1-year course; I-semester 2024/2025
TOPICS OF PRACTICAL CLASSES AND SEMINAR
I. Practical class: Cell structure and various cell types. Short test from cell structure and histological technique (07.10; 09.10; 10.10. 2024).
Slide no 1. Cells of epithelium from oral cavity - H+E
Slide no 10. Slide no 2. Nerve cells (spinal cord gray matter).- H+E staining (Hematoxylin and Eosin).
Slide no 103. Egg cell ( ovary) - H+E
Slide no 4. Carbohydrate deutoplasm (deuteroplasm) – section of the liver, showing intracellular deposits of glycogen. The glycogen appears as fine or coarse dark-carmine granules. - Best’s Carmine stain.
Slide no 5. Lipids deutoplasm – section of the unilocular (yellow or white) adipose tissue. The fat is stained black with osmic acid in the adipose cells cytplasm. - Osmic acid stain.
Slide no 6. Lipids lixiviated deutoplasm.- H+E
Slide no 7. Melanocytes ( cells from fish epidermis) – not stained
Slide no 8. The Golgi complex occupies a characteristic apolarized position in the cytoplasm by nerve cells (pseudounipolar neurons). The sensory ganglion (the posterior root ganglion of spinal nerve)– nerve cells impregnated with silver nitrate. - Silver stain
II. Practical class: Epithelial tissue. Simple epithelium. (14.10; 16.10; 17.10. 2024).
Slide no 10. Simple squamous epithelium - H+E
Slide no 11. Simple squamous epithelium, - Sliver stain.
Slide no 12. Simple(one layer) cuboidal epithelium from kidney collecting tubules. - Azan stain.
Slide no 13. Simple columnar epithelium that covers the internal surface of a small intestinum.- H+E stain.
Slide no 14. Pseudostratified (pseudo two-layer)columnar epithelium that covers the internal surface of the ductus epidimidis. This epithelium is composed of rounded basal and long columnar calls. The epithelium surface is covered by long, branched microvilli called stereocilia. - H+E stain.
Slide no 15. or 36. Pseudostratified columnar ciliated epithelium (respiratory tracts epithelium) in the trachea. - H+E stain.
III. Practical class: Stratified epithelium. Principal types of exocrine glands. (21.10; 23.10; 24.10. 2024).
Slide no 89. Nonkeratinized stratified squamous epithelium in the esophagus. – H+E.
Slide no 101. Stratified transitional epithelium in the urinary bladder. - Azan stain.
Slide no 116. Stratified squamous keratinized epithelium of thick skin. Epidermis. - H+E stain.
Slide no 58. Serous gland ( parotid gland) - Azan stain.
Slide no 95. Mucous gland ( large intestine) - H+E
Slide no 20. Unicellular, intraepithelial exocrine glands. Goblet cells of the intestines. These cells are secreting mucous to the extracellular space. - Azan stain.
IV. Practical class: Blood cells. (28.10; 30.10; 31.10. 2024).
Slide no 21. Frog’s blood. A demonstrational histological specimen. - Pappenhaim’s method stain.
Slide no 22. Human blood. - Pappenhaim’s method stain.
Slide no 23. Reticulocytes – young erythrocytes may have a few cytoplasmic granules and netlike structure. - Cresol blue stain.
V. Practical class: Proper connective tissue. (04.11; 06.11; 07.11. 2024).
Slide no 24. Mucous tissue ( Wharton’s jelly), umbilical cord. - Azan stain.
Slide no 68. Reticular connective tissue (lymph node), - H+E.
Slide no 27. Reticular connective tissue. Reticular fibers in the medulla and cortecs of a lymph node (or in the white pulp of spleen). The reticular fibers are seen as a network of dark, thin, wavy fibers. Silver stain.
Slide no 28. Loose connective tissue – elastic fibers. Total preparation (this is a whole mount specimen of rat mesentery; the mesentery is lying on the slide) showing blue fuchsin-stained (or eosin-stained) no anastomosing bundles of collagen fibers; while the elastic fibers appear as thin, brown-black, branching (dark anastomosing) resorcin-stained filaments. Collagen bundles of various thicknesses are observed. Fuchsin (or eosin) + resorcin stain.
Slide no 29. Loose connective tissue ( elastic and collagen fibers) - Orcein and resorcin-fuchsin, aniline blue)
Slide no 30. Fibroblasts and acidophilic cells of connective tissue, H+E
Slide no 6. and 33. Lipids “lixiviated” deutoplasm – section of the unilocular (yellow or white) adipose tissue. Fat has been dissolved out of the section during the preparation of the slide, shoving large, empty spaces in the adipose cell cytoplasm. H+E stain.
Slide no 34. Longitudinal section of dense regular connective tissue from a collagen tendon. Thick bundles of parallel collagen fibers fill the intercellular spaces between fibroblasts. H+E stain.
Slide no 35. Longitudinal section of dense regular elastic tissue from an elastic. - Van Gieson’s method stain.
VI. Practical class: Structure of bone and cartilage. (11.11 (to be agreed); 13.11; 14.11. 2024).
Slide no 36. Hyaline cartilage. Section of trachea. Chondrocytes are located in matrix lacunae. The cartilage interstitial growth is reflected by the chondrocyte pairs and clusters that are responsible for the formation of isogenous groups. The intense hematoxylin-staining of the matrix around each chondrocytes is visible. H+E stain. The more darkly stained area is a zone of the cartilage matrix that is rich in glycosoaminoglycans. - H+E stain.
Slide no 37. Elastic cartilage from the auricle of the ear. Elastic fibers is orcein in brown, or resorcin in red stained. Orcein or resorcin stain.
Slide no 38. Cancellous (spongy) bone. Areas with numerous trabecule of bone and interconnecting cavities – corresponding to cancellous bone (it is internal structure of bones). The outher portion of the bone has a solid structure and represents compact (dense) bone.
Slide no 39. Structure of compact bone, both transverse and longitudinal sections (two fragments of compact bone). The Haversian systems are visible, as are the osteocytes in their lacunae. Decalcified compact bone.
VII. Practical class: Structure of muscle tissue. (18.11; 20.11; 21.11. 2024).
Slide no 41. Isolated muscle cells
Slide no 42. Skeletal muscle fibers – longitudinal and cross section (tongue), - H+E
Slide no 43 or 94. Smooth muscle – transverse (cross) section and longitudinal section. - H+E stain.
Slide no 44. Skeletal muscle – cross section and longitudinal section of transversal striated skeletal muscle (tounge), showing many muscle fibers with peripherally located nuclei. - Silver stain.
Slide no 45. Longitudinal and transverse section of cardiac muscle, showing numerous muscle fibers with centrally placed nuclei. - H+E stain.
VIII. Practical class: Nerve tissue and glial cells and nerve system. (25.11; 27.11; 28.11. 2024).
Slide no. 47. Dorsal root ganglion (spinal ganglion) =sensory ganglion. The sensory ganglion comprise large neuronal cell bodies (pseudounipolar neurons), which are surrounded by abundant small satellite cells. H+E stain. Nerve fibers passing to the center of the ganglion, the ganglion cells being located peripherally, each cell body is seen to be surrounded by a layer of flattened satellite cells. Satellite cells are represented by the very small nuclei at the periphery of the neuronal cell bodies.- H+E stain.
Slide no. 8. Silver-impregnated dorsal rot ganglion.
Slide no. 119. Cerebrum – cerebral cortex (gray mater) – has 6 layers of nerve cells. Pyramidal cells are typical of the cerebral cortex. H+E stain.
Slide no. 120. Cerebellum – cerebellar cortex: molecular layer + Purkinje cell layer + granular layer. Purkinje cells are typical of the cerebellar cortex. - H+E stain.
Unnumberd slide. - Silver impregnated cerebellum. Draw the Purkinje cell Demonstrational slide.
Slide no. 2 or 123. Spinal cord. Draw the following: a) gray matter and white matter in the diagram, b) motor neurons with their basophilic bodies (Nissl bodies). Toluidine blue stain.
Slide no 52. Isolated nerve fiber (peripheral nerve), longitudinal section – impregnated with OsO4 . (osmic acid stain).
Slideno 53. Nerve fibers (peripheral nerve), cross section – impregnated with OsO4 . (osmic acid stain).
IX. Practical part of partial test I: histological technique, cell structure, epithelial tissue, and blood, Proper connective tissue, structure of bone and cartilage, structure of muscle tissue, nerve tissue and glial cells and nerve system . (02.12; 04.12; 05.12. 2024).
Theoretical part 02.12. 2024 at 11.30-12.15 (lecture time)
X. I Seminar - General Embryology ( 09.12.; 11.12.; 12.12.2024 )
XI. II Seminar - General Embryology ( 13.01.; 15.01; 16.01.2025 )
XII. III Seminar - Embryology ( 20.01. ; 22.01; 23.01.2025 )
XIII. Partial test III - Embryology 27.01.2025 at 11.30-12.15 (lecture time)
XIV. Practical class: The circulatory system (The blood vascular system). (03.02.; 05.02.; 06.02.2025)
Slide no. 45. –Heart. H+E staining (Hematoxylin and Eosin).
Slide no. 61. - Capillaries, postcapillaries and precapillaries H+E stain.
Slide no. 62. – Artery + vein, stained with H+E stain.
Slide no. 63. – Artery and vein stained orcein and resorcin-fucsin (elastic membranes and fibers).
Slide no. 64. – Aorta stained H+E.
Slide no. 65. – Aorta stained with orcein and resorcin-fucsin (elastic membranes and fibers).
Slide no. 66.– Endothelium; valves H+E.
XVII. Practical class: The immune system; lymphoid organs + bone marrow. (10.02.; 12.02.; 13.02.2025)
Slide no. 68. - Lymph node. H+E stain.
Slide no. 69. - Palatine tonsil.H+E stain.
Slide no. 70. - Spleen. H+E stain.
Slide no. 71. - Bone marrow.H+E stain.
Slide no. 72. – Thymus. H+E stain.
Or slide no. 126. – Thymus+ thyroid gland + trachea + esophagus. H+E stain.
1-year course; I-semester 2024/2025
TECHNIQUE DETAILED DRAFT OF CLASSES
Histology, Cytology and its methods of study
PREPARATION OF TISSUES FOR LIGHT MICROSCOPIC EXAMINATION
Tissue ProcessingTissues from the body taken for diagnosis of disease processes must be processed in the histology laboratory to produce microscopic slides that are viewed under the microscope by pathologists. The techniques for processing the tissues, whether biopsies, larger specimens removed at surgery, or tissues from autopsy, are described below. The persons who do the tissue processing and make the glass microscopic slides are histotechnologists.
Fixation - types of fixatives.The purpose of fixation is to preserve tissues permanently in as life-like a state as possible. Fixation should be carried out as soon as possible after removal of the tissues (in the case of surgical pathology) or soon after death (with autopsy) to prevent autolysis. There is no perfect fixative, though formaldehyde comes the closest. Therefore, a variety of fixatives are available for use, depending on the type of tissue present and features to be demonstrated.There are five major groups of fixatives, classified according to mechanism of action:Aldehydes; Mercurials; Alcohols; Oxidizing agents; Picrates.
Aldehydesinclude formaldehyde (formalin) and glutaraldehyde. Tissue is fixed by cross-linkages formed in the proteins, particularly between lysine residues. This cross-linkage does not harm the structure of proteins greatly, so that antigenicity is not lost. Therefore, formaldehyde is good for immunoperoxidase techniques. Formalin penetrates tissue well, but is relatively slow. The standard solution is 10% neutral buffered formalin. A buffer prevents acidity that would promote autolysis and cause precipitation of formol-heme pigment in the tissues.
Glutaraldehydecauses deformation of alpha-helix structure in proteins so is not good for immunoperoxidase staining. However, it fixes very quickly so is good for electron microscopy. It penetrates very poorly, but gives best overall cytoplasmic and nuclear detail. The standard solution is a 2% buffered glutaraldehyde
Mercurialsfix tissue by an unknown mechanism. They contain mercuric chloride and include such well-known fixatives as B-5 and Zenker's. These fixatives penetrate relatively poorly and cause some tissue hardness, but are fast and give excellent nuclear detail. Their best application is for fixation of hematopoietic and reticuloendothelial tissues. Since they contain mercury, they must be disposed of carefully.
Alcohols, including methyl alcohol (methanol) and ethyl alcohol (ethanol), are protein denaturants and are not used routinely for tissues because they cause too much brittleness and hardness. However, they are very good for cytologic smears because they act quickly and give good nuclear detail. Spray cans of alcohol fixatives are marketed to physicians doing PAP smears, but cheap hairsprays do just as well.
Oxidizing agentsinclude permanganate fixatives (potassium permanganate), dichromate fixatives (potassium dichromate), and osmium tetroxide. They cross-link proteins, but cause extensive denaturation. Some of them have specialized applications, but are used very infrequently.
Picratesinclude fixatives with picric acid. Foremost among these is Bouin's solution. It has an unknown mechanism of action. It does almost as well as mercurials with nuclear detail but does not cause as much hardness. Picric acid is an explosion hazard in dry form. As a solution, it stains everything it touches yellow, including skin.
Fixation – factors affecting fixation. There are a number of factors that will affect the fixation process: Buffering; Penetration; Volume; Temperature; Concentration Time interval.
Fixation isbest carried out close to neutral pH, in the range of 6-8. Hypoxia of tissues lowers the pH, so there must be buffering capacity in the fixative to prevent excessive acidity. Acidity favors formation of formalin-heme pigment that appears as black, polarizable deposits in tissue. Common buffers include phosphate, bicarbonate, cacodylate, and veronal. Commercial formalin is buffered with phosphate at a pH of 7. Penetration of tissues depends upon the diffusability of each individual fixative, which is a constant. Formalin and alcohol penetrate the best, and glutaraldehyde the worst. Mercurials and others are somewhere in between. One way to get around this problem is sectioning the tissues thinly (2 to 3 mm). Penetration into a thin section will occur more rapidly than for a thick section. The volume of fixative is important. There should be a 10:1 ratio of fixative to tissue. Obviously, we often get away with less than this, but may not get ideal fixation. One way to partially solve the problem is to change the fixative at intervals to avoid exhaustion of the fixative. Agitation of the specimen in the fixative will also enhance fixation.
Increasing the temperature, as with all chemical reactions, will increase the speed of fixation, as long as you don't cook the tissue. Hot formalin will fix tissues faster, and this is often the first step on an automated tissue processor.
Fixatives - general usage. There are common usages for fixatives in the pathology laboratory based upon the nature of the fixatives, the type of tissue, and the histologic details to be demonstrated. Formalin is used for all routine surgical pathology and autopsy tissues when an H and E slide is to be produced. Formalin is the most forgiving of all fixatives when conditions are not ideal, and there is no tissue that it will harm significantly. Most clinicians and nurses can understand what formalin is and does and it smells bad enough that they are careful handling it. Zenker's fixatives are recommended for reticuloendothelial tissues including lymph nodes, spleen, thymus, and bone marrow. Zenker's fixes nuclei very well and gives good detail. However, the mercury deposits must be removed (dezenkerized) before staining or black deposits will result in the sections. Bouin's solution is sometimes recommended for fixation of testis, GI tract, and endocrine tissue. It does not do a bad job on hematopoietic tissues either, and doesn't require dezenkerizing before staining. Glutaraldehyde is recommended for fixation of tissues for electron microscopy. The glutaraldehyde must be cold and buffered and not more than 3 months old. The tissue must be as fresh as possible and preferably sectioned within the glutaraldehyde at a thickness no more than 1 mm to enhance fixation. Alcohols, specifically ethanol, are used primarily for cytologic smears. Ethanol (95%) is fast and cheap. Since smears are only a cell or so thick, there is no great problem from shrinkage, and since smears are not sectioned, there is no problem from induced brittleness.
For fixing frozen sections, you can use just about anything--though methanol and ethanol are the best. Concentration of fixative should be adjusted down to the lowest level possible, because you will expend less money for the fixative. Formalin is best at 10%; glutaraldehyde is generally made up at 0.25% to 4%. Too high a concentration may adversely affect the tissues and produce artefact similar to excessive heat. Also very important is time interval from of removal of tissues to fixation. The faster you can get the tissue and fix it, the better. Artefact will be introduced by drying, so if tissue is left out, please keep it moist with saline. The longer you wait, the more cellular organelles will be lost and the more nuclear shrinkage and artefactual clumping will occur.
Tissue Processing. Once the tissue has been fixed, it must be processed into a form in which it can be made into thin microscopic sections. The usual way this is done is with paraffin. Tissues embedded in paraffin, which is similar in density to tissue, can be sectioned at anywhere from 3 to 10 microns, usually 6-8 routinely. The technique of getting fixed tissue into paraffin is called tissue processing. The main steps in this process are dehydration and clearing.Wet fixed tissues (in aqueous solutions) cannot be directly infiltrated with paraffin. First, the water from the tissues must be removed by dehydration. This is usually done with a series of alcohols, say 70% to 95% to 100%. Sometimes the first step is a mixture of formalin and alcohol. Other dehydrants can be used, but have major disadvantages. Acetone is very fast, but a fire hazard, so is safe only for small, hand-processed sets of tissues. Dioxane can be used without clearing, but has toxic fumes.The next step is called "clearing" and consists of removal of the dehydrant with a substance that will be miscible with the embedding medium (paraffin). The commonest clearing agent is xylene. Toluene works well, and is more tolerant of small amounts of water left in the tissues, but is 3 times more expensive thanxylene. Chloroform usedto be used, but is a health hazard, and is slow. Methyl salicylate is rarely used because it is expensive, but it smells nice (it is oil of wintergreen). There are newer clearing agents available for use. Many of them are based on limolene, a volatile oil found in citrus peels. Another uses long chain aliphatic hydrocarbons (Clearite). Although they represent less of a health hazard, they are less forgiving with poorly fixed, dehydrated, or sectioned tissues.
Finally, the tissue is infiltrated with the embedding agent, almost always paraffin. Paraffins can be purchased that differ in melting point, for various hardnesses, depending upon the way the histotechnologist likes them and upon the climate (warm vs. cold). A product called paraplast contains added plasticizers that make the paraffin blocks easier for some technicians to cut. A vacuum can be applied inside the tissue processor to assist penetration of the embedding agent. The above processes are almost always automated for the large volumes of routine tissues processed. Automation consists of an instrument that moves the tissues around through the various agents on a preset time scale. The "technicon" tissue processor is one of the commonest and most reliable (a mechanical processor with an electric motor that drives gears and cams), though no longer made. Newer processors have computers, not cam wheels, to control them and have sealed reagent wells to which a vacuum and/or heat can be applied.
Tissues that come off the tissue processor are still in the cassettes and must be manually put into the blocks by a technician who must pick the tissues out of the cassette and pour molten paraffin over them. This "embedding" process is very important, because the tissues must be aligned, or oriented, properly in the block of paraffin. Alternatives to paraffin embedding include various plastics that allow thinner sections. Such plastics include methyl methacrylate, glycol methacrylate, araldite, and epon. Methyl methacrylate is very hard and therefore good for embedding undecalcified bone. Glycol methacrylate has the most widespread use since it is the easiest to work with. Araldite is about the same as methacrylate, but requires a more complex embedding process. Epon is routinely used for electron microscopy where very thin sections are required. Plastics require special reagents for deydration and clearing that are expensive. For this reason, and because few tissues are plastic embedded, the processing is usually done by hand. A special microtome is required for sectioning these blocks. Small blocks must be made, so the technique lends itself to small biopsies, such as bone marrow or liver.
Sectioning.Once the tissues have been embedded, they must be cut into sections that can be placed on a slide. This is done with a microtome. The microtome is nothing more than a knife with a mechanism for advancing a paraffin block standard distances across it. There are three important necessities for proper sectioning: (1) a very sharp knife, (2) a very sharp knife, and (3) a very sharp knife. Knives are either of the standard thick metal variety or thin disposable variety (like a disposable razor blade). The former type allows custom sharpening to one's own satisfaction, but is expensive (more than $100 per blade). The latter cost about $1 per blade and are nearly as good. The advantage of the disposable blade becomes apparent when sectioning a block in which is hidden a metal wire or suture. Plastic blocks (methacrylate, araldite, or epon) are sectioned with glass or diamond knives. A glass knife can section down to about 1 micron. Thin sections for electron microscopy (1/4 micron) are best done with a diamond knife which is very expensive ($2500).
Microtomes have a mechanism for advancing the block across the knife. Usually this distance can be set, for most paraffin embedded tissues at 6 to 8 microns. The more expensive the microtome ($15,000 to $20,000), the better and longer-lasting this mechanism will be. Sectioning tissues is a real art and takes much skill and practice. Histotechnologists are the artists of the laboratory. It is important to have a properly fixed and embedded block or much artefact can be introduced in the sectioning. Common artefacts include tearing, ripping, "venetian blinds", holes, folding, etc. Once sections are cut, they are floated on a warm water bath that helps remove wrinkles. Then they are picked up on a glass microscopic slide. The glass slides are then placed in a warm oven for about 15 minutes to help the section adhere to the slide. If this heat might harm such things as antigens for immunostaining, then this step can be bypassed and glue-coated slides used instead to pick up the sections.
Frozen Sections.At times during performance of surgical procedures, it is necessary to get a rapid diagnosis of a pathologic process. The surgeon may want to know if the margins of his resection for a malignant neoplasm are clear before closing, or an unexpected disease process may be found and require diagnosis to decide what to do next, or it may be necessary to determine if the appropriate tissue has been obtained for further workup of a disease process. This is accomplished through use of a frozen section. The piece(s) of tissue to be studied are snap frozen in a cold liquid or cold environment (-20 to -70 Celsius). Freezing makes the tissue solid enough to section with a microtome.Frozen sections are performed with an instrument called a cryostat. The cryostat is just a refrigerated box containing a microtome. The temperature inside the cryostat is about -20 to -30 Celsius. The tissue sections are cut and picked up on a glass slide. The sections are then ready for staining.
Staining. The embedding process must be reversed in order to get the paraffin wax out of the tissue and allow water soluble dyes to penetrate the sections. Therefore, before any staining can be done, the slides are "deparaffinized" by running them through xylenes (or substitutes) to alcohols to water. There are no stains that can be done on tissues containing paraffin. The staining process makes use of a variety of dyes that have been chosen for their ability to stain various cellular components of tissue. The routine stain is that of hematoxylin and eosion (H and E). Other stains are referred to as "special stains" because they are employed in specific situations according to the diagnostic need.
2. DETAILED DRAFT OF CLASSES
Cell structure (Cell components): The cell is composed of two basic components (parts) – cytoplasm and nucleus.
1. The cytoplasm is composed of a cytosol (matrix), in which the organelles are embedded, the cytoskeleton, and various deposits (carbohydrates, lipids, and pigments). The cytoplasm is surrounded with a plasma membrane (plasmalemma) – separating the cytoplasm from its extracellular environment. The plasmalemma is the external limit (selective barier) of the cell, there is a continuum between the interior of the cell and the extracellular environment.
2. Cytosol – structure and functions.
3. Plasma membranes are composed of phospholipids, cholesterol, proteins, and chains of oligosaccharides covalently linked to phospholipids and protein molecules.
4. The plasmalemma – structure and functions. The plasmalemma contains proteins called integrins (integral proteins that are linked to cytoskeletal filaments and extracellular molecules), and peripheral proteins exhibit a looser association with menbrane surfaces. The fluid mosaic model for mosaic membrane proteins is dispositioned in the plasmalemma structure. The 2-layered lipids structure of all plasma membranes – called Unit Membrane. Plasmalemma receptors.
5. Structure and functions of organelles: Endoplasmic Reticulum (rough and smooth); Golgi Complex (Golgi Apparatus); Lysosomes: Phagocytic vacuoles, Phagolysosomes, Heterophagolysosomes, Autophagolisosomes, Residual bodies; Early (peripheral) endosomes; Late (perinuclear) endosomes – endosomal compartment; Pinocytotic vesicles; Coated vesicle; Transporting vesicles; Peroxisomes (Microbodies); Proteasomes; Ribosomes; Polyribosomes; Mitochondria :external and internal mitochondrial membranes, mitochondrial cristae and matrix, the cristae are covered with globular units that participate in the formation of ATP, Mitochondrial DNA (mtDNA); Centrosome (Cell center); Centrioles and Microtubule – Organizing Cell Centers; Secretory granules.
6. The Cytoskeleton: Microtubules; Actin microfilaments (this protein is present in all cells) and Myosin (this protein is present in muscle cells – Thick myosin filaments); Intermediate filaments: Keratins; Vimentin; Desmin; Glial filaments; Neurofilaments.
8. The Cell Nucleus: Heterochromatin and Euchromatin; Nuclear matrix; Nucleolus;
Nuclear Envelope – is made of 2 membranes.
9. Cellular transport: Transport across the cell membrane. Passive transport; Facilitated transport; Active transport; Ionic pomp; Ion channels and membrane potentials. The action potential (the role of Na+ – K+ channel). Exocytosis; Endocytosis; Pinocytosis (fluid-phase endocytosis); Transcytosis; Potocytosis; Phagocytosis (heterophagocytosis, autophagocytosis); Secretion. Cytoskeleton (structure). Cell adhesion and extracellular matrix. Intercellular signaling (endocrine and paracrine signaling).
3. DETAILED DRAFT OF CLASSES
Epithelial Tissue. Simple (one layer) epithelium
1. Tissues: Concept and Classificatio. 2. The forms and characteristics of epithelial cells. Stem cells = undifferentiated cells. 3. The function of epithelial tissue: Surface epithelium. Endothelium. Mesothelium (mesodermal epithelium). Sensory epithelium. Secretory epithelium. Epithelial-reticular cells of thymus (stellate cells). Epithelization = covering by the epithelium. Ion-pumping epithelial cells. Supporting cells (sustentacular cells). Shedding of epithelium (exfoliation; desquamation). Renewal and regeneration of one layer epithlial tissues. 4. Basal lamina and term Basement membrane. 5. Intercellular junctions.
6. Specializations of the cell surface:Glycocalyx (cell coat), Microvilli; Stereocilia; Cilia and Flagella. 7 Principal types of epithelia: Covering epithelia and Glandular epithelia. 8.Common types of covering epithelia: Simple epithelium and Stratified epithelium. 9.Simple (one layer) Epithelium: Squamous; Cuboidal and Columnar epithelium. 10. Simple pseudostratified (Ciliated pseudostratified) epithelium (layers of cells with nuclei at different levels; all cells adhere to basal lamina). Ciliated columnar cells. Basal (short) cells. Goblet cells. Ciliated cells. Neuroepithelial cells. Neuroendocrine cells.
4. DETAILED DRAFT OF CLASSES
Stratified Epithelium (two or more layers). Principal types of Exocrine Glands.
1. Stratified (multilayer) squamous keratinized epithelium. 2. Stratified squamous nonkeratinized epithelium. 3. Stratified cuboidal epithelium. 4. Stratified transitional epithelium. 5. Stratified columnar epithelium.
6. Principal types of exocrine unicellular intraepithelial glands and exocrine multicellular glands – simple and compound glands: a) simple tubular, b) simple coiled tubular,
c) simple branched tubular, d) simple branched acinar, e) compound tubuloacinar,
f) compound tubular, g) compound acinar.
7. Function of exocrine gland cells: a) serous cells, b) mucous secreting cells, c) cells that transport ions (The cell uses the energy stored in ATP); d) cells that transport by pinocytosis.
8. Myoepithelial cells. 9. Renewal and regeneretion of stratified epithelium.
5. DETAILED DRAFT OF CLASSES
I. Blood. (Composition of plasma, blood cells, hematopoiesis).
1. Definition of blood.
2. Composition of blood: a/ cells: Erythrocytes (Red blood cells, RBC); White blood cells (WBC), b/ plasma, serum, c/ normal percentages of the different types of blood cells.
3. Structure and function of red blood cells (erythrocytes)
4. Granulocytes: a/ characteristic of granulocytes, b/ structure and function of neurotrophils, c/ structure and function of basophils, d/ structure and function of eosinophils.
5. Agranulocytes: a/ characteristic of agranulocytes, b/ structures and function of monocytes, c/ types of limphocytes: B lymphocytes, T lymphocytes (Tc lymphocytes = T cytotoxic cells = killer cells; Th lymphocytes = T helper cells; LGL = Large granular lymphocytes; LAK = Lymphokine-activated killer).d/ Structures and function of lymphocytes.
6. Trombocytes (Blood plateles) – structures and function.
II. Hematopoiesis
1. Theory of hematopoiesis: a) monophyletic (or unitarian) theory of hematopoiesis, b). polyphyletic theory of hemopoiesis.
2. Hematopoiesis in early embryonic development: a) “Blood islands” in the wall of the yolk sac of the embryo, b) Hepatic phase. c) Bone marrow phase.
3. Development of erythrocytes (erythropoiesis): Proerythroblast → Basophilic erythroblast → Polychromatophilic erythroblast → Normoblast → Reticulocyte →Erythrocyte.
4. Kinetics of erythropoiesis.
5. Megakaryocyte development: Megakaryoblast→ Promegakaryocyte → Megakaryocyte → Blood platelets.
6. Development of granulocytes (granulopoiesis)
Myeloblast → Promyelocyte → Eosinophilic myelocyte → Eosinophilic metamyelocyte → Eosinophil
Promyelocyte → Neutrophilic myelocyte → Neutrophilic metamyelocyte → Band cell → Neutrophil
Promyelocyte → Basophilic myelocyte → Basophilic metamyelocyte → Basophil
7. Kinetics of granulopoiesis.
8. Monocyte development: Monoblast → Promonocyte → Monocyte → Macrophage
9. Lymphopoiesis.
10. Bone marrow: a). Red bone marrow. b). Yellow bone marrow.
11. Number and percentage of blood corpuscles (blood count).
6. DETAILED DRAFT OF CLASSES
Proper connective tissue.
I. Types of the connective tissue: 1. Immature gelationous tissue; 2. Mature gelationous tissue = Mucous connective tissue (Wharton gelatinum from the umbilical cord); 3. Connective tissue proper [Loose (areolar); Dense (regular and irregular)].
4. Connective tissue proper with special properties: a) Reticular connective tissue (Hematopoietic tissue); b) Adipose tissue (White = yellow adipose tissue; Brown adipose tissue = Multilocular fat); c) Elastic tissue].
II . Functions of the connective tissue.
III. Extracellular matrix: 1. Ground substance, Tissue fluid. 2. Fibres: Collagen fibres; Elastic fibres, and Oxytalan fibres; Reticular fibres (Argyrophylic fibres); Elaunin fibres.
IV. Cells of proper connective tissue: Mesenchymal cell; Fibroblast; Fibrocyte; Myofibroblast; Pericyte (Advential cell, Perivascular cell); Histiocyte (Mononuclear cell), Mononuclear Phagocyte System (MPS); Multinuclear giant cell; Plasma cell; Mast cell (Mastocyte, Labrocyte): a) Mucosal mast cell (MMC), b) Connective tissue mast cell (CTMC); Melanophore and Melanocyte.
7. DETAILED DRAFT OF CLASSES
Bone and cartilage (Supporting connective tissue).
1. Structure of Cartilage: perichondrium, chondroblastic layer (cambial layer), matrix (ground substance and fibres), territorial matrix (basophilic peripheral zone), chondron, chondrocyte (cartilage) lacuna, chondroblasts, chondrocytes, isogenous (isogenic) group, chondroclasts. Cartilage as an organ. Chondrogenesis.
2. Types of Cartilage: - Hyaline Cartilage; - Elastic Cartilage; - Fibrocartilage (Intervertebral disks). 3) Localisation and function of the cartilage.
4. Types of Bone: I. Woven bone (primary = immature = primitive = primordial = nonlamellated = fibrous); II. Lamellar bone (secondary = mature = adult).
Ad II. Lamellar bone: a). Compact bone. b). Cancellous bone (spongy = cancellated).
5. Bone cells and their function ( Osteoblasts, Osteocytes, Osteoclasts). 6. Structure of Bone: Bone matrix. Periosteum and endosteum. Osteon (Haversian system). Intercalated lamellae system. Bone trabecula. Bone lamella. Osteoid (Demineralized bone matrix). Ossein fibers. Lamella ossea (Haversian). Intermediate lamellae. Osteoni (Havers) canales. Nourishing (Volkmann’s) canals. Bone canalicular network. Osteocytic lacunas. Osteocyte processes. 7. Histogenesis (intramembranous ossification, endochondral ossification). 8. Bone growth and remodeling. Periosteal (apposition) osteogenesis. 9. Metabolic role of bone tissue (calcium reservoir). Joints.
8. DETAILED DRAFT OF CLASSES
Muscle tissue. Formation and classification of muscle tissue: I. Striated skeletal muscle tissue: 1. Structure of the striated muscle fiber: a/ myofibriles, b/ sarcomere, c/ smooth endoplasmic reticulum (SER) and T – tubule (transverse tubule) – muscle triad. 2. Types of the striated muscle fibers: a/ fibers type 1 – muscle fiber red, b/ fibers type 2 – muscle fiber white. 3. Satellite cells.
4. Structure of the skeletal muscle: a/ epimysium, b/ perimysium, c/ endomysium.
II. Striated cardiac muscle tissue: 1. Myocardium = cardiac muscle. 2. Structure of cardiac working muscle cells (cardiocytes = cardiac cells = cardiac muscle fibers); – dyad; the intercalated discs represent the junctional complex; gap junctions provide ionic continuity between adjacent cardiac muscle cells. 3. Structure of the conducting system of the heart. Atrioventricular node. Atrioventricular bundle of His. Purkinje cells (fibers).
4. Cardiac myoendocrine cells . atrial natriuretic factors (ANF, BNP, CNP) = atrial natriuretic peptides (ANP, BNP, CNP).
III. Smooth muscle tissue
1. Smooth muscle cells. 2. Structure and localization of the smooth muscle tissue.
9. DETAILED DRAFT OF CLASSES
Nerve tissue and glial cells and nerve system.
1. Structure of nerve tissue. 2.Neurons (nerve cells) classification: a) Golgi type I neurons, b) Golgi type II neurons; or: a) multipolar neurons, b) bipolar neurons, c) pseudounipolar neurons, d) unipolar neurons; or a) motor (efferent) neurons, b) sensory (afferent) neurons, c) interneurons, d) secretory neurons.
3. Most neurons consist of 3 parts: a) the dendrites, which are elongated processes specialized in receiving stimuli from other neurons, sensory epithelial cells and ennvironment; b) the neuronal body (perikaryon + nukleus); and c) axon, which is a single long process specialized in generating or conducting nerve impulses to other cells.
4. Structure of nerve cell: neuroplasm, axon hillock, myelin sheath, telodendron, dendritic spine, neurofibrils, neurofilaments, microtubules (neurotubules), Nissl substance (granules), neurosecretory substance (granules), lipofuscin, neuromelanin, neuropil, synapses (chemical, electrical, mixed; axodendritic, axosomatic, axoaxonic, somatodendritic, somatosomatic, dendrodendritic synapse), axon terminal, presynaptic part, presynaptic membrane, synaptic vesicles, synaptic cleft, intersynaptic substance, postsynaptic part (membrane), excitatory post-synaptic potential = EPSP, inhibitory post-synaptic potential = IPSP, chloride-zinc ion channel (transport channels for chloride and zinc), voltage-gated Ca2+ channels.
5. Neurotransmitters: acetylcholine = Ach, epinephrine (adrenaline), norepinephrine (noradrenaline), serotonin 5-hydroxytryptamine = 5-HT substance P, somatostatin, vasoactive intestinal peptide = VIP.
6. Chemically-defined cells in central nervous system (CNS): a/ aminergic cells: noradrenergic (norepinephric) cells, adrenergic (epinephric) cells, dopaminergic cells, serotoninergic cells; b/ cholinergic cells; c/ polipeptidergic cells: somatostatinergic cells, calcitonin gene-related peptidergic (CGRP) cells.
7. Neuroglia – glial cells of central nervous system [glial body, glial filaments (glial fibrillary acid protein), glial processes]: a/ ependymal cells (lining cavities of central nervous system) – (planar ep. c., cuboid ep. cells, columnar ep. c., tanycyte); b/ astrocytes – structural support, repair processes, blood-brain barrier, metabolic exchanges (protoplasmic astrocytes, fibrous astr., fibroprotoplasmic astr.); c/ oligodendrocytes (myelin production, electric insulation), d/ microglia cells (macrophagic activity).
8. Neuroglia – Glial cells of peripheral nervous system: a/ ganglionic glial cells (smsll satellite cells), b/ Schwann (neurolemmal) cells (nerve myelin production, electric insulation);
9. Nerves: cranial nerves, spinal nerves. Nerves are bundles of nerve fibers surrounded by connective tissue sheaths.
10. Nerve structure. Endoneurium; Perineurium; Epineurium. Nerve fibers. Myelinated fibers. Unmyelinated fibers. Neurolema. Neurolema cells (Schwann cells). Myelin sheath. Myelin lamelle. Mesoaxon. Node of Ranvier. Intrnodal segment. Schmidt-Lanterman cleft. Nerve ending. Free nerve ending.
11. Myelinogenesis (myelinization – myelin production). Oligodendrocytes produce the myelin in the central nervous system. The same oligodendrocyte forms myelin sheaths for several nerve fibers. Schwann cell forms myelin around a segment of one axon in the peripheral nervous system.
12.The central and peripheral nervous system – development and histological structure.