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.
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.
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.
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.
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. –
2. N.V.Bhagavan. Medical Biochemistry. –
3. Reginald H Garret, Charles M. Grisham. Biochemistry
with a Human Focus. –
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. –
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). –
2. Michael L. Bishop, Edward P. Fody,
Larry E. Schoeff
Clinical chemistry: principles, procedures, correlations. –
3. John W. Suttie. Introduction
to Biochemistry. –
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. –
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