Course 1- recap

Properties Primary cells

What is histopathology? The microscopic study of diseased tissue

The most used apparatus in the histology labs to cut Microtome/Cryotome
tissue sections
What Haematoxylin-eosin staining is showing?
Hematoxylin shows the ribosomes, chromatin (genetic
material) within the nucleus a deep purple-bluish color.
Eosin gives cytoplasms a pink color

Haematoxylin-eosin staining of a glomerulus of a

kidney.
 Course 1- recap
Properties
Staining is the process for coloring tissues by using dyes.
What contains a dye molecule?
Special techniques used in Histopathology
Immunostains are composed of specific antibodies that
should only bind to the corresponding antibody in the
tissues.
There are two main methods of immunodetection:
The commonest used chromogen which results in
characteristic ‘brown’ staining.
Primary cells
A dye molecule has two domains: the chromogen provides
the color and the auxochrome makes possible the binding to
the tissue. Chromophore is the molecular structure of the
chromogen that absorbs a particular light wavelength;
- the direct method
- the indirect method involves the primary antibody (P) binding to a
labelled secondary antibody (SA); The secondary antibody is
labelled with an enzyme (E) (chromogen), which converts a
substrate to a coloured compound (Co) which allows visualisation
of the reaction in the tissues.
is horseradish peroxidase,
Ki-67 staining
Hemodynamic disorders:
edema, hyperemia,
congestion, hemostasis and
hemorrhagic disorders
 Hemodynamics

•“Hemodynamics” often refers to basic measures of cardiovascular

function, such as arterial pressure or cardiac output. 

•The health of cells and tissues depends on the circulation of

blood, which delivers oxygen and nutrients and removes wastes
generated by cellular metabolism.

•Under normal conditions, as blood passes through capillary beds,

proteins in the plasma are retained within the vasculature, and
there is little net movement of water and electrolytes into the
tissues.
 Edema
Disorder that perturb cardiovascular, renal, or hepatic function are often marked by the
accumulation of fluid in tissues (edema) or body cavities (effusions).
Elevated hydrostatic pressure or diminished colloid osmotic pressure disrupts this balance and results in
increased movement of fluid out of vessels.

Factors influencing fluid movement across capillary walls. Normally,

hydrostatic and osmotic forces are nearly balanced so that there is
little net movement of fluid out of vessels. Many different pathologic
disorders are associated with increases in capillary hydrostatic
pressure or decreases in plasma osmotic pressure that led to the
extravasation of fluid into tissues. Lymphatic vessels remove much of
the excess fluid, but if the capacity for lymphatic drainage is
exceeded, tissue edema results.
Edema
Edema fluids and effusions may be inflammatory or noninflammatory. The protein-rich
exudates accumulate due to increases in vascular permeability caused by inflammatory mediators. In
contrast, noninflammatory edema and effusions are protein-poor fluids called transudates.

Causes of edema
Increased Hydrostatic Pressure
- mainly caused by disorders that impair venous return.

Reduced Plasma Osmotic Pressure

Under normal circumstances albumin accounts for almost
half of the total plasma protein. Inadequate synthesis or
increased loss of albumin from the circulation are common
causes of reduced plasma oncotic pressure.
Causes of edema

Sodium and Water Retention

Increased salt retention—with obligate retention of
associated water—causes both increased hydrostatic pressure
(due to intravascular fluid volume expansion) and
diminished
vascular colloid osmotic pressure (due to dilution).

Lymphatic Obstruction
Trauma, fibrosis, invasive tumors, and infectious agents
can all disrupt lymphatic vessels and impair the clearance
of interstitial fluid, resulting in lymphedema in the affected
part of the body.
Mechanisms of systemic edema in heart failure, renal failure,
malnutrition, hepatic failure, and nephrotic syndrome
Hyperemia and congestion
Hyperemia and congestion both stem from
increased blood volumes within tissues but
have different underlying mechanisms and
consequences.
Hyperemia - active process in which arteriolar dilation;
- leads to increased blood flow;
- tissues turn red (erythema).

Congestion - a passive process resulting from reduced venous

outflow of blood from a tissue;
- an abnormal blue-red color (cyanosis)
Hyperemia and congestion - morphology

Conjunctival hyperemia Skin hyperemia

Liver with chronic passive congestion and
hemorrhagic necrosis. (A) Central areas are red
and slightly depressed compared with the
surrounding tan viable parenchyma, forming a
“nutmeg liver” pattern (so-called because it
resembles the cut surface of a nutmeg); (B)
Centrilobular necrosis with degenerating
hepatocytes and hemorrhage.
Hyperemia and congestion

Cyanosis of the hand of a patient with low oxygen saturation

An 86-year-old female patient from northeast Mexico presented with diffuse lepromatous leprosy
(DLL). Sequence analysis of four genes (rrs, rpoB, sigA, and hsp65) from the skin biopsy
specimen identified “Mycobacterium lepromatosis.” This is the first independent confirmation
of a case of DLL due to M. lepromatosis.
Hyperemia and congestion

Histopathology: (a) Skin biopsy specimen of the cyanotic lesions, showing vasculitis and
thromboembolism. (b) Many acid-fast bacilli were seen upon Fite-Faracco staining (the routinely used
method to demonstrate Mycobacterium leprae in tissue sections).

Source: Vera-Cabrera et al., Case of Diffuse Lepromatous Leprosy Associated with “Mycobacterium lepromatosis”, J. Clin. Microbiol., p.
4366–4368, 2011, doi:10.1128/JCM.05634-11.
Hemostasis
Hemostasis can be defined simply as the process by which blood clots form at sites of vascular injury.

Hemostasis is a precisely orchestrated process involving

platelets, clotting factors, and endothelium that occurs at
the site of vascular injury and culminates in the formation
of a blood clot, which serves to prevent or limit the extent
of bleeding. It is a multistep process:
1. Arteriolar vasoconstriction occurs immediately and
markedly reduces blood flow to the injured area.

2. Primary hemostasis: the formation of the platelet plug.

Hemostasis
3. Secondary hemostasis: deposition of fibrin.

4. Clot stabilization and resorption.

Endothelial cells are central regulators of hemostasis;

the balance between the antithrombic and prothrombotic
activities of endothelium determines whether thrombus
formation, propagation or dissolution occur.
Platelets play a critical role in hemostasis by forming
the primary plug that initially seals vascular defects and by
providing a surface that binds and concentrates activated
coagulation factors.
 https://www.youtube.com/watch?v=cy3a__OOa2M
Hemostasis
Coagulation Cascade
The coagulation cascade is a series of amplifying enzymatic reactions that lead to the deposition of an insoluble
fibrin clot.
Hemorrhagic disorders
Disorders associated with abnormal bleeding inevitably stem from primary or secondary defects in vessel walls,
platelets, or coagulation factors, all of which must function properly to ensure hemostasis.

(A) Punctate petechial hemorrhages of the colonic mucosa, a consequence of thrombocytopenia. (B) Fatal
intracerebral bleed.
Hemorrhagic disorders
Diseases associated with sudden, massive
hemorrhage include: aortic dissection and
aortic abdominal aneurysm, myocardial
infarction complicated by rupture of the aorta or
the heart.
General principles related to abnormal bleeding
and its consequences:
• Defects of primary hemostasis (platelet defects or von
Willebrand disease)
• Defects of secondary hemostasis (coagulation factor defects)
• Generalized defects involving small vessels

Also, there are subtle defects in clotting that

only become evident under conditions of
hemostatic stress, such as surgery, childbirth,
dental procedures, menstruation, or trauma.
 Hemorrhagic disorders

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5374099/

Results of hematoxylin and eosin (H&E), periodic acid-Schiff (PAS), and immunohistochemical Results of hematoxylin and eosin (H&E), periodic acid-Schiff (PAS), and immunohistochemical
staining (3,3'-diaminobenzidine and hematoxylin, DAB&H) of renal tissue from control staining (3,3'-diaminobenzidine and hematoxylin, DAB&H) of renal tissue from rats in which HS
rats. A. H&E staining. D, distal tubules; G, glomerulus; P, proximal tubules. B. PAS staining. was induced. A. H&E staining. Arrowheads, pyknotic nuclei; Arrows, detached tubular cells; Di,
Arrows, brush borders; BM, tubular basement membranes; D, distal tubules; P, proximal dilated renal tubules; N, tubular necrosis; S, swollen tubular cells; Star, blood containing renal
tubules. C. iNOS immunostaining (DAB&H). D. BAX immunostaining (DAB&H).  tubules. B. PAS staining. BM, tubular basement membranes. C. iNOS immunostaining
(DAB&H). D. BAX immunostaining (DAB&H). 
Hemorrhagic disorders
Thrombosis
The primary abnormalities that lead to thrombosis are:
(1) endothelial injury;
(2) stasis or turbulent blood flow;
(3) hypercoagulability of the blood (the so-called Virchow
triad).
Hemorrhagic disorders

Pulmonary embolism. Light micrograph showing a blood clot (round, dark pink) completely
blocking the lumen (inner space) of the pulmonary artery in the lung. This is known as a
pulmonary embolism (PE) and is caused when a fragment of a thrombus (blood clot) elsewhere in
the circulatory system, breaks off and travels in the blood to the arteries in the lungs. It most
commonly occurs as a complication of deep vein thrombosis (DVT) and can cause shortness of
breath and even lead to sudden death.
 What are PT and PTT assays for?

Two laboratory tests are used commonly to evaluate

coagulation disorders: Prothrombin Time (PT) which
measures the integrity of the extrinsic system as well as
factors common to both systems and Partial
Thromboplastin Time (PTT), which measures the integrity
of the intrinsic system and the common components.