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Pre-release title. On Special. Firm Sale. Internationally Sourced. Stress results in activation of the sympathetic nervous system, which in turn leads to increased thermogenesis and vasoconstriction of skin vessels resulting in divergence of core and surface temperature 25 — In the present study, the perianal region was used for its accessibility and need for only minimal animal handling during surface temperature acquisition. Thus, the difference between core and surface temperatures can be attributed to variations in ambient temperature, measurement location and thermoregulatory responses, handling stress, and the intentional use of standard non-contact infrared thermometers without a calibration feature.

Nevertheless, we found a strong correlation between core and surface temperatures. Genotype-dependent effects on body temperature were most pronounced in homozygous Cd11b knockout animals whose hypothermic response to LPS was significantly reduced following the second injection, which might be attributable to a conditioning effect of repeated inflammatory stimuli. In addition, Cd11b knockout animals had a lower surface temperature following the second LPS injection which might be due to an altered thermoregulatory response of this genotype.

A threshold determined by body temperature was used as a refined humane endpoint in previous studies.

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These methods only allow a parameter search with large increments 0. To identify the parameter that could be used as the humane endpoint in the present study, we applied an automatized parameter search with finer increments in the predictor 0. In addition, previous studies refrained from conducting a comprehensive assessment of prediction models e. To our knowledge, this is the first study to determine a temperature-based threshold and to compare the prediction accuracies of core vs.

Factors other than variation of measurements need to be considered when assessing the relative merits of these two methods. Regarding surface temperature measurements, a standard non-contact infrared thermometer and only minimal training and technical expertise are sufficient. In contrast, core temperature measurements using temperature transponders require a dedicated RFID system consisting of passive usually non-reusable RFID transponders and a reader device; transponder implantation needs to be performed by an experimenter with previous surgical experience followed by regular checks for transponder functionality and dislodgement.

Thus, in the hands of a skilled animal technician, using infrared thermometry is likely to reduce handling time and animal distress. However, surface temperature measurements are prone to higher inter- and intra-subject variation and highly investigator-dependent, whereas transponder-based readings are more robust and less investigator-dependent. Advantages and limitations of the two temperature measurement techniques assessed in the present study. One important conclusion from our study is therefore that, given the above constraints of using implantable RFID temperature transponders, infrared thermometry is acceptable as surrogate whenever variation of measurements can be counterbalanced with multiple measurements or large numbers of animals.

However, to quantify subtle changes in temperature requires the use of the former despite being more cumbersome, expensive, and time consuming.

As the result, both core and surface temperature are equally suited to predict death allowing for termination of experiments at earlier time points to reduce unnecessary distress. Limitations of our study include the use of only one disease model, namely LPS-induced hypothermia. We chose an endotoxin model for its reproducibility and high translational relevance, as hypothermia is a common feature during severe illness in mice 28 — It would nevertheless be of interest to compare surface and core temperature measurements during hyperthermia as produced by stress or pharmacological intervention.

Brown adipose tissue is an important heat generator in mice and thus might have a confounding effect on core temperature readings Only female mice were used for this study. However, the stage of the oestrous cycle was not determined. Thus, physiological temperature fluctuations with the oestrous cycle may have had a confounding effect on temperature measurements. Both core and surface temperature gave rise to comparable accuracy in death prediction.

However, due to the small number of animals that died in the present study 6. Finally, because of transponder malfunction and technical errors during temperature acquisition, we had to exclude core temperature measurements from 22 animals partially or in their entirety from further analysis. In conclusion, this is the first study to apply systematic assessment of two distinct methods of temperature measurement in mice following an endotoxin challenge and to compare their predictive strengths towards death as an outcome event.

We find that both methods are adequately suited for the prediction of death, and hence that the less expensive and cumbersome non-contact infrared thermometry can serve as a reliable alternative for implantable transponder-based systems. This finding is of practical importance as it encourages adoption of simple temperature measurement tools to monitor disease progression and apply humane endpoints in mouse models of acute disease.

Animals were kept in specific-pathogen-free SPF conditions according to FELASA regulations and group-housed with ad libitum access to food and water in type III polycarbonate cages equipped with environmental enrichment tools red transparent plastic nest box and brown paper towels. Room temperature was maintained at To minimize confounding effects, injections and temperature measurements were scheduled at the same time each day and experimenters wore single-use coveralls Microgard , Ansell Microgard, Kingston Upon Hull, UK , gloves and surgical masks whenever in contact with animals.

Information on strain and treatment group assignment was concealed from experimenters until the end of the study. There were no specific exclusion criteria. Severity of disease was scored on a scale from 0 to 5 normal score, 0; maximum severity: 5. When the general activity and response to stimuli of an animal matched criteria from different severity levels, the average of the two severity levels was assigned as the sickness score.

When the passive RFID transponder is within read range, its internal antenna draws energy from the radio waves emitted by the reader. This energy powers the chip, which then sends data back to the reader. Three weeks prior to injection, temperature transponders were implanted subcutaneously in the region between the scapulae as described previously 13 , Illustration of the measurement modalities used to obtain core and surface temperature data. Red circle depicts perianal region used as the site for surface temperature acquisition. Non-contact infrared thermometers measure the infrared energy emitted by an object for estimating its temperature and can only be used to monitor surface temperatures.

During subsequent temperature acquisition, the base of the tail was fixed with two fingers and then gently lifted while the animal gripped a metal rod on the cage lid with its front paws, thus allowing for exposure of the ano-genital area.

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Temperature was measured in the perianal region, with a searchlight indicating the measured area Fig. To minimize confounding effects by urination or defecation, temperature measurements were only taken when the measurement area was clean. The sequence of core or surface temperature measurements was randomized. Handling was minimized to reduce stress and discomfort. To induce a systemic inflammatory response, animals were treated with lipopolysaccharide LPS , a cell wall component of Gram-negative bacteria. Louis, USA at a dose of 1.

Survival times were determined to fulfil the objectives of another study, for which these animals were used. For all animals, temperature measurements were repeated in triplicate. Results are expressed as mean SD unless otherwise specified. To assess the reliability of surface temperature in predicting the corresponding core temperature, a random intercept mixed effects model 36 , 37 with 3 levels was used to fit the data 1st level, temperature measures; 2nd level, time points where measures from different temperature monitoring methods were combined; 3rd level, animals , for its advantage in dealing with missing values caused by different survival times and measurement intervals.

Risk of death as an outcome event was examined with the scikit-learn toolkit sklearn 38 for both core and surface temperature. Electronic supplementary material. Supplementary information accompanies this paper at Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. National Center for Biotechnology Information , U. Sci Rep. Published online Feb Author information Article notes Copyright and License information Disclaimer. Julius Valentin Emmrich, Email: ed.

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Corresponding author. Received Nov 13; Accepted Feb This article has been cited by other articles in PMC. Abstract Body temperature is a valuable parameter in determining the wellbeing of laboratory animals. Introduction The Three Rs Replacement, Reduction and Refinement were introduced almost 60 years ago as guiding principles for humane animal research 1.

Open in a separate window. Figure 1. Figure 2. Table 1 Number of dead and transponder-implanted animals, time of death and last recorded core and surface temperatures before death following lipopolysaccharide LPS injection per genotype. MEI - English / Studying & Workbooks / Education Studies & Teaching: Books

Comparison of measurement modalities Surface temperature of the heating pad measured with infrared thermometer 1 was 1. Figure 3. Using core temperature or surface temperature to predict death To compare the accuracy of both core and surface temperature in the prediction of death as an outcome event, we developed a temperature-based death prediction model using machine learning algorithms.

Discussion The aim of this study was to compare and assess the respective merits of two commonly used procedures to assess body temperature in mice during acute illness following an endotoxin challenge. Figure 4. Table 2 Advantages and limitations of the two temperature measurement techniques assessed in the present study. Exclusion criteria and humane endpoints There were no specific exclusion criteria. Transponder implantation, anaesthesia, and temperature measurement For temperature acquisition using radio-frequency identification RFID technology, passive RFID transponders were implanted subcutaneously.

Figure 5. Non-contact infrared thermometry and temperature measurement Non-contact infrared thermometers measure the infrared energy emitted by an object for estimating its temperature and can only be used to monitor surface temperatures. Endotoxin-induced systemic inflammatory response To induce a systemic inflammatory response, animals were treated with lipopolysaccharide LPS , a cell wall component of Gram-negative bacteria.

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Data analysis and statistics For all animals, temperature measurements were repeated in triplicate. Electronic supplementary material Supplementary Materials K, pdf. Author Contributions J. Notes Competing Interests The authors declare no competing interests.