J. Clin. Invest. 118(1): 161-172 (2007)
Scoring spontaneous pain. Wild-type mice (C57BL/6J; n = 5/group) were subjected to CCI, placed in plexiglass cylinders (19 × 31 cm), and allowed to habituate. Baseline response measurements were determined for each animal prior to testing. One animal at a time was continuously observed for 2 min. This was repeated 2 more times within the next 2 h. Various positions of the injured hind paw were continuously observed and rated according to the following numerical scoring system: 0, paw placed normally on the floor; 1, paw placed lightly on the floor and toes ventroflexed; 2, only internal edge of the paw contacted the floor; 3, contact restricted to the heel; 4, total paw elevation. During each 2-minute observation period, scoring was done at 5-s intervals by an investigator who was blinded to the experimental groupings. The spontaneous pain index for each observation period was calculated by multiplying the interval number by the interval score and dividing by the total number of intervals (25 in all/session) (80). The overall pain index for each mouse on each day was obtained by averaging the scores obtained during the 3 observation periods. In general, the 3 scores obtained on a single day with 1 mouse varied by less than 20% relative to the mean. Results at each day are presented as the mean ± SD for the cohort.
Am J Epidemiol. 2009 June 15; 169(12): 1437–1444.
Baseline measurements Methods for blood processing in the ARIC Study have been described (10). Participants were asked to fast for 12 hours before their morning clinic appointments. The measurement of serum magnesium was based on the procedure of Gindler and Heth and used the metallochromic dye, calmagite (1-(1-hydroxy-4-methyl-2-phenylazo)-2-napthol-4-sulfonic acid). The laboratory coefficient of variation, based on blinded split samples sent 1 week apart to the laboratory, was 3% (1), and repeated testing of 40 individuals over several weeks yielded a reliability coefficient of 0.69 (11). Plasma fibrinogen and the von Willebrand factor antigen were measured by the thrombin time titration method and enzyme-linked immunosorbent assay, respectively. Serum glucose was measured by a hexokinase/glucose-6-phosphate dehydrogenase method. Body mass index was calculated as weight (kg)/height (m)2. We defined prevalent coronary heart disease and stroke at baseline, for exclusion, as a self-reported history of a physician-diagnosed heart attack, prior myocardial infarction by electrocardiogram, prior cardiovascular surgery, prior coronary angioplasty, or prior stroke or transient ischemic attack identified by a standardized interview (12). Dietary information over the last year was collected by using Willett's 61-item food frequency questionnaire, adapted for interviewer administration and otherwise modified only slightly (13). Dietary magnesium intake was computed by multiplying the magnesium content of each food item by the frequency of its daily consumption and summing over all items. In the ARIC Study, serum magnesium was remeasured for 91% of the participants in 1990–1992, and dietary magnesium was remeasured for 82% in 1993–1995. The Spearman correlation coefficients between 2 visits were 0.45 for serum magnesium and 0.54 for dietary magnesium. The Spearman correlation coefficient between serum and dietary magnesium at baseline was 0.04.
Gynecol Oncol. 2012 April; 125(1): 136–140.
A phase II study single agent of aflibercept (VEGF Trap) in patients with recurrent or metastatic gynecologic carcinosarcomas and uterine leiomyosarcoma. A trial of the Princess Margaret Hospital, Chicago and California Cancer Phase II ConsortiaEvaluation of toxicity and tumor responseBaseline evaluations included medical history, physical examination, laboratory tests (hematology, urinalysis, coagulation, blood chemistry), ECG (if indicated), and pregnancy test and were performed within 7 days of administration of protocol therapy. Physical exam, lab tests (hematology, biochemistry and urinalysis), evaluation of toxicity according to NCI Common Terminology Criteria for Adverse Events (CTCAE) Version 3 were performed on day 1 of each cycle. Blood pressure and pulse were measured every cycle with measurements made prior to commencing aflibercept, at 30 minutes into the infusion and 30 minutes post infusion during cycle 1 and 2. Radiologic assessment of measureable disease was performed by computed tomography (CT) or magnetic resonance imaging (MRI) within 28 days prior to registration and every 4 cycles (8 weeks). Response was defined using Response Evaluation Criteria in Solid Tumors (RECIST) [23].
EMBO J. 2011 February 16; 30(4): 719–730.
Image acquisition and analysisFixed-cell images were acquired using a Zeiss LSM510 confocal microscope. Z-stacks of 6–12 images were taken at 2048 × 2048 resolution, optical slice depth of 1 μm per image, and z-step of 0.37 μm. At the time of acquisition, laser power was adjusted so that all spines were below the threshold of saturation. Analysis was by ImageJ software (NIH). Maximum intensity projections were processed to smooth contours and a binary mask was obtained after edge detection. The cross-sectional area, and number of spines was calculated. For each condition, 90–100 μm sections of secondary dendrite from each neuron were analysed, 4–6 neurons per experiment, from three separate experiments, resulting in 700–1400 spines per condition. Experiments were both imaged and analysed with the experimenter blind to the experimental conditions. Statistical analyses were performed in Excel (Microsoft). The Student's t-test was performed on spine density data. Cumulative plots were analysed using Kolmogorov–Smirnov test (K–S test).Time lapse live confocal images of dendritic spines were acquired using a Nikon Eclipse Ti-E microscope. Z-stacks of 15–20 images were taken at various time points at 512 × 512 resolution with a z-step size of 0.4 μm. Neurons were continually perfused at 35°C with HBS at a flow rate of 4 ml/min. For chem-LTD, buffer was switched to HBS containing 20 μM NMDA and 20 μM glycine for 3 min, followed by return to normal HBS. For PMA treatment, perfusion buffer was HBS containing 50 μM D-AP5, 5 μM NBQX and 0.5 μM PMA. Baseline perfusion buffer was the same minus PMA. Glutamate receptor antagonists were present to prevent any PKC-stimulated release of endogenous glutamate from activating postsynaptic glutamate receptors (Calabrese and Halpain, 2005). Analysis was by ImageJ. Maximum intensity projections were produced for each time point and the images were processed and analysed as above.
BMC Fam Pract. 2013; 14: 45.
Clinical outcome measuresFor the cost-utility analysis, the EuroQol-5D with three levels for each of the five health dimensions (EQ-5D-3L) was used to assess quality of life at baseline, and at 6, 12 and 24month follow-up [10]. Health utilities were estimated with the Dutch tariff [11]. QALYs were calculated by multiplying the utilities with the amount of time a participant spent in a particular health state. Transitions between health states were linearly interpolated.Outcome measures of the cost-effectiveness analyses were the estimated risk of developing T2DM and the estimated risk of CVD mortality. The 9-year risk of developing T2DM was estimated with the risk formula derived from the Atherosclerosis Risk In Communities (ARIC) Study, based on ethnicity, parental history of diabetes, systolic blood pressure, waist circumference, and height [12]. This formula was selected because of its potential applicability in primary care, since it includes only non-invasive methods to assess participants’ T2DM risk. The 10-year risk of CVD mortality was estimated with the formula developed by the Systematic COronary Risk Evaluation (SCORE) project [9] which includes sex, smoking status, total cholesterol, and systolic blood pressure. The application of the SCORE formula was considered to be the most useful for the calculation of CVD mortality risk. Former comparative analyses of the SCORE, the Framingham and the UKPDS formulae demonstrated that the Framingham function may overestimate an individual’s absolute chronic heart disease risk, and the UKPDS formula was not chosen based on its specific inclusion of T2DM as variable [13]. Baseline risks and follow-up risks were estimated while standardizing the age of each participant at 60years. The formulas and the estimated risks are further explained in Additional file 1. Data was collected by means of physical measurements and questionnaires. A detailed description of the data collection has previously been published [5].
