THE RELEVANCE OF ROULEAUX FORMATION

Research by: Muoka Chibuzor Gospel Bsc Biochemistry

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Table of content

1.0 THE HUMAN BLOOD 

1.1 THE BLOOD AS A MIXTURE

2.0 ROULEAUX FORMATION 

2.1 PLASMA PROTEINS AND ROULEAUX FORMATION

2.2 AUTOAGGLUTINATION VS ROULEAUX FORMATION

2.3 THE RELEVANCE OF ROULEAUX FORMATION AND ERYTHROCYTE SEDIMENTATION RATE

IN DISEASE CONDITION 

2.4 THE CELL INEFFECTIVENESS DUE TO ROULEAUX FORMATION

3.0 PULSED ELECTROMAGNETIC FIELD (PEMF) THERAPY AS A REMEDY FOR ROULEAUX EFFECT

1.0    The HUMAN BLOOD

The Blood is a body fluid that is found in the circulatory system of humans and animals. It is responsible for various biological processes that are essential for the continuation of life. It is often considered as the ‘fluid of life’ because it carries oxygen (O₂) from lungs to all parts of the body and carbon dioxide (CO₂) from all parts of the body to the lungs from where it is expelled out (respiratory function).

It is known as ‘fluid of growth’ because it carries nutritive substances like glucose, amino acids, lipids and vitamins derived from digested food in the digestive system to various parts of the body (nutritive function). It also transports hormones released from endocrine gland (ductless glands) directly into the blood to all the tissues, and also enzymes to where they are of great importance (transport function).

The blood is also called the ‘fluid of health’ because it protects the body against the diseases (defensive function) and gets rid of the waste products and unwanted substances by transporting them to the excretory organs like kidneys (excretory function). It also involves in the maintenance of a constant internal environment (homeostatic control).

Blood is mostly liquid, with numerous cells and proteins suspended in it, making blood "thicker" than pure water. The average person has about 5 litres (more than a gallon) of blood. Liquid called plasma makes up about half of the content of blood. Plasma contains proteins that help blood to clot, transport substances through the blood, and perform other functions. Blood plasma also contains glucose and other dissolved nutrients.

About half of blood volume is composed of blood cells, of which includes red blood cells, which carry oxygen to the tissues, white blood cells, which fight infections, and platelets, smaller cells that help blood to clot. Blood is conducted through blood vessels (arteries and veins). Blood is prevented from clotting in the blood vessels by their smoothness, and the finely tuned balance of clotting factors.

Figure 1.0

The blood vessel and its constituents (plasma proteins, lymphocytes, red blood cells, platelets, eosinophil, basophil, macrophage, neutrophil).

1.1    THE BLOOD AS A MIXTURE

According to the online biology dictionary, a mixture is a combination of substances in which the individual substances do not change or combine chemically but instead retain their individual properties or can be any combination of them. With respect to the definition of a mixture, the blood when examined critically is a mixture and can be physically separated into its various components by using a centrifuge.

The blood proteins also known as the plasma proteins are proteins present in the blood plasma. The plasma contains globulins (including antibodies) mol wt = 56,000 g/mol, albumin (the chief protein constituent) mol wt = 69,000 g/mol, and fibrinogen (responsible in part, for the clotting of blood) mol wt = 400,000 g/mol.

Plasma serves a variety of functions, from maintaining a satisfactory blood pressure and volume to supplying critical proteins for blood clotting and immunity.  It also serves as the medium for exchange of vital minerals such as sodium and potassium and helps to maintain a proper pH (acid-base) balance in the body, which is critical to cell function. Plasma is obtained by separating the liquid portion of blood from the cells using an anticoagulant. The serum is simply gotten when fibrinogen is removed from blood plasma.

The Blood cells comprises of the red blood cells (erythrocytes), the white blood cells (leucocytes), and the platelets (thrombocytes).

The Red blood cells: RBCs are also known as erythrocytes and are the most common type of blood cell. RBCs are anucleate, have a biconcave (dumb-bell) shape, red in colour, rich in haemoglobin, they develop from mesenchyme cells in embryos, and in adults they form from the reticuloendothelial cells of the liver and also bone marrow. They have a lifespan of 100-120 days after which they become susceptible to macrophages for destruction (senescence of RBCs).

The rate of settling of RBCs in anticoagulated blood can be estimated using the Erythrocyte Sedimentation Rate (ESR) technique. The Hematocrit (HCT) or Packed Cell Volume (PVC) is used to estimate the percentage of blood volume filled or occupied by erythrocytes, of which is a measure of the oxygen-carrying capacity of the blood. Properties of RBCs includes: Rouleaux formation, Specific gravity (SG) of about 1.092-1.101 which is greater than the SG of water which is 1, and high suspension stability.

The White blood cells: WBCs also known as leucocytes are cells of the immune system that are involved in protecting the body against infectious diseases and foreign invaders. They are nucleated, they include – the neutrophiles, eosinophiles, basophils, lymphocytes, and monocytes. All WBCs are derived from multipotent cells in the bone marrow known as hematopoietic stem cells.

The Platelets: Platelets also known as thrombocytes are anucleate blood cells whose sole aim is to partake in stopping bleeding by clumping and clotting blood vessel injuries. The mechanical properties of a blood clot are essential for proper haemostasis and wound healing.  A blot clot needs to be strong in order to withstand physical and chemical abrasion, but it also needs to be elastic enough not to crack or split with body movement. The binding of fibrinogen to platelets is an important part of wound healing by aggregating to form a haemostatic plug and by stimulating the activation of blood coagulation factors.

It is worthy to note that the blood proteins (plasma proteins) should not be confused with nor interchanged with blood cells. Although through their physiological functions, they often work together as a team for the overall performance of the human circulatory system of which also refers to the cardiovascular system.

2.0    ROULEAUX FORMATION

Figure 2.0 Rouleaux formation effect 

Rouleaux formation is the arrangement of red blood cells (erythrocytes) in fluid blood with their biconcave surfaces in apposition, thereby forming groups that resemble stacks of coins (false agglutination, psuedoagglutination). Rouleaux formation is a reversible phenomenon that occurs during low blood flow and small shearing forces in circulation.

The flat surfaces of the discoid (biconcave) red blood cells give them a large surface area to make contact with and stick to each other, thereby forming a rouleau. They occur mostly when the concentration of plasma proteins is high, and as result of this, the erythrocyte sedimentation rate (ESR) is increased.

2.1    PLASMA PROTEINS AND ROULEAUX FORMATION

The supply of oxygen and nutrients and the disposal of metabolic waste in the organs depend strongly on how blood, especially red blood cells, flow through the microvascular network. Macromolecular plasma proteins such as fibrinogen cause red blood cells to form large aggregates, called rouleaux, which are usually assumed to be disaggregated in the circulation due to the shear forces present in bulk flow. 

This leads to the assumption that rouleaux formation is only relevant in the venule network and in arterioles at low shear rates or stasis. Globulin and fibrinogen accelerate the tendency of rouleaux formation by the red blood cells.

2.2    AUTOAGGLUTINATION VS ROULEAUX FORMATION

The observation of red blood cell agglutination (also referred to as autoagglutination) must be distinguished from rouleaux formation which is a physiological phenomenon. The presence of antibodies (usually IgM) on the surface of red blood cells is responsible for the phenomenon of autoagglutination.

Cold agglutinin disease is an autoimmune disease characterized by the presence of high concentrations of circulating antibodies, usually IgM, directed against red blood cells. It is a form of autoimmune hemolytic anemia, specifically one in which antibodies only bind red blood cells at low body temperatures, typically 28–31 °C. Cold agglutinin disease was first described in 1957.

Figure 3.0 Rouleaux Vs Autoaggulutination

Agglutination can be observed during immune-mediated hemolytic anemia, but also during cryoglobulinemia (a far more rare condition). Agglutinating red blood cells resemble grapelike clusters whereas red blood cells in rouleaux formation resemble a stack of coins. In order to clearly distinguish erythrocyte agglutination from rouleaux formation, a simple saline test can be performed, whereby RBCs in rouleaux formation dissolve immediately but agglutinated RBCs do not.

Note: Hemolytic anemia is a condition in which red blood cells are destroyed and removed from the bloodstream before their normal lifespan is over.

Cryoglobulinemia is a medical condition in which the blood contains large amounts of cryoglobulins – proteins (mostly immunoglobulins themselves) that become insoluble at reduced temperatures. This should be contrasted with cold agglutinins, which cause agglutination of red blood cells.

2.3    THE RELEVANCE OF ROULEAUX FORMATION AND ERYTHROCYTE SEDIMENTATION RATE (ESR) IN DISEASE CONDITION

Erythrocyte Sedimentation Rate (ESR) is the rate at which the erythrocytes settle down. Normally, the red blood cells (RBCs) remain suspended uniformly in circulation. This is called suspension stability of RBCs. If blood is mixed with an anticoagulant and allowed to stand on a vertical tube, the red cells settle down due to gravity with a supernatant layer of clear plasma.

ESR is also called sedimentation rate, sed rate or Biernacki reaction. It was first demonstrated by Edmund Biernacki in 1897. Rouleaux formation is responsible for ESR, which is an important diagnostic and prognostic tool.

The ESR is a commonly performed test, the ESR does not diagnose any disease, the same way rouleaux formation is a non-specific indicator of the presence of disease. A high ESR indicates the presence of infection, inflammation or malignancy. The higher the ESR, the more certain are the presence of one of these diseases.

Conditions which cause rouleaux formation include infections, multiple myeloma, Waldenstrom's macroglobulinemia, inflammatory and connective tissue disorders, and cancers. It also occurs in diabetes mellitus and is one of the causative factors for microvascular occlusion in diabetic retinopathy.

2.4    THE CELL INEFFECTIVENESS DUE TO ROULEAUX EFFECT

This fact is alarming enough, but another factor must be taken into account.  Capillaries are so small (5-10 μm) that blood cells can only pass through them in single file. The Rouleaux effect is not just about the stacking cells, but also the formation of Rouleaux stacks into combined branches. Cells that are stacked in this way cannot pass through the capillaries as capillaries can only accept free flowing singular and independent red blood cells. 

Figure 4.0 The Circulatory system

Blood that cannot pass through the capillaries cannot absorb or dispel toxins or carbon dioxide, neither can it absorb and dispel nutrients or oxygen. 

Blood affected by the Rouleaux effect recirculates without having performed the task it is sent to undertake. If the blood cells are subjected to the Rouleaux effect, disease results because the flow of blood throughout the body is restricted, and the oxygen level in the blood is drastically reduced.

Bacteria are anaerobic, meaning they thrive without oxygen, so disease will thrive in blood deprived of oxygen. Parasites and viruses also thrive in anaerobic acidic environments. This problem affects all the major systems in the body, as the blood absorbs and diffuses oxygen and nutrients as well as picking up and dispelling toxins and carbon dioxide at a capillary level. 

3.0   PULSED ELECTROMAGNETIC FIELD THERAPY AS A REMEDY FOR ROULEAUX EFFECT

The Pulse electromagnetic field (PEMF) therapy is basically, the application of a magnetic field, generated from low and high frequencies for therapeutic purposes. Every single process in the human body takes place due to the electromagnetic exchange between cells. Our internal chemistry is controlled by natural electromagnetic energy.

Figure 5.0 Man and Electromagnetism

In modern world people are exposed to all kinds of artificial electric and magnetic fields – from high voltage power lines surrounding the cities we live in, to our cell phones that we don’t seem to be able to live without. Even something as habitual as a microwave oven, can be considered potentially harmful.

All those devices tend to interrupt natural electromagnetic processes in the body, and make our body’s systems less efficient. PEMF seeks to tune up your body’s electromagnetic system to counteract the negative effects of the environment’s “electrosmog” also known as “dirty electricity.” This is the type emitted from our mobile phones, computers, and cordless phones etc., (which all typically come with warnings about safe exposure limits.)

3.1       How blood flow is affected by electrosmog: Viewing blood under a microscope in a body badly affected by adverse electromagnetism shows red blood cells (erythrocytes) clumping together, sticking and stacking thereby decreasing the surface area of the cell itself.  This reduces its ability to function at an optimum level to absorb nutrients and heal areas of weakness. The sticking and stacking is known as the Rouleaux Effect. If two cells stack the combined surface area is reduced by 71 %. Their efficiency is reduced further as the number of cells stack up. A stack of eight or more cells reduces their total cellular efficiency by 50 %.

Another dangerous effect of such stacking is the fact that those cells cannot pass through the capillaries as capillaries can only accept free flowing singular and independent red blood cells. Erythrocyte aggregation obviously affects hemodynamic mechanisms. Hemodynamics is the word used to describe the processes dealing with blood flow through our body. Hemodynamics is one of the most essential elements of metabolism, as it determines whether body’s organs are sufficiently supplied with nutrients and effectively relieved from toxins. 

3.2       PEMF works at a cellular level: Before PEMF therapy emerged there had already been numerous scientific reports supporting the fact that electrical currents can induce significant acceleration of repair in blood and tissue. PEMF systems solve these problems by using magnetic fields as opposed to direct electrical fields. The specific magnetic field is used to generate an electrical field at all levels of the tissues in the right range of field strength and duration to cause a cellular response without creating any risk.

REFERENCES AND FURTHER READINGS

1. Autoagglutination and Rouleaux formation. Retrieved February 1st, 2018. From http://www.medical-labs.net/autoagglutination-vs-rouleaux-formation-2605/ Web

2. Blood. Undated. Retrieved February 1^st, 2018. From https://en.wikipedia.org/wiki/Blood_cell Web

3. Erythrocyte agglutination. Retrieved February 3rd, 2018. From http://www.medvet.umontreal.ca/clinpath/banq-im/hematology/agglutinationE.htm Web

4. Henry Suarez. (2016).  Rouleaux. Retrieved February 4^th, 2018. From https://sonographictendencies.wordpress.com/2016/11/05/rouleaux/ Web

5. Jonathan Bowen. (2014). Optimizing blood flow. Retrieved February 4th, 2018. From https://www.pemf-tech.com/topic/Optimizing-Blood-Flow Web

6. K Sembulingam and Prema Sembulingam. Essentials of Medical Physiology (THE RED BLOOD CELLS). 6^th edition. India.

7. M. Brust, O. Aouane, M. Thiébaud, Et al. (2014). The plasma protein fibrinogen stabilizes clusters of red blood cells in microcapillary flows.  Sci Rep. doi: 10.1038/srep04348 PMCID: PMC3949293

8. PEMF Therapy. Retrieved February 4th, 2018. From http://medistars.ru/pemf-therapy Web

9. Plasma. Undated. Retrieved February 2^nd, 2018. From https://www.redcrossblood.org/learn-about-blood/blood-components/plasma Web

10. Rouleaux formation. Undated. Retrieved February 2^nd, 2018. From https://en.wikipedia.org/wiki/Rouleaux Web

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