METHODOLOGICAL INSTRUCTIONS

METHODICAL INSTRUCTION

for 2nd year BSN (4 year)  foreign students to be taught in field – 1101 Medicine Specialty 6.110100 “Nursing”

Educational – Qualification level – Bachelor

LESSON № _20_ (PRACTICAL – 6 HOURS)

Theme:

Non-protein nitrogen containing and nitrogen not containing organic components of blood. Biochemical composition of  blood in norm and pathology: acute phase proteins, enzymes of blood plasma.  Residual nitrogen. Lipoproteins of blood plasma.

 

Aim: To be able to determine the residual nitrogen content in blood and use this data for diagnostics of liver, kidney and endocrine glands diseases.

 

Professional orientation of students:

The knowledge of the residual nitrogen in blood is necessary for future doctors to diagnose and treat the diseases of liver, kidney and endocrine glands.

 

Methodology of Practical Class (900-1200 hour).

 

Work 1. The measurement of residual nitrogen content in blood.

The principle of method.

Residual nitrogen is determined in protein free filtrate after the sedimentation of blood proteins. Protein free filtrate is mineralized by the concentrated sulfuric acid. Nitrogen (ammonia) is bounded with sulfuric acid and ammonia sulfate is formed. Ammonia sulfate reacts with Nessler reagent and yellow colored compound is produced. The intensity of color is proportional to  nitrogen concentration.

M a t e r i a l s    a n d    r e a g e n t s. Blood mineralizate, 50 % NaOH, Nessler reagent, standard solution of residual nitrogen.

 P r o t o c o l.

Add into the two tubes: 1-st 1 ml of blood mineralizate, 2-nd – 1 ml of standard solution of nitrogen. Add into the both tubes  10 ml of distilled water, 6 drops of 

50% NaOH and 0,5 ml of Nessler reagent. Measure the extinction using blue light filter.

Calculation:

X = (Eexp x Cst )/ Est  (g/l).

Eexp – extinction of sample containing mineralizate;

Est - extinction of sample containing standard solution;

Cst – residual nitrogen concentration in standard solution.

 

Individual Students Program.

You should be  prepared for the practical class using the existing textbooks and lectures. Special attention should be paid to the following questions:

 

Non-protein nitrogenous containing and nitrogen not containing organic components of blood. Lipoproteins of blood plasma.

1.  Non-protein nitrogenous containing and nitrogen not containing organic components of blood.

2.  The pathways of conversion of carbon skeleton of amino acids. Glycogenic and ketogenic amino acids.

3.  Residual nitrogen, its components, ways of formation, content in blood.

4.  The principle of method of residual nitrogen measurement in blood; clinical significance. The kinds of azotemia.

 

Seminar discussion of theoretical issues (1230 – 1400 hour).

Test evaluation and situational tasks.

Multiple Choice. Choose the correct answer/statement.

1. The main way of ATP production in RBC is:

A. Krebs cycle.

B. Glycolysis.

C. Ketolysis.

D. Tissue respiration.

2. What kinds of polypeptide chains are there in hemoglobin A?

A. 2a2d.

B. 2a2β.

C. 2a2g.

D. 3a2β.

3. Complex of hemoglobin with CO is:

A. Oxyhemoglobin.

B. Carbhemoglobin.

C. Carboxyhemoglobin.

D. Methemoglobin.

4. The total contents of proteins in blood plasma is:

A.65-85 g/l.

B. 55-75 g/l.

C. 45-65 g/l.

D.85-95 g/l.

5. The main function of fibrinogen is:

A.Transport.

B. Participation in blood clotting. 

C. Defense.

D.Depot of amino acids.

6. Ceruloplasmin is the protein of:

A.b-globulin fraction.

B. a1-globulin fraction.

C. a2-globulin fraction.

D.g-globulin fraction.

 

Real-life situations to be solved

1.  In β-polypeptide chain of hemoglobin glutamate in 6th position is substituted by valine. What are the causes and the consequences of such a change?

2.  Patient  lost consciousness as result of coal-gas breathing. What derivative of hemoglobin is formed in this case?

3.  The content of albumins in blood is 20 g/l. The possible causes and consequences of such state.

4.  The patient suffers from liver cirrhosis. Which changes on electrophoregramm can be observed?

 

Individual student work (1415-1500 hour)  are checked by solving situational tasks for each topic, answers in test evaluations and constructive questions (the instructor has tests & situational tasks).

 

Students should know:

1.  The biochemistry and functions of blood in the human body.

2.  The structure and function of blood proteins

3.  The pathways of amino acids metabolism, regulation of protein metabolism.

4.  The transport forms of lipids.

 

Students should be able to:

1.  Determine the hemoglobin derivates by spectral analysis

2.  Determine the fractions of blood proteins by electrophoresis method.

3.  Determine the residual nitrogen contents in blood and use this analysis for diagnostics of liver, kidney and endocrine glands diseases

4.  Determine the percentage ratio of a- and b-lipoproteins.

 

 

Answers to the self-Assessment.

1.  B.  2. B. 3. C. 4. A.  5. B. 6. C.

2.   

Real-life situations to be solved

1. The cause is defect of gene responsible for hemoglobin synthesis. Hemoglobin S is synthesized as result, and sickle cell anemia is developed. This form of hemoglobin can be crystallized very easy, causing hemolysis.

2. In the case of connection of hemoglobin with carbon monoxide CO carboxyhemoglobin is formed. It i causes hypoxia due to high affinity of  hemoglobin to CO and violation of oxyhemoglobin formation and dissociation.

3. The contents of albumins in blood is decreased. Causes: kidney or liver diseases, starvation. Consequences: disorders of transport and buffer function of blood, edema. 

4. The fraction of albumins will be decreased and fraction of g-globulins will be increased.

 

 

References.

Basic:

1.  Donald Voet, Judith G. Voet, Charlotte W.Pratt. Fundamentals of Biochemistry. – New YorkToronto, 2008. – 931 p.

2.  N.V.Bhagavan. Medical Biochemistry. – San DiegoTokyo, 2008. – 938 p.

3.  Reginald H Garret, Charles M. Grisham. Biochemistry with a Human Focus. – AustraliaUnited States, 2007. – 893 p.

4.  Thomas M. Devlin. Textbook of Biochemistry with Clinical Correlations. - Wiley-Liss, 2006. - 1248 p.

5.  Robert K. Murray, Daryl K. Grander. Harper’s illustrated Biochemistry. – India: International Education, 2003.- 693 p.

6.  University Web – site > Intranet > Students' facilities> Practical classes materials > Practical classes materials

7.  University Web – site > Intranet > Students facilities > Lecture presentations > Biochemical composition of blood in norm and pathology: protein plasma, acute phase proteins, enzymes of blood plasma. Non-protein nitrogen containing and nitrogen not containing organic components of blood. 

Additional:

1.  Geoffrey Beckett, Simon Walker, Peter Rae, Peter Ashby  Clinical biochemistry (Lecture notes). – Oxford: Blackwell Publishing Ltd, 2005.- 319 p.

2.  Michael L. Bishop, Edward P. Fody, Larry E. Schoeff   Clinical chemistry: principles, procedures, correlations.   USA: Blackwell Publishing Ltd, 2005.- 730 p.

3.  John W. Suttie. Introduction to Biochemistry. – New York: Holt, Rinehart and Winston, Inc., 1992.- 364 p.

4.  John Mc Murry, Mary E. Castellion. General, Organic and Biological Chemistry.- New Jersy: Prentice Hall, 1992.- 764 p.

5.  Robert K. Murray, Daryl K. Granner. Harper’s illustrated Biochemistry. – India: International Education, 2003.- 693 p.

 

 

 

The methodical instruction has been worked out by: as.-prof. Kuzmak I.P.

 

 

Methodical instruction was discussed and adopted at the Department sitting

 

11.06.2013. Minute № 13

 

Methodical instruction was adopted and reviewed at the Department sitting

 

29.08.2013. Minute № 2