A review of the control of breathing during exercise

During the past 100 years many experimental investigations have been carried out in an attempt to determine the control mechanisms responsible for generating the respiratory responses observed during incremental and constant-load exercise tests. As a result of these investigations a number of different and contradictory control mechanisms have been proposed to be the sole mediators of exercise hyperpnea. However, it is now becoming evident that none of the proposed mechanisms are solely responsible for eliciting the exercise respiratory response. The present-day challenge appears to be one of synthesizing the proposed mechanisms, in order to determine the role that each mechanism has in controlling ventilation during exercise. This review, which has been divided into three primary sections, has been designed to meet this challenge. The aim of the first section is to describe the changes in respiration that occur during constant-load and incremental exercise. The second section briefly introduces the reader to traditional and contemporary control mechanisms that might be responsible for eliciting at least a portion of the exercise ventilatory response during these types of exercise. The third section describes how the traditional and contemporary control mechanisms may interact in a complex fashion to produce the changes in breathing associated with constant-load exercise, and incorporates recent experimental evidence from our laboratory.     

Teaching acid/base physiology in the laboratory

Acid/base homeostasis is one of the most difficult subdisciplines of physiology for medical students to master. A different approach, where theory and practice are linked, might help students develop a deeper understanding of acid/base homeostasis. We therefore set out to develop a laboratory exercise in acid/base physiology that would provide students with unambiguous and reproducible data that clearly would illustrate the theory in practice. The laboratory exercise was developed to include both metabolic acidosis and respiratory alkalosis. Data were collected from 56 groups of medical students that had participated in this laboratory exercise. The acquired data showed very consistent and solid findings after the development of both metabolic acidosis and respiratory alkalosis. All results were consistent with the appropriate diagnosis of the acid/base disorder. Not one single group failed to obtain data that were compatible with the diagnosis; it was only the degree of acidosis/alkalosis and compensation that varied.     

Does the type of exercise affect tryptophan catabolism in horses?

Tryptophan (Trp) is an essential amino acid which metabolises via the kynurenine pathway to generate a number of bioactive substances referred to as kynurenines. Among those are 3-hydroxykynurenine (3-HKyn) and quinolinic acid, which are neurotoxic, as well as kynurenic acid (Kyna) and xanthurenic acid (XA), which, similarly to nicotinamide (NAm), show neuroprotective and anti-depressive effects. Routine exercise is known to modulate Trp metabolism in skeletal muscle and is thus believed to reduce the risk of depressive states in humans and laboratory animals. Analogously, it was hypothesised that exercise can influence Trp metabolism in horses as well. The aim of this study was to evaluate the influence of two different types of exercise on Trp metabolism in horses of the same breed. A total of 32 purebred Arabian horses were involved in the study. The 22 three-year-old racehorses were subjected to short-time intense exercise. Ten other horses were made to perform endurance competitions at a distance of 80 km. Blood samples were collected at rest and following the end of the exercise period. Plasma concentrations of Trp, kynurenine (Kyn), Kyna, 3-HKyn, XA and NAm were determined using Ultra-High Performance Liquid Chromatography-Electrospray Ionisation-Tandem Mass Spectrometry. Short-time intense exercise led to an increase in plasma concentrations of Kyn, Kyna and XA. The endurance effort induced an increase in Kyna and a decrease in Trp and NAm levels. Both types of exercise, short-time intensive exercise and endurance exercise induced an increase in Trp metabolites, especially Kyna, and did not induce an increase in Trp level. Thus, from a pathophysiological perspective of the kynurenine pathway’s influence on mental state, both types of exercise induced beneficial effects in horses.     

A computer simulation of free-range exercise in the laboratory.

We present a technique for simulating dynamic field (free-range) exercise, using a novel computer-controlled cycle ergometer. This modified cycle ergometer takes into account the effect of friction and aerodynamic drag forces on a 70-kg cyclist in a racing position. It also affords the ability to select different gear ratios. We have used this technique to simulate a known competition cycle route in Cape Town, South Africa. In an attempt to analyze the input stimulus, in this case the generated power output of each cyclist, eight subjects cycled for 40 min at a self-selected, comfortable pace on the first part of the simulated route. Our results indicate that this exercise input excites the musculocardiorespiratory system over a wide range of power outputs, both in terms of amplitude and frequency. This stimulus profile thereby complies with the fundamental requirement for nonlinear (physiological) systems analysis and identification. Through a computer simulation, we have devised a laboratory exercise protocol that not only is physiologically real but also overcomes the artificiality of most traditional laboratory exercise protocols.     

Teaching baroreflex physiology to medical students: a comparison of quiz-based and conventional teaching strategies in a laboratory exercise

Quiz-based and collaborative teaching strategies have previously been found to be efficient for the improving meaningful learning of physiology during lectures. These approaches have, however, not been investigated during laboratory exercises. In the present study, we compared the impact of solving quizzes individually and in groups with conventional teaching on the immediate learning during a laboratory exercise. We implemented two quizzes in a mandatory 4-h laboratory exercise on baroreflex physiology. A total of 155 second-year medical students were randomized to solve quizzes individually (intervention group I, n = 57), in groups of three to four students (intervention group II, n = 56), or not to perform any quizzes (control; intervention group III, n = 42). After the laboratory exercise, all students completed an individual test, which encompassed two recall questions, two intermediate questions, and two integrated questions. The integrated questions were of moderate and advanced difficulty, respectively. Finally, students completed an evaluation form. Intervention group I reached the highest total test scores and proved best at answering the integrated question of advanced difficulty. Moreover, there was an overall difference between groups for student evaluations of the quality of the teaching, which was highest for intervention group II. In conclusion, solving quizzes individually during a laboratory exercise may enhance learning, whereas solving quizzes in groups is associated with higher student satisfaction.     

HSP70 expression: does it a novel fatigue signalling factor from immune system to the brain?

Integrative physiology studies have shown that immune system and central nervous system interplay very closely towards behavioural modulation. Since the 70-kDa heat shock proteins (HSP70s), whose heavy expression during exercise is well documented in the skeletal muscle and other tissues, is also extremely well conserved in nature during all evolutionary periods of species, it is conceivable that HSP70s might participate of physiologic responses such as fatigue induced by some types of physical exercise. In this way, increased circulating levels of extracellular HSP70 (eHSP70) could be envisaged as an immunomodulatory mechanism induced by exercise, besides other chemical messengers (e.g. cytokines) released during an exercise effort, that are able to binding a number of receptors in neural cells. Studies from this laboratory led us to believe that increased levels of eHSP70 in the plasma during exercise and the huge release of eHSP70 from lymphocytes during high-load exercise bouts may participate in the fatigue sensation, also acting as a danger signal from the immune system.     

Laboratory demonstration of vascular smooth muscle function using rat aortic ring segments

This laboratory exercise uses a simple preparation and a straightforward protocol to illustrate many of the basic principles of vascular biology covered in an introductory physiology course. The design of this laboratory allows students to actively participate in an exercise demonstrating the regulation of arterial tone by endothelial and extrinsic factors. In addition, this hands-on laboratory allows students to gather data using well-known basic biomedical research techniques. Specifically, students are introduced to an isolated organ-chamber technique that is widely used to study cellular mechanisms of many tissues including vascular smooth muscle contraction and dilation. On the basis of student evaluations, participation in the experiments and interpreting data reinforce lecture materials on smooth muscle and endothelial cell function and illustrate mechanisms regulating vascular tone. Students come away with a greater understanding of vascular biology, a deeper appreciation of integrative physiology, and an understanding of the process of conducting tissue-chamber experiments.     

Fish on Prozac: a simple, noninvasive physiology laboratory investigating the mechanisms of aggressive behavior in Betta splendens

The neuromodulator serotonin is an important regulator of aggressive behavior in vertebrates. Experimentally increasing synaptic levels of serotonin with fluoxetine, a selective serotonin reuptake inhibitor, has been shown to reliably decrease the expression of aggressive behavior. Here, we describe a method by which fluoxetine can be noninvasively administered to male Betta splendens (an attractive model for the study of aggressive behavior) and describe a simple laboratory exercise that allows students to experimentally investigate the physiological mechanisms of aggressive behavior. We demonstrate that relatively short-term exposure (3 h) of male bettas to as little as 3 microg/ml of fluoxetine-treated aquarium water is sufficient to reduce the expression of specific aggressive behaviors. We emphasize the physiological concepts that can be addressed with this exercise, including the role of the serotonergic system in regulating aggression, and the interplay of environmental contaminants and physiology in regulating the expression of behavior. We also highlight important aspects of experimental design. This exercise can be flexibly altered to accommodate one or several laboratory periods. It is also low cost, is low impact to the animals, and requires minimal preparation time for instructors.     

跑台运动在动物实验研究中的应用

跑台在实验动物研究中的应用越来越多,主要集中在建立运动模型以及探究运动对生理和病理的作用,以促进运动训练的科学化、竞技体育成绩的提高、运动疗法在疾病康复中合理有效的应用等,同时也是研究运动与代谢,运动与心脑血管病等的研究工具。本文对动物跑台的基本构造、使用方法和研究进展进行了分析和总结,为利用跑台进行研究提供一定的参考。     

The use of scenario-based-learning interactive software to create custom virtual laboratory scenarios for teaching genetics

Mutagenesis screens and analysis of mutant phenotypes are one of the most powerful approaches for the study of genetics. Yet genetics students often have difficulty understanding the experimental procedures and breeding crosses required in mutagenesis screens and linking mutant phenotypes to molecular defects. Performing these experiments themselves often aids students in understanding the methodology. However, there are limitations to performing genetics experiments in a student laboratory. For example, the generation time of laboratory model organisms is considerable, and a laboratory exercise that involves many rounds of breeding or analysis of many mutants is not often feasible. Additionally, the cost of running a laboratory practical, along with safety considerations for particular reagents or protocols, often dictates the experiments that students can perform. To provide an alternative to a traditional laboratory module, we have used Scenario-Based-Learning Interactive (SBLi) software to develop a virtual laboratory to support a second year undergraduate course entitled "Genetic Analysis." This resource allows students to proceed through the steps of a genetics experiment, without the time, cost, or safety constraints of a traditional laboratory exercise.     

The pentose phosphate pathway in the yeasts Saccharomyces cerevisiae and Kloeckera apiculata,an exercise in comparative metabolism for food and wine science students

Comparative cellular metabolism can be a difficult area of biochemistry to teach in the undergraduate laboratory class. Student practicals involving animal tissues generally require approval from animal ethic committees, and the relevance for students whose primary interest in biochemistry is in the area of food and wine sciences, is often questioned. In this report, we present an undergraduate practical exercise in which glucose catabolism via the pentose phosphate pathway is compared in two types of yeast with direct relevance to the wine and food industries, Saccharomyces cerevisiae and Kloeckera apiculata. The exercise is carried out as a demonstration to second year undergraduate students, studying metabolic biochemistry. It is of some value in that it illustrates comparative cellular metabolism in wine yeasts and introduces the students to the safe use of radioisotopes.     

Glucose transport in cultured animal cells: an exercise for the undergraduate cell biology laboratory

Membrane transport is a fundamental concept that undergraduate students of cell biology understand better with laboratory experience. Formal teaching exercises commonly used to illustrate this concept are unbiological, qualitative, or intricate and time consuming to prepare. We have developed an exercise that uses uptake of radiolabeled nutrient analogues by attachment-dependent animal cells cultured on multiwell trays. This system can readily be manipulated within a typical 3-h laboratory period to yield reproducible, biologically relevant, quantitative data regarding key aspects of membrane transport. Each 24-well tray of cultures allows a group of two to four students to compare eight conditions in triplicate. If different groups of students test different conditions or different types of cells, data can be shared for an even broader experience. The exercise is also readily adaptable for open-ended student projects. Here we illustrate the exercise measuring uptake of the nonmetabolizable glucose analogue [(3)H]-2-deoxy-D-glucose. Students successfully tested the effects of competing sugars, putative inhibitors of the GLUT1 transporter, and changes in cell physiology that might be expected to affect glucose transport in epithelial cells and fibroblasts. In this exercise students find the nutritional and medical implications of glucose transport and its regulation intriguing. They also learn to handle radioisotopes and cultured cells.     

Greenbeards in yeast: an undergraduate laboratory exercise to teach the genetics of cooperation

Recent years have seen a dramatic increase in our understanding of the social behaviour of microbes. Here, we take advantage of these developments to present an undergraduate laboratory exercise that uses the cooperative flocculating behaviour of yeast (Saccharomyces sp.) to introduce the concept of inclusive fitness and teach the genetics of cooperation. Students generate their own data using co-cultures of various yeast strains and perform statistical analyses to test whether kin selection or greenbeard effects determine the cooperative flocculating behaviour. The lab has run successfully for two consecutive years in a second year course with some 1, 200 students per year at the University of Toronto, Canada. We discuss the benefits of using microbes to teach social evolution, describe the set-up and learning outcomes of the laboratory exercise, and then outline possible extension and variants of the lab. In addition to providing students with the opportunity to use a model organism to study social behaviour, students are also taught common laboratory skills, such as replica plating and sterile techniques. Ultimately, while the genetics of cooperation has traditionally been taught through computer simulations and evolutionary games, this exercise demonstrates a way to experimentally introduce the topic.     

Different cells make different proteins: a laboratory exercise illustrating tissue-specific protein expression in animals

All the cells of higher organisms have the same DNA but not the same proteins. Each type of specialised cell that forms a tissue has its own pattern of gene expression and, consequently, it contains a particular set of proteins that determine its function. Here, we describe a laboratory exercise addressed to undergraduate students that aims to reinforce the following concept: what makes cells different is they make different proteins. The students performed a denaturing polyacrylamide gel electrophoresis (SDS–PAGE) to separate the proteins extracted from different animal tissues, and then obtained and compared the corresponding protein profiles. In addition, the students performed a Western blot analysis to detect the presence or absence of a tissue-specific protein in the different tissue extracts. This laboratory exercise allowed students to understand the basis of the cell differentiation better and also provided them the opportunity to learn a variety of analytical laboratory techniques.     

Body fat of mice and men: A class exercise in theory or practice

An adaptation of some methods used in obesity research is presented as a teaching example to illustrate the use of 'animal models' in medical research. From sixth-form level upwards, it serves as a theoretical exercise in the analysis and interpretation of methods and data. For undergraduates it is also suitable as a laboratory exercise in dissection and measurement techniques.

Four simple means of altering fat levels in laboratory mice are described, contrasting invasive injection techniques with non-invasive dietary and behavioural means. Several measures of body fatness can be evaluated, using either the experimental mice or, more simply, untreated mice from outbred stocks. Guidelines are given for practicals in which objective and subjective measures of fatness in human subjects can be collected. Sample data for mice are given that, alone or together with class-collected data, allow graphical and statistical analysis. Throughout, the exercise lends itself to discussion of the assumptions both of the methods used on mice and men, and of the relation of such investigations to the problems of overweight in man. Obesity has many manifestations: the study of a variety of ‘animal models’ is one approach in the search for relevant physiological knowledge.     

Beta-blockade and oxygen delivery to muscle during exercise.

The most commonly observed effect of beta-blockade on cardiovascular function has been a reduction in heart rate both at rest and during exercise. The body attempts to compensate by increasing stroke volume and (or) increasing the extraction of O2 from the blood to maintain O2 delivery to the muscle. This paper examines the roles of muscle mass involved in the exercise as well as the time course of change in cardiac output and peripheral blood flow in an attempt to understand whether O2 supply is limited by beta-blockade. Experiments are reported in which the kinetics of cardiac output response at the onset of submaximal cycle exercise were slowed in subjects taking oral propranolol. Taken in consideration with other data from our laboratory and with data in the literature, it was concluded that beta-blockade does impair O2 transport. The degree of impairment is dependent on the total muscle mass involved and the metabolic demand.     

Thinking about evolution: combinatorial play as a strategy for exercising scientific creativity

An enduring focus in education on how scientists formulate experiments and ‘do science’ in the laboratory has excluded a vital element of scientific practice: the creative and imaginative thinking that generates models and testable hypotheses. In this case study, final‐year biomedical sciences university students were invited to create and justify a taxonomy of selected vertebrates on the basis of their brain organisation, as part of an exercise exploring the evolution of embryonic development. While raising a number of issues surrounding the context and methods of comparative zoology, this exercise also invoked a set of cognitive processes that can neither be adequately characterised as role‐play nor critical thinking. By contrast, the act of formulating and justifying taxonomy identifies a style of creative thought that is a prerequisite for hypothesis formation. A defining characteristic of this exercise is that it engages activities that are independent of disciplinary perspective. This flexibility in approach may provide a route through to defining what qualifies as a creative teaching exercise in science.     

Using "spinal shrinkage" as a trigger for motivating students to learn about obesity and adopt a healthy lifestyle

Obesity is a global problem; however, relatively little attention is directed toward preparing and inspiring students of medicine and allied medical sciences to address this serious matter. Students are not routinely exposed to the assessment methods for obesity, its overall prevalence, causative factors, short- and long-term consequences, and its management by lifestyle modification. This physiology laboratory exercise involving students of medicine (n = 106) was developed to 1) introduce medical students to methods of obesity assessment and to differentiate between general and abdominal obesity, 2) generate an interest and sensitivity about obesity, and 3) stimulate thinking about modification of their lifestyle in relation to eating habits, weight control, and physical activity. Spinal shrinkage (the difference between the standing height of a person and his/her recumbent length) was used as an immediate observable parameter to demonstrate the effect of adiposity. Spinal shrinkage is recognized as an index of the compressive forces acting on the spine and is related to body mass index. A positive correlation (r = 0.365, P < 0.05) was observed between body mass index and spinal shrinkage. A questionnaire was used to assess student responses to this exercise. Students were motivated to engage in more physical activity (74%), adopt healthier eating (63%), and enhance their knowledge about obesity (67%). They expressed keen interest in the laboratory exercise and found the sessions enjoyable (91%). The laboratory exercise proved to be a success in motivating the students to actively learn and inquire about obesity and to adopt a healthier lifestyle.     

Wheel running in the wild

The importance of exercise for health and neurogenesis is becoming increasingly clear. Wheel running is often used in the laboratory for triggering enhanced activity levels, despite the common objection that this behaviour is an artefact of captivity and merely signifies neurosis or stereotypy. If wheel running is indeed caused by captive housing, wild mice are not expected to use a running wheel in nature. This however, to our knowledge, has never been tested. Here, we show that when running wheels are placed in nature, they are frequently used by wild mice, also when no extrinsic reward is provided. Bout lengths of running wheel behaviour in the wild match those for captive mice. This finding falsifies one criterion for stereotypic behaviour, and suggests that running wheel activity is an elective behaviour. In a time when lifestyle in general and lack of exercise in particular are a major cause of disease in the modern world, research into physical activity is of utmost importance. Our findings may help alleviate the main concern regarding the use of running wheels in research on exercise.     

Integration of laboratory exercises in developmental biology and neurobiology courses using the Xenopus oocyte expression system

This collaborative laboratory exercise integrates two upper division laboratory courses (Developmental Biology and Neurobiology) offered to biology majors at Wake Forest University. The laboratory exercise involves the use of the Xenopus oocyte expression system to study the function of specific membrane receptors and ligand-activated channels. cDNA or mRNA for receptor proteins is injected into Xenopus oocytes. The oocytes are assayed for expression of receptor proteins and two-electrode voltage clamping is done to determine whether the expressed proteins are functional in the oocyte system. This series of laboratory exercises is innovative in its interdisciplinary and collaborative approach to undergraduate teaching, and in its use of sophisticated molecular biological and physiological techniques in the undergraduate teaching laboratory. Students learn first-hand how these techniques have been used to achieve a new level of understanding of both development and neurobiology. Journal of Industrial Microbiology & Biotechnology (2000) 24, 353–358. Received 02 April 1999/ Accepted in revised form 10 November 1999