LESSON 5: CARBOHYDRATES

QUESTIONS FOR RESEARCH:

1. Define the following terms: carbohydrate, reducing sugar, glycogenesis, gluconeogenesis,

glycolysis, glycogenolysis, hyperglycemia, insulin, hypoglycemia, glucagon, OGTT, diabetes
mellitus, and ketosis.

Carbohydrates are one of the most common and numerous organic molecules in nature since
they are necessary for all living things to function. Through the process of photosynthesis,
carbohydrates are created from carbon dioxide and water in green plants. In addition,
carbohydrates, which are both essential structural components and energy sources in living
organisms, make up a portion of the structure of nucleic acids, which carry genetic information.

Reducing sugar is the kind of sugar that functions as a reducing agent and can successfully give
electrons to another molecule by oxidizing it. According to another definition, reducing sugar is
any sugar that has the propensity to serve as a reducing agent because it either has an
aldehyde group (-CHO) or a ketone group (-CO-).

Glycogenesis, or the production of glycogen from glucose, is the process through which
animals' livers and muscles store their main source of stored carbohydrates. When blood
glucose levels are high enough, the liver and muscle cells can store extra glucose, which
triggers glycogenesis.

Gluconeogenesis, as opposed to glucose produced through the breakdown of the large storage
molecule glycogen, is the process by which new glucose molecules are created within the body.
Although the kidney and small intestine can also experience it to a lesser extent, the liver is
where it occurs most frequently. The breakdown of glucose molecules into their constituent
parts occurs during the process of glycolysis, which is the reverse of gluconeogenesis.

The method by which glucose is metabolized to produce energy is known as glycolysis. It

generates water, ATP, NADH, and two pyruvate molecules. It doesn't need oxygen to happen
because it happens in a cell's cytoplasm. In both aerobic and anaerobic organisms, it happens.

Glycogen is a stored source of energy found in muscle and liver tissue that is broken down into
glucose, which the body may use right away, in a process known as glycogenolysis. The control
of blood glucose is one of its key functions.

High blood sugar is formally referred to as hyperglycemia (blood sugar). When the body can't
properly use its insulin supply or has too little of it, high blood sugar results.

The hormone insulin has a variety of functions in the metabolism of the body. The body's use
and storage of glucose and fat are controlled by insulin.

Blood sugar (glucose) levels that are below the normal range are known as hypoglycemia.

The hormone glucagon is produced by the body naturally, primarily in the pancreas. It has the
opposite impact of what insulin does, which is to lower blood glucose levels.

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 A screening test for diabetes mellitus called the oral glucose tolerance test (OGTT) measures
plasma glucose levels after the patient swallows an oral glucose load.

Diabetes mellitus is a condition of carbohydrate metabolism characterized by inadequate

production or response to insulin by the body, which makes it difficult to maintain healthy blood
sugar (glucose) levels.

When your body is unable to burn enough carbs for energy, a condition known as ketosis takes
place. Rather, it burns fat to create something called ketones that it can utilize as fuel.

2. If a person eats a large, balanced meal containing more calories than needed for immediate
energy demands, how does the body handle the carbohydrates? How are carbohydrates
metabolized?

Because the cells in our muscles, brain, and other tissues use monosaccharides directly for
energy, carbohydrates that have been broken down to mostly glucose are the preferred source
of energy for our body. The digestive tract also contains a number of hormones, such as
glucagon and insulin. By adjusting the amount of glucose in the bloodstream, they keep our
blood sugar levels in check. If glucose is not used immediately, the body transforms it into
glycogen, a polysaccharide similar to starch that is stored in the liver and muscles as a readily
available source of energy. Our body turns glycogen back into glucose when necessary to
maintain a stable blood sugar level, such as in the intervals between meals, at night, during
bursts of physical activity, or during brief fasting intervals.

When humans eat more carbohydrates than they require at the time, the body stores part of
them (as glycogen) in the cells and turns the excess into fat. The body can quickly and
efficiently convert the complex carbohydrate glycogen into energy. The glycaemic response is
the process by which blood glucose levels increase after consuming a diet high in
carbohydrates and subsequently fall again. It shows how quickly glucose is absorbed and
digested, as well as how well insulin works to keep blood sugar levels normal.

The process of breaking down complex sugars into monosaccharides by the salivary amylase
enzyme is the first step in the metabolism of carbohydrates. These may then penetrate the
intestinal membrane and enter the bloodstream before reaching bodily tissues. Foods are
broken down into sugars, sometimes referred to as saccharides, through the digestion or
metabolism of carbohydrates. The body uses these chemicals, which start digesting in the
mouth, for everything from regular cell function to cell growth and repair.

3. What hormones are involved in glucose homeostasis and how do they act? What factors are
involved in the regulation of glucose and how do they specifically affect blood glucose
concentration?

Through a system known as the Feed Mechanism, hormones keep the internal environment of
the human body in a state of homeostasis. In a similar way, the Thermostat of various domestic
appliances causes them to start or stop operating at a certain temperature (it maintains the
balance of machine working).

The amount of blood glucose sugar, or "homeostasis," is significantly influenced by the

hormones insulin and glucagon.

When raising blood glucose levels over their typical levels. The pancreas reacts by releasing the
hormone insulin, which causes the liver to store a rise in blood glucose levels and stimulates the

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 body's cells to take up glucose from the blood.

After lowering blood glucose levels below what is considered normal. The pancreas reacts by
releasing the glucagon hormone, which influences the liver and causes it to convert glycogen
stored there more quickly into glucose sugar, raising blood glucose levels until they return to
normal.

When the pancreas releases less of the hormone insulin. Increases in blood glucose levels or
the development of diabetes in a person.

4. What is diabetes mellitus and what are the clinical laboratory assays that are used to diagnose
and confirm this disorder?

Hyperglycemia arises from a collection of metabolic illnesses known as diabetes mellitus, in

which glucose is not properly utilized.

Plasma glucose and HbA1c are two laboratory tests that are used in the diagnosis and follow-up
of diabetic patients. With glucose, information from numerous assay techniques can be merged
to produce a longitudinal patient record that uses standardized assays and reference ranges
that are widely accepted. On the other hand, due to variability in interference from different
hemoglobins, patient findings for HbA1c should not be pooled from different techniques.
Consideration should also be given to the red blood cell (RBC) survival times, with shorter
survival durations leading to an artificial reduction in the HbA1c.

Using the assumption that RBCs have an average lifespan of 120 days while in circulation, the
HbA1c result is utilized to offer an indication of the patient's glycemic control during the previous
two to three months. During that time, the Amadori rearrangement creates HbA1c from the wild
type (or typical) HbA, permanently binding blood glucose to hemoglobin in red blood cells. An
average estimated glucose level (eAG), which may be determined from the percentage of
HbA1c, can be found by multiplying the amount of circulating glucose by the percentage of
HbA1c.

5. What are the underlying principles for each of the following methods and what specific
interferences might lead to the misinterpretation of results from these assays: glucose oxidase,
glucose dehydrogenase, and hexokinase.

Glucose oxidase
o The emission of hydrogen peroxide occurs simultaneously with the oxidation of -D-
glucose to D-glucono-lactone, which is catalyzed by glucose oxidase. This hydrogen
peroxide (H2O2) engages in a second reaction with 4-aminoantipyrine and p-
hydroxybenzoic acid in the presence of peroxidase (POD), which results in the
quantitative synthesis of a quinoneimine dye complex that is detected at 510 nm.

Glucose dehydrogenase
o The following reaction is used to determine the change in absorbance at 340 nm.

GLUCOSE ↔ D-GLUCONO-δ-LACTONE
NAD⁺↔NADH + H⁺

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 Hexokinase
o Glucose +ATP ↔ Glucose 6 phosphate +ADP
Glucose 6 Phosphate + NAD ↔ 6-
Phosphogluconate + NADH+H⁺
Conversion of NADH from NAD at 340nm
increase in O.D. is measured at fix interval
Increase O.D. /min is directly conc. of glucose in
the specimen = ∆ O.D.

The conversion of glucose to glucose-6-phosphate (G-6-P) and adenosine diphosphate

(ADP) is catalyzed by the enzyme hexokinase (HK) (ADP). The enzyme glucose-6-
phosphate dehydrogenase (G-6-PD) oxidizes G-6-P to 6-phosphogluconate and
reduced nicotinamide adenine dinucleotide in the presence of nicotinamide adenine
dinucleotide (NAD) (NADH). At 340 nm, the increase in NADH concentration may be
detected spectrophotometrically and is directly proportional to the rise in glucose
concentration.

6. What are the differences between type 1, type 2, and gestational diabetes mellitus? How do
clinical laboratory assay values compare between the three types?

TYPE 1 DIABETES

The failure of a pancreas to make insulin causes type 1 diabetes. Type 1 diabetes arises as a
result of the immune system incorrectly classifying pancreatic cells as foreign intruders and
eliminating them. Type 1 diabetics who want to stay healthy must inject themselves with insulin
every day. Although type 1 diabetes can develop at any age, it is more frequently found in
children or young people. Although no one is certain of what causes type 1 diabetes, experts
think a combination of genetic and environmental factors are to blame despite the fact that the
exact etiology is likewise unknown.

Your doctor will use the random plasma glucose test to measure your blood glucose at a single
point in time if you have obvious signs of diabetes. The glycated hemoglobin test, also known as
the A1C test, which measures your average blood sugar levels over the previous three months,
may be performed if your blood sugar is high when the test is delivered. These tests can
establish that you have diabetes, but they cannot identify the kind for your doctor. An
autoantibody test can be used to determine the kind. Autoantibodies are antibodies that wrongly
target your healthy cells and organs. In type 1 diabetes, certain autoantibodies are frequently
found, but not in type 2 diabetes.

TYPE 2 DIABETES

You could get type 2 diabetes, which is the most common type, if your pancreas doesn't
generate enough insulin or if your body doesn't use insulin properly. Despite typically affecting
middle-aged and older persons, type 2 diabetes can strike anyone at any age.

The A1C test is most frequently used to identify type 2 diabetes. Other diagnostic procedures
include a random blood sugar test, which is carried out regardless of when you last had a meal,
a fasting blood sugar test, which is given after an overnight fast, and an oral glucose tolerance
test (OGTT), which entails fasting the previous night, drinking a sweet beverage, and having
blood sugar levels monitored repeatedly for several hours.

A routine type 2 diabetes screening is advised for persons 45 years of age and older, as well as
individuals who are overweight or obese and who have at least one significant diabetes risk

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 factor. Women who have gestational diabetes should have regular type 2 diabetes testing.

GESTATIONAL DIABETES

Some pregnant women acquire gestational diabetes, which usually, but not always, resolves on
its own after the baby is born. Gestational diabetes can create health concerns for both the
mother and the baby, so it is critical to manage the illness for both maternal and child health.
Women who have gestational diabetes are more likely to develop type 2 diabetes later in life.
Pregnant women who are diagnosed with gestational diabetes may also have type 2 diabetes.

Pregnant women typically are screened for gestational diabetes around weeks 24 and 28 of
their pregnancy. A glucose test, also known as a glucose screening test, is done an hour after
they consume a sugary liquid containing glucose, and if their blood glucose level is high, then
their doctor may request that they return for an oral glucose tolerance test (OGTT).

7. What are some of the causes of hypoglycemia, and how would you test for them?

Hypoglycemia happens when the blood sugar level falls below a healthy level. It is also known
as low blood glucose.

COMMON CAUSES:

o Taking in greater doses of diabetes drugs such as insulin, sulfonylureas (for example,
glibenclamide and gliclazide), and prandial glucose regulators (for example, repaglinide,
nateglinide).
o Antimalarial medications, for instance.
o Certain hormones important with glucose metabolism, such as cortisol, are deficient.

DIAGNOSIS
Hypoglycemia is easily detected by measuring glucose levels.
o Blood glucose test
 A blood test to measure the blood sugar levels at that particular point of time is
the only diagnostic method for hypoglycemia.

8. What are some examples of inherited disorders of carbohydrate metabolism?

An inherited or acquired disorder that affects the metabolism of the carbohydrates.

Representative examples include diabetes mellitus, glycogen storage disease,
mucopolysaccharidoses, and lactose intolerance.

9. What is the principle of glucose measurement in a hand-held glucose meter? What can affect
these values?

The glucometer's principal function is to measure the amount of "sugar" in the blood. This

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 activity can be performed in two ways. The first choice is photometric determination, whereas
the second is electromechanical determination.
Although blood glucose measurement is commonly performed, the use of a whole-blood sample
introduces complications and compromise in terms of the assay principle, the method of
calibration and the expression of results.

Most point-of-care devices are calibrated against a method chosen by the manufacturer for
reference purposes, and assumptions are made that blood samples from different people will
behave similarly in both the reference and point-of-care procedures, which are not necessarily
right.

Whereas most standard laboratory procedures assess blood glucose concentration in plasma or
whole blood, direct-reading electrode systems measure it as molality in mmol/kg water, which is
quantitatively larger, but results have been frequently factored and presented as plasma glucose
concentration, for instance.

Nevertheless, there is inconsistency, and the range of methodologies and concepts makes
comparing findings of blood glucose tests by different methods challenging.

It has been hypothesized that by stating all data as plasma glucose concentration, regardless of
specimen type or analytical method utilized, some uncertainty could be avoided.

Variation in the location of blood sampling can potentially create mistakes, particularly in point-
of-care testing.

10. What are the specimen requirements for fasting glucose, insulin, glycated hemoglobin, glucose
meter analyses, and urine glucose?

FASTING GLUCOSE
o Specimen type: Plasma
o Preferred collection container: 4 mL grey-top (Na fluoride/K oxalate) tube
o Specimen required: 2 mL plasma; minimum 0.5 mL.
o Special notes: Place specimen on ice, or centrifuge and separate plasma from cells.
o Patient preparation: 8-hour fasting is required.

INSULIN
o Specimen type: Serum or plasma
o Preferred collection container: 3.5 mL gold-top (serum separator) tube
3 mL green/yellow-top (plasma separator) tube
o Specimen required: 2 mL aliquot of serum or plasma; minimum 0.5 mL. Please submit a
separate frozen aliquot for this test. It may not be combined with any other test requiring
a frozen aliquot.

GLUCOSE METER ANALYSES

o Specimen type: Plasma or Serum
o Preferred collection container: Stat/Line draws: 3 mL green/yellow-top (plasma
separator) tube
o Routine requests/off-site specimens: 3.5 mL gold-top (serum separator) tube
4 mL grey-top (Na fluoride/K oxalate) tube

o Specimen required: 2 mL of serum or plasma; minimum 0.5 mL.

o Special notes: Outreach clients: Specimen may also be collected in grey-top tubes.
Place specimen on ice, or centrifuge and separate plasma from cells.

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 o Patient preparation: Fasting specimen is not required. For fasting glucose, please order
Fasting Plasma Glucose (GLUFP).

URINE GLUCOSE
o Specimen type: Random urine
o Preferred collection container: Non-sterile specimen container
o Specimen required: 5 mL of urine; minimum 1 mL.
o Special notes: Urine containing preservatives is not acceptable.

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