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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 ANATOMY & PHYSIOLOGY II Table of Content Experiments: 1.

Cardiovascular System 2. Cardiovascular Physiology 3. Lymphatic System 4. Respiratory System 5. Lung Volume and Capacities 6. Digestive System 7. Urinary System 8. Renal Physiology 9. Reproductive System Appendix A: Heart Appendix B: Respiratory Pathways Appendix C: Digestive System Appendix D: Urinary System Appendix E: Reproductive Systems References

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Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Experiment 1: Cardiovascular System HEART The heart is a pump with four chambers and valves that maintain a one-way flow of blood. The wall of the heart includes: (1) cardiac musculature for contraction to propel the blood out of each chamber and into the major vessels, (2) a fibrous skeleton for attachment of the valves and (3) an internal conducting system for synchronization of muscle contraction. Purkinje Fibers Purkinje fibers are modified cardiac muscle fibers found in the subendocardium of the ventricles. They constitute part of the specialized impulse conducting system, which connects to the right and left bundle branches and regulates the heartbeat. These are large muscular fibers with a vacuolated cytoplasm due to the high glycogen content. Other characteristics that help distinguish Purkinje fibers from typical cardiac muscle fibers are that they contain fewer myofibrils, and more sarcoplasm. Part A: Gross Anatomy Identify the heart, its chambers and the associated blood vessels. Label the heart in Appendix A. From the human heart model, notice how much thicker the myocardium of the left ventricle is than that of the right ventricle. Compare the shape of the left ventricular cavity to the shape of the right ventricular cavity. Record your observation. Part B: Histology The cardiovascular system consists of the heart and blood vessels. The heart is a specialized blood vessel that acts as a pump to circulate the blood. Knowledge of the structure and function of blood vessels is important in understanding vascular diseases, the leading cause of death in this country. Blood vessels are divided into three groups: arteries, veins, and capillaries. Two other types of atypical blood vessels are sinusoidal capillaries and sinusoids. Arteries and veins are further divided, according to size, into large, medium, and small blood vessels. The vascular system is subjected to varying degrees of hydrostatic pressure, and the structure of vessels varies in an adaptive fashion. Blood vessels are thickest and their walls more complex in the immediate vicinity of the heart, where hydrostatic pressure is greatest. As blood vessels decrease in size their wall becomes thinner and less complex. Arteries and veins of large or medium caliber have three tunics: tunica intima, tunica media, and tunica adventitia. The tunica adventitia contains small blood vessels (vasa vasorum), which supply nutrients to tissues in the outer one-half of the wall of the blood vessel. Small nerves, representing fibers of the autonomic nervous system, are also present in the tunica adventitia and they innervate the smooth muscle of the vessel. Tabulate the important identifying characteristics you can use to differentiate between the types of medium blood vessels. Slides: Heart: observe the cardiac muscle. Identify the nucleus, striations, intercalated discs, and sarcolemma of the individual cells. Aorta, Vena Cava Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science 1

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Part C: Physiology 1. Heart sound The beating of human heart usually produces four sounds, but only two may be detected with a stethoscope. The sounds of the heartbeat are usually described as lub-dup (pause) lub-dup, and so forth. These sounds are produced by blood turbulence caused by the closure of the heart valves. The first sounds results from the closing of the atrioventricular valves during ventricular systole. The second sound results from the closing of the semilunar valves during ventricular diastole. If any of the heart valves are defective and do not close properly, an additional sound known as a heart murmur, may be heard. Use of stethoscope Procedure: 1. The stethoscope should be used in a quite room. 2. The earpiece of the stethoscope should be cleaned with alcohol just before using and should also be pointed slightly forward when placed in the ear. They will be more comfortable and easier to hear through them. 3. Listen to the heart sound of your laboratory partner by placing the diaphragm of the stethoscope at several positions on the chest wall. 4. The first sound is best heard at the apex of the heart. 5. The second sound is best heard in the second intercostals space. 6. CAUTION! Assuming that your partner has no known or apparent cardiac or other health problems and is capable of such an activity, ask him/her to run in place about 25 steps. Listen to the heart sounds again. a. Which heart sounds is the loudest? b. Did you hear a third sound? c. Where does the first sound originate? d. Where does the second sound originate? e. After you had exercised, how did the heart sounds differ from before? f. Did they differ in rate and intensity? g. Did the first or second increase in loudness?

Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 2. Pulse Rate A wave of pressure, called pulse, is produced in the arteries due to ventricular contraction. Pulse rate and heart rate are essentially the same. Pulse can be felt readily where an artery is near the surface of the skin and over the surface of a bone. Pulse rate vary considerably in individuals because of time of the day, temperature, emotions, stress, and other factors. The normal adult pulse rate of the heart at rest is about 75 beats per minute. Procedure: Radial Pulse 1. Using your index and middle finger palpate your laboratory partners radial artery. 2. The thumb should never be used because it has its own prominent pulse. 3. Palpate the area behind your partners thumb just inside the bony prominence on the lateral aspect of the wrist. 4. CAUTION. Do not apply too much pressure. 5. Count the pulse, change position, and record your results. Carotid Pulse 1. Using the same finger that you used for the radial pulse, place them on either side of your partners larynx. 2. Gently press downward and toward the back until you feel the pulse. You must feel the pulse clearly with at least two fingers, so adjust your hand accordingly. 3. The radial and the carotid pulse can be compared under the following conditions: a. Sitting quietly b. Standing quietly c. Right after walking 60 steps d. Right after running in place 60 steps 4. Note how long it takes the pulse to return to normal after walking and running exercise. 5. Compare your radial and carotid pulse in the table provided. Radial Pulse Rate Sitting quietly Standing quietly After walking After running Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science 3 Carotid Pulse Rate

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 3. Blood Pressure Clinically, the term blood pressure refers to the pressure within the large arteries. Arterial blood pressure can be measured either directly, by the insertion of a needle or catheter into the artery in such a way that the needle is pointing upstream or against the flow of the blood, or indirectly by a sphygmomanometer. Blood pressure is recorded in millmeters of mercury (mmHg) and is normally taken in the brachial artery. The highest pressure in the artery occurs during ventricular systole is termed systolic blood pressure. The lowest occurring during ventricular diastole is called diastolic blood pressure. The average blood pressure of a young adult is about 120mmHg systolic and 80mmHg diastolic, abbreviated to be 120/80. The difference between systolic and diastolic pressure is called pulse pressure. Auscultation method Sounds of blood flow are heard with a stethoscope. Blood flow in an artery is impeded by increasing pressure within a sphygmomanometer. When the cuff of the sphygmomanometer applies sufficient pressure to completely occlude blood flow, no sounds can be heard distal to the cuff because no blood can flow through the artery. When cuff pressure drops below the maximal (systolic) pressure in the artery, blood is heard passing through the vessel. When cuff pressure drops below the lowest (diastolic) pressure in the vessel, the sound becomes muffled and usually disappears. The sounds heard through the stethoscope via this procedure are termed korotkoff sounds. Procedure 1. Either you or your laboratory partner should be comfortably seated, at ease, with your arm bared, slightly flexed, abducted, and perfectly relaxed. Rest the forearm on a table in the supinated position. 2. Wrap the deflated cuff of the sphygmomanometer around the arm with the lower edge about 2.5 cm above the antecubital space. Close the valve of the neck of the rubber bulb. 3. Clean the earpiece of the stethoscope with alcohol before using it. Using the diaphragm of the stethoscope, find the pulse in the brachial artery just above the bend of the elbow, on the inner margin of the biceps brachii muscle. 4. Inflate the cuff by squeezing the bulb until the air pressure within it just exceeds 170mmHg. At this point the wall of the brachial artery is compressed tightly, and no blood should be able to flow through. 5. Place the diaphragm of the stethoscope firmly over the brachial artery and, while watching the pressure gauge, slowly turn the valve, releasing air from the cuff. Listen carefully for Korotkoff sounds as you watch the pressure fall. The first loud, rapping sound you hear is the systolic pressure. 6. Continue listening as the pressure falls. The pressure recorded on the mercury column when the sounds become faint or disappear is the diastolic pressure reading. 7. Repeat this procedure twice to see if you get consistent results. Allow a few minute between readings. 8. Have your partner stand and record his/her blood pressure twice. 9. Now, assuming that your partner has no known or apparent cardiac or other health problems, and is capable of such activity, have your partner do some exercise such as running in place for 40 steps and measure the blood pressure again immediately after the completion of the exercise. Record the pulse pressure in the table provided below.

Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Blood pressure-Auscultation Method Systolic Pressure Left Arm Sitting Standing After running Right Arm Diastolic Pressure Left Arm Right Arm Pulse Pressure

Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Experiment 2: Cardiovascular Physiology A. Cardiac Conduction System and Electrocardiogram (ECG or EKG) The heart contains specialized muscle tissue that functions like neural tissue. It spontaneously forms impulses and transmits them to the myocardium to initiate contraction. This specialized tissue forms the conduction system of the heart, which consists of the sinoatrial (S-A) node, atrioventricular (A-V) node, A-V bundle and Purkinje fibers. The origination and transmission of impulses through the conduction system of the heart generates electrical currents that may be detected by electrodes placed on the body surface. An instrument called an electrocardiograph is used to transform the electrical currents picked up by the electrodes into a recording called an electrocardiogram (ECG or EKG). A normal ECG is shown below.

An electrocardiogram has three distinct waves: The P wave, the QRS wave and the T wave. The P wave is produced by the depolarization of the atria. The QRS wave is produced by the depolarization of the ventricles. The greater size of the QRS wave is due to the greater muscle mass of the ventricles. The T wave is produced by the repolarization of the ventricular myocardium. Repolarisation of the atria is not detected because it is masked by the stronger QRS wave. Special Electric Properties of Cardiac Muscle: Automaticity & Rhythmicity Cardiac muscle differs from skeletal muscle both functionally and in its fine structure. Skeletal muscle must be electrically stimulated to contract. In contrast, heart muscle can and does depolarize spontaneously in the absence of external stimulation. This property, called automaticity, is due to plasma membranes that have reduced permeability to potassium ions but still allow sodium ions to slowly leak into the cells. This leakage causes the muscle cells to slowly depolarize until the action potential threshold is reached and fast calcium channels open, allowing Ca2+ entry from the extra cellular fluid. Shortly thereafter, contraction occurs. The spontaneous depolarization-repolarization events occur in a regular and continuous manner in cardiac muscle, a property referred to as rhythmicity. Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science 6

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 In the following experiment, you will observe these properties of cardiac muscle in a computer simulation.

Nervous Stimulation of the Heart Both the parasympathetic and sympathetic nervous systems innervate the heart. Stimulation of the sympathetic nervous system increases the rate and force of contraction of the heart. Stimulation of the parasympathetic nervous system (vagal nerves) decreases the depolarization rhythm of the sinoatrial node and slows transmission of excitation through the atrioventricular node. If vagal stimulation is excessive, the heart will stop beating. After a short time, the ventricles will begin to beat again. This is referred to as vagal escape and may be the result of sympathetic reflexes or initiation of a rhythm by the Purkinje fibers The heart's effectiveness as a pump is dependent both on intrinsic (within the heart) and extrinsic (external to the heart) controls. In the first experimental series, Activities 1-3, you will investigate some of these factors. The nodal system, in which the "pacemaker" imposes its depolarization rate on the rest of the heart, is one intrinsic factor that influences the heart's pumping action. If its impulses fail to reach the ventricles (as in heart block), the ventricles continue to beat but at their own inherent rate, which is much slower than that usually imposed on them. Although heart contraction does not depend on nerve impulses, its rate can be modified by extrinsic impulses reaching it through the autonomic nerves. Cardiac activity is also modified by various chemicals, hormones, ions, and metabolites. The effects of several of these chemical factors are examined in the next experimental series, Activities 4-9. The frog heart has two atria and a single, incompletely divided ventricle. The pacemaker is located in the sinus venosus, an enlarged region between the vena cava and the right atrium. The sinoatrial (SA) node of mammals may have evolved from the sinus venosus. Choose Frog Cardiovascular Physiology from the main menu. The opening screen will appear in a few seconds (Figure 6.1). When the program starts, you will see a tracing of the frog's heartbeat on the oscilloscope display in the upper right part of the screen. Because the simulation automatically adjusts itself to your computer's speed, you may not see the heart tracing appear in real time. If you want to increase the speed of the tracing (at the expense of tracing quality), click the Tools menu, choose Modify Display, and then select Increase Speed. The oscilloscope display shows the ventricular contraction rate in the Heart Rate window. The heart activity window to the right of the Heart Rate display provides the following messages: Heart Rate Normal-displayed when the heart is beating under resting conditions. Heart Rate Changing-displayed when the heart rate is increasing or decreasing. Heart Rate Stable-displayed when the heart rate is steady, but higher or lower than normal. For example, if you applied a chemical that increased heart rate to a stable but higher-than-normal rate, you would see this message. The electrical stimulator is below the oscilloscope display. In the experiment, clicking Single Stimulus delivers a single electrical shock to the frog heart. Clicking Multiple Stimulus delivers repeated electrical shocks at the rate indicated in the Stimuli/sec window just below the Multiple Stimulus button. When the Multiple Stimulus button is clicked, it changes to a Stop Stimulus button that allows you to stop electrical stimulation as desired. Clicking the (+) or (-) buttons next to the Stimuli/sec window adjusts the stimulus rate. The voltage delivered when Single Stimulus or Multiple Stimulus is clicked is displayed in the Voltage window just below the Single Stimulus button. The simulation automatically adjusts the voltage for the experiment. The post like apparatus extending upward from the electrical stimulator is the electrode holder into which you will drag and- drop electrodes from the supply cabinet in the bottom left corner of the screen. Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science 7

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 The left side of the screen contains the apparatus that sustains the frog heart. The heart has been lifted away from the body of the frog by a hook passed through the apex of the heart. Although the frog cannot be seen because it is in the dissection tray, its heart has not been removed from its circulatory system. A thin string connects the hook in the heart to the force transducer at the top of the support bracket. As the heart contracts, the string exerts tension on the force transducer that converts the contraction into the oscilloscope tracing. The slender white strand extending from the heart toward the right side of the dissection tray is the vagus nerve. In the simulation, room-temperature (23oC) frog Ringer's solution continuously drips onto the heart to keep it moist and responsive so that a regular heart beat is maintained. The two electrodes you will use during the experiment are located in the supply cabinet beneath the dissection tray. The Direct Heart Stimulation electrode is used to stimulate the ventricular muscle directly. The Vagus Nerve Stimulation electrode is used to stimulate the vagus nerve. To position either electrode, click and drag the electrode to the two-pronged plug in the electrode holder and then release the mouse button.

Figure 6.1 Activity 1: Recording Baseline Frog Heart Activity 1. Before beginning to stimulate the frog heart experimentally, watch several heartbeats. Be sure you can distinguish atrial and ventricular contraction (Figure 6.2a). 2. Record the number of ventricular contractions per minute displayed in the Heart Rate window under the oscilloscope. ____________________________________bpm (beats per minute) Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science 8

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Activity 2: Investigating the Refractory Period of Cardiac Muscle Repeated rapid stimuli could cause skeletal muscle to remain in a contracted state. In other words, the muscle could be tetanized. This was possible because of the relatively short refractory period of skeletal muscle. In this experiment you will investigate the refractory period of cardiac muscle and its response to stimulation. 1. Click and hold the mouse button on the Direct Heart Stimulation electrode, and drag it to the electrode holder. 2. Release the mouse button to lock the electrode in place. The electrode will touch the ventricular muscle tissue. 3. Deliver single shocks by clicking Single Stimulus at each of the following times. You may need to practice to acquire the correct technique. near the beginning of ventricular contraction at the peak of ventricular contraction during the relaxation part of the cycle Watch for extra systoles, which are extra beats that show up riding on the ventricular contraction peak. Also note the compensatory pause, which allows the heart to get back on schedule after an extra systole (Figure 6.2b).

During which portion of the cardiac cycle was it possible to induce an extra systole?

4. Attempt to tetanize the heart by clicking Multiple Stimulus. Electrical shocks will be delivered to the muscle at a rate of 20 stimuli/sec. What is the result?

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 5. Considering the function of the heart, why is it important that the heart muscle cannot be tetanized?

6. Click Stop Stimulus to stop the electrical stimulation.

Activity 3: Examining the Effect of Vagus Nerve Stimulation The vagus nerve carries parasympathetic impulses to the heart, which modify heart activity. 1. Click the Direct Heart Stimulation electrode to return it to the supply cabinet. 2. Click and drag the Vagus Nerve Stimulation electrode to the electrode holder. 3. Release the mouse button to lock the electrode in place. The vagus nerve will automatically be draped over the electrode contacts. 5. Adjust the stimulator to 50 stimuli/sec by clicking the (+) or (-) buttons. 6. Click Multiple Stimulus. Allow the vagal stimulation to continue until the heart stops momentarily and then begins to beat again (vagal escape), and then click Stop Stimulus. What is the effect of vagal stimulation on heart rate? ________________________________________________________________________ ________________________________________________________________________ The phenomenon of vagal escape demonstrates that many factors are involved in heart regulation and that any deleterious factor (in this case, excessive vagal stimulation) will be overcome, if possible, by other physiological mechanisms such as activation of the sympathetic division of the autonomic nervous system (ANS).

Assessing Physical and Chemical Modifier of Heart Rate Now that you have observed normal frog heart activity, you will have an opportunity to investigate the effects of various modifying factors on heart activity. After removing the agent in each activity, allow the heart to return to its normal rate before continuing with the testing. Choose Modifiers of Heart Rate from the Experiment menu. The opening screen will appear in a few seconds (Figure 6.3). The appearance and functionality of the oscilloscope display is the same as it was in the Electrical Stimulation experiment. The solutions shelf above the oscilloscope display contains the chemicals you'll use to modify heart rate in the experiment. You can choose the temperature of the Ringer's solution dispensed by clicking the appropriate button in the Ringer's dispenser at the left part of the screen. The doors to the supply cabinet are closed during this experiment because the electrical stimulator is not used. When you click Record Data in the data control unit below the oscilloscope, your data is stored in the computers memory and is displayed in the data grid at the bottom of the screen; data displayed include the solution used and the resulting heart rate. If you are not satisfied with a trial, you can click Delete Line. Click Clear Table if you wish to repeat the entire experiment.

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Activity 4: Assessing the Effect of Temperature 1. Click the 5oC Ringer's buffer to bathe the frog heart in cold Ringer's solution. Watch the recording for a change in cardiac activity. 2. When the heart activity window displays the message Heart Rate Stable, click Record Data to retain your data in the data grid. What change occurred with the cold (5oC) Ringer's solution?

3. Now click the 23oC Ringer's button to flood the heart with fresh room-temperature Ringer's solution. 4. After you see the message Heart Rate Normal in the heart activity window, click the 32 oC Ringer's button. 5. When the heart activity window displays the message Heart Rate Stable, click Record Data to retain your data. What change occurred with the warm (32oC) Ringer's solution?

Record the heart rate at the two temperatures below. _______________________________bpm at 5oC; _______________________________bpm at32oC What can you say about the effect of temperature on heart rate?

6. Click the 23oC Ringer's button to flush the heart with fresh Ringer's solution. Watch the heart activity window for the message Heart Rate Normal before beginning the next test.

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2

Activity 5: Assessing the Effect of Pilocarpine 1. Click and hold the mouse on the pilocarpine dropper cap. 2. Drag the dropper cap to a point about an inch above the heart, and release the mouse. 3. Pilocarpine solution will be dispensed onto the heart, and the dropper cap will automatically return to the pilocarpine bottle. 4. Watch the heart activity window for the message Heart Rate Stable, indicating that the heart rate has stabilized under the effects of pilocarpine. 5. After the heart rate stabilizes, record the heart rate in the space provided below, and clicks Record Data to retain your data in the grid. ______________________________________bpm What happened when the heart was bathed in the pilocarpine solution? ________________________________________________________________________ 6. Click the 23oC Ringer's button to flush the heart with fresh Ringer's solution. Watch the heart activity window for the message Heart Rate Normal, an indication that the heart is ready for the next test Pilocarpine simulates the effect of parasympathetic nerve (hence, vagal) stimulation by enhancing acetylcholine release; such drugs are called parasympathomimetic drugs. Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science 12

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Activity 6: Assessing the Effect of Atropine 1. Drag-and-drop the atropine dropper cap to a point about an inch above the heart. 2. Atropine solution will automatically drip onto the heart, and the dropper cap will return to its position in the atropine bottle. 3. Watch the heart activity window for the message Heart Rate Stable. 4. After the heart rate stabilizes, record the heart rate in the space below, and click Record Data to retain your data in the grid. ____________________________ bpm What is the effect of atropine on the heart?

Atropine is a drug that blocks the effect of the neurotransmitter acetylcholine, liberated by the parasympathetic nerve endings. Do your results accurately reflect this effect of atropine?

Are pilocarpine and atropine agonists or antagonists in their effects on heart activity?

5. Click the 23oC Ringer's button to flush the heart with fresh Ringer's solution. Watch the heart activity window for the message Heart Rate Normal before beginning the next test.

Activity 7: Assessing the Effect of Epinephrine 1. Drag-and-drop the epinephrine dropper cap to a point about an inch above the heart. 2. Epinephrine solution will be dispensed onto the heart, and the dropper cap will return to the epinephrine bottle. 3. Watch the heart activity window for the message Heart Rate Stable. 4. After the heart rate stabilizes, record the heart rate in the space provided below, and clicks Record Data to retain your data in the grid. ____________________________ bpm

What happened when the heart was bathed in the epinephrine solution?

Which division of the autonomic nervous system does its effect imitate?

5. Click the 23C Ringer's button to flush the heart with fresh Ringer's solution. Watch the heart activity window for the message Heart Rate Normal, meaning that the heart is ready for the next test.

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Activity 8: Assessing the Effect of Digitalis 1. Drag-and-drop the digitalis dropper cap to a point about an inch above the heart. 2. Digitalis solution will automatically drip onto the heart, and then the dropper will return to the digitalis bottle. 3. Watch the heart activity window to the right of the Heart Rate window for the message Heart Rate Stable. 4. After the heart rate stabilizes, record the heart rate in the space provided below, and clicks Record Data to retain your data in the grid. ____________________________ bpm What is the effect of digitalis on the heart?

5. Click the 23oC Ringer's button to flush the heart with fresh Ringer's solution. Watch the heart activity window for the message Heart Rate Normal, and then proceed to the next test. Digitalis is a drug commonly prescribed for heart patients with congestive heart failure. It slows heart rate, providing more time for venous return and decreasing the workload on the weakened heart. These effects are thought to be due to inhibition of the sodium-potassium pump and enhancement of Ca2+ entry into myocardial fibers.

Activity 9: Assessing the Effect of Various Ions To test the effect of various ions on the heart, apply the desired solution using the following method. 1. Drag-and-drop the calcium ions dropper cap to a point about an inch above the heart. 2. Calcium ions will automatically be dripped onto the heart, and the dropper cap will return to the calcium ions bottle. 3. Watch the heart activity window for the message Heart Rate Stable 4. After the heart rate stabilizes, record the heart rate in the space provided below, and clicks Record Data to retain your data in the grid. 5. Click the 23oC Ringer's button to flush the heart with fresh Ringer's solution. Watch the heart activity window for the message Heart Rate Normal, which means that the heart is ready for the next test. 6. Repeat steps 1 through 5 for sodium ions and then potassium ions. Effect of Ca2+ Does the heart rate stabilize and remain stable?

Describe your observations of force and rhythm of the heartbeat.

Effect of Na+ Does the heart rate stabilize and remain stable?

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Describe your observations of force and rhythm of the heartbeat.

Potassium ion concentration is normally higher within cells than in the extra cellular fluid. Hyperkalemia decreases the resting potential of plasma membranes, thus decreasing the force of heart contraction. In some cases, the conduction rate of the heart is so depressed that ectopic pacemakers (pacemakers appearing erratically and at abnormal sites in the heart muscle) appear in the ventricle, and fibrillation may occur. Was there any evidence of premature beats in the recording of potassium ion effects?

Was arrhythmia produced with any of the ions tested? _________________________ If so, which? __________________________________________ Click Tools >Print Data to print your recorded data for this experiment

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Experiment 3: Lymphatic System Lymphatic tissue, often referred to as lymphoid tissue, is defined as reticular connective tissue that is infiltrated with lymphocytes. Lymphatic tissue might be compared to a sponge with the framework of the sponge consisting of reticular fibers (except for the thymus which has a cytoreticulum) and the holes of the sponge containing lymphocytes distributed as the prominent cell type. Lymphatic tissue is widely distributed in the body, either as lymphatic organs or as diffuse, dense, or nodular collections of lymphocytes, and it collectively constitutes the lymphatic system. A lymphatic organ is a mass of lymphatic tissue that is surrounded by a connective tissue capsule or covered by an epithelium. Lymphatic organs include (1) lymph nodes, (2) thymus, (3) spleen, (4) the two palatine tonsils, (5) the pharyngeal tonsil, and (6) the lingual tonsil. All lymphatic organs have efferent lymph vessels, but only lymph nodes have afferent lymph vessels. Much lymphatic tissue of the body is not part of lymphatic organs and is found in many locations, including the wall of the gastrointestinal tract, and the wall of the air passages of the respiratory system. The abbreviations GALT and BALT stand for gut associated lymphoid tissue and bronchial associated lymphoid tissue, respectively. Lymphatic tissue is called diffuse lymphatic tissue if only a few lymphocytes are present or dense lymphatic tissue if many lymphocytes are present. Dense lymphatic tissue may appear as cords or nodules. A nodule is a spherical mass of lymphocytes, measuring 2-3 mm in diameter. An active nodule is one that is producing lymphocytes and consists of two zones. The outer zone, also called peripheral zone or corona, stains dark and is crowded with small lymphocytes. The inner zone, also called germinal center, central zone, secondary zone, or reaction center, appears pale and contains several cell types, including large lymphocytes, medium lymphocytes, reticular cells, and macrophages. An inactive nodule is not actively producing lymphocytes, does not contain a germinal center, and is made up primarily of small lymphocytes. Keep in mind that a nodule is not permanent, and it may disappear and reappear as well as fluctuate between periods of activity and inactivity.

Part A: Gross anatomy Locate the position of lymph nodes, thymus, spleen and tonsils in the human model.

Part B: Histology Be able to identify the different lymphatic organs with the scanning lens or low power objective. Tabulate the characteristics that serve to identify each lymphatic organ.

Slides: Lymph node Thymus Spleen Tonsil

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Experiment 4: Respiratory System The respiratory system consists of the lungs and the passages that carry air to and from the lungs. The system is composed of three functional parts: conducting, respiratory, and ventilating portions. The conducting portion consists of the nasal cavities and associated sinuses, nasopharynx, oropharynx (which conducts both air and food), larynx, trachea, bronchi, bronchioles, and terminal bronchioles. The olfactory mucosa is associated with the conducting portion, being found in the superior part of each nasal cavity. The respiratory portion is specialized for the rapid exchange of gases between blood and air. It includes the respiratory bronchioles, alveolar ducts, alveolar sacs, and alveoli. Components of the ventilating portion include the thoracic cage, intercostal muscles along with certain other muscles, muscular diaphragm, and elastic tissue of the lungs. These components assist the conducting and respiratory portions of the respiratory system in performing their functions.

Part A: Gross anatomy Observe the organs involved in respiratory system from the human model and label the respiratory pathway in Appendix B.

Part B: Histology Identify the tissues of the respiratory system with the slides given.

Slides: Trachea Lung

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Experiment 5: Lung Volume and Capacities Pulmonary ventilation is the process by which gases are exchanged between the atmosphere and the alveoli. Air moves throughout the respiratory system as a result of pressure gradients between the external environment and the respiratory system. Likewise, O2 and CO2 then move between the respiratory alveoli and the pulmonary capillaries of the cardiovascular system, and the peripheral capillaries of the cardiovascular system and the tissue of the body as a result of diffusion gradients. We breathe in (inhale) when the pressure inside the lungs is less than the air pressure in the atmosphere; similarly, we breathe out (exhale) when the pressure inside the lungs is greater than the pressure in the atmostsphere. Measurement of respiratory volume In clinical practice, respiration refers to one complete respiratory cycle that is one inhalation and one exhalation. At rest, a healthy adult averages 12 breaths per minute. The lung volume changes at different temperature and pressure as well as water saturation. The air in the body is at different temperature than the air contained in the spirometer; body air is also saturated with water vapor. Hence, lung volumes and capacities are corrected for these differences by converting to the body temperature, pressure (atmospheric), saturated (water vapor) (BTPS) factor. Forced Expiratory Volume (FEVT) The T indicates that the volume of air is timed, FEV1 is the volume of air forcefully expired in 1 sec; FEV2 is the volume of air forcefully expired in 2 sec, and so on. A normal healthy individual should be able to expel 83% of the total capacity during the first second, 94% in the second sec, and 97% in the third sec. For individuals with disorder such as asthma and emphysema, the percentage can be considerably lower.

Procedure using a spirogram 1. CAUTION! When using the spirometer, the disposable mouthpiece is discarded after each use by each subject, and the hose is then detached and rinsed with70% alcohol. 2. Before starting, a little practice may be necessary to learn to inhale and exhale only through the mouth and into the hose. A nose clip may be necessary to prevent leakage from the nose. 3. Have your lab partner close the free-breathing valve and turn on the spirometer to the slow speed. 4. Perform the following exercise and record your results. 5. Inspire normally and then exhale normally three times. This volume is your tidal volume. Repeat twice, record the values and take the average. 6. Expire normally then exhale as much air as possible three times. Record this volume. This value is your expiratory reserve volume. Repeat twice, record the values and take the average. 7. After taking a deep breath, exhale as much air as possible three times. This volume is your vital capacity. Repeat twice, record the values and take the average. 8. Calculate your inspiratory reserve volume. Inspiratory reserve volume = vital capacity - (tidal volume + expiratory reserve volume)

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Results: Tidal Volume (ml) First Time Second Time Third Time Average Normal Value Expiratory Reserve (ml) Vital Capacity (ml) Inspiratory reserve (ml)

Normal lung volumes and capacity for healthy Malaysian at standing position: Male (L) 0.54 2.22 2.76 1.62 1.54 3.16 4.38 5.92 Female (L) 0.43 1.75 2.21 1.30 1.23 2.53 3.50 4.74

Tidal Volume (TV) Inspiration Reserve Volume (IRV) Inspiration Capacity (IC) Expiratory Reserve Volume (ERV) Residual Volume (RV) Functional Residual Capacity (FRC) Vital Capacity (VC) Total Lung Capacity (TLC)

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Experiment 6: Digestive System Body cells required a continuous supply of nutrients in order to carry out their vital functions. Nutrients come from the food we eat. However, most food molecules are too large to pass through directly into the blood; therefore, they must be digested to break them down into absorbable molecules. Digestion of food and absorption of nutrients are the major functions of the digestive systems. Digestion involves both mechanical and chemical processes. Mechanical digestion is the physical breakdown of food into smaller pieces, which provides a greater surface area for contact with digestive secretions. Chemical digestion is the splitting of complex nonabsorbable food molecules into small, absorbable nutrient molecules. The digestive systems consist of the alimentary canal, which includes the mouth, pharynx, esophagus, stomach, small intestine, large intestine, and anus. The major accessory organs are the teeth, salivary glands, liver, gall bladder, and pancreas.

Part A: Gross anatomy Label the organs involved in digestive system (Appendix C)

Part B: Histology The histological structure of the alimentary canal wall consists of four basic tunics: the mucosa (and its three sub-layers), the submucosa (connective tissue layer deep to the mucosa), the muscularis externa (composed of circular and longitudinal smooth muscle layers), and the serosa (the outermost layer). Examine the large leaf-like villi, which increase the surface area for absorption.

Slides: 1. 2. 3. 4. 5. Esophagus Small intestine (Jejunum, Deodenum) Large Intestine Stomach Liver- observes the central canals and how the liver cells form cords that radiate from those canals. Identify a triad, a region containing a branch of the hepatic artery, a branch of the hepatic portal vein and a bile duct. The liver units, called lobules, are 6-sided and a triad is found at each corner. 6. Pancreas 7. Gall bladder 8. Appendix

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2

See Table below for a comparison of structure of parts of the GI tract

Components of the GI tract and their relative characteristic features.

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(a)

(h)

(b)

(c)

(d)

(e)

(f)

(g)

Micrographs of various parts of the GI tract. Note that (h) is out of order!! (a) = esophagus, (b) = fundus of the stomach, (c) = pylorus, (d) = duodenum, (e) = jejunum and ileum, (f) = colon and rectum, (g) = appendix and (h) = anus

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Experiment 7: Urinary System The urinary system consists of the two kidneys and the excretory passages, which convey urine from the kidneys to the exterior of the body. Excretory passages include the minor calyces, major calyces, renal pelvis (one for each kidney), the two ureters, the urinary bladder, and the urethra. Urine formed in either kidney is emptied into the minor calyces and then passes via the major calyces, renal pelvis, and ureter to the urinary bladder where it is stored until conducted by the urethra to the exterior of the body. Part A: Gross anatomy Identify the components in the urinary system and in the kidney: Appendix D Part B: Histology Kidney Before examining the sections of kidney, study the text illustrations below and become familiar with the large subdivisions of the kidney as seen in a radial section.

Schematic representation depicting the general organization of the kidney and a juxtamedullary nephrons with its collecting duct. Ureter The two ureters are the part of the excretory passageway, which conduct urine from the kidneys to the bladder. The wall of the ureter is formed of three layers: the mucosa, muscularis, and fibrosa or serosa. Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science 23

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Urinary bladder The urinary bladder is an expandable, hollow, muscular sac, which receives urine from the two ureters and temporarily stores it until discharged via the urethra. Like the ureter, the bladder has three layers (mucosa, muscularis, and fibrosa or serosa) with each layer being much thicker than the corresponding layer of the ureter.

Slides: 1. Kidney: observe the structure of nephrons. Identify the glomerulus, renal tubule and glomerular capsule. 2. Ureter 3. Urinary bladder

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Experiment 8: Renal Physiology Kidneys play a vital role in homeostasis or the maintenance of constant conditions within the body. They regulate the chemical content, the pH and the osmotic pressure of the blood. Kidneys form urine through the process of (1) glomerular filtration, (2) tubular reabsoprtion and (3) tubular secretion. Small molecules such as water and some chemical substances are filtered out of the glomeruli, moved through nephron and into the collecting ducts. As the filtrate passes through the nephrons, substances such as sodium, calcium, glucose and amino acids are reabsorbed, whereas other substances such as potassium and hydrogen ions are secreted by cells of the nephron tubule. The reabsorption of glucose is usually complete that sugar is not found in the urine. However, the capacity of kidneys to conserve glucose is not unlimited. If the blood sugar level exceeds a value known as renal threshold, the kidneys cannot reabsorb all the glucose in the glomerular filtrate and some sugar will appear in the urine. Water is reabsorbed both in the nephrons and collecting ducts of the kidneys to an extent that the volume of urine excreted represents less than 1% of the volume glomerular filtrate. Water reabsorption is enhanced by a hormone, antidiuretic hormone (ADH), which is released from the posterior pituitary. A change in fluid intake or a loss from the body can change the rate of ADH release and consequently change the rate of water excretion. Although the kidneys are not solely responsible for regulation of blood pH, they aid in maintenance of the blood at pH7.4. A change in kidney function with a resultant change in pH of the urine excreted can allow blood pH to remain unchanged despite altered metabolic activity or the intake of acidic or alkaline substances. For the composition of the blood to be held relatively constant despite the varied intake and utilization of substances by the body, it is apparent that urine composition must vary. This exercise will demonstrate the alteration in urine composition and the rapidity with which kidney function can change following consumption of a large volume of hypotonic solution, isosmotic solution or a bicarbonate solution.

Materials Water 300mOsm NaCl solution (isosmotic to ECF) 3%, 350mM NaHCO3 20% potassium chromate solution 2.9% silver nitrate solution Hydrometer (urinometer) pH meter Thermometer

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Procedure 1. Starting at dinner the night before lab, attempt to consume normal amount of liquid. On the day of lab, try to have a glass of drinks one to two hours before lab. Try not to take in unusual amount of caffeine or alcohol in the few hours before lab. 2. Four subjects with normal kidney function will be required for the experiment, with each drinking pure water or a solution or as control. The four treatment are: a. Subject 1: drink 700ml of water b. Subject 2: drink 350ml of 300mOsm NaCl solution (isosmotic to ECF) c. Subject 3: drink 350ml of 3% NaHCO3 d. Subject 4: drink 350ml of water 3. Urine samples will be taken before and after these treatments and the samples analyzed to determine renal function. 4. Subject 4 acts as a control for the group. However, each individual will also provide his or her own control by sampling urine production before and after the above treatments. 5. Urine sample should be collected every 30 minutes as described below. Analyze each sample as soon as possible to avoid samples accumulation. 6. At the beginning of the lab session, each subject should empty his or her bladder and note the time. DO NOT COLLECT THIS sample. 7. After 30 minutes, again empty the bladder. This time collect the full sample. This sample will serve as a pre-treatment control. 8. Immediately after step 7, subject 1-4 should drink the fluid appropriate to their treatments as quickly as possible. If the solution cannot be finished before the first post-treatment sample is taken, record the volume that was drunk. 9. 30 minutes after step 8, each subject should again empty his or her bladder and collect the full sample. 10. Repeat step 9 after another 30 minutes.

Measurements For each urine sample, you will collect data allowing you to determine the rate of urine formation, the concentration of urinary solids and pH of the urine. 1. Volume measurement: Use a graduated cylinder to measure the volume of each urine sample collected. 2. Urine pH determination: Check the pH of each urine sample using a pH meter. Be sure to rinse the probe with distilled water after use. 3. Specific gravity measurement: Use a hydrometer (urinometer) to measure the specific gravity of the urine sample. a. Pour the sample to be tested into the cylinder. There must be sufficient volume so that the hydrometer will float. b. Place the hydrometer in the urine sample and spin it slowly. Be sure that the hydrometer does not touch the side of the cylinder. c. When the hydrometer stops spinning, note the point at which the meniscus of the urine intersects the scale and read the specific gravity indicated on the scale. All the numbers on the scale represent a specific gravity of 1.000 or higher. Only the last two digits of the reading may be seen on the scale. For example, if the meniscus intersects the line indicated as 23, the specific gravity should be recorded as 1.023 Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science 26

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 d. A temperature correction factor is necessary when determining specific gravity of a urine sample because hydrometers are calibrated for use at 15oC and urine is usually tested at higher temperature. Measure the temperature of each sample at the time that its specific gravity as tested. For every 3oC above 15oC, add 0.001 to the specific gravity reading obtained from the scale of the hydrometer. e. Rinse the hydrometer with distilled water and dry it after each use. Failure to wash and dry the hydrometer before using it again can cause error in the next specific gravity measurement. f. If the urine volume is not great enough to float the hydrometer, make all the other required tests using urine sample. 4. Chloride concentration: Chloride concentration will be measured directly, while sodium ion concentration is simply assumed to be equal to the chloride ion concentration. a. Place 10 drops of urine in a test tube using a standard glass pipet b. Add one drop of 20% potassium chromate solution c. Add 2.9% of silver nitrate solution drop-by-drop, mixing the fluid continuously. Count the number of drops required to change the bright yellow solution to brown. Calculation Calculate the following variables: 1. Urine production rate in ml/min 2. Urinary solids (mass of solute in the urine) in g/L Mass= (specific gravity-1)X 1000 X 2.66g 3. Sodium chloride content. Each drop of silver nitrate added during the titration represents 1.0g/L NaCl

Weight , kg (lb) 45.36 (100) 49.9 (110) 54.4 (120) 59.0 (130) 63.5 (140) 68.0 (150) 72.6 (160) 77.1 (170) 81.6 (180) 86.2 (190) 90.7 (200) 95.3 (210) 99.8 (220) 104.3 (230) 108.9 (240)

Intake Volume Normal volume (ml) 225 250 275 300 325 325 350 375 400 425 450 475 500 525 550

High Volume (ml) 450 500 550 600 625 650 700 750 800 850 900 950 1000 1050 1100

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Experiment 9: Reproductive System The human male and female reproductive systems are specially adapted for their roles in reproduction. The gonads, the ovaries and the testes, form the gametes. Other reproductive organs nurture or transport male and female sex cells to sites where they may unite. The fertilized egg develops within the female reproductive system and culminates in the birth of a baby. Sexual maturation and pregnancy in females are regulated by hormones secreted by the pituitary gland and the gonads. The organs of male reproductive system include: 1. paired testis which produce sperms and male hormone (testosterone) which regulates male differentiation, both physical and physiological as well as psychic. 2. ducts that store and transport sperms (epididymis, ductus deferens and urethra); 3. accessory glands (paired seminal vesicles, a single prostate gland and bulbourethral glands) 4. external accessory structures including the scrotum and penis. The reproductive role of the female is much more complex compared to that of the male. Not only must she produce the female gamates, the reproductive system also provides a suitable environment for the development of the offspring and is actively involve in birthing process. The organs of the female reproductive system include: 1. paired ovaries which produce female sex cells (ova) and female hormones (estrogen and progesterone) 2. paired uterine tubes, which transport the female sex cells 3. a uterus where internal development of the offspring occurs 4. a vagina which serves as the female copulatory organ and birth canal 5. accessory glands and external organs

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Part A: Gross anatomy Female reproductive organs & Male reproductive organ: Appendix E Part B: Histology Mammary Gland Mammary glands, found only in mammals, are specialized to provide nourishment for the offspring. They are located in the subcutaneous connective tissue. In humans, the glands are sometimes unequal in size, the right more often larger and lower than the left. Inactive glands are compound tubular, but active glands (in pregnancy and lactation) are compound tubuloalveolar. See Figure below.

Schematic diagram of the changes in the mammary gland Mammary glands are ectodermal derivatives. There is one lactiferous duct for each lobe and 1525 lobes can be present in each gland. Each lobe is actually an independent gland with its own duct system. The ducts end blindly and have no alveoli at their ends. Slides: Testis, Sperm Smear Ovary (Graafian follicles) Uterus Mammary gland Lab Manual Version 01-09 Bachelor of Science (Hons) Biomedical Science 29

UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2

Appendix A: Heart

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Appendix B: Respiratory Pathways

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Appendix C: Digestive System

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Appendix D: Urinary System

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 Appendix E: Reproductive Systems

Female reproductive organ

Male reproductive organ

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UTAR UMSD1213 Anatomy & Physiology II Year 1 Trimester 2 References 1. Zao P, Stabler T, Peterson G, Smith L. PhysioExTM 6.0 for Anatomy & Physiology, 2006. Pearson Benjamin Cummings. 2. Waugh A & Grant A. Ross and Wilson Anatomy & Physiology, 2004. Churchill Livingstone. 3. Martin TR. Ed. Shier D, Buttler J, Lewis R. Holes Human Anatomy & Physiology Laboratory Manual, 2004. McGraw-Hill http://www.mc.vanderbilt.edu/histology/

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