PRACTICAL GUIDANCE Introduction of the practical The teaching of pharmacology given to medical faculty students is incomplete, whether using

the classical methodology or the more newer Problem Based Learning (PBL) methodology, without a conducting a pharmacological practice. This practice isn't aimed to increase the practical skill of the students its purpose is to help the participants obtain a direct explanation from observing the effects and responses of a drug in various levels of an organisms: organs, tissues and even towards cell cultures. The participants would actively give a predetermined treatment to the organs or tissues, observe and not the changes or responses that occur, report in groups and in class, and, more importantly, conduct an analysis an discuss any results found deviating from the existing theories. For that purpose, and also as a guide in completing the practice and a practice report, is this module As for the practice itself, participants are expected to prepare and learn beforehand the pharmacological effects of the certain drug used in the forthcoming practice, as it would ensure that the practice runs smoothly. Participants should work earnestly, seriously, with discipline and peacefully, so the practice would give benefit to the teaching of pharmacology.

PRACTICAL 1A: THE EFFECTS OF DRUGS ON THE BLOOD VESSELS INTRODUCTION Blood vessels disorders are mostly found in the pathomechanisms of several diseases, such as hypertension, vascular angiopathy on diabetes, etc. In order to solve this problem, many drugs have been developed - some target the smooth muscle of the blood vessels (alpha-1 blocker, isosorbide dinitrate), others the endothelial cells indirectly (help to maintain/increase the quantity and quality of the endothelial cells). To observe or develop drugs and even explain the pathomechanism of the blood vessel disorder, an experimental method takes place, in which is used a separated blood vessels. The blood vessels used are artery and aorta with or without endothelial cell. OBJECTIVES: 1) Observe the vasoconstriction and vasodilatation effects from the separated aorta with and without endothelial cell. 2) Understand the mechanism of a drug that has a vasodilatation effect which works at the smooth muscle of blood vessels or endothelial cell. METHODS: 1. Experiment Animal: Marmot 2. The drugs used are: Phenylephrine 0,5 ml 10-4M Phenylephrine followed by isosorbide dinitrate 0,5 ml 10-4M Phenylephrine followed by acetylcholine 0,5 ml 10-4M 3. Equipment: organ bath, Mac-lab computer 4. Experimental methods: Isolate the marmot's aorta from it's thorax cavity Prepare the whole blood vessel with and without endothelial cell (after wiped with a cotton) in a length of more than 1 cm long. Fixate the aorta with 2 hooks in to the organ bath with Krebs solution inside, and maintain the temperature at 37oC. Ensure the organ bath content is more or less 5 ml Ensure the weight of aorta is 1 g, and connect to an isotonic contraction recorders using isotonic transducers. Insert the drugs in the organ bath and observe the response from the recorder. Figure: A technique diagram of an Aortic ring preparation Details: a) Tissue/ Organ b) Henselheit Kreb's Solution c) Double Jacketed Chamber d) Transducer DISCUSSION AND ASSIGNMENT Phenylephrin's effect on the blood vessels a) What effect did you observe after phenylephrine was administered to the stock? b) How is the mechanism of action? c) Name the same type of drugs which has the same effect. d) Based on the effect, what are the side-effects of this drug? 2. Isosorbid's effect on the blood vessels a) What effect did you observe after isosorbid was administered to the stock?

b) How is the mechanism of action? c) Name the same type of drugs which has the same effect. d) Based on the effect, what are the side-effects of this drug? 3. Acetylcholine's effect on the blood vessels a) What effect did you observe after acetylcholine administered to the stock? b) How is the mechanism of action? c) Name the same type of drugs which has the same effect d) Based on the effect, what are the side-effects of this drug?

PRACTICAL 1B: THE EFFECT OF HISTAMIN AND ISOPRENALIN TO THE SEPARATED TRACHEA STOCK INTRODUCTION Bronchus spasm in asthma is a result of an inflammation process and a hyperactive bronchus. Asthma therapy, besides using anti-inflammation drugs to manage the underlying process of asthma also uses bronchodilators which functions mainly as a symptomatic drug. To observe or develop drugs which would function as a bronchodilator or bronchoconstrictor, an experimental method takes place in which a separated trachea stock is used. In this isolated tissue method, the trachea is more often used than bronchus smooth muscle because it is structurally and functionally (the distribution and number of receptors) similar to bronchus smooth muscles, but it is easier to prepare. OBJECTIVES: 1. To observe the bronchocontriction and bronchodilatasi effects on the separated stock of respiratory tract smooth muscles. 2. To understand the mechanism of bronchocontriction and bronchodilatation. METHOD: 1. Experiment Animal: Marmot 2. Drugs used: histamine and isoprenaline 3. Equipment used: organ bath, Mac-Lab computer 4. Experimental method: a) Isolate the trachea of the marmot from the thorax, and make a connected ring stock. b) Fixate stock using two strings with one end hooked with an isotonic transducer. c) Use Kreb's solution as the media solution, with the volume of the organ bath up to 25 ml. d) The trachea load is about 0.500 grams. e) Insert the drugs in to the organ bath and observe the response on the recorder. 5. Drugs used: a) 10-5 M histamine (it would give 80% of the maximum effect which is 0.1 ml 2.5x10-3). b) After the effect of histamine reaches maximum, administer isoprenalin with succesive concentration 10-8 M, 10-7 M, 10-6 M, and 10-5M. 6. Observe the changes of contraction on the recorder following the administration of drugs Figure: Technique Diagram of Separated Trachea Preparation DISCUSSION: 1. The effect of histamine on the respiratory tract smooth muscle a) What effect did you observe after histamine is administered to the stock? b) How is the drugs mechanism of action? c) Name the same type of drugs with the same effects. d) Based on the effect, what are the side effects of this drug? e) Explain the mechanism of histamine release following an allergic reaction. 2. The effect of Isoprenalin on the respiratory tract smooth muscle a) What effect did you observe after isoprenalin is administered to the stock? b) How is the drugs mechanism of action? c) Name the same type of drugs with the same effects. d) Based on the effect, what are the side effects of this drug?