HOMEOSTASIS:

CONCEPT AND APPLICATION IN

CLINICAL CHEMISTRY

By

BASIL, B – MBBS (Nig), FMCPath.

Consultant Chemical Pathologist/Metabolic Medicine Physician.

July 2022.
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 Let’s try these:
1. Because they are 2. Ion pumps and
embedded within the phagocytosis are both
membrane, ion channels examples of ________.
are examples of _____. a) Endocytosis
a) receptor proteins b) passive transport
b) integral proteins c) active transport
c) peripheral proteins d) facilitated diffusion
d) glycoproteins

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 Let’s try these:
3. What volume of a 4. Calculate the amount
6.0M glucose stock in moles of sodium
solution is necessary to hydroxide (NaOH)
prepare 2.5 L of 0.4M needed to make 2.5L of
solution? 0.1M NaOH.
a) 333mL a) 0.125 mol
b) 166mL b) 0.250 mol
c) 666mL c) 0.500 mol
d) 83mL d) 1.000 mol

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 Introduction:
• Systems attain and exist in “stable state” unless altered
by an “external” force.
• The source of the external impact may be from within
or without the organism, but certainly not a regular
part of the system affected.
• In humans, the body has the tendency and ability to
make adjustments so as to contain the external impact
and reestablish a stable state.
• Frequently, the body’s attempt to stabilize the affected
system, or failure to do so, results in diseased states.
• Distortions of this stable state usually commence at the
molecular level prior to overt clinical manifestations.
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 What is Homeostasis?
• Homeostasis is the body’s tendency to resist change in
order to maintain a stable, relatively constant internal
environment.
• It can be influenced by either internal or existing
conditions (intrinsic factors), or external or
environmental conditions (extrinsic factors).
• Components of all homeostatic mechanisms include:
• Sensor or receptor – detects changes in the internal or
external environment.
• Integrating center or control center – receives information
from the sensors and initiates the response to maintain
homeostasis.
• Effector – organ or tissue that receives information from the
control center and acts to bring about the changes needed to
maintain homeostasis
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 Mechanisms of Control:
• Homeostatic control can be achieved via feedback
mechanisms – negative or positive feedback.
• Negative feedback (commoner):
• Uses one of the products of the reaction to reduce the output or
activity of the process and return the system to its normal range of
functioning.
• Set-point or normal range may be altered by certain conditions,
which will be “unexpectedly” maintained by this mechanism.
• Positive feedback:
• Activated component enhances or further upregulates the process
that gave rise to itself in order to create an even stronger response.
• Designed to push levels out of normal ranges – not used as often in
homeostatic responses.

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Positive vs Negative Feedback:

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 Disease – A Homeostatic
Imbalance:
• Disease is any failure of normal physiological function
that leads to negative symptoms.
• While disease is often a result of infection or injury,
most diseases involve the disruption of normal
homeostasis.
• Anything that prevents positive or negative feedback
system from working correctly could lead to disease if
the mechanisms of disruption become strong enough.
• Many diseases are caused by a broken feedback loop
• Specific examples of homeostatic imbalance would be
discussed during the course of your pathology program.

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Evaluating Homeostasis to Assess
Health:
• Since homeostatic imbalances can lead to disease
states or even death, homeostasis has been identified
as one of the eight core concepts of biology.
• The American Association of Medical Colleges reports
that ability to identify and apply knowledge about
homeostasis should be regarded as one of the key
competencies of physicians.
• Thus, physicians need a way to evaluate the
homeostatic health of their patients.
• They need to be able to evaluate the mixtures of
compounds that are found within the human body –
vital to the Chemical Pathologist.
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 Some Useful Terminologies in
Homeostatic Assessment:
• Mixture – substance that is composed of two or more
substances.
• Heterogeneous – composition is not uniform throughout the
mixture
• Homogeneous – combine intimately and observed as a single
substance
• Colloid – contains particles with diameters ranging from 2 to 500
nm; and are cloudy or opaque
• Solution – have particle sizes of a typical ion or small molecule
(~0.1 - 2 nm in diameter), are transparent, and may be colored
• More emphasis would be laid on solutions which are the
main type of mixture encountered in the clinical lab
• The major component of the solution is called the solvent,
and the minor component(s) are called the solute.
• Solutions – may be gas-gas solutions, liquid solutions or
solid solutions… Examples???
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 Determining Concentration of
Solutions:
• Concentration – amount of solute in a given volume of a
solution.
• The most common ways of presenting solution
concentration are Molarity and Parts Per Solutions.
• Molarity – number of moles of solute present in exactly 1L
of solution.
• The units of molarity are therefore moles per liter of
solution (mol/L), abbreviated as M (read as molar).

• The equation above can be used to calculate how much

solute is required to make any amount of a desired solution.
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Example 1 – Problem:
• Calculate the number of moles of sodium hydroxide
(NaOH) needed to make 2.50 L of 0.100 M NaOH.
• Given:
(1) identity of solute = NaOH,
(2) volume of solution = 2.50 L, and
(3) molarity of solution = 0.100 mol/L
Note: when calculating problems always write out the units of
molarity as mol/L, rather than M. This will allow you to cancel out
your units when doing the calculation.
• Asked for:
• Amount of solute in moles.
• Strategy:
• (1) Rearrange the equation above to solve for the desired unit, in
this case for moles.
• (2) Double check all the units in the equation and make sure they
match. Perform any conversions that are needed so that the units
match.
• (3) Fill in values appropriately and do the math.
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Example – Solution:
• (1) Rearrange the equation above to solve for moles.

• (2) Double check all the units in the equation and make sure they
match.
• Volume = 2.50 L; molarity = 0.100 mol/L.
• The volume units for both are in Liters (L) and thus, match. Therefore,
no conversions need to be made.
• (3) Fill in values appropriately and do the math:

? mol of NaOH = 0.250 mol of NaOH

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Example – Preparing the Solution:
• Convert the amount in moles of the sample into the amount in
grams represented by that number.
• (1) Calculate the molecular mass of NaOH:
? g/mol of NaOH = 23 + 16 + 1 = 40g/mol of NaOH

• (2) Convert moles of NaOH to grams of NaOH:

0.25 mol of NaOH x 40 g NaOH
1 mol NaOH

? g of NaOH = 10g NaOH

• Therefore, 10g of NaOH is needed to make 2.5L of a 0.1M

solution.
• To actually make the solution, dissolve the solute in a small
amount of the solvent and then once the solute is dissolved, the
final volume can be brought up to 2.5 L.
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Example – Preparing the Solution:

Diagram showing the preparation of a solution of known concentration by

dissolving a measurable amount (10.0g in this case) of the solute in a specified
volume (500mL) of solution.
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 Determining Concentration of
Solutions – contd:
• Parts Per Solutions – expresses the concentration based on
the quantity of solute in a fixed quantity of solution.
• The “quantities” referred to here can be expressed in mass,
in volume, or both (i.e., m/m, v/v or m/v).
• In clinical chemistry, (m/v) is commonly used, with mass
expressed in grams and volume in mL, as in g/mL or g/dL
• It may also be expressed as “parts per 100” (i.e., “percent”).
• Also, recall that the solution is the sum of both the solvent
and the solute when you are performing percent
calculations.

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Example 2 – Problem/solution:
• A 7.0% v/v solution of ethanol in water, would
contain 7 mL of ethanol in a total of 100 mL of
solution. How much water is in the solution?
• In this problem, we know that the:
Solution = Solute + Solvent

• Thus, we can fill in the values and then solve for the
unknown.
100 mL = 7 mL + X mL of Solvent (in this case water)

• shifting the 7 over to the other side, we can see that:

100 mL – 7 mL = 93 mL H2O

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Example 3 – Problem/Solutions:
• What is the (m/v)% of a solution if 24.0 g of sucrose is
dissolved in a total solution of 243 mL?

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Example 4 – Problem/Solutions:
• How many grams of NaCl are required to make 625
mL of a 13.5% solution?

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 Dilutions:
• Homeostatic changes may occur in response to changes in
the concentrations of solutions (e.g plasma, urine) in the
body.
• A solution of a desired concentration can be prepared by
diluting a small volume of a concentrated solution with
additional solvent.
• Stock solution – solution of known concentration, often
used to prepare solutions of desired concentration.
• Diluting a stock solution is preferred when making solutions
of very weak concentrations, because the alternative
method, weighing out tiny amounts of solute, can be
difficult to carry out with a high degree of accuracy.
• Dilution is also used to prepare solutions from substances
that are sold as concentrated aqueous solutions, such as
strong acids.
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Preparing a solution of known concentration:
• Involves determining the amount of solute desired in the
final volume of the more dilute solution, and then
calculating the volume of the stock solution that contains
this amount of solute.
• The law of mass action explains the relationship between
the volume and concentration of the stock solution and the
volume and concentration of the desired diluted solution
can be expressed mathematically as:
MsVs = MdVd
Where:
• Ms is the concentration of the stock solution,
• Vs is the volume of the stock solution,
• Md is the concentration of the diluted solution, and
• Vd is the volume of the diluted solution.
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Preparation of a Solution of Known
Concentration by Diluting a Stock Solution:

(a) A volume (Vs) containing the desired amount of solute (Ms) is measured from a
stock solution of known concentration. (b) The measured volume of stock solution
is transferred to a second volumetric flask. (c) The measured volume in the second
flask is then diluted with solvent up to the volumetric mark [(Vs)(Ms) = (Vd)(Md)].
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Example:
• What volume of a 3.00 M glucose stock solution is
necessary to prepare 2500 mL of 0.400 M solution?
• Given: volume and molarity of dilute solution, and
molarity of stock solution
• Asked for: volume of stock solution
• For Dilution problems, as long as you know 3 of the
variables, you can solve for the 4th variable.
• Start by rearranging the equation to solve for the variable
that you want to find.
• In this case, you want to find the volume of the stock
solution, Vs

.
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Example – contd:
• Next, check to make sure that like terms have the same
units.
Vs = 333mL of Stock Solution
• Note that if 333mL of stock solution is needed, then you can
also calculate the amount of solvent needed to make the
final dilution.
• Total volume – volume of stock solution = volume of solvent
needed for the final dilution.
• In this case 2,500 mL – 333 mL = 2,167 mL of water needed
to make the final dilution.
• This should be done in a graduated cylinder or volumetric
flask.

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 Movement of substances across
membranes:
• Cell membranes have the ability to regulate intracellular
concentration of substances and are selectively permeable.
• Movement of substances across the membrane occurs via
• Passive transport – without the expenditure of cellular energy
• Active transport – using energy from ATP
• Passive transport occurs via diffusion down a concentration
gradient, and involves substances like O2, CO2, etc., capable
of freely moving across certain membranes.
• This mechanism of molecules moving across a cell
membrane from the side where they are more concentrated
to the side where they are less concentrated is a form of
passive transport called simple diffusion

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 Movement of substances across
membranes – contd:

Simple Diffusion across the Cell (Plasma) Membrane:

• The structure of the lipid bilayer allows small, uncharged substances
such as oxygen and carbon dioxide, and hydrophobic molecules such as
lipids, to pass through the cell membrane, down their concentration
gradient, by simple diffusion.
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 Movement of substances across
membranes – contd:
• Protein channels and specialized transport mechanisms are
needed for substances that cannot cross the lipid bilayer
due to their size, charge, and/or polarity – Facilitated
diffusion.
• This may be via use of:
• Channel proteins – less selective, or
• Carrier proteins – more selective.
• NB: facilitated diffusion is a passive process.
• Water also can move freely across the cell membrane of all
cells, either through protein channels or by slipping
between the lipid tails of the membrane itself.
• This may be by osmosis – Define Osmosis?

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 Movement of substances across
membranes – contd:

Facilitated Diffusion:
(a) Facilitated diffusion of substances crossing the cell (plasma) membrane takes place with the help
of proteins such as channel proteins and carrier proteins. Channel proteins are less selective than
carrier proteins, and usually mildly discriminate between their cargo based on size and charge. (b)
Carrier proteins are more selective, often only allowing one particular type of molecule to cross.28
 Movement of substances across
membranes – contd:

Osmosis:
• Osmosis is the diffusion of water through a semipermeable membrane
down its concentration gradient. If a membrane is permeable to water,
though not to a solute, water will equalize its own concentration by
diffusing to the side of lower water concentration (and thus the side of
higher solute concentration). In the beaker on the left, the solution on
the right side of the membrane is hypertonic. 29
 Movement of substances across
membranes – contd:
• Movement of water molecules is not itself
regulated by cells but by the concentration of
solutes in the ICF vs ECF – Tonicity.
• The relative solute concentration of a cell to that of
the ECF surrounding the cells can be
• Hypotonic
• Isotonic, or
• hypertonic

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 Movement of substances across
membranes – contd:

States of Tonicity.
• A hypertonic solution has a solute concentration higher than another
solution. An isotonic solution has a solute concentration equal to
another solution. A hypotonic solution has a solute concentration lower
than another solution.
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 Movement of substances across
membranes – contd:
• Active transport:
• ATP is required to move a substance across a membrane,
often with the help of protein carriers, and usually against
its concentration gradient.
• Types:
• Pumps – e.g., Na-K pump
• Symporters – secondary active transporters moving two
substances in the same direction usually by taking
advantage of concentration gradient created by pumps e.g
Na-glucose symporter
• Antiporters – also a secondary active transporter e.g., Na-
H antiporter
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 Movement of substances across
membranes – contd:

Sodium-Potassium Pump:
• The sodium-potassium pump is found in many cell (plasma) membranes. Powered by ATP,
the pump moves sodium and potassium ions in opposite directions, each against its
concentration gradient. In a single cycle of the pump, three sodium ions are extruded from
and two potassium ions are imported into the cell.
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 Movement of substances across
membranes – contd:
• Other forms of active transport do not involve membrane
carriers:
• Endocytosis:
• Phagocytosis
• Pinocytosis
• Receptor-mediated endocytosis
• Exocytosis
• Phagocytosis and pinocytosis take in large portions of
extracellular material, and they are typically not highly
selective in the substances they bring in.
• Receptor-mediated endocytosis is endocytosis by a portion
of the cell membrane that contains many receptors that are
specific for a certain substance.
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 Movement of substances across
membranes – contd:

Three Forms of Endocytosis:

• Endocytosis is a form of active transport in which a cell envelopes extracellular
materials using its cell membrane. (a) In phagocytosis, which is relatively
nonselective, the cell takes in a large particle. (b) In pinocytosis, the cell takes in
small particles in fluid. (c) In contrast, receptor-mediated endocytosis is quite
selective. When external receptors bind a specific ligand, the cell responds by
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endocytosing the ligand.
 Movement of substances across
membranes – contd:
Exocytosis.
• Exocytosis is much like
endocytosis in reverse. Material
destined for export is packaged
into a vesicle inside the cell. The
membrane of the vesicle fuses
with the cell membrane, and the
contents are released into the
extracellular space.
• Some endocrine cells utilize this
transport method for release of
their hormones.

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 Now you can do better!
1. Because they are 2. Ion pumps and
embedded within the phagocytosis are both
membrane, ion channels examples of ________.
are examples of _____. a) Endocytosis
a) receptor proteins b) passive transport
b) integral proteins c) active transport
c) peripheral proteins d) facilitated diffusion
d) glycoproteins

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 Now you can do better!
3. What volume of a 4. Calculate the amount
6.0M glucose stock in moles of sodium
solution is necessary to hydroxide (NaOH)
prepare 2.5 L of 0.4M needed to make 2.5L of
solution? 0.1M NaOH.
a) 333mL a) 0.125 mol
b) 166mL b) 0.250 mol
c) 666mL c) 0.500 mol
d) 83mL d) 1.000 mol

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THANKS!!!
QUESTIONS???

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