Blood

1. Blood

2. Main features • Blood cells and blood plasma • Plazma proteins cause blood clotting and form thrombs. • Serum does not contain white or red blood cells nor a clotting factor; it is the blood plasma with the fibrinogens removed. • Blood cells are: erythrocytes, leukocytes and platelets http://en.wikipedia.org/wiki/Blood

3. Formed elements

4. Blood plasma Blood plasma - 55% of blood volume. 90% - 92% water, 7%-9% proteins, 1% inorganic elements, glucose, hormons, vitamins, a.o. Tissue liquid – similar concentration of electrolyte, smaller concentration of proteīns a.o. organic substances. Border: basal membrane and wall of blood vessels with Continuous or Fenestrated endothelium. Ions

5. Proteins

6. Complement Cascade of protein activation, to attach them to antigens and cause destruction of foreign cells in human body. http://highered.mcgraw-hill.com/sites/0072507470/student_view0/chapter22/animation__activation_of_complement.html

7. Blood clotting http://www.youtube.com/watch?v=xNZEERMSeyM

9. Erytrocytes http://www.pbrc.hawaii.edu/~kunkel/gallery. Erythrocytes (red), leukocytes (green) un platelets (yellow). (SEM x 9,900). Erythrocytes can move through capillaries with diameter smaller than diameter of erythrocyte.

10. carbohydrates carbohydrates Extracellular matrix Protein 3 Phosfolipids Glycophorin Cholesterol Glicolipids -Spektrin ß -Spectrin Actin -Ankirin Protein 4,1 Protein 4,1 Protein 4,2 Tropomiosin Plsmatic membrane of an erythrocyte Band 3 protein - Cl-/HCO3 antiporter Glikophorin – negative charge on membrane surface

11. Erythrocytes change their shape during the movement. • Colour depends on concentration of CO2 • Hemoglobin - 4 polipeptide chains. Functional group – heme, contains Fe atoms. • Content: 65% water, 1% organelles, 9% hemoglobine. Fe http://en.wikipedia.org/wiki/File:Erytrocyte_deoxy_to_oxy_v0.7.gif

12. Human blood group system • Glycolipids A and B (aglutinogens) • 0-I group, A -II groupa, B-III groupa, AB-IV groupa. • Plazma contain proteins – aglutins. Can be attached to aglutinogens.

13. Sickle cell anemia

14. Rhesus factor • 50 aglutinogens • most important - C,D,E aglutinogens • Cause hemolysis of fetus if in the blood of “Rh-” mother antibodies against Rh becomes synthesized after first labour.

15. Classification of leukocytes • Granulocytes – contain specifc and unspecific granules • Agranulocytes – do not contain granules Cell structure after staining can be: eosinophilic (acidophilic), basophilic (stain with methilene blue) and azurophilic (purple), light purple (weakstaining with eosin un methilene blue, specific granules)

16. Leukocyte differencial analysis • Number of leukocytes in addults: • Neutrophilic granulocytes - 2.0–7.0×109/l (40–80%) • Limphocytes - 1.0–3.0×109/l (20–40%) • Monocytes - 0.2–1.0×109/l (2–10%) • Eosinophilic granulocytes - 0.02–0.5×109/l (1–6%) • Basophilic granuloccytes- 0.02–0.1×109/l (< 1–2%) • Numbers can differ in different populations and in different laborotories.

17. Immature granulocyte (IG) • Immature granulocytes (IGs) encompass immature cells of granulocytic lineages, including metamyelocytes, myelocytes, and promyelocytes, which are easily recognized morphologically and are reported by automated analyzer as IG altogether. IG normally absent from peripheral blood. • Increased IG occurs accompanied by an increase in neutrophils in the following conditions:[2] • Bacterial infections • Acute inflammatory diseases • Cancer (particularly with marrow metastasis) • Tissue necrosis • Acute transplant rejection • Surgical and orthopedic trauma • Myeloproliferative diseases • Steroid use • Pregnancy (mainly during the third trimester)

18. Neutrophil granulocytes — May indicate bacterial infection. May also be raised in acute viral infections. Because of the segmented appearance of the nucleus, neutrophils are sometimes referred to as "segs." The nucleus of less mature neutrophils is not segmented, but has a band or rod-like shape. Less mature neutrophils — those that have recently been released from the bone marrow into the bloodstream — are known as "bands" or "stabs". Stab is a German term for rod.[3] • Lymphocytes — Higher with some viral infections such as glandular fever and. Also raised in chronic lymphocytic leukemia (CLL). Can be decreased by HIV infection. In adults, lymphocytes are the second most common WBC type after neutrophils. In young children under age 8, lymphocytes are more common than neutrophils.[3] • Monocytes — May be raised in bacterial infection, tuberculosis, malaria, Rocky Mountain spotted fever, monocytic leukemia, chronic ulcerative colitis and regional enteritis [3] • Eosinophil granulocytes — Increased in parasitic infections, asthma, or allergic reaction. • Basophil granulocytes — May be increased in bone marrow related conditions such as leukemia or lymphoma.[3]

19. Monocytes un macrophages

20. Monocytes and macrophages are phagocytes.[3] Macrophages function in both non-specific defense (innate immunity) as well as help initiate specific defense mechanisms (adaptive immunity) of vertebrate animals. • Macrophages have the unique ability to metabolize one amino acid, arginine, to either a "killer" molecule (Nitric Oxide) or a "repair" molecule (Ornithine). • Macrophages predominantly expressing the killer or repair phenotype are now mainly called M1 or M2 macrophages because these 2 types of macrophages also stimulate T cell responses that further activate the killer macrophages or T cell phenotype (Th1), or stimulate antibody production (Th2)

21. Monocytes • Large number of lysosomes.

22. Makrophages • Beta glikans and other signalling moleculesactivatemacrophages

23. Macrophages 1. Erithrocytes; 2. – cytoplasm; 3. - nucleus, 4. – bacteria containing endosomes; 5. - pseudopodia. Scale bar: 10 μm. Specific ekxpresion ofproteinsCD14, CD40, CD11b, CD64, F4/80 (pele)/EMR1 (cilvēks), lysozyme M, MAC-1/MAC-3 and CD68 by flow cytometry or immunohistochemical staining.[4]

24. Macrophages www.beta-glucan-13d.com/ http://www.life.umd.edu/classroom/bsci422/mosser/macrophage.jpg

25. Macrophagesand cytoskeleton • http://www.svi.nl/products/software/fluvr/

26. When a monocyte enters damaged tissue through the endothelium of a blood vessel, a process known as the leukocyte extravasation, it undergoes a series of changes to become a macrophage. • Monocytes are attracted to a damaged site by chemical substances through chemotaxis, triggered by a range of stimuli including damaged cells, pathogens and cytokines released by macrophages already at the site. • At some sites such as the testis, macrophages have been shown to populate the organ through proliferation. • Unlike short-lived neutrophils, macrophages survive longer in the body up to a maximum of several months. http://en.wikipedia.org/wiki/Macrophage

27. Role in adaptive immunity • Macrophages are versatile cells that play many roles. As scavengers, they rid the body of worn-out cells and other debris. • Along with dendritic cells, they are foremost among the cells that "present" antigen, a crucial role in initiating an immune response. • As secretory cells, monocytes and macrophages are vital to the regulation of immune responses and the development of inflammation; they produce a wide array of powerful chemical substances (monokines) including enzymes, complement proteins, and regulatory factors such as interleukin-1. • At the same time, they carry receptors for lymphokines that allow them to be "activated" into single-minded pursuit of microbes and tumour cells. http://en.wikipedia.org/wiki/Macrophage

28. After digesting a pathogen, a macrophage will present the antigen (a molecule, most often a protein found on the surface of the pathogen and used by the immune system for identification) of the pathogen to the corresponding helper T cell. • The presentation is done by integrating it into the cell membrane and displaying it attached to an MHC class II molecule, indicating to other white blood cells that the macrophage is not a pathogen, despite having antigens on its surface. • Eventually, the antigen presentation results in the production of antibodies that attach to the antigens of pathogens, making them easier for macrophages to adhere to with their cell membrane and phagocytose. In some cases, pathogens are very resistant to adhesion by the macrophages. http://en.wikipedia.org/wiki/Macrophage

29. Antigen presentation • The antigen presentation on the surface of infected macrophages (in the context of MHC class II) in a lymph node stimulates TH1 (type 1 helper T cells) to proliferate (mainly due to IL-12 secretion from the macrophage). • When a B-cell in the lymph node recognizes the same unprocessed surface antigen on the bacterium with its surface bound antibody, the antigen is endocytosed and processed. • The processed antigen is then presented in MHCII on the surface of the B-cell. • T cells that express the T cell receptor which recognizes the antigen-MHCII complex (with co-stimulatory factors- CD40 and CD40L) cause the B-cell to produce antibodies that help opsonisation of the antigen so that the bacteria can be better cleared by phagocytes. • Macrophages provide yet another line of defense against tumor cells and somatic cells infected with fungus or parasites. • Once a T cell has recognized its particular antigen on the surface of an aberrant cell, the T cell becomes an activated effector cell, producing chemical mediators known as lymphokines that stimulate macrophages into a more aggressive form. http://en.wikipedia.org/wiki/Macrophage

30. In spite of a spectrum of ways to activate macrophages, there are two main groups designated M1 and M2. • M1 macrophages, as mentioned earlier (previously referred to as classically or alternatively activated macrophages),[7] M1 "killer" macrophages are activated by LPS and IFN-gamma, and secrete high levels of IL-12 and low levels of IL-10. • In contrast, the M2 "repair" designation broadly refers to macrophages that function in constructive processes like wound healing and tissue repair, and those that turn off damaging immune system activation by producing anti-inflammatory cytokines like IL-10. • M2 is the phenotype of resident tissue macrophages, and can be further elevated by IL-4. • M2 macrophages produce high levels of IL-10, TGF-beta and low levels of IL-12. Tumor-associated macrophages are mainly of the M2 phenotype, and seem to actively promote tumor growth.[8] http://en.wikipedia.org/wiki/Macrophage

31. Role in muscle regeneration • The first step to understanding the importance of macrophages in muscle repair, growth, and regeneration is that there are two “waves” of macrophages with the onset of damageable muscle use – subpopulations that do and do not directly have an influence on repairing muscle. • The initial wave is a phagocytic population that comes along during periods of increased muscle use that are sufficient to cause muscle membrane lysis and membrane inflammation, which can enter and degrade the contents of injured muscle fibers.[9][10][11] • These early-invading, phagocytic macrophages reach their highest concentration about 24 hours following the onset of some form of muscle cell injury or reloading.[12] • Their concentration rapidly declines after 48 hours.[10] • The second group is the non-phagocytic types that are distributed near regenerative fibers. These peak between two and four days and remain elevated for several days during the hopeful muscle rebuilding.[10] • The first subpopulation has no direct benefit to repairing muscle, while the second non-phagocytic group does. http://en.wikipedia.org/wiki/Macrophage

32. It is thought that macrophages release soluble substances that influence the proliferation, differentiation, growth, repair, and regeneration of muscle, but at this time the factor that is produced to mediate these effects is unknown.[12] • It is known that macrophages' involvement in promoting tissue repair is not muscle specific; they accumulate in numerous tissues during the healing process phase following injury.[13] • A study conducted in 2006 showcased macrophage influences on muscle repair of soleus muscle on mice.[14] • The first procedural step was to make sure macrophages are present in the muscle after onset of muscle injury, and then decrease their presence to see what effects were had on the muscle. • By using anti-F4/80 to bind to macrophages and render them useless, it was seen that when the second wave of macrophages were depleted, there were many more lesions in the muscle cell membrane between the second and fourth day – showing muscle damage when repairing is supposed to occur. • After testing for membrane lesions in both the total amount of muscle fibers present, it was noticed that most of the damage occurred in muscle cells that did not have the second subpopulation of macrophages present. Macrophages depletion prevents muscle membrane repair. http://en.wikipedia.org/wiki/Macrophage

33. When examining muscle regeneration, a significant reduction was found in the amount of myonuclei. • Depletion of macrophages was found to cause, between the second and fourth day of repair, much less muscle regeneration compared to muscle with macrophage population.[14] • Macrophages promote muscle regeneration between the second and fourth day. • To determine the influence of macrophages in muscle growth, muscle cross-sectional area in macrophage-depleted muscle area was measured against two muscle sets: muscle that was injured and had macrophage presence and muscle that was not injured and had macrophage presence. • The macrophage-depleted muscle showed less growth after four days, and injured muscle with macrophages nearly grew back to the level of uninjured muscle.[14] http://en.wikipedia.org/wiki/Macrophage

34. Role in wound healing • Macrophages replace Polymorphonuclear neutrophils as the predominant cells in the wound by two days after injury.[16] • Attracted to the wound site by growth factors released by platelets and other cells, monocytes from the bloodstream enter the area through blood vessel walls.[17] • Numbers of monocytes in the wound peak one to one and a half days after the injury occurs. Once they are in the wound site, monocytes mature into macrophages. The spleen contains half the body's monocytes in reserve ready to be deployed to injured tissue.[18][19] • The macrophage's main role is to phagocytize bacteria and damaged tissue,[15] and they also debride damaged tissue by releasing proteases.[20] • Macrophages secrete a number of factors such as growth factors and other cytokines, especially during the third and fourth post-wounding days. These factors attract cells involved in the proliferation stage of healing to the area.[21] • Macrophages may also restrain the contraction phase.[22] Macrophages are stimulated by the low oxygen content of their surroundings to produce factors that induce and speed angiogenesis[23] and they also stimulate cells that reepithelialize the wound, create granulation tissue, and lay down a new extracellular matrix.[24][25] http://en.wikipedia.org/wiki/Macrophage

35. Neutrophilic granulocytes11.11.2013

39. Eozinophilic granulocytes • Eosinophils also have lobed nuclei (two to four lobes). The number of granules in an eosinophil can vary because they have a tendency to degranulate while in the blood stream.[15] • Eosinophils play a crucial part in the killing of parasites (e.g., enteric nematodes) because their granules contain a unique, toxic basic protein and cationic protein (e.g., cathepsin[12]);[16] receptors that bind to IgE are used to help with this task.[17] • These cells also have a limited ability to participate in phagocytosis,[18] they are professional antigen-presenting cells, they regulate other immune cell functions (e.g., CD4+ T cell, dendritic cell, B cell, mast cell, neutrophil, and basophil functions),[19] they are involved in the destruction of tumor cells,[15] and they promote the repair of damaged tissue.[20] A polypeptide called interleukin-5 interacts with eosinophils and causes them to grow and differentiate; this polypeptide is produced by basophils.[16] http://en.wikipedia.org/wiki/Granulocyte

41. Basophylic granulcytes • Basophils are one of the least abundant cells in bone marrow and blood (occurring at less than two percent of all cells). Like neutrophils and eosinophils, they have lobed nuclei; however, they have only two lobes, and the chromatin filaments that connect them are not very visible. • Basophils have receptors that can bind to IgE, IgG, complement, and histamine. The cytoplasm of basophils contains a varied amount of granules; these granules are usually numerous enough to partially conceal the nucleus. Granule contents of basophils are abundant with histamine, heparin, chondroitin sulfate, peroxidase, platelet-activating factor, and other substances. http://en.wikipedia.org/wiki/Granulocyte

42. When an infection occurs, mature basophils will be released from the bone marrow and travel to the site of infection.[21] When basophils are injured, they will release histamine, which contributes to the inflammatory response that helps fight invading organisms. • Histamine causes dilation and increased permeability of capillaries close to the basophil. • Injured basophils and other leukocytes will release another substance called prostaglandins that contributes to an increased blood flow to the site of infection. Both of these mechanisms allow blood-clotting elements to be delivered to the infected area (this begins the recovery process and blocks the travel of microbes to other parts of the body). • Increased permeability of the inflamed tissue also allows for more phagocyte migration to the site of infection so that they can consume microbes.[18] http://en.wikipedia.org/wiki/Granulocyte

43. Heparinocytes • Mast are now considered to be part of the immune system. • Mast cells are very similar to basophil granulocytes (a class of white blood cells) in blood. • Both are granulated cells that contain histamine and heparin, an anticoagulant. Both cells also release histamine upon binding to immunoglobulin E.[3] • These similarities have led many to speculate that mast cells are basophils that have "homed in" on tissues. Furthermore they share a common precursor in bone marrow expressing the CD34 molecule. • Basophils leave the bone marrow already mature, whereas the mast cell circulates in an immature form, only maturing once in a tissue site. The site an immature mast cell settles in probably determines its precise characteristics.[2] • The first in vitro differentiation and growth of a pure population of mouse mast cells has been carried out using conditioned medium derived from concanavalin A-stimulated splenocytes.[6] • Later, it was discovered that T cell-derived interleukin 3 was the component present in the conditioned media that was required for mast cell differentiation and growth.[7] http://en.wikipedia.org/wiki/Mast_cell

44. http://en.wikipedia.org/wiki/Mast_cell

46. Megakariocytes produce thrombocytes

47. Megakariocytein red bone marrow. (left side) • Megakarioc form thrombocytes. Gimsza stain. (right side)

48. Thrombocytes lack major organelles. Thrombocytes of nonmammalian vertebrates have a nucleus and resemble B lymphocytes. Mammalian thrombocytes are anucleated cells called platelets which additionally aggregate in response to ADP, serotonin, and adrenaline.

49. www.ipfdd.de/research/ res16/a18/a18.html

50. Trombocīti veido agregātus. www.explorepub.com/articles/darkfield_charts/ fungus5.html