Laboratory work in biology identifying catalase activity. Detection of organic substances

Annotation

The lecture notes presented to your attention are intended to prepare students medical universities to pass tests and exams. The book includes full course lectures on anesthesiology and resuscitation, written by accessible language and will be an indispensable assistant for those who want to quickly prepare for the exam and pass it successfully.

Marina Aleksandrovna Kolesnikova

Lecture No. 1. The concept of resuscitation

Lecture No. 2. Basic manipulations in intensive care

1. Tracheostomy

2. Conicotomy

3. Puncture of the pleural cavity

4. Puncture and catheterization of the epidural space

5. Lumbar puncture

Lecture No. 3. Acute disturbances of consciousness

Lecture No. 4. Cardiopulmonary resuscitation

Lecture No. 5. Emergency conditions in pulmonology

1. Laryngospasm

2. Pulmonary embolism

3. Bronchial asthma

Lecture No. 6. Emergency conditions in cardiology

1. Myocardial infarction

2. Cardiogenic shock

3. Hypertensive crisis

4. Heart rhythm disturbances, life-threatening

Lecture No. 7. Acute renal failure

Lecture No. 8. Acute liver failure

Lecture No. 9. Shock

Lecture No. 10. Acute poisoning

1. Poisoning methyl alcohol

2. Poisoning ethyl alcohol

3. Ethylene glycol (antifreeze) poisoning

4. Dichlorethane poisoning

5. Poisoning with poisonous mushrooms (fly agaric, false mushrooms, morels, toadstool)

6. Snake poisoning

7. Poisoning with concentrated acids (nitric, acetic, sulfuric)

8. Poisoning with arsenic and its compounds

9. Alkali poisoning

10. Atropine poisoning

11. Cannabis poisoning (hashish, marijuana, marijuana, plan)

12. Poisoning by cocaine and dicaine

13. Poisoning with narcotic analgesics (morphine, omnopon, droperidol)

Lecture No. 11. Pain and analgesics

2. Analgesics medicines

Lecture No. 12. Anesthesia. Types and stages of anesthesia

1. Theories of anesthesia

2. Preparing the patient for anesthesia

3. Intravenous anesthesia

4. Inhalation anesthesia

5. Stages of anesthesia

6. Methods for monitoring anesthesia

7. Complications of anesthesia

Lecture No. 13. Local anesthesia

1. Superficial anesthesia

2. Regional anesthesia

3. Cervical plexus anesthesia (CPA)

4. Brachial plexus anesthesia (BPA)

5. Anesthesia of peripheral nerves in the wrist area

6. Anesthesia of the lower extremities

7. Epidural anesthesia

8. Lumbar anesthesia

9. Cervical vagosympathetic blockade according to A. V. Vishnevsky

Lecture No. 14. Assessment of patient severity and monitoring

Lecture No. 15. Artificial ventilation

Lecture No. 16. Infusion therapy

1. Blood transfusion

2. Parenteral nutrition

3. Detoxification therapy

Marina Aleksandrovna Kolesnikova

Anesthesiology and resuscitation. Lecture notes

Lecture No. 1. The concept of resuscitation

Reanimatology is a section clinical medicine, studying the problems of revitalizing the body, developing principles for the prevention of terminal conditions, methods of resuscitation and intensive care. Practical methods revitalization of the body is united by the concept of “resuscitation”.

Resuscitation (from the Latin “revitalization” or “animation”) is a system of measures aimed at restoring severely impaired or lost vital important functions organism and its removal from the terminal state and clinical death. Effective resuscitation measures include chest compressions and artificial ventilation. If they are ineffective within 30 minutes, biological death is declared.

Intensive care is a set of measures used to treat severe, life-threatening conditions and provides for use according to indications large complex therapeutic measures, including intravenous infusions, long-term artificial ventilation, cardiac pacing, dialysis methods, etc.

A critical condition is the impossibility of maintaining the integrity of the body’s functions as a result of an acute dysfunction of an organ or system, requiring medicinal or hardware-instrumental replacement.

A terminal state is a borderline state between life and death, a reversible extinction of body functions, including the stages of preagony, agony and clinical death.

Clinical death is a terminal condition in which there is no blood circulation and breathing, the activity of the cerebral cortex ceases, but metabolic processes are preserved. In case of clinical death, it remains possible effective implementation resuscitation measures. The duration of clinical death is from 5 to 6 minutes.

Biological death- this is an irreversible cessation physiological processes in organs and tissues, in which resuscitation is impossible. It is determined by a combination of a number of signs: the absence of spontaneous movements, contractions of the heart and pulse in large arteries, breathing, reaction to painful stimuli, corneal reflex, maximum dilation of the pupils and the absence of their reaction to light. Reliable signs of death are a decrease in body temperature to 20 ° C, the appearance of cadaver spots and muscle rigor.

Lecture No. 2. Basic manipulations in intensive care

Percutaneous puncture and catheterization of the main vein (subclavian). Indications: large volumes of infusion-transfusion therapy, parenteral nutrition, detoxification therapy, intravenous antibiotic therapy, cardiac probing and contrast, measurement of central venous pressure, implantation of a pacemaker, impossibility of catheterization of peripheral veins. Contraindications: violation of the blood coagulation system, inflammatory and purulent process at the site of puncture and catheterization, trauma in the clavicle area, superior vena cava syndrome, Paget-Schretter syndrome. Instruments and accessories for puncture and catheterization: puncture needle, set of plastic catheters, set of conductors, syringe for intramuscular injections volume 10 ml, scissors, needle holder, surgical needle and silk ligature, adhesive plaster. Technique. Catheterization is carried out in compliance with the rules of asepsis and antisepsis, cleaning the operator’s hands, the surgical field and covering with sterile material. The patient is positioned horizontally on the back with the arms brought to the body and the head turned in the opposite side. Local anesthesia is used - 0.5–1% solution of novocaine. It is better to perform the puncture on the right, since when puncturing the left subclavian vein there is a danger of damaging the thoracic lymphatic duct. The puncture point is on the border of the inner and middle third of the clavicle, 2 cm below it. The needle is passed slowly at an angle of 45° to the collarbone and 30–40° to the surface chest between the clavicle and the first rib in the direction of the upper edge of the sternoclavicular joint. When passing the needle, the syringe piston is periodically pulled up to determine whether it has entered the vein, and novocaine is injected along the needle. When a vein is pierced, there is sometimes a sinking feeling. After entering the vein, the syringe is disconnected from the needle and the cannula is closed with a finger. Then a guidewire is inserted through the needle to a length of 15–20 cm and the needle is removed. A catheter of the appropriate diameter is passed along the conductor and, together with the conductor, is inserted into the vein 6–8 cm, after which the conductor is carefully removed. To check that the catheter is positioned correctly, a syringe is attached to it and 2-3 ml of blood is drawn into it, after which a plug is placed or infusion therapy is started. The catheter is fixed to the skin with a silk ligature. To do this, a sleeve is made from an adhesive plaster on the catheter 3–5 mm from the skin, on which silk is tied, then passed through the ears of the catheter and tied again. After fixing the catheter, the puncture site is covered with an aseptic sticker. Complications: puncture of the subclavian artery, air embolism, puncture of the pleural cavity, damage to the brachial plexus, damage to the thoracic lymphatic duct, damage to the trachea, thymus and thyroid gland, suppuration at the puncture site.

1. Tracheostomy

Indications: obstruction of the larynx and upper trachea due to obstruction by a tumor or foreign body, paralysis and spasm vocal cords, sharp swelling of the larynx, acute respiratory distress, aspiration of vomit, prevention of asphyxia in severe chest injuries. Instrumentation: 2 scalpels, 2 anatomical and surgical tweezers each, several hemostatic clamps, an elevator, a grooved probe, 2 blunt and 1 single-tooth sharp hook, a Trousseau or Deschamps dilator, surgical needles with a needle holder.

Technique

The patient lies on his back, with a cushion under his shoulders, and his head thrown back. If the patient is in a state of asphyxia, the cushion is placed only at the last moment, before opening the trachea. Local infiltration anesthesia is performed with a 0.5–1% solution of novocaine with the addition of adrenaline. In case of acute asphyxia, surgery can be performed without anesthesia. Identification points: the angle of the thyroid cartilage and the tubercle of the cricoid cartilage. An incision in the skin, subcutaneous tissue and superficial fascia is made from the lower edge of the thyroid cartilage to the jugular notch strictly along midline neck. The median vein of the neck is pushed back or ligated, finding white line, along which the muscles are bluntly pushed apart and the isthmus of the thyroid gland is exposed. The edges of the incision are spread apart with a Trousseau expander, ligatures are applied to the edge of the wound and carefully inserted...

Laboratory work № 1

ROLE OF ENZYMES IN ACCELERATING REACTIONS IN THE CELL

(DETECTION OF CATALASE ACTIVITY)

Target: detect the action of the catalase enzyme in plant and animal cells, compare the enzymatic activity of natural and boiling-damaged cells.

Equipment: 3% hydrogen peroxide solution, pieces of raw and boiled potatoes and meat (liver, lungs), test tubes.

Detection of catalase activity

Catalase is an enzyme that catalyzes the decomposition of hydrogen peroxide to form molecular oxygen, released in the form of gas bubbles:

catalase

2 H 2 O 2 _ → 2H 2 O + O 2

Hydrogen peroxide is produced in some plant and animal cells as a byproduct of redox reactions. This compound is toxic to cells, and catalase ensures its effective removal. Catalase is one of the fastest working enzymes: one molecule of catalase decomposes up to 200,000 molecules of hydrogen peroxide in one second. Catalase is localized in membrane vesicles of cells - microbodies and peroxisomes.

Progress

Take 4 clean test tubes and place in the first of them a small amount of finely grated potatoes, in the second - some boiled potatoes, in the third - finely chopped pieces of meat (liver, lung), in the fourth - a little chopped boiled meat. Add 3-4 ml to each test tube. 3% hydrogen peroxide solution. Observe what happens in the test tubes. Record the observation results in the table.

Enzymatic activity of natural and damaged cells

Explain your results. Draw a conclusion about the catalytic activity of catalase in living and dead cells.

Conclusion: ________

_______________

_______________

_____________

Control questions:

1. What are enzymes? What structure of proteins creates their activity?___________

2. What properties do enzymes have?______

3. What is called active center enzyme? How many such centers can there be in an enzyme?_

4. Due to what enzymes accelerate chemical reactions?___________________________

5. *Alcohol, phenol, chloramine and other antiseptics are used in medicine to treat areas of the body contaminated with pathogenic flora. Explain why._____

Gr.___3

Laboratory work No. 2

DETECTION OF ORGANIC SUBSTANCES

Target: identify organic substances in tissues (starch, protein, fats) and study their properties.

Equipment: gauze bag, ground wheat grains (wheat flour), 5% iodine solution, sunflower seeds (or any other oilseed crop: cotton, flax, peanuts, soybeans, etc.).

Organic compounds– carbon-containing substances, characteristic of living nature, constitute on average 20-30% of the mass of cells of living organisms. The main properties of cells and organisms are determined by organic polymers: proteins, carbohydrates, nucleic acids, as well as complex connections– fats and a number of molecules of hormones, pigments, individual nucleotides, in particular ATP. Besides organic matter the cells contain minerals and water, but the content of organic substances is always higher. The amount of organic matter may vary.

Proteins – irregular, or informational, polymers whose monomers are amino acids.

Based on their composition, proteins are divided into:

- simple– consist of only amino acids. For example, plant proteins are prolamins, they are contained in the gluten of cereal seeds and do not dissolve in water;

- complex– in addition to amino acids, they contain other organic compounds (nucleic acids, lipids, carbohydrates), phosphorus compounds, and metals. Accordingly, they are called: nucleoproteins, lipoproteins, glycoproteins, phospho- and metalloproteins.

Carbohydrates - compounds containing carbon, hydrogen and oxygen. They are divided into mono-, di- and polysaccharides. Polysaccharides are high molecular weight carbohydrates consisting of large number monosaccharides, their molecular mass is large, the molecules have a linear or branched structure. In functional terms, polysaccharides for reserve and structural purposes are distinguished. Insoluble in water starch– main reserve polysaccharide plant cells(polymer ά - glucose); when exposed to iodine it turns blue; contained in large quantities in potato tubers, fruits, seeds. Glycogen- a polysaccharide found in the tissues of the human and animal body, as well as in mushrooms and yeast, - plays important role in the transformation of carbohydrates in cells. Fiber (cellulose)– main structural polysaccharide cell membranes plants.

Lipids and lipids– fats and fat-like substances – organic compounds with different structure. They do not dissolve in water, but dissolve well in organic compounds: ether, gasoline, chloroform, etc.

By chemical structure lipids - compounds of glycerol - trihydric alcohol - with high molecular weight organic acids(bold), do not have a polymer structure.

Seed composition

Appendix No. 1.

Instructions for conducting laboratory work.

Laboratory work No. 3

Subject:

Target: develop knowledge about the role of enzymes in cells; find out the enzymatic properties of protein peroxidases; strengthen the ability to work with a microscope; conduct experiments and explain the results of the work.

Equipment: fresh 3% hydrogen peroxide solution, test tubes, tweezers, plant tissue (pieces of raw and boiled potatoes) and animal tissue (pieces of raw and boiled meat), sand, mortar and pestle.

additional information: Hydrogen peroxide is formed in the cell during metabolism and has a mutagenic effect. H2O2 is a chemically unstable substance and can spontaneously decompose to form stable compounds: 2 H2O2 = 2 H2O + O2

Progress.

1. Prepare four test tubes with a fresh 3% hydrogen peroxide solution, then place a piece of raw potato in the first test tube, a piece of boiled potato in the second, a piece of raw meat in the third, and a piece of boiled meat in the fourth. Observe what happens in each test tube.

2. Make a table showing the activity of each tissue under different treatments.

3. Grind a piece of raw potato in a mortar with a small amount sand. Transfer the crushed potatoes along with the sand into a test tube and drop a little hydrogen peroxide into it. Compare the activity of crushed and whole plant tissue.

4. Explain your results.

Answer the questions:

How does enzyme activity manifest itself in living and dead tissues?

Does enzyme activity differ in plant and animal tissues?

How does grinding tissue affect enzyme activity?

How would you propose to measure the rate of decomposition of hydrogen peroxide?

Do you think all living organisms contain the enzyme peroxidase?

ensuring the decomposition of hydrogen peroxide?

Sample lab report

« Catalytic activity enzymes in living tissues"

Conclusion: the action of peroxidase enzymes is similar in plant and animal cells, the commonality of physiological processes is one of the proofs family ties between plant and animal organisms.