 Wiley Periodicals, Inc. Environ Toxicol 54: 549-554, 2009.

Summary The role of genetic risk factors in genetic screening for

Summary The role of genetic risk factors in genetic screening for prevention of heart disease

is yet to be defined. The technology is already available, but functional analysis may be a prerequisite for their clinical application.”
“Four kinds of biorefinery residues (the ethanol, the butanol, the xylitol and the lactic acid biorefinery residue) were utilized to modify the phenol-formaldehyde (PF) resin, respectively by copolymerization. Prior to the reaction, the LY3023414 composition and functional group of each biorefinery residue were characterized and used to evaluate their suitability as replacement for the preparation of PF resins. It was found that ER (bio-ethanol biorefinery residue) exhibited high reactivity due to high content of hydroxyl group. The influence of each biorefinery residue for the replacement of phenol on the properties of residue-modified phenol formaldehyde (RPF) resins was also investigated using Fourier transform infrared spectroscopy (FT-IR), solid state (CNMR)-C-13 and thermo gravimetric analyzer (TGA). ER was found to be the best biorefinery residue for the modification of PF resin. 50% phenol could be replaced by ER without influencing the properties of adhesives and plywoods. In industrial production, the plywoods bonded by ER-derived PF resins met the requirement of both first grade and E-0 grade (GB/T 9846.3-2004). (C) 2012 Elsevier B.V. All rights reserved.”
“BACKGROUND: Minimally

invasive surgical P005091 mouse procedures

such as tension-free vaginal tape sling should not imply that a minimal preoperative evaluation is all that is required.

CASE: A 52-year-old multiparous perimenopausal woman presented with postoperative nausea, vomiting, and vague abdominal-pelvic discomfort after placement of tension-free Birinapant in vitro vaginal tape. The final diagnosis was invasive molar pregnancy.

CONCLUSION: This case raises the awareness of the rare diagnosis of gestational trophoblastic disease in perimenopausal women and emphasizes the importance of a complete preoperative evaluation in those scheduled to undergo minimally invasive procedures. (Obstet Gynecol 2011; 117: 487-8) DOI: 10.1097/AOG.0b013e3182055efc”
“Purpose of review We will review the role of microRNAs (miRNAs), small noncoding RNAs with regulatory function, in myocardial infarction (MI). Specifically, we will examine the effect of MI on miRNAs’ expression in the heart, the effect of MI on circulating miRNAs, which miRNAs’ overexpression or downmodulation appears to have a therapeutic role in MI and which cardiac miRNAs are modulated by drugs/experimental molecules/cell transplantation strategies which have an established or potential therapeutic role in MI.

Recent findings A rapidly increasing number of studies are showing that cardiac and circulating miRNAs are markedly altered in MI. These novel findings shed new light on the mechanisms that lead to MI complications, post-MI ventricular remodeling and cardiac repair.

g buccal, sublingual and nasal routes The evaluation of the sys

g. buccal, sublingual and nasal routes. The evaluation of the systemic

plasma concentration and the derivation of the pharmacokinetic parameters of candidate compounds in preclinical studies are essential for drug development. The effect of site of blood sampling on the measured drug concentration, in both animals and humans, is to some extent known but it is not always taken into consideration in the design of pharmacological and toxicological studies. Methods: Blood Epigenetics inhibitor samples were collected both from leg and jugular veins from beagle dogs following a single sublingual dosing of Compound A in order to determine the impact of different sites of blood sampling on plasma pharmacokinetics. Plasma was prepared by centrifugation and plasma concentrations

of Compound A were determined by protein precipitation and liquid chromatography followed by mass spectrometric detection. The pharmacokinetic parameters were calculated by non-compartment methods. Results: Sampling from the jugular vein resulted in higher and more variable exposure during the absorption phase compared to sampling from a leg vein. The plasma exposure in the jugular vein, in terms of C-max, was 4-fold compared to that in the leg vein and an approximately 2-fold bioavailability was observed. Discussion: The aim of this investigation was to determine the impact of the different sites of blood sampling on assessing systemic plasma exposure and pharmacokinetic parameters for Compound A following sublingual dosing to dogs. The results demonstrate the significant impact that the site of blood sampling has on PK selleck compound parameters, and raise concerns of using the jugular vein as a site of sampling check details after sublingual and other transmucosal routes of dosing in the head region. (c) 2012 Elsevier Inc. All rights reserved.”
“Background and Purpose: Open radical inguinal lymphadenectomy is reported to have

morbidity as high as 50%. We describe our endoscopic inguinal lymphadenectomy that aims at decreasing the morbidity of the procedure without compromising the oncologic outcomes.

Patients and Methods: Eleven groin dissections were undertaken in seven male patients. The procedure was performed via three ports. The first one was a 10-mm incision 3 mm distal to the apex of the femoral triangle. Two additional trocars (10 mm and 5 mm) were positioned 6 cm medially and laterally to the apex of the triangle, respectively. Taking the great saphenous vein as a landmark, the superficial and deep components were dissected. The boundaries of dissection were the same as those of radical inguinal lymphadenectomy. The numbers of lymph nodes harvested were recorded. The morbidity was retrospectively analyzed.

Results: The mean operative time was 126 minutes. The mean number of lymph nodes was 12.3. The averaged output of drainage per leg was 50.8 mL each day.

The nanoscale metastable (Fe,Co)(23)B(6) phase precipitated in th

The nanoscale metastable (Fe,Co)(23)B(6) phase precipitated in the glassy matrix after annealing, while the two-stage-like glass transition disappeared, indicating the two-stage-like glass transition results from the overlap of the endothermic reaction for the glass transition with the exothermic reaction for the formation of the (Fe,Co)(23)B(6) phase in the supercooled liquid region. The (Fe(0.9)Co(0.1))(67.5)Nb(4)Gd(3.5)B(25) glassy

alloy exhibits high glass-forming ability, enabling the formation of glassy alloy rods with diameters exceeding 3.0 mm, rather high saturation magnetization of 0.91 T, low coercive force of 2.5 A/m, and high fracture strength of 3870 MPa.”
“BACKGROUND: Hydroponic greenhouse effluent has high concentrations of total phosphorus (30100 mg PO4-P L1) and nitrates (200300 buy FK228 mg NO3-N L1). Current technologies for effluent treatment have limitations of performance

and high maintenance costs. The goals of this study buy ISRIB were to investigate strategies which combine alkali treatment and microalgae cultivation for removal of nutrients from hydroponic greenhouse effluent. RESULTS: Treatment with strong alkali was found to effectively remove 97% of total phophorous especially in the form of phosphate, without affecting the nitrate ion concentration in the greenhouse effluent. After alkali treatment, marine algae Dunaliella salina (UTEX 1644) cultivation on treated hydroponic effluent (pH 7.5) showed > 80% decrease in nitrate content in the effluent within 4 days of cultivation. In the same period, the

carotene content of the micro-algal system was in the range 0.5 +/- 0.02 mu g mg1 (dry cell weight) which was 1.5 times higher than in the control. CONCLUSION: This study demonstrated that combination of a conventional alkali precipitation method with a microalgae treatment system is a highly efficient approach for the removal of excess nutrients from hydroponic greenhouse effluent in a short treatment time. The microalgae can provide a source of value in the form of carotene. (c) 2012 Society of Chemical GSK J4 datasheet Industry”
“Conducting clinical pharmacology research studies in pediatric patients is challenging because of ethical and practical constraints but necessary to ensure that drugs are used safely and effectively in this population. Developments in laboratory analytical techniques, such as improved assay sensitivity and the use of alternative sample matrices, can reduce blood loss and offer less invasive blood sampling, causing less trauma to the patient and fewer ethical concerns. Recent advances in data analysis techniques, which aim to extract the maximum amount of useful information from small sample numbers, should be considered when planning a clinical trial and incorporated into the study design.