ONLY INTRODUCTION
Assignment: The analysis and presentation of Promethion metabolic cage derived data Task: You are required to write up a mock-journal article that includes an introduction, methods, results, discussion and conclusion. Introduction: A brief background summary. Methods: i.e. EchoMRI scan and Promethion metabolic cage. Results: Using the given data you will be required to analyze, run statistics and produce graphs. Done Discussion & conclusion: Talk about data with reference to literature and have strong, concise concluding remarks. Required for submission: · Word document of mock journal article · Excel spreadsheet/(s) of your own data analysis, statistical tests and graphs. Background: 4 Wild type control (C57b/10) mice 4 mdx mice 6 day stay in Prometheon EchoMRI pre and post for body composition All groups provided with the same pre- and post-treatment echoMRI body composition data and Prometheon data. All Groups will be required to analyze activity and body composition parameters: · Energy expenditure and gas parameters · Cage (non)Activity · Body composition All Groups will be required to make comparison of three items: · Acclimatisation to the Prometheon cages · Diurnal variation · Dystrophic vs non-dystrophic (main comparison) In vivo Analysis of Acclimatisation, Metabolism and Body Composition of wild type (C57b/10) mice and mdx mice inside Prometheon cage system. Abstract Introduction: Additionally, we aimed to investigate Methods: Results: Conclusion: Introduction: Materials & Methods: Results: Table 1: Average Energy expenditure of each group of mice during the 6-day period WT (C57b/10) 0.653 mdx 0.564 Fig. 1: Comparison of total energy expenditure of WT and mdx mice groups taken inside the prometheon cages with a total duration of 6 days Table 2: averaged wheel running comparison for WT and mdx mice Distance (m) WT 7290.9 mdx 6465.6 Fig.2: comparison of the distance ran by each mice group using the running mill inside the prometheon cage. Table 3: averaged pedestrian distance of each mice group inside the prometheon cage. Pedestrian distance refers to the activity of the mouse inside the cage (eg, wandering, walking around). Distance (m) WT 139.8 mdx 196.4 Fig 3: pedestrian distance travelled by each mouse group inside the prometheon cage, representing their activeness inside the cage. Measured during nocturnal hours. Table 4: averaged non-activity of each WT and mdx group during nocturnal hours x-beam reading WT 34.97 mdx 42.08 Fig 4: graph showing the non-activity (lounging) of each mouse group inside prometheon cage Table 5: measured average of duration of sleep (diurnal) WT 87.14 mdx 80.77 Fig 5: comparison of WT and mdx mice and their sleep cycle Table 6: Averaged gas parameters of WT and mdx mice inside the prometheon cage. VO2 VCO2 mdx 1.88544513 1.77779433 WT 2.18943846 2.05363658 Figure 6: VO2 & VCO2 reading of each mouse group during the 6-day period Fig 7: Fat percentage of WT and mdx mice before and after the 6-day duration inside the prometheon cage system Fig 8: lean muscle mass measurements using echoMRI, comparing before and after the 6-day period inside the prometheon cages Fig 9: water composition of both WT and mdx mice before and after the 6-day period inside the prometheon cage system Fig 10: graph that shows the weight difference before and after the 6-day period in prometheon cage system Discussion: Conclusion: Sleep (diurnal) WTmdx87.13578099999999480.772418000000002 x beam Gas Parameters mdxVO2VCO21.88544512999999991.7777943300000001WTVO2VCO22.18943845999999992.05363658 mL/min WT vs mdx Energy Expenditure (Nocturnal) WT (C57b/10)mdx0.653070524999999960.56380677899999998 x-beam reading WT vs. mdx Wheel Meters WTmdx7290.90177800000046465.5850829999999 distance (m) wt vs mdx ped distance WTmdx139.831163196.42083299999999 meters Still WTmdx34.97193332999999942.081790419999997 x beam Week 4 – Workshop and Lab in vivo Analysis of Physical Activity, Metabolism and Body Composition Learning objectives - Learn how to conduct live animal investigations (EchoMRI and Promethion Metabolic System) - The principle of both systems - Determine primary outcomes of experimental design and interrogate the differences obtained o Mouse phenotypes (control strain versus mdx) o Time-point measures EchoMRITM: Body composition analysis The EchoMRI™ Analyzers deliver precise body composition measurements of fat, lean, free water, and total water masses in live animals weighing up to 500, 700, 900 and 1100 grams. Scanning takes 0.5 – 3.2 minutes, depending on the precision options. The live animals need no anesthesia and no special preparation before measurement. EchoMRI™ Analyzers are exceedingly easy to operate. A training period of one hour is sufficient for individuals without prior knowledge. Numeric results are stored and can be viewed again later at any time, as well as extracted either into Excel and ASCII files or into Access database. Figures above are from two different research papers from our lab. The lean and fat mass index results are collated and finalised from raw EchoMRI data. Here, the average values of the triplicate scans are taken and divided by the animal’s total body mass. Promethion Metabolic Cage System The Promethion cage system allows for in vivo measurement of many physical activity and metabolic parameters. Promethion uses pull-mode air flow generators, allowing the use of an unsealed “Home” cage environment for your animal. Cage activity and monitoring functions, including running wheel, food hopper, water bottle and scale, are built into the cage lid. You simply lift the lid to clean the cage, leaving the animal in the security of its home cage. There is no handling of the mouse, no unfamiliar new environment. This is a significant advancement over conventional systems sealed cage systems, which can cause stress effects and require long acclimation times. In our studies, we typically house our mice for up to 1 week to allow for acclimitisation and familiarisation. Specifically, there are many aspects of physical activity, behaviour and metabolism that we can elucidate from this system including: Energy expenditure (measured in kilocalories) during various levels of physical activity Wheel activity (distance, time, velocity) Pedestrian meters (non-wheel activity) Respiratory exchange ratios (RER)/substrate utilisation Food and water intake Sedentary/inactivity/sleep Changes in body mass Faecal and urine collection Activity monitoring (location in cage) With data recorded in real time, specific measures can be correlated and contrasted appropriately. For example: RER can be correlated with running wheel intensity or sedentary/sleep at that given time point – indicating substrate utilisation at that work rate. Mouse cage The standard configuration model 3721 Mouse Cage comes with 2 cage tubs, cage lid with 3 accessory holes, air flow manifold with filters and tubing, an acrylic block and filter cover. Items include food/water hopper, body mass bracket and running wheel capacities and are fixed in specific locations. Total Activity Monitoring The BXYZ Beambreak Activity Monitor enables real time analysis of total activity with a 0.25cm calculated centroid. Designed to be modular, the BXYZ can easily be used stand alone, or synchronized with metabolic measurement and other elements of the Promethion system. It is capable of ignoring fixed objects, allowing flexibility to use standard or customized cages without interfering with activity monitoring. Using Sable data analysis software, you’ll have graphic representation of position within the cage, calculation of total distance travelled, rearing information, and storage of raw position vs. time data for other traceable, quantifiable analyses of activity level. The black ‘beam breaks’ are displayed below and surround the cage for accurate detection of raw positioning. Wheel Activity Monitoring The Promethion wheel monitoring system incorporates a durable, stainless steel wheel into the cage for 180 or 360 degree monitoring of voluntary wheel revolutions. Designed to integrate with calorimetry, the wheel monitoring data can easily be synchronized with RQ, VO2, beambreak activity, or any other parameter of the Promethion system. The in- cage enrichment wheel provided with the Promethion wheel monitoring system is designed not to restrict flow around the wheel ensuring the most accurate and repeatable calorimetry data. The system provides superior temporal resolution with real time monitoring of revolutions per minute, permitting energy expenditure during running and resting periods to be easily differentiated. Food and Water Intake Monitoring The Promethion food intake monitoring system features high precision sensors capable of measuring real time food intake for mice and rats. The MM-1 Load Cell with 3mg resolution can be used with the food hopper or any other Promethion mass measurement device, allowing maximum flexibility in feeding such as varying the location of the hopper within the cage and feeding liquid diets. The AC-2 Access Control Module can be setup for paired feeding, yoked feeding, time and duration limited feeding, and quantity limited feeding providing unmatched flexibility in experimental design. The Promethion food intake monitoring system can be incorporated into existing cage systems allowing the use of standard cages and requiring no cage modifications reducing animal stress during acclimation. The Promethion water intake monitoring system includes a standard water bottle and a universal MM-1 Load Cell with a 3mg resolution. The MM-1 load cell can be used with many different water bottle sizes and shapes allowing the use of existing cleaning, sterilizing and filling supplies. The AC-2 Access Control Module allows controlled access to liquid diets or water consumption. The system provides real time monitoring, allowing for drip mitigation that differentiates between drinking bouts and dripping events. Promethion also provides air flow through the cage keeping bedding dry leading to healthier, happier animals. The Promethion water intake monitoring system can be incorporated into existing cage systems allowing the use of standard cages and requiring no cage modifications reducing animal stress during acclimation. Food hopper Water hopper Body Mass Monitoring Promethion body mass monitors are in-cage enrichment devices attached to a Promethion universal MM-1 load cell. The body mass monitor allows the real time recording of body mass when the animal interacts with the device. The real time data from the Promethion body mass monitor provides automated, voluntary weighing of the animal – up to 800 mg – resulting in frequent measurements of body mass and no handling stress on the animal during weighing. Recording of body mass can be combined with food and water intake, and synchronized with metabolic measurement, giving the investigator access to a more complete picture of energy expenditure. Data extraction and acquisitioning ExpeData allows the user to explore acquired data in a rich graphic environment where each parameter of the system has been recorded once per second enabling real time analysis or user-selectable temporal resolution for comparison to previous research. ExpeData automated analysis scripts can be run