LESSON 1:

Circulatory System

Systemic circulation - composed of the heart (propels blood to other parts of the body)

- provides the functional blood supply to all body tissue. It carries oxygen and nutrients to the
cells and picks up carbon dioxide and waste products. Systemic circulation carries oxygenated
blood from the left ventricle, through the arteries, to the capillaries in the tissues of the body.

Capillaries – where exchange of gases happens; expels the waste products

Lungs - excretes the waste especially carbon dioxide
Plants - photosynthesis product = oxygen

Pulmonary circulation

special exception from the rule:

pulmonary vein - oxygenated blood
pulmonary artery - deoxygenated blood

1 RBC = contains 3,000,000 Hgb (Hemoglobin)

Blood Vessels:

Arteries and veins - injured = surgery

Arterioles and venules – injured = natural hemostasis
Capillaries – injured = natural hemostasis

Primary and Secondary Hemostasis

Endothelium - non-contact surface, single layer, simple squamous

negative charge = platelet, endothelium

Tunica intima – innermost layer,

Tunica media – made up of smooth muscle
Tunica externa/adventitia – fibrous, thinner than media, supplies blood, this is where nerve endings are
located

Lumen - central space of the vessel, where blood flows

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Peripheral Nervous System
Neuroepithelium
- epithelium containing sensory nerve endings and found in certain sense organs
- for senses

Note: It is normal to see small amount pus cells in human secretions; product of phagocytosis.

OVERVIEW OF HEMOSTASIS AND THROMBOSIS

Thrombosis (clotting)

- blood should be kept at a liquid state

- should not clot when inside the blood vessel

Atherosclerosis - lumen become filled with cholesterol build-ups sticking into the walls of arteries

Emboli/embolus - an embolus is often a small piece of a blood clot that breaks off

4 Major Components of Hemostasis

1. Vascular System
2. Platelets (Thrombocytes)
3. Blood Coagulation Factors - main player in secondary hemostasis
4. Fibrinolysis and ultimate tissue repair

Fibrinolysis -  is the enzymatic breakdown of fibrin in blood clots. Plasmin cuts the fibrin mesh at various
places, leading to the production of circulating fragments that are cleared by other proteases.
Primary fibrinolysis is a normal body process.

Enzyme = plasmin
Liver = plasminogen

Plasmin – an important enzyme present in blood that degrades many blood plasma proteins, including
fibrin clots.

Processes Involved in Hemostasis Following Injury to Small Blood Vessels:

1. Blood vessel spasm

- caused by irritation (foreign substances), protective mechanism by the nervous system,

neurologic response (usually lasts for 1 minute)

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2. Formation of platelet plug

- Injured sub-endothelium becomes positive charged, a contact surface  platelets will be

attracted = adhesion of platelets

Serotonin = prolongs vasoconstriction

3. Contact among damaged blood vessel, blood platelets, and coagulation proteins

4. Development of a blood clot around the injury

5. Fibrinolytic removal of excess hemostatic material

Blood Vasculature

Types of Blood Vessels:

- Arteries and veins
- Arterioles and venules
- Capillaries

Tissue Zones:
1. Tunica adventitia
2. Tunica media
3. Tunica intima

Note: veins have wider lumen than arteries because they only received deoxygenated blood while
arteries received propelled blood (with pressure), that is why structurally they have thicker walls.

Vasculature Physiology

The Role of Vasoconstriction in Hemostasis

- Vasoconstriction is a reflex in which blood vessels narrrow to increase blood pressure

Aspirin
- a blood thinner
- used by elderly people to maintain to lower their blood pressure
- aspirin paralyzes cyclooxygenase

Note: Epinephrine and serotonin promotes vasoconstriction

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The Role of Endothelium

- Regulates the permeability of the inner vessel wall and provides the principial stimulus to thrombosis
following injury to a blood vessel.

- Involved in the clotting process by producing or storing clotting components.

Dengue fever - toxin - dilation of gap junctions

Petechiae - are small (1–3 mm), red, nonblanching macular lesions caused by intradermal capillary
bleeding
Purpura - larger, typically raised lesions resulting from bleeding within the skin (purpura may be
petechiae that have spread and joined together.)

- Rich with plasminogen activator, which, if appropriately stimulated, is released and activates
plasminogen, which ensures rapid lysis of fibrin clots.

Weibel Palade Bodies, Megakaryotic cells

Weibel Palade Bodies - are the storage granules of endothelial cells, the cells that form the inner lining
of the blood vessels and heart. They store and release two principal molecules, von Willebrand factor
and P-selectin, and thus play a dual role in hemostasis and inflammation.

Substance in Endothelial Cells

Prostaglandin  Prostacyclin

Endothelins - are produced in a variety of tissues, where they act as modulators of vasomotor tone, cell
proliferation, and hormone production.

3 Members of Endothelin Family:

1. Endothelin 1 - produced in endothelial cells, vascular smooth muscle cells

2. Endothelin 2 - produced in the kidney, smaller amounts in myocardium, placenta, and uterus
3. Endothelin 3 - high concentrations in the brain and may regulate important functions. Proliferation
and development of neurons and astrocytes. Also found in the gastrointestinal tract and in the lung and
kidney.

White matter – myelin, axons

Grey matter - neurons

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THE ENDOTHELIAL DYSFUNCTION

Endothelial Dysfunction - play an important role in the initiation, progression, and clinical complications
of various forms of inflammatory and degenerative vascular disease.

Stimuli of Endothelial Dysfunction:

1. Immunoregulatory Substances (Tumor Necrosis Factor/TNF & Interleukin-1/IL1)

2. Viral Infection and Transformation
3. Bacterial Toxins
4. Cholesterol / Oxidatively modified lipoproteins

• Disruption of the endothelium directly activates all four

components of hemostasis.
• After this event, the following events take place:

1. Initially, rapid vasoconstriction for up to 30 minutes reduces blood flow and promotes contact
activation of platelets and coagulation factors.

2. In the second phase, platelets adhere immediately to the exposed

sub-endothelial connective tissue, particularly collagen.

The aggregated platelets enhance sustained vasoconstriction by releasing

thromboxane A2 and vasoactive amines, including serotonin and epinephrine.

3. In the third phase, coagulation is initiated through both the intrinsic and
extrinsic systems.

4. Finally, fibrinolysis occurs following the release of tissue plasminogen

activators (t-PAs) from the vascular wall. Fibrinolytic removal of excess
hemostatic material is necessary to reestablish vascular integrity.

MAINTENANCE OF VASCULAR INTEGRITY

• Essentail Factors for Vascular Integrity

1. Circulating functional platelets
2. Adrenocorticosteroids
3. Ascorbic Acid

A lack of these factors produces fragility of the vessels, which makes them prone to disruption.

Note: normal range of platelets/thrombocytes: 150,000 - 450,000

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Note: Vitamin C = food of fibroblasts  collagen synthesis

- The integrity of arterioles and venules depends on vasoconstriction, the formation of a plug of fused
platelets over the injury, and the formation of a fibrin clot.

- Vasoconstriction is of ultimate importance in damaged arteries. Veins, which contain 70% of the blood
volume, may rupture with a slight increase in hydrostatic pressure.

Fibrin clot - Fibrin clots are lysed by plasmin, a serine protease that circulates in the blood as the inactive
proenzyme, plasminogen.

Fibrin polymer - an end product of the enzymatic cascade of blood clotting. In vivo formation of the
polymeric fibrin network, along with platelet adhesion and aggregation, are the key events in salutary
stopping of bleeding at the site of injury (hemostasis) as well as in pathological vascular occlusion
(thrombosis)

LESSON 2:

MEGAKARYOPOIESIS

THE MEGAKARYOCYTIC CELL SERIES

• Megakaryocytopoiesis proceeds initially through a phase

characterized by mitotic division of a progenitor cell,
followed by a wave of nuclear endoreduplication.

• Endoreduplication is the process in which chromosomal

material (DNA) and the other events of mitosis occur without
subsequent division of the cytoplasmic membrane into
identical daughter cells.

erythropoiesis
lymphopoiesis

Sinusoid - irregular tubular space for the passage of blood, taking the place of capillaries and venules in
the liver, spleen, and bone marrow. The sinusoids form from branches of the portal vein in the liver and
from arterioles (minute arteries) in other organs.

example: bone marrow sinusoid

monocytes – are also known as blood macrophages

macrophages - mobile or stationary (non-mobile)
Note: The fastest during chemotaxis - neutrophils

Thrombopoietin - the hormone thought to stimulate the

production and maturation of megakaryocytes, which in
turn produce platelets, has recently been purified and

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cloned.

• Thrombopoietin activity results from several different

cytokines: erythropoietin, IL-3, and granulocyte-
macrophage colony-stimulating factor (GM-CSF). These
substances have been shown to be able to increase
megakaryocyte size, maturational stage, and ploidy.

MEGAKARYOBLAST

SIZE: 10-24 mm
NUCLEUS: Round
Nucleoli: 2-6
Chromatin: Homogeneous, loosely
organized
CYTOPLASM: Basophilic
Granules: Absent by Wright stain
N/C RATIO: 3:1
REFERENCE INTERVAL:
Bone Marrow: 20% of megakaryocyte
precursors in bone marrow
Peripheral Blood: 0%

Most important criteria for determining the

age of cell
- nature of chromatin material
- no clumpiness/clumping = new, young cells

PROMEGAKARYOCYTE

SIZE: 15-40 mm
NUCLEUS: Indented
Nucleoli: Variable
Chromatin: Condensed
CYTOPLASM: Basophilic
Granules: Present
N/C RATIO: 1:2
REFERENCE INTERVAL:
Bone Marrow: 25% of megakaryocyte
precursors in bone marrow
Peripheral Blood: 0%

MEGAKARYOCYTE

• Megakaryocytes are the largest bone marrow

cells, ranging up

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to 160 mm in size. The nuclear- cytoplasmic (N:C) ratio can be as high as 1:12.
• Nucleoli are no longer visible.
• Distinctive feature of the megakaryocyte is
that it is multilobular, not multinucleated.
• The fully mature lobes of the megakaryocyte shed platelets
from the cytoplasm on completion of maturation.

PLATELETS

- Fragments of cytoplasm of megakaryocytes

• Platelets have an average diameter of 2 to 4 mm (5mm maximum), with younger platelets being larger
than older ones.

6mm = giant thrombocytes

• In contrast to megakaryocytes, platelets have no nucleus.

• The cytoplasm is light blue, with evenly dispersed, fine red-purple granules (Azurophilic - easily takes a
stain with azure dyes

• An inactive or unstimulated platelet circulates as a thin, smooth-surfaced disc.

• Platelets circulate at the center of the flowing bloodstream through endothelium-lined blood vessels
without interacting with other platelets or with the vessel wall.

• Stronger stimulation causes platelets to become sticky without losing their discoid shape; however,
changes in shape to an irregular sphere with spiny pseudopods will occur with additional stimulation.

• This alteration in cellular shape is triggered by an increase in the level of cytoplasmic calcium.

LESSON 3:

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a) Cellular ultrastucture of a mature Platelet

b) Platelet kinetics, function, life span, normal values

aggregration - energy-dependent process, needs ATP, through glycolytic pathway

SOL-GEL ZONE - maintain the disk shape of the platelet

- microfilaments, mucoprotein = actin, myosin
- microtubules

Alpha granules - most prominent granules

CELLULAR ULTRASTRUCTURE OF A MATURE PLATELET

THE GLYCOCALYX or a FLUFFY COAT - these surrounds the cellular membrane of the platelet externally.
• unique among the cellular components of the blood.
• composed of plasma proteins and carbohydrate molecules that are
related to the coagulation, complement, and fibrinolytic systems.

MICROFILAMENTS & MICROTUBULES

• located directly beneath the cell membrane of platelet
• provide structure of the platelet to maintain its discoid shape
• maintains the posin of the organelles
• secondary system of microfilaments is functional in internal organization and secretion of blood
coagulation products, such as fibrinogen.

GRANULES
Alpha Granules - most abundant contains:
• heparin-neutralizing platelet factor 4 (PF 4)
• beta-thromboglobulin
• platelet-derived growth factor
• platelet fibrinogen
• fibronectin
• von Willebrand factor (vWF)
• thrombospondin

GRANULES
Dense or Delta Granules contains:
• serotonin

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• adenosine diphosphate (ADP)
• adenosine triphosphate (ATP)
• calcium.
• Lysosomes - store hydrolase enzymes

Other Cytoplasmic Constituents

• Contractile proteins, including actomyosin (thrombosthenin), myosin, and filamin
• Glycogen
• Enzymes of the glycolytic and hexose pathways

PLATELET KINETICS, LIFESPAN & NORMAL VALUES

• An average megakaryocyte produces about 1,000 to 2,000 platelets.

• Marrow transit time / maturation period of megakaryocyte - 5 days
• Platelets initially enter the spleen & remains for 2 days.
• Platelets are in either the circulating blood or the active splenic pool
• Approximately 2/3 of the total number of platelets are in systemic circulation
• The remaining 1/3 exists as pool of platelets in the spleen that free exchange with
the general circulation
• A normal person has an average of 250 × 109/L (range, 150 ×
109/L to 450 × 109/L) platelets in the systemic circulation.
• Platelet turnover or effective thrombopoiesis averages 35 ×
109/L ± 4.3 × 109/L/day.
• The life span of a mature platelet is 9.0 days ± 1 day.
• At the end of their life span, platelets are phagocytized by the
liver and spleen and other tissues of the mononuclear phagocytic
system.

Clot retraction - thrombosthenin

Normal size of spleen - platelet count = 200,000 per microliter of blood

2/3 - located at blood

1/3 - located at the spleen (cycles between the blood and spleen during 9-11 days)

No spleen = 3/3 of platelets is in the blood (meaning: all platelets are in the blood flow)

Thrombocyte number concentration

Average = 250 x 10^9/L

Range = 150 x 10^9L to 450x10^9

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