1. General Pathology Cellular and Organ Pathology Disorders of Glycogen Degradation. Pathology of Calcification. Jaroslava Dušková Inst. Pathol.,1st Med. Faculty, Charles Univ. Prague

2. Glycogen u linear and branched polymer u cca 60 000  - D-glucose molecules – monoparticles (beta) - muscle – complex particles (alpha) - hepatocyte

3. Enzymes Involved in Glycogen Metabolism u g.-synthase - brancher u phophorylase kinase - debrancher u g-6-phosphatase u  -glucosidase

4. Storage Diseases Def.: inborn errors of metabolism (mostly single gene abnormality) leading to an enzyme defect with subsequent accumulation of the substrate (& lack of the product) in tissues or organs „thesaurismoses“

5. E defect - gl-6 - phosphatase Organ damage - liver, kidney Glycogenosis I – von Gierke

6. E -defect - alfa1, 4 - glycosidase Organ damage - heart Glycogenosis II – Pompe

7. Glycogen Storage Diseases -1. DiseaseE- defSeverity of Disease Tissues Involved von Gierke Glucose-6 phosphatase SevereLiver,kidney, gut Pompe  1,4 glucosidase LethalHeart (+systemic) CoriAmylo-1,6 Glucosidase (debrancher) LethalLiver (+systemic)

8. Glycogen Storage Diseases -2. DiseaseE- defSeverity of Disease Tissues Involved Andersen Amylo-1,4-1,6 transglucosidase (brancher) LethalLiver (+systemic) Mc Ardle Muscle phosphorylase MildSkeletal muscle Hers Liver phosphorylase MildLiver

9. Clear Intracellular Vacuoles & adjunct techniques u accumulations of water neg. u lipides SUDAN, OIL RED u polysaccharides PAS, A-PAS

10. Glycogen water soluble easily lost in long lasting water based fixative solutions

11. Calcification Def.: depositions of Ca (mostly phosphate salts) in tissues or organs Classification: dystrophic metastatic

12.  Matrix vesicles - osteoblasts- nidus calcification  Non collagen proteins - osteopontin, osteonektin, osteokalcin, Gla protein, sialoprotein  Alkalic phosphatase  Phospholipids  Collagen I  Hydroxyapatite Calcification - physiology

13. Calcification  dystrophic  metastatic Calcinosis  localized  generalized Chondrocalcinosis - pseudogout Pathology Conditions with Calcium Deposits

14.  Basophilic  Von Kossa - Ag impregnation  Alizarine red +  Tetracyclin fluorescence  Polarized light birefringence Calcification - Microscopy

15. Calcification Dystrophic Serum Ca level: normal Tissues/Organs status dystrophic changes (necrosis, scar…, low metab. turnover) Metastatic Serum Ca level: Tissues/Organs status normal, local alcalisation (acid secretion - urine, stomach juice, sweat…)

16. Ca in mitochondria and GER in cytoplasm bound to proteins Released after cell damage Activaton of protein kinases Phosphorylation of protein and chromatin phragmentation Activaton of phospholipid degradation and loss Membrane damage Activaton of proteases Cytoskeletal disassembly

17. Necrosis or degeneration of tissue Release of enzymes Breakdown of organic phophates Increased deposition of calcium Alteration of pH Dystrophic Calcification

18. Calcification Dystrophic Serum Ca level: normal Tissues/Organs status dystrophic changes (necrosis, scar…, low metab. turnover) Metastatic Serum Ca level: Tissues/Organs status normal, local alcalisation (acid secretion - urine, stomach juice, sweat…)

19. Ca phophate Ca3(PO4)2 Ca diphosphate (Ca2P2O7) Hydroxyapatite (Ca5 (PO4)3.OH Ca Salts in the Calcified Foci

20. Calcification  dystrophic  metastatic Calcinosis  localized  generalized Chondrocalcinosis - pseudogout Pathology Conditions with Calcium Deposits

21. Frequent  necrotic tissue  connective tissue  vessels  kalkospherites  intracellular calcifiction of lysosoms Less frequent tendon, cartilage, elastics, bas. membranes Dystrophic Calcification

22. Forms - localised generalised Localisation – connective tissue, muscles Calcinosis

23. pyrophophate and hydroxyapatite deposits localisation – synovial membrane, cortilage, bone Chondrocalcinosis crystal deposit disease (pseudogout)

24. Calcium pyrophosphate deposition disease (Chondrocalcinosis, pseudogout) Clinic: may simulate different diseases Arthroscopy - chalky white deposits Tophaceus deposits with crystalline aggregates Crystals stain with von Kossa technique Foreign body type multinucleated giant cell reaction Deposits are in synovium, cartilage, bone