HISTOPATHOLOGIC AND CYTOLOGIC DISEASE PROCESS: ETIOLOGY

TECHNIQUES LECTURE
Two major classes of etiologic factors.
GENERAL PATHOLOGY: AN INTRODUCTION
1. Genetic – inherited mutations,
polymorphisms, etc.)
INTRODUCTION
2. Acquired – infection, nutritional, chemical,
physical
- PATHOLOGY is the study of disease
- Logos (study)
One etiologic factor – One disease
- Pathos (disease)
- (Koch’s postulate or Germ Theory of Disease):
- study of the structural, biochemical, and
NOT applicable to majority of diseases
functional changes in cells, tissues, and organs
that underlie disease
Most diseases are multifactorial
- attempts to explain the signs and symptoms
manifested by patients by using molecular,
DISEASE PROCESS: PATHOGENESIS
microbiologic, immunologic, and morphologic
techniques, providing a rational basis for
- Refers to the sequence of events in the
clinical care and therapy.
response of cells or tissues to the etiologic
- General Pathology vs. Systemic Pathology
agent, from the initial stimulus to the ultimate
expression of the disease
Anatomic pathology
- Studies gross and microscopic morphological
changes in cells, tissues and organs to diagnose
disease

Clinical pathology
- Studies changes in cells and tissues using
biochemical, immunological, microbiological - One of the main domains of pathology
and molecular techniques to diagnose disease. - Molecular pathology

Molecular pathology
- Is defined as the analysis of nucleic acids and
proteins for the following purposes
• To diagnose disease
• To predict the occurrence of disease
• To predict the prognosis of diagnosed
disease, and
• To guide therapy such as selecting - A cell from amniotic fluids that is positive for
treatments and monitoring the trisomy 21 by FISH (red signals)
effectiveness of therapies.

INTRODUCTION: Disease Process

- ETIOLOGY cause
- PATHOGENESIS mechanism of development
- MOLECULAR/MORPHOLOGIC CHANGES
biochemical and structural alterations induced
in the cells and organs of the body.
- CLINICAL MANIFESTATIONS functional
consequences of these changes.
- The pathologist is on the case starting with the
H&E all the way to the FISH electronic sign –
out.
 DISEASE PROCESS: MOLECULAR/MORPHOLOGIC disease, as well as its progress (clinical course
CHANGES and outcome)

IMMUNOHISTOCHEMISTRY APPLICATION

FATHER OF MODERN PATHOLOGY

Who is Rudolf Virchow?

- From a soft tissue tumor arising from the leg
of a 15-year-old male
- EWINGS SARCOMA VS. PNET
- The tumor cells exhibit CD99 + membranous
staining.
- German pathologist and stateman, one of the
most prominent physicians of the 19th century.
- He is considered as the “FATHER OF MODERN
PATHOLOGY”
- Pioneered the modern concept of pathological
processes by his application of the cell theory
to the organs and tissues of the body.
▪ Emphasized that diseases arose, not in
organs or tissues in general, but primarily in
their individual cells.
- October 13, 1821 – September 5, 1902

“Virtually all forms of tissue injury starts with

molecular or structural alterations in CELLS.”

“The body is a cell state in which every cell is a citizen.

Disease is merely the conflict of the citizens of the
state brought about by the action of external forces”

- Rudolf Virchow

DISEASE PROCESS: FUNCTIONAL DERANGEMENTS

AND CLINICAL MANIFESTATIONS

- End results of genetic, biochemical, and

structural changes in cells and tissues which
are functional abnormalities.
- These abnormalities then lead to the clinical
manifestations (symptoms and signs) of
 REVIEW OF NORMAL CELL STRUCTURE FUNCTIONS OF MEMBRANE PROTEINS
- Protein molecules “float” among the
phospholipid molecules
- Functions
▪ Marker molecules
▪ Attachment proteins (cadherins and
integrins)
▪ Transport proteins
▪ Receptor proteins
▪ Enzymes

CYTOPLASM
- the material between the plasma membrane
and the nucleus
• Half cytosol
✓ Consists of a fluid part (the site
- The CELL is the basic structural and functional of chemical reactions), the
unit of life cytoskeleton and cytoplasmic
- Each cell is a highly organized unit inclusions.
▪ Plasma membrane – forms the outer ✓ The cytoskeleton supports the
boundary of the cell cell and enables cell
▪ Cytoplasm – the material between the movements.
plasma membrane and nucleus ❖ Microtubules –
▪ Nucleus - contains the cell’s genetic provide support, aid in
material and directs cell activities cell division, and are
▪ Cellular organelles – each performs components of
specific functions organelles
❖ Actin filaments –
PLASMA MEMBRANE support the plasma
- Plays a dynamic role in cellular activity membrane and define
▪ Encloses cell the shape of the cell.
▪ Supports the cell contents ❖ Intermediate
▪ A selective barrier that regulates what filaments – provide
goes into and out of the cell. mechanical support to
▪ Plays a role in communication the cell.
between cells
- Separates intracellular substances from - The material between the plasma membrane
extracellular substances and the nucleus
▪ Intracellular: inside cells Half organelles
▪ Extracellular (intercellular): between Cytoplasmic Inclusions are
cells aggregates of chemicals either
produces by the cell or taken in by the
cell (lipids, glycogen, hemoglobin,
melanin)
NUCLEUS CYTOPLASMIC ORGANELLES
- Specialized subcellular structures with specific
- The nuclear envelope consists of two separate functions
membranes with nuclear pores. - Membranous
• Encloses jellylike nucleoplasm, which • Mitochondria
contains essential solutes • Peroxisomes
• Lysosomes
- DNA and associated proteins are found inside • Endoplasmic reticulum
the nucleus • Golgi apparatus
• DNA is the hereditary material of the - Non-membranous
• Centrioles
cell and controls the activities of the cell
• Ribosomes
• Contains the genetic library with
blueprints for nearly all cellular proteins NUCLEOLI and RIBOSOMES
• Dictates the kinds and amounts of
proteins to be synthesized Nucleoli
• Between cell divisions DNA is - Dark staining spherical bodies within the nucleus
organized as chromatin. • Consist of RNA and proteins
• During cell division chromatin • Produces ribosomal ribonucleic acid
condenses to form chromosomes consisting of (rRNA)
two chromatids connected by a centromere. • Site of ribosomal subunit assembly

Ribosomes
- Sites of protein synthesis
• Free ribosomes are not attached to any
organelles
Synthesize proteins used inside
the cell
• Attached ribosomes are part of a network
of membranes called the Rough Endoplasmic
Reticulum (RER)
Produce proteins that are
secreted from the cell.

PRODUCTION OF RIBOSOMES

1. Ribosomal proteins, produced in the cytoplasm, are

transported through nuclear pores into the
nucleolus.
2. rRNA, most which is produced in the nucleolus, is
assembled with ribosomal proteins to form small
and large ribosomal subunits.
 3. The small and large ribosomal subunits leave the
nucleolus and the nucleus through nuclear pores
4. The small and large subunits, now in the cytoplasm
combine with each other and with mRNA during
protein synthesis.
ENDOPLASMIC RETICULUM (ER)
- Series of membranes forming sacs and tubules that
extend from the outer nuclear membrane into the
cytoplasm
- Two varieties: Rough Endoplasmic Reticulum and
Smooth Endoplasmic reticulum
✓ Rough endoplasmic Reticulum
➢ Studded with ribosomes
➢ Major site of protein synthesis
✓ Smooth endoplasmic Reticulum
➢ Does not have ribosomes
attached
➢ Major site of lipid and
carbohydrate synthesis
➢ Catalyzes the following reactions
in various organs of the body.
❖ Liver: lipid and
cholesterol metabolism
breakdown of glycogen
and along with the
kidneys, detoxify drugs
❖ Testes: synthesis of
steroid – based
hormones.
❖ Intestinal cells:
absorption, synthesis,
and transport of fats
❖ Skeletal and cardiac
muscle: storage and
release of calcium.
GOLGI APPARATUS
- Series of closely packed membranous sacs that
collect, package, and distribute proteins and lipids
by the Endoplasmic Reticulum
• Secretory vesicles: small, membrane –
bound sacs that transport material from the Golgi
apparatus to the exterior of the cell
FUNCTION OF THE GOLGI APPARATUS
1. Some proteins are produces at ribosomes on the
surface of the RER and are transferred into the
cisterna as they are produces
2. The proteins are surrounded by a vesicle that forms
from the membrane of the ER
3. This transport vesicle moves from the ER to the
Golgi apparatus, fuses with its membrane, and
releases the proteins into its cisterna
4. The Golgi apparatus concentrates and, in some
cases, modifies the proteins into glycoproteins or
lipoproteins
5. The proteins are packaged into vesicles that form
from the membrane of the Golgi apparatus
6. Some vesicles, such as lysosomes, contain enzymes
that are used within the cell.
 7. Secretory vesicles carry proteins to the plasma CENTRIOLES AND SPINDLE FIBERS
membrane, where the proteins are secreted from
the cell by exocytosis. Centrioles
8. Some vesicles contain proteins that become part of - Cylindrical organelles located in the centrosome
the plasma membrane. • Pinwheel array of nine triplets of
microtubules
LYSOSOMES • Centrosome: a specialized zone of
cytoplasm
- Spherical membranous bags containing digestive ❖ The site of microtubule
enzymes formation
• Digest ingested bacteria, viruses, and • Microtubules called spindle fibers extend
toxins out in all directions from the centrosome
• Degrade nonfunctional organelles Spindle fibers are involved in the
• Breakdown glycogen and release thyroid separation of chromosomes
hormone during cell division.
• Breakdown non – useful tissue • Form the bases of cilia and flagella
• Breakdown bone to release Ca2+
• Secretory lysosomes are found in white
blood cells, immune cells and melanocytes.

PEROXISOMES

- Membranous sacs containing oxidases and

catalases
• Breakdown fatty acids, amino acids, and
hydrogen peroxide
• Detoxify harmful or toxic substances
• Neutralize dangerous free radicals
➢ Free radicals: highly reactive chemical
with unpaired electrons (i.e., O2-)

MITOCHONDRIA

- the major sites of

CILIA, FLAGELLA AND MICROVILLI
the production of
ATP (the major
- Cilia: move substances over the surface of cells
energy source for
- Flagella: are much longer that cilia and propel
cells) via aerobic
sperm cells
cellular respiration
- Microvilli: increase the surface are of cell and aid in
- Have a smooth
absorption and secretion.
outer membrane
and an inner
membrane that is
enfolded to
produce cristae
- Contain their own
DNA, can produce
some of their own
proteins, and can
replicate independently of the cell.
DISCUSSION RECORDED LECTURE NOTES:
SCOPE
- How is the disease attained or acquired?
- How does it progress
- What are the changes it causes into the human
body?
- How do we diagnose them in the laboratory?
DISCUSSION
- To manage or treat a certain type of disease
you need to have a correct diagnosis.
- Two major approaches in studying pathology
▪ General pathology
▪ Systemic pathology (per system)
TWO MAIN BRANCHES OF PATHOLOGY
- Clinical pathology, machines is needed for
biochemical changes is already branched out.
• Molecular pathology
• Serologic pathology
• Microbiologic pathology
• Blood banking
- Anatomic pathology is Study the morphologic
changes in the busy, the structure, cell, tissue
changes. You do this to give diagnosis.
Anatomic pathology is already branched out.
• Surgical pathology
• Diagnostic cytology
• Etc.
- Molecular pathology
• Monitoring the effectiveness of
therapies which is applicable in cancer
management, not only for infectious
diseases.
• Management of cancers particularly
lymphomas and breast cancer
INTRODUCTION: DISEASE PROCESS
- How do we get the disease?
- How does it progress
- What are the changes it causes to the body?
- 4 major elements/aspects of disease process.
▪ Etiology: the cause, pinang galingan
bakit siya nagkasakit, what causes the
disease
▪ Pathogenesis: the making, paano from
the cause napunta sa effect, journey of
a healthy cell/tissue from a diseased
cell/tissue.
▪ Molecular/Morphologic changes:
visible changes na pwedeng itest para
malaman yung etiology.
▪ Clinical Manifestation: pag
symptomatic na yung patient example
pag may cancer sa breast naka kapa na
ng tumor or breast mass.
ETIOLOGY
- Genetic: inherited, ex. Your mother, lolo and
your aunt has diabetes, it’s in the genes that is
why it can be inherited.
- Acquired: external, not eating properly,
ingestion of chemical, extreme heat, extreme
cold
- Early days of pathology one scientist has
postulated that there is only one etiologic
factor per disease “Koch’s postulate/ Germ
Theory of Disease”
- Germ Theory of Disease: for every disease
there is only one cause.
- Koch’s postulate not applicable to majority of
diseases.
- Most diseases are multifactorial nowadays,
hindi lang isa ang cause ng certain type of
disease.
PATHOGENESIS
- Rudolf Virchow
- A journey, exposure progression symptoms.
- How does a certain type of disease happen or
progress, a journey from a healthy to
unhealthy cell/tissue?
- By studying the pathogenesis of disease, you
will know what you can do to control in order
for the disease to be prevented.
- Treatment for prevention.
- Most disease nowadays has a molecular or
genetic underlying cause na pwede nating ma
solusyonan.
- Thermocycler: equipment used in molecular
lab for Polymerase Chain Reaction (process).
MOLECULAR/MORPHOLOGIC CHANGES
Example:
morphologic
changes =
tuberculosis
- Granuloma: group of epithelioid cells which is
histocytes
- Histocytes: transformed by etiologic agent
which is tuberculosis (Mycobacterium
tuberculosis)
- Fascination necrosis: Necrosis central area
- Chronic inflammation: all lymphocytes, at the
periphery of the granuloma
- Presence of Giant cells, lung hand giant cells
(horseshoe shape cells)
- This group of morphologic changes =
mycobacterium tuberculosis
- If you see this group of morphologic changes in
any organ the possible disease is
“Tuberculosis” can be in the lungs, lymph
nodes, fallopian tube, colon, etc.
- Treatment: TB treatment
- Diagnostic pathology: term used that
pathology is used for diagnosing diseases.
Under anatomic pathology and diagnostic
pathology.
• Encompasses molecular, biologic and
immunologic approaches for analyzing
disease states. Hindi nalang basta
surgical pathology. Kasi nagkaron ng
advancement sa cytology, pwedeng
aspiration, fluids etc.
• Surgical pathology: uses morphologic
or histologic study of the diseased
organs to give a diagnosis, studying
gross specimen, end point is yung
slides, tissue slides.
• Gross specimen: mismong organ ang
inaaral, ginagawa primarily sa
histopath lab.
• Diagnostic cytology: ginagawa sa
anatomic pathology.
• Studying morphologic or structural
changes.
- True case from a 15 – year – old male.
- Histopath diagnosis: Ewings sarcoma vs. PNET
(Peripheral Nerve tumor)
- Tumor cells upon submitting to molecular
testing exhibit CD99 + membranous staining
- CD99: protein
- Cell membrane is stained
- CD99 protein is concentrated in the cell
membrane
- This is a case of EWINGS SARCOMA
- PNET will not stain with CD99
FUNCTIONAL DERANGEMENT AND CLINICAL
MANIFESTATIONS
- This is what clinicians are often encountered.
- Mga nararamdaman
- End result of the of the disease process from
etiology -> pathogenesis -> molecular changes
-> clinical manifestation
- Abnormalities
- Signs and symptoms, outcome
- Symptoms: subjective
- Subjective: Di natetest with your 5 senses
- Healthy organs -> exposure to things that can
harm us can be internal or external -> disease
Cardiac muscle disease
- Etiology: hypertension
- Pathogenesis: increase mitochondria
- Molecular changes: increase of individual
muscle cells (adaptation)
- Clinical manifestation: space almost closes
cardiac hypertrophy (thickening of the heart
muscle) heart compensates kaya bumibilis
yung tibok ng heart or mabilis mapagod ang
possible symptoms.
Cardiac muscle cell injury
- Nawalan ng blood supply, cell death, nawalan
ng oxygen (used for ATP production), cardiac
cell death, causing morphologic change, heart
muscle wall becomes fibrotic (yellowish),
fibrous tissue does not contracts, resulting
clinical manifestation to the patient kasi di na
efficient yung pumping ng blood.
RUDOLF VIRCHOW
- Cell theory: diseases arose not in the organs or
tissues in general but primarily on their
individual cells.
NORMAL CELL STRUCTURE
- Cell
- Plasma membrane
- Cytoplasm
- Nucleus
- Cellular organelles
PLASMA MEMBRANE
- Cell membrane: a phospholipid bilayer, polar
region are usually proteins, fatty acids, lipids
inside.
- Some proteins are transmembrane: crosses
the membrane
- Proteins: Some are surfaced inside, surfaced
outside, transmembrane
- Cadherins and integrins: very prominent in
inflammatory cells and in endothelium
- Intracellular
- extracellular
CYTOPLASM
- in between plasma membrane and nucleus
- inclusions: nakaka penetrate ng plasma
membrane
- cytoskeleton: responsible for the cell
movements and supports the cell
HALF CYTOSOL
- microtubules: contains tubulin subunits 25 um
- actin filaments: 8um
- intermediate filaments: 10um
HALF ORGANELLES
- cytoplasmic inclusions: membrane bound
produced due to phagocytosis
NUCLEUS
- contains two layers of membrane for the
nuclear envelope
- DNA transcription and replication occurs
within the cell, but the translation occurs in the
rough endoplasmic reticulum
- So, the mRNA/translation has to travel through
that pore into the listeria (Endoplasmic
Reticulum) where your ribosomes are located
for translation
- Histone protein: post packaging of DNA after
replication
CYTOPLASMIC ORGANELLES - The nucleus down to the GA is connected via
Membranous membrane
- Mitochondria - The mRNA produced in the nucleus goes
- Peroxisomes outside the nuclear pore and goes into the RER
- Lysosomes while the ribosomes are going to be translated
- Endoplasmic Reticulum resulting to protein product
- Golgi Apparatus - The protein product will travel into the SER if
Non-membranous its needed to be packaged with a carbohydrate
- Centrioles moiety para maging glycoprotein or if it has to
- Ribosomes be packaged with lipid, all happens in SER

NUCLEOLI AND RIBOSOMES GOLGI APPARATUS

Nucleoli
- Dark staining spherical bodies
Ribosomal subunits.
If the protein needed to be form or synthesized
is for the use of the cell usually it occurs in the
free ribosomes
If the proteins is something to be secreted out
or packaged well and has to go through Golgi
apparatus this happens in the attached
ribosome.
Production of ribosomal subunits occurs in the
nucleoli
Since the rRNA are proteins that are used for
the cell usually they are translated in the
ribosomal site and enters the cell through the
nuclear pores into the nucleoli where they are
assembled.
The assembly of the ribosomal subunits
happens in the nucleoli
When they are already assembled that’s the
time they go back out into the cytoplasm for
translation of other proteins
ENDOPLASMIC RETICULUM
- RER: Major site for protein synthesis
- SER: Major site for lipids and carbohydrates
synthesis
- All those proteins and lipids produced in ER
arrived in Golgi apparatus where they are
packaged and is enclosed in secretory vesicles
- Secretory vesicle: sacs that transport material
from GA to the exterior of the cells.
- Plasma cell: a cell that produces
immunoglobulin
- Immunoglobulin: protein product
- Periphery has a lighter staining portion, kasi
ganon ka well developed and Golgi apparatus
- Any cell that has a function of secreting protein
such as glands are very prominent
LYSOSOMES
- Frontline defense
- Contains digestive enzymes for those frontline
cells (macrophages, etc.)
PEROXISOMES
- Membranous sacs with oxidase and catalase
- Detoxify harmful substances

MITOCHONDRIA
- Powerhouse of the cell
- Major site of the production of ATP
- ATP: major energy source
- Produces 36 chromosomes
- Contains the enzyme used in Kreb cycle or the
electron transport chain
- Nagkakaron ng hypertrophy because of the
increase load of mitochondria because of
 replication as they are considered
independent kasi may sarili silang DNA

CENTRIOLES AND SPINDLE FIBERS

- Centrioles
- Centrosome
- Spindle fibers

CILIA
- Move substance over the surface of cells
Example is the respiratory epithelial cell to
move mucus pataas, pa out
- Flagella much longer than cilia and propel
sperm cells
- Only the sperm cells in the human body has a
flagella, so in the female you can say that there
is no flagellated cell

MICROVILLI
- Increase the surface area of cell and aid in
absorption and secretion
- Primarily seen in organs that are active in
absorption such as the Gastrointestinal Tract,
as well as in organs that secrete like your
epididymis.