Bryan Janier

C1
Experiment 5 Tests to Diagnose Diabetes Mellitus
1. Give the characteristic differences between the glucose tolerance curve of a diabetic person
and that of a normal individual. (Ayens answer)
After ingestion of glucose, diabetic patients would exhibit a much greater than normal
rise in blood glucose level, as demonstrated by the upper curve in Figure 1, and the glucose level
falls back to the control value only after 4 to 6 hours. Furthermore, it fails to fall below the control
level. The slow fall of this curve and its failure to fall below the control level demonstrate that
either (1) the normal increase in insulin secretion after glucose ingestion does not occur, or (2)
the person has decreased sensitivity to insulin. A diagnosis of diabetes mellitus can usually be
established on the basis of such a curve, and type 1 and type 2 diabetes can be distinguished from
each other by measurements of plasma insulin with plasma insulin being low or undetectable in
type 1 diabetes and increased in type II diabetes.

Figure 1 - Glucose tolerance curve in a normal person and in a person with diabetes

2. What is the principle involved in the blood glucose determination used in this experiment?
In the presence of the glucose oxidase, D-glucose is specifically oxidized to gluconic acid and
hydrogen peroxide. Hydrogen peroxide is then converted to O-diansidine, through the catalytic
action of peroxidase to the red brown semiquinone. The colour intensity is directly proportional
to the glucose concentration and is measured photometrically.
3. What is the importance of the following especially regarding interpretation of results?
A. Doing an eight-hour overnight fasting versus an eight hour fast anytime of the day
B. Separation of the plasma within 60 minutes of blood collection

Bryan Janier
C1
Fasting overnight versus an eight hour fast anytime of the day
-

Variation in results from various factors like stress, strenuous activities and smoking which
are experience during the day can be avoided when doing an 8-hour overnight fasting.
Diurnal variations in cortisol levels can also affect glucose levels since cortisol increase blood
glucose.

Within 60 minutes of blood collection

- Serum or plasma must be separated from the cells within one hour to prevent losses of
glucose (preferably within 30 minutes)
- At room temperature 20-25oC, glycolysis decreases glucose by 5-7% per hour (5-10 mg/dL) in
normal uncentrifuged coagulated blood
- At refrigerated temperature (4oC), glucose is metabolized at the rate of about 1-2 mg/dL per
hour
- WBC and RBC metabolize glucose resulting to decrease value in clotted, uncentrifuged blood
(leukocytosis can lead to excessive glycolysis)
4. Give 2 examples of point of care testing (POCT) glucometers and differentiate the principles and
results from whole blood determinations.
For inpatient care, central laboratory testing of plasma glucoseextremely accurate and
precise remains the gold standard. Its main disadvantage is that this analysis is remotely located
from the patient, thereby increasing the time for starting treatment based on test results.
POCT glucose devices deliver rapid and relatively precise glucose measurements performed on
whole blood at the patients bedside. Glucose POCT devices are regarded as satisfactorily precise
for the monitoring of many glycemic disorders but not sufficiently precise to establish the initial
diagnosis of the condition.

Variance seen in the plasma concentration may be as much as 12% to 15% higher than whole
blood values. This is a reflection of the water content of red blood cells, which is lower than
plasma, thus resulting in dilution of the glucose concentration. Fasting glucose concentrations in
capillary blood (POCT) are slightly higher than in venous blood, but the disparity between the two
can translate into statistically significant differences (up to 70 mg/dl) in postprandial specimens.
This occurs because the postprandial capillary specimens are glucose-rich due to not having
delivered their glucose load to the tissues, while, conversely, the venous specimens are glucosepoor post-systemic samples.

Bryan Janier
C1
5. Explain the rationale of why glycosylated hemoglobin is a good test of glycemic control? What
conditions can falsely elevate or increase glycosylated hemoglobin values?
The HbA1c test measures the amount of sugar that is attached to the hemoglobin in red
blood cells, with results given as a percentage. Red blood cells live in the bloodstream for about
four months, the HbA1c test shows the average blood sugar for the past several months Unlike
your regular blood sugar test, the HbA1c test is not affected by short-term changes.
Glycation of hemoglobin is permanent. A buildup of glycated hemoglobin within the red
cell, therefore, reflects the average level of glucose to which the cell has been exposed during
its life-cycle. Measuring glycated hemoglobin assesses the effectiveness of therapy by monitoring
long-term serum glucose regulation. The HbA1c level is proportional to average blood glucose
concentration over the previous four weeks to three months. Normal levels of glucose produce a
normal amount of glycated hemoglobin. As the average amount of plasma glucose increases, the
fraction of glycated hemoglobin increases in a predictable way. This serves as a marker for average
blood glucose levels over the previous months prior to the measurement.
Not enough iron in bloodstream may lead to false positive in A1C test results. Most people
have only one type of hemoglobin, called hemoglobin A. If you have an uncommon form of
hemoglobin (known as a hemoglobin variant), your A1C test result may be falsely high or falsely
low. Hemoglobin variants are most often found in blacks and people of Mediterranean or
Southeast Asian heritage. Patients with abnormal hemoglobin, Hb S and F can also cause false
postives. The hemoglobin A1c level can be falsely high in patients who have end-stage renal
disease (ESRD), as both uremia and dialysis can complicate glycemic control by affecting the
secretion, clearance, and peripheral tissue sensitivity of insulin.
6. How is glucose/Hba1c testing results interpreted? Based on your result, what is your samples
value in the glucose continuum?
For someone who doesn't have diabetes, a normal A1C level can range from 4.5 to 6 percent.
Someone who's had uncontrolled diabetes for a long time might have an A1C level above 9
percent. When the A1C test is used to diagnose diabetes, an A1C level of 6.5 percent or higher on
two separate tests indicates you have diabetes. A result between 5.7 and 6.4 percent is
considered prediabetes, which indicates a high risk of developing diabetes.

Bryan Janier
C1
Experiment 6 Establishing a Lipid Profile
1. How does a dietary cholesterol enter the circulation?
Digestion of triacylglycerols
-

Dietary fat (triacylglycerol) leaves the stomach and enters the small intestine where it is
emulsified (suspended in small particles in the aqueous environment) by bile salts.
Pancreatic lipase hydrolyzes fatty acids of all chain lengths from positions 1 and 3 of the
glycerol moiety of the triacylglycerol, producing free fatty acids and 2-monoacylglycerol.

Absorption of dietary lipids

-

The fatty acids and 2-monoacylglycerols produced by digestion are packaged into micelles,
tiny microdroplets that are emulsified by bile salts.
The micelles travel through a layer of water (the unstirred water layer) to the microvilli on the
surface of the intestinal epithelial cells, where the fatty acids, 2-monoacylglycerols, and other
dietary lipids are absorbed, but the bile salts are left behind in the lumen of the gut.
Short- and medium-chain fatty acids (C4 to C12) do not require bile salts for their absorption.
They are absorbed directly into intestinal epithelial cells. Because they do not need to be
packaged to increase their solubility, they enter the portal blood (rather than the lymph) and
are transported to the liver bound to serum albumin.

Synthesis of chylomicrons
-

Within the intestinal epithelial cells, the fatty acids and 2-monoacylglycerols are condensed
by enzymatic reactions in the smooth endoplasmic reticulum to form triacylglycerols.
Triacylglycerols are transported in lipoprotein particles because they are insoluble in water. If
triacylglycerols entered the blood directly, they would coalesce, impeding blood flow.

Transport of dietary lipids in the blood

-

By the process of exocytosis, nascent chylomicrons are secreted by the intestinal epithelial
cells into the chyle of the lymphatic system and enter the blood through the thoracic duct.
Nascent chylomicrons begin to enter the blood within 1 to 2 hours after the start of a meal;
as the meal is digested and absorbed, they continue to enter the blood for many hours.

2. How does it regulate de novo cholesterol synthesis in the liver?

(Ayens answer)

3. What are the various types of hyperlipidemias?

Type I
- Due to accumulation of chylomicrons
- Two genetic forms are known
o Lipoprotein lipase deficiency and ApoCII deficiency

Bryan Janier
C1
-

Patients with type I hyperlipidemia have exceedingly high plasma triacylglycerol levels (over
1000 mg dL-1) and suffer from eruptive xanthomas (triacylglycerol deposits in the skin) and
pancreatitis

Type II
-

Characterized by elevated LDL levels

Due to genetic defect sin the synthesis, processing, or function of the LDL receptor
o Heterozygotes have elevated LDL levels
o Homozygous patients have very high LDL
May suffer myocardial infarctions before age 20

Type III
-

Due to abnormalities of ApoE which interfere with the uptake of chlymicron and VLDL
remnants
Hypothyroidism can produce a very similar hyperlipidemia
These patients have increased risk of atherosclerosis

Type IV
-

Most common abnormality

VLDL levels are increased often due to
o Obesity
o Alcohol abuse
o Diabetes
Familial forms are also known but the molecular defect is unknown

Type V
-

Similar to type I
Associated with high chylomicron triacylglycerol levels, pancreatitis, and eruptive xan-thomas