A. LABORATORY RULES
a. Attendance for laboratory sessions is compulsory.
b. Lab coat must be worn at all time while working in the laboratory.
B. GROUPING
a. A class representative/leader to be selected among the students to
coordinate class activities.
b. Students are grouped into 4 equal groups.
c. Each of 4 groups is subdivided into 3 sub-groups.
d. A sub-group leader is to be appointed among group members and he/she
is responsible for the safety of the equipments and other facilities to be
used in the experiments.
C. REPORT WRITING
a. Group report:
Each sub-group is required to submit a laboratory report.
b. Content of the report:
The laboratory report must contain the following section;
i.
Introduction:
a. Background of the study
b. Problem of the study
c. Objectives of the study
ii.
iii.
Results:
a. Tables of means, statistical analysis, etc
b. Graphs
iv.
Discussion:
a. Summary of the results
b. Literatures to support the findings
v.
Conclusion:
- Summary of the findings
1
vi.
References:
- List of literatures cited
SBP 3107
Introduction:
Glucose is an essential fuel for neurons, erythrocytes and contracting skeletal
muscle cells. The higher centers of the brain are highly sensitive to even a short
term deficiency of glucose.
Glucose enters most of the cells via active process but it is necessary to
maintain adequate concentration of glucose in the blood. It is not surprising to see
that the levels of several hormones in the plasma are changed with the small
changes in blood glucose concentration. Normal value of blood glucose for normal
person is between 70-100 mg/ml (3.9-6.1 mmol/L). Two hours after meals, blood
glucose levels should be less than 140 mg/ml (7.8 mmol/L).
Blood glucose level has been used as one of the indicators to determine the
health status of an individual.
This kit
provides a simple direct method for determination of blood glucose using only a
drop of whole blood.
Glucose ingestion is to
stimulate the condition after meal. The effect of increasing the concentration of
ingested glucose also will be studied in this experiment.
Principle of the technique:
Each blood glucose ACCU-CHECK tm Test Strip contains the following reagents:
Potassium ferricyanide (43.7%), Glucose dehydrogenase (1.2%), buffer (24.7%),
stabilizer (19.4%) and non-active ingredient (11.0%).
When a drop of blood is applied to the curve at the edge of the ACCUCHECKtm test strip, the glucose dehydrogenase enzyme in the strip converts the
glucose in the blood sample to gluconolactone. This reaction creates a harmless
electrical current which is in proportional with the magnitude of the reaction. The
meter will interpret the current produces into the amount of glucose in the blood.
Students will use this kit to determine their blood glucose levels during
fasting and study the effect of glucose loading on the level of blood glucose during
fasting.
Materials:
1.
2.
3.
4.
5.
Methodology:
A. Measurement of blood glucose
1. If you are using a new pack of ACCU-CHECKtm Test Strip, take out the
chip code included in the pack and insert it into the code chip slop in
the meter with meter turned off.
2. Switch on the glucose meter.
3. Insert a test strip to the glucose meter.
4. Clean your finger tip using alcohol and punch using lancet needle to
take a few drops of blood.
5. Touch the drop of blood to the curve at the edge of the test strip. Hold
finger to the curve of test strip and the yellow window of the strip is
completely filled up with blood (no yellow color is visible). Blood will
be drown into the strip automatically. Do not place the blood drop on
the top of the test strip.
6. Wait for about 30 seconds and take the reading.
7. Remove the test strip from the meter and discard the strip.
B. Blood glucose levels during fasting and after glucose ingestion
1. Divide the class into 8 groups equally and allocated the group to the
treatment as shown in table 1.
2. Choose 4 members from each group of 1-4 and asked them to fast
(without food) overnight but allowed to take plane water.
3. Choose 4 members from each group of 5-8 and asked them to take
normal meal two hours before class.
4
Nutritional Status
Fasting
Normal Feeding
Data analysis:
1. Pooled the data of the blood glucose level from your group and others.
2. Calculate the means and standard deviation of the blood glucose level in
each time of sampling for each concentration of glucose given.
Time
Fasting
0
30
60
90
Total
Normal
feeding
Total
0
30
60
90
(gm/50ml)
20
40
G2
G3
G6
G7
0
G1
G5
80
G4
G8
Total
Analysis of variance:
Source
df
Student
Time
Student*time
MS
Group
Nutritional status
Error
24
Total
35
Discussion:
The discussion will be at least on the following questions:
a. What is the normal glucose levels in the blood during fasting?
b. What is the effect of glucose ingestion during fasting and no fasting?
c. How does the glucose in blood change following glucose
infusion/injection?
Grou
p
Subjec
t
Matric
No.
Glucose
ingestion
(gm)
Nutrition
al Status
FS
FS
FS
FS
20
Time after
ingestion
(min)
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
Blood
glucose
level
(mmol/L)
FS
20
FS
20
FS
40
FS
40
FS
40
FS
80
FS
80
FS
80
NF
NF
NF
NF
20
NF
20
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
NF
20
NF
40
NF
40
NF
40
NF
80
NF
80
NF
80
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
0
30
60
90
SBP 3107
Introduction:
Food taken up everyday consists of carbohydrate, protein, fat, minerals and
water.
digestive tract is very rich in nutrients (glucose, amino acids, fatty acids, vitamins,
minerals) in which the level of nutrients have to be reduced to a physiological level
before the blood is circulated throughout the body.
Insulin, secreted by the beta cell of the islet of Langerhans in the pancreas,
regulates sugars level in the circulatory blood.
secretion of insulin by its secretary cells. One hour after taken 50 gm of glucose by
mouth, the plasma insulin level increased from fasting level (16uU/ml) to a
maximum level (58uU/ml) whereas after 100gm of glucose, the rise was from 17
uU/ml to 158 uU/ml.
Insulin causes rapid uptake, storage and use of glucose by almost all tissues
of the body especially by the liver, muscles and adipose tissue. In the liver, insulin
inhibits phophorylase, the enzyme that causes the breakdown of liver glycogen to
into glucose. Insulin also enhances the uptake of glucose from the blood in the liver
cells by increasing the activity of the enzyme glucokinase.
The activities of
The condition of
diabetes mellitus will be induced by injection of streptozotocin (STZ) drug and the
effect can be observed within 3-4 days later. Insulin is then injected to see whether
the glucose levels in the blood change following infusion of glucose solution.
Reagents:
1.
2.
3.
4.
Streptozotocin (STZ)
Citrate buffer (ph 4.5)
Insulin
Glucose solution
Materials:
1.
2.
3.
4.
Insulin
No Insulin
0
G1
G1
Normal
Diabetic
Glucose Solution
Glucose Solution
20
G2
G2
40
G3
G3
80
G4
G4
0
G5
G5
20
G6
G6
40
G7
G7
80
G8
G8
n = 3 rats
Experimental Animals:
1. Keep the rats in the cages according to their group in the animal house.
Feed and water to be given accordingly.
2. The experimental period lasts for about one week.
Treatments:
1. At day -4 (Tuesday), 0.2ml of STZ solution (50mg/kg bw) is injected
subcutaneously to all diabetic-induced rats.
2. At day -1 (Thursday), all the rats are fasted over night but water is given
as usual.
3. At day 0 (Friday), 0.2ml of glucose solution at various concentration is
injected subcutaneously to the respective groups.
4. 30 minutes after glucose injection, 0.2ml insulin (1 IU/kb) is injected
subcutaneously to the rats in normal insulin and diabetic insulin groups
while the no insulin groups are injected with the buffer only.
12
Measurements:
1. Blood glucose level is determined using glucose kit as describe in the
previous laboratory exercise.
following schedule.
a. Just before glucose infusion
b. Just before insulin or buffer injection
c. 2 times at 30 minutes interval after insulin or buffer injection
Tim
e
Insulin
-30
0
30
60
Total
No
Insulin
-30
0
30
60
Total
Grand Total
Normal
Glucose Solution
20
40
80
Diabetic
Glucose Solution
20
40
Total
80
13
Source
df
Rat
Time
Rat*Time
12
Diabetic Status
MS
Injection
Error
37
Total
60
Results:
Plot the means of glucose level vs time of experiment.
Use time = 0 as time before glucose injection.
Discussion:
The discussion will be at least on the following questions:
a. What is the normal glucose levels in the blood of rats during fasting?
b. What is the effect of STZ injection on glucose levels in the blood?
c. How does the glucose in blood change following glucose
infusion/injection?
d. What is the effect if insulin injection on the blood glucose levels?
e. How does the insulin injection influence the blood glucose level?
f. How do you conclude this experiment.
14
Group
(Animal
)
Animal
No.
Diabetic
Status
Glucose
Concent.
Injection
Normal
Insulin
Normal
Insulin
Normal
Insulin
Normal
No
Insulin
Normal
No
Insulin
Normal
No
Insulin
Normal
20
Insulin
Normal
20
Insulin
Normal
20
Insulin
15
Time
(min)
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
Glucose Level
(mg/ml)
Normal
20
No
Insulin
Normal
20
No
Insulin
Normal
20
No
Insulin
Normal
40
Insulin
Normal
40
Insulin
Normal
40
Insulin
Normal
40
No
Insulin
Normal
40
No
Insulin
Normal
40
No
Insulin
Normal
80
Insulin
Normal
80
Insulin
16
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
Normal
80
Insulin
Normal
80
No
Insulin
Normal
80
No
Insulin
Normal
80
No
Insulin
Diabetic
Insulin
Diabetic
Insulin
Diabetic
Insulin
Diabetic
No
Insulin
Diabetic
No
Insulin
Diabetic
No
Insulin
Diabetic
20
Insulin
10
11
17
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
12
13
14
Diabetic
20
Insulin
Diabetic
20
Insulin
Diabetic
20
No
Insulin
Diabetic
20
No
Insulin
Diabetic
20
No
Insulin
Diabetic
40
Insulin
Diabetic
40
Insulin
Diabetic
40
Insulin
Diabetic
40
No
Insulin
18
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
15
Diabetic
40
No
Insulin
Diabetic
40
No
Insulin
Diabetic
80
Insulin
Diabetic
80
Insulin
Diabetic
80
Insulin
Diabetic
80
No
Insulin
Diabetic
80
No
Insulin
Diabetic
80
No
Insulin
16
SBP 3107
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
-30 (before
glu)
0 (before ins)
30 min after
60 min after
Introduction:
Heart is an important organ of the body. It function as a pump to circulate
blood from the liver enriched in nutrients and from the lung enriched in oxygen, to
the organs throughout the body and circulate back the blood containing metabolic
by products of the cells to the execratory organs for excretion of these metabolic by
products before the blood is circulate back to the body.
The heart function is regulated by system many factors through nervous and
hormonal systems to meet the immediate or long term supply of the blood by the
organs which varies from time to time or from one physiological condition to others.
One of the hormones that involved in regulation of heart function is adrenalin
hormone.
mainly related to stress. Adrenalin stimulates the heart function by increasing the
heart rate, force of contraction and volume of contraction with the purpose to
increase blood flow and supply to the organs. These effects will be studied in this
experiment using frog heart as a model.
20