Chapter 6

PATHOLOGICAL PIGMENTATIONS

 

 

 

                   

6.1  AN OVERVIEW

Pigments are colored substances, some of which are normal constituents of cells (melanin), while others are abnormal and collect in cells under special circumstances. The various pigments (intracellular accumulations) discussed in this section differ greatly in origin, chemical composition and biological significance. Traditionally, pigments are classified as exogenous (coming from outside the body) and endogenous (synthesized within the body itself). 

6.2 TERMINAL OBJECTIVES

Once this section is completed, the student should be able to perform the following tasks.

6.3 EXOGENOUS PIGMENTS 

6.3.1 CARBON PIGMENT

Carbon or coal dust pigment is the most common exogenous pigment encountered in animals and man. It is virtually a ubiquitous air pollutant of urban life and occurs primarily in the lungs and related lymph nodes. When inhaled, carbon is picked up by alveolar macrophages and transported through lymphatic channels to the regional lymph nodes. Accumulations of this pigment blacken lung tissues and the related lymph nodes. Anthracosis is the condition that occurs when carbon particles are found as a black pigment in the lungs. (Pneumoconiosis is a general term which refers to the condition that develops in the lungs subsequent to inhaling any of the exogenous pigments.)

Microscopically, lesions in the lungs are centered around small bronchioles as collection of black granules which may be found in macrophages or free in the tissue. In sections, carbon can be distinguished from other pigment by its black color and by its resistance to all solvents and bleaching agents. Grossly, the lungs may be gray or mottled in color. The regional lymph nodes may be black, particularly in the medullary region where the particles are held in sinusoidal macrophages.

In general, anthracosis does not interfere with normal respiratory function, nor does it predispose to infection. However, slight lung fibrosis may occur if excessive amounts are inhaled over long periods. Regardless, carbon is the only exogenous pigment of concern in animals. 

6.3.2 MINERAL DUST

There are several mineral dusts (including carbon) that may cause visible discolorations in the lungs and related lymph nodes. These pigments are of considerable concern in human occupational health safety, but they are not of concern in animals.

The student should read and understand the material in the textbook (Jones and Hunt, page 76) relative to:

Remember, pneumoconiosis is the term that refers to the condition caused by any of the mineral dusts. 

6.3.3 CARTENOID OR LIPOCHROME PIGMENTS

These are a closely related group of exogenous pigments which are yellow or brown and soluble in fat solvents. They include carotene and xanthophyll which collect in various body tissues. The amount absorbed and retained in the body varies with the breed and species. (The student should be able to distinguish hepatic carotenosis from a liver filled with retained bile.) 

6.4 ENDOGENOUS PIGMENTS

6.4.1 LIPOFUSCIN

Lipofuscin is an insoluble pigment which represents the indigestible residue of autophagic vacuoles within cells formed during aging or atrophy. The pigment appears to be composed of polymers of lipids and phospholipids complexed with protein. The following is the manner in which lipofuscin is formed:

"During atrophy and aging, degenerating cellular organelles are enclosed in autophagic vacuoles. Subsequently, lysosomes discharge their hydrolytic enzymes into these membrane bounded vacuoles and the cellular organelles are digested by autophagy. However, some of the organelle components may resist digestion or be incompletely digested. (Lipoproteins and other lipids make up most of the indigestible debris and their accumulation reflect the lack of sufficient quantities of lipase in most lysosomes.) When organelles are not digested completely, the debris persists as membrane-bounded residual bodies. Some of these residual bodies may be extruded from the cytoplasm, or may be eventually digested. However, in some instances, the residual bodies persist in the cytoplasm of atrophic or aging cells. Microscopically, lipofuscin pigment appears as minute yellow-brown granules. In cardiac muscle cells, the granules are located chiefly near the "poles" of the nuclei. In other organs and tissues, they are distributed throughout the cytoplasm. Grossly, the lipofuscin pigment may impart a brownish discoloration to tissues when present in sufficient amounts (brown atrophy). Lipofuscin itself is not injurious to the cell or to its function."

Lipofuscin occurs in a variety of organs and tissues, but it is especially prominent in the brain neurons, myocardial cells and in the adrenal and thyroid glands. Vitamin E deficiency may accentuate lipofuscin formation. In vitamin E deficient dogs, for example, large amounts of lipofuscin is found in the smooth muscle of the intestine. The involved gut becomes brown in color and the condition is oftentimes referred to as "brown dog gut. 

6.4.2 CEROID

is an acid-fact variant of lipofuscin which is commonly associated with disturbances in vitamin E and fatty acid metabolism. Sometimes, for obscure reasons, lipofuscin (or a pigment indistinguishable from lipofuscin) undergoes physical or chemical transformation so that it becomes autofluorescent and stains positively with acid-fast stains. 

6.4.3 MELANIN

Melanin is a normal endogenous brown-black pigment which gives color to the skin, hair, leptomeninges and choroid of the eyes. The pigment is pathological when it occurs in places or amounts that are not considered normal for the species concerned.

Melanin is a high molecular weight biochrome bound to protein. It is synthesized by melanocytes where it resides in characteristic granules called melanosomes. Melanocytes stem from melanoblasts which are non-pigmented precursor cells of neural crest origin. Melanophages are phagocytic cells in the dermis which accept and store melanin, but do not synthesize it. Melanophores, or contractile cells, are melanocytes of some lower vertebrate that participate in the rapid color changes by intracellular aggregation and dispersion of melanosomes. The formation of melanin begins with the enzyme tyrosinase, a copper-protein substance that facilitates the oxidation of tyrosine to dihydroxyphenylalanine (DOPA) and DOPA to dopaquinone in the initial stages of synthesis.

Remember, the biosynthesis of melanin occurs in melanocytes. In the process of normal pigmentation in the skin, for example, the basal epithelial cell of the epidermis acquires melanin from the melanocytes. The dendrites of the melanocytes form "bridges" between the epithelial cells and the melanocytes and actually inject the pigment-containing melanosomes into the epidermal cells. In domestic mammals, the only known function of melanin is protection against solar ultraviolet radiation.

Under normal or physiologic conditions, melanin may be observed under the conditions listed in the textbook (Chapter 2, page 79)

6.4.4 MELANOSIS

Refers to the presence of melanin in an abnormal location and represents a congenital mislocation of melanocytes. It is usually found incidentally during necropsy or at the time of slaughter. Grossly, lesions are irregular in size and shape and they appear black in color. There is no change in texture or consistency of the involved organ or tissue. Microscopically, there are scattered melanocytes mixed with fibroblasts (lesions are usually on the surface of organs). 

6.4.5 MELANOMAS

Are neoplasms derived from melanoblasts and melanocytes. These neoplasms may be benign or malignant. Melanomas can occur in many tissues, but they usually originate where precursor cells are numerous (skin, etc.). 

6.4.6 ALBINISM

Is a congenital defect characterized by an absence of melanin (hereditary inability to synthesize melanin). Apparently, melanocytes are unable to synthesize sufficient tyrosinase.

The dihydroxyphenylalanine (DOPA) test is used to identify cells that have the capability to make melanin. Melanin is made from tyrosine (DOPA is a closely related chemical). When a suitable solution of DOPA is placed on tissue containing cells capable of producing melanin, a black granular precipitate forms. Fontana's silver solution may be employed to detect melanin granules.  

6.5 HEMOGLOBIN-DERIVED PIGMENTS 

6.5.1 HEMOGLOBIN:

Hemoglobin, the oxygen-carrying pigment of erythrocytes, is a combination of globin and the pigment complex heme. During normal and pathologic breakdown of hemoglobin, different types of pigment complexes are formed. Most of these are heterogeneous and are not chemically defined. Ferratin and hemosiderin are the principal iron storage compounds in the body. Hemoglobin itself is usually not visible in tissues. However, it may become visible in renal tubules as a reddish-orange color if released from erythrocytes in large quantities. Free hemoglobin in the blood is referred to as hemoglobinemia. Hemoglobinuria refers to free hemoglobin in the urine. 

6.5.2 HEMOSIDERIN:

Hemosiderin is a brown, granular iron-containing pigment which forms when erythrocytes are lysed. It develops within macrophages anywhere in the body, but is particularly common in the spleen, liver and in foci of hemorrhage.

In the body, iron is absorbed in the ferrous form and changed to the ferric form in the bloodstream where it is normally carried by transferrins (transport protein). In cells, iron is normally stored in association with a protein, apoferratin, to form ferritin micelles. Ferritin is diffuse in cells and is not visible. However, when there is a local or systemic excess of iron, ferratin forms hemosiderin granules which are easily observed with the light microscope. Thus, hemosiderin pigment represents aggregates of ferritin micelles. Under normal conditions, small amounts of hemosiderin can be seen in reticuloendothelial cells of the bone marrow, liver and spleen (all actively engaged in erythrocyte breakdown).

In general, hemosiderin is seen in tissues whenever there is excess breakdown of erythrocytes (these situations are outlined in your textbook).

Microscopically, hemosiderin is usually found within macrophages in the form of yellow to brown, sharply circumscribed masses. The Prussian blue reaction can be employed to confirm its presence in both gross and microscopic sections. Grossly, large accumulations of hemosiderin impart a brownish color to the organs or tissues.

Hemosiderin itself is not harmful; however, its presence indicates previous hemorrhage or a hemolytic disease (destruction of red blood cells). 

6.5.3 GAMNA GANDY BODIES

(Fibrosiderotic plaques) are organized focal hemorrhages found along the edges of the spleen which contain a heavy concentration of hemosiderin. In addition to hemosiderin laden macrophages, these foci contain calcium salts encrusted over fibrotic connective tissue and elastic fibers. 

6.5.4 "HEART FAILURE CELLS"

Are hemosiderin laden alveolar macrophages that occur subsequent to chronic left heart failure and passive congestion of the lungs. 

6.5.5 HEMOCHROMATOSIS

Refers to a group of disorders, primarily occurring in man, which are characterized by cirrhosis and fibrosis of the liver and widespread hemosiderosis. Occasionally, similar conditions occur in animals. 

6.5.6 HEMATIN

is a pigment formed from the action of acid or alkali on hemoglobin after death (it is not a metabolite or precursor of hemoglobin). Brownish crystals (similar to hemosiderin) are formed but they are extracellular whereas hemosiderin is intracellular. Hematin is most commonly encountered when unbuffered formalin is used during the fixation of tissues. It does not stain blue with Prussian blue stain because the iron is too tightly bound to react. 

6.5.7 BILIRUBIN

Is an orange-yellow pigment derived primarily from the breakdown of hemoglobin (hemoglobin is derived from the breakdown of erythrocytes after their normal life span of approximately 120 days). After the breakdown of hemoglobin into globin and heme, the heme (iron-porphyrin compound) is separated into iron and porphyrin components. The porphyrin ring is split by heme oxygenase to form biliverdin. The biliverdin is then reduced by a reductase to bilirubin. Bilirubin is formed by reticulo-endothelial cells any place in the body, but especially in the spleen. following its formation, bilirubin goes through the following steps leading to the formation of bile pigments:

Initially, bilirubin is transported from reticuloendothelial cells (spleen, etc.) to the liver as a complex with plasma albumin. This bilirubin-albumin complex is known as non-conjugated bilirubin (indirect reacting bilirubin, hemo-bilirubin). At the liver cell plasma membrane, bilirubin dissociates from albumin and is subsequently conjugated within liver cells with glucuronic acid. This bilirubin-glucuronic complex is known as conjugated bilirubin (direct reacting bilirubin, cholebilirubin). The conjugated bilirubin is secreted by liver cells into the bile canaliculi. It is subsequently transported to the duodenum via the biliary tree. In the colon, conjugated bilirubin is reduced by bacterial enzymes to urobilinogen. Eighty to ninety percent of the urobilinogen is oxidized to urobilin and excreted in the feces. The remaining urobilinogen is reabsorbed into the portal circulation and returned to the liver (constituting the so-called enterohepatic circulation), while some reaches the general circulation and is excreted by the kidney (this urobilinogen imparts a sight yellow color to urine as it is oxidized to urobilin on standing).

Remember, the steps described above occur subsequent to the breakdown of erythrocytes after their normal life span. However, under certain pathologic conditions (excessive destruction of erythrocytes, liver diseases, bile duct obstruction), non-conjugated and/or conjugated bilirubin reach such high concentrations in the circulating blood until tissues are tinged with yellow. Staining of tissues with either non-conjugated or conjugated bilirubin is referred to as jaundice or icterus. 

6.5.8 ICTERUS (JAUNDICE):

Icterus is the condition in which tissues are stained (yellowish) with either non-conjugated or conjugated bilirubin. It implies hyperbilirubinemia. Grossly, the yellowish discoloration is best observed in the sclera or other tissues which are pale normally. Microscopically, bilirubin is not observed under ordinary conditions since it is soluble. However, in some situations, the pigment may collect in large enough quantities to be seen in the bile capillaries of the liver as yellowish-brown to slightly green material.

Traditionally, icterus is classified according to the causative mechanism into:

6.5.9 THE DIAGNOSIS OF ICTERUS

Is adequately discussed in your textbook. Some of the methods and procedures employed in the diagnosis of icterus include:

6.5.9.1 ICTERUS INDEX TEST:

This test is used to determine the presence or absence of icterus by comparing in a colorimeter the color of the blood serum with the yellow tint of a solution of potassium dichromate of standard strength. A high icterus index indicates the presence of icterus.

6.5.9.2 THE VAN DEN BERGH REACTION:

A positive direct Van den Bergh reaction indicates the presence of conjugated bilirubin (obstructive icterus), whereas a positive indirect Van den Bergh reaction indicates the presence of non-conjugated bilirubin (hemolytic icterus). If a positive direct and indirect Van den Bergh reaction is attained, this indicates the presence of both conjugated and non-conjugated bilirubin which is characteristic of toxic icterus.

6.5.9.3 URINE UROBILINOGEN:

A small amount of urobilinogen in the urine is normal. Its absence indicates obstructive icterus. Large amounts of urobilinogen in the urine indicate hemolytic icterus or it may indicate the presence of toxic icterus.

6.5.9.4 COLOR OF FECES:

In obstructive icterus (complete), the feces are pale. In hemolytic icterus, the feces are darker than normal.

Remember, icterus is not a disease, but an important clue to any one of several different disorders. 

COMPARISON OF THE TYPES OF ICTERUS 

6.5.10 PHOTOSENSITIZING PIGMENTS

Photosensitizing pigments are fluorescent pigments which accept light rays of one wavelength and transform them into rays of a longer wavelength. Photosensitization is the condition that results when light acts on a fluorescent pigment in tissues.

The three sets of circumstances which give rise to the presence of photosensitizing pigments in tissues are:

6.6 POST-INSTRUCTIONAL SELF-EXAMINATION

Questions

After completing section 6, each student should be in a position to provide appropriate answers for the following questions.

  • 1. What is the most important exogenous pigment encountered in animals? Explain.
  • 2. What is the interrelationship between anthracosis and pneumoconiosis?
  • 3. What significant gross and microscopic lesions are associated with cases of anthracosis?
  • 4. In tissue sections, how would you distinguish carbon pigment from other inhaled pigments?
  • 5. What special procedures, stains, etc., would you employ in order to make a tentative diagnosis of the following:
    • -- Ceroid
    • -- Cells with the capability of producing melanin
    • -- Detection of melanin granules
    • -- Hemosiderin
    • -- Amyloid
    • -- Glycogen
    • -- Lipids
  • 6. What are carotenoids or lipochrome pigments?
    • -- Under what circumstances are these pigments normal within the body?
  • 7. What is hepatic carotenosis? How would you distinguish this condition from a liver filled with retained bilirubin?
  • 8. What is lipofuscin? Ultrastructurally, in what cellular component would you expect to observe this pigment? What is the most likely composition of lipofuscin?
  • 9. Briefly outline the manner in which lipofuscin is formed in cells.
  • 10. Give the gross and microscopic features of lipofuscin.
  • 11. Under what circumstances would you expect lipofuscin to develop in cells?
  • 12. How would you describe and use the following terms: (1) "brown dog gut," (2) grown atrophy and (3) "wear and tear pigment?"
  • 13. How would you distinguish lipofuscin from ceroid? What is an acid-fast stain?
  • 14. How would you distinguish carbon-containing macrophages from melanophages in lung sections?
  • 15. In what body sites would you expect to find melanin as a normal endogenous pigment?
  • 16. Under what circumstances would you expect melanin to appear as a pathologic pigment?
  • 17. How would you distinguish melanocytes from melanophages in tissue sections?
  • 18. Matching:
    • ____ Melanophages A. Hereditary absence of melanin
    • ____ Melanophores B. Oxidizes tyrosine to DOPA
    • ____ Tyrosinase C. Phagocytic cells
    • ____ Hemoglobinuria D. Precursors of melanocytes
    • ____ Heart failure cells E. Melanin-containing neoplasms
    • ____ Melanosomes F. Melanin in abnormal sites
    • ____ Melanoblasts G. Melanin granules
    • ____ Melanosis H. Melanocytesof lower vertebrates
    • ____ Melanoma I. Hemoglobin in the urine
    • ____ Albinism J. Hemosiderin laden macrophages
    • ____ Gamna Gandy bodies K. Fibrosiderotic plaques of
  • spleen
  • 19. Briefly discuss the occurrence of melanin as outlined in your textbook (page 79).
  • 20. Briefly describe the test employed in an attempt to identify cells with the capability of producing melanin.
  • 21. Briefly describe the process by which normal skin receives its melanin pigment.
  • 22. How would you distinguish between hemoglobin and porphyrin?
  • 23. What is "imbibition of hemoglobin?"
  • 24. What important pigments are found in the heme portion of hemoglobin?
  • 25. What are the principal iron storage compounds in the body?
  • 26. What is the interrelationship between ferratin and hemosiderin?
  • 27. Describe the microscopic changes associated with hemosiderosis including any special procedures, etc., that you would employ to identify it.
  • 28. Describe the manner in which Gamna Gandy bodies are formed.
  • 29. What are "heart failure cells?"
  • 30. How would you distinguish hemosiderosis from hemochromatosis?
  • 31. Describe the circumstances in which you would expect hematin pigment to form.
  • 32. In tissue sections, how would you distinguish hemosiderin from hematin?
  • 33. How would you characterize bilirubin?
  • 34. Outline the important steps in the formation of urobilinogen, beginning with the death of erythrocytes at the end of their normal life span.
  • 35. What is the interrelationship between non-conjugated bilirubin, conjugated bilirubin and urobilinogen?
  • 36. A six-year-old Coonhound was submitted to the clinic with signs of icterus. Examination revealed clay-colored feces and an absence of urobilinogen in the urine. What type icterus would you suspect?
  • 37. What is the interrelationship between hemobilirubin, cholebilirubin and urobilinogen?
  • 38. A four-year-old German Shepherd was submitted to your veterinary clinic with intense yellow mucous membranes and increased pigmentation of the feces. An indirect Van den Bergh reaction was attained. What type icterus would you suspect?
  • 39. Under what circumstances would you expect obstructive, toxic and hemolytic icterus to occur in a 12-year-old Coonhound?
  • 40. Why would you expect to find non-conjugated and conjugated bilirubin in a dog with toxic icterus?
  • 41. Why would you expect non-conjugated bilirubin to be lacking in the bloodstream of a Coonhound with obstructive icterus?
  • 42. Distinguish between hemolytic, toxic and obstructive jaundice on the basis of the following:
    • -- Pigments that stain tissues
    • -- Color of feces
    • -- Consistency of feces
    • -- Color of urine
    • -- Icterus index
    • -- Van den Bergh reaction
  • 43. What is a photosensitizing pigment? What is photosen-sitization?
  • 44. Under what circumstances would you expect photosensitizing pigments to occur in animals?
  • 45. Distinguish between hepatotoxic photosensitization and primary photosensitization.
  • 46. What is the fundamental cause of congenital porphyria?
  • 47. Explain how congenital porphyria develops subsequent to defective hemoglobin synthesis. What factor is believed to be lacking when Type I porphyrin accumulates in the bloodstream?
  • 48. What gross changes are expected in bones, teeth and urine in cases of congenital porphyria?
  • 49. How would you make a tentative diagnosis of congenital porphyria at the time of necropsy?
  • 50. Discuss the manner in which congenital porphyria is transmitted in cats, swine and cattle.
  • 51. Briefly discuss the skin lesions expected in swine with congenital porphyria.
  • 52. Briefly explain the manner in which hepatotoxic photosensitization develops in animal.
  • 53. What fluorescent pigment is incriminated in cases of hepatotoxic photosensitization?
  • 54. Under what circumstances would you expect primary photosensitization to develop in a 6-year-old Holstein cow?
  • 55. What is the so-called Dubin-Johnson pigment?
  • 56. What is the so-called Cloisonne' kidney?
  • 57. Distinguish between acholuric and choluric icterus.
  • 58. Under what circumstances would you expect to observe biliverdin in tissues?