Introduction and Cellular injury

Patho- logy
• Pathology
Study of disease by scientific methods such as disease study
molecular, microbiologic, immunologic, and
morphologic techniques

Core of Pathology

• Cause: etiology 病因
• Mechanism: pathogenesis 疾病機轉
• Morphologic change:形態學或分子改變
• Clinical significance:臨床功能上的意義
clinical features, course, and prognosis
Why learn pathology
• Pathology serves as a bridge between the basic science
and clinical medicine, and it is of central importance to
the medical students
• General pathology
basic reactions of cells and tissues to abnormal Clinical
stimuli that underlie all diseases.
• Systemic pathology
specialized organs and tissues to more or less
well-defined stimuli. Basic
The Hospital Pathology
• Anatomical pathology 解剖病理
examine tissue by means of biopsy, exfoliated or needle aspiration cytology,
necropsy, frozen section, etc.
• Clinical pathology 臨床病理
examine non-tissue specimen from patient, including biochemistry, hematology,
microbiology, immunology, general microscopy etc.
The cell as a unit of health and disease

Genome

• Human • Worm
- 3.2 billion base pairs - 0.1 billion base pairs
- 20,000 genes - 20,000 genes
- 98.5% Noncoding DNA
→regulating gene expression
- person-to-person variation in < 0.5% of DNA
- single-nucleotide polymorphism (SNP)
copy number variations (CNV)
different numbers of contiguous 103-106 base pairs
Non–protein-coding sequences

• Promoter & enhancer regions

• binding sites for maintaining
chromatin structures

• Noncoding regulatory RNAs → micro-RNAs & long noncoding RNAs

• Mobile genetic elements (transposons)
• Telomere and Centromere

Epigenetic (above genetics) factors

Histones and histone modifying factors
Gene regulation
• Micro-RNA (miRNA) (2006 Nobel Prize)
– 22 nucleotides on average
– Posttranscriptional silencing mRNA
– One miRNA regulates multiple protein-coding genes

Long non-coding RNAs (lncRNAs)

>200 nucleotides

– promote gene activation

– gene suppression

– promote chromatin modification

– influence chromatin architecture

Gene editing
• CRISPRs/Cas9 system

• Clustered regularly interspaced short palindromic

repeats (CRISPRs) → guide RNA

• Cas (or CRISPR-associated genes) → endonuclease

No homologous DNA
Basic subcellular constituents NonHomologous
End Joining Homologous aDNA
Recombination

compartmentalize within membrane-bound

intracellular organelles
 • Protection and nutrient acquisition
Plasma membrane • Bilayers phospholipids, cholesterol, and proteins
• hydrophilic head face the aqueous environment
• hydrophobic lipid tails to form a barrier

Cytoskeleton and cell-cell interactions

Actin microfilament: 5-9 nm
Intermediate filament: 10 nm
Microtubule: 25 nm
Tight (occluding) junction
Desmosome
Anchoring junction
Gap (communicating) junction
Tyrosine kinase-based receptor signaling

Ligand binding
Receptor dimerization→autophosphorylation→adapter
(bridging) proteins →Activate GDP-bound RAS →interacts with
RAF & PI3K→Activate MAPK & mTOR→cellular responses

• RAS mutation delayed GTP hydrolysis will augment

proliferative signaling

Interaction with the extracellular matrix

• Integrins interact with the cytoskeleton at focal

adhesion complexes. Cell integrate signals from
ECM components and growth factors receptors
Maintaining cell populations Cyclin, Cyclin-dependent kinase
Cell cycle CDK inhibitor
(DNA synthesis) (premitotic growth)

(presynthetic growth)

Mechanisms regulating cell populations

Self-renewal
Asymmetric division
 Stem cell niches

CK7

• Skin stem cells

bulge area of the hair follicle, sebaceous glands, lower epidermis
• Small intestine stem cells
near the base of the crypt, above Paneth cells
• Liver stem cells (oval cells)
in the canals of Hering (thick arrow) structures that connect bile ductules (thin arrow)
to parenchymal hepatocytes
Cellular Adaptation, Injury and Death
• Cellular adaptation • Necrosis & Apoptosis
• Causes of cell injury • Cellular Accumulation
• Reversible & Irreversible cell injury • Cellular Aging

Cellular Responses to Injury

• Adaptations 調適
• Altered physiologic or Nonlethal injurious stimuli
• Cell injury 細胞損傷
• reduced oxygen; chemical; microbial infection
• Cellular accumulation 細胞堆積
• Metabolic alterations, genetic or acquired
• Cellular aging 老化
• Cumulative sublethal injury over long life span
Cellular response to stress and injurious stimuli

調適 損傷
Cellular responses to stress

Injury損傷
Adaptation調適

Death
壞死 凋亡
Cellular Adoptions調適

• Cause (etiology)
• Altered physiologic stimuli

• Mechanism (Pathogenesis)
• stimulation of cells by factors, activate receptors and downstream signaling
pathways

• Morphological (pathologic) changes

• hyperplasia, hypertrophy, atrophy, metaplasia

• Effect (clinical significance)

Cellular Adaptation調適
依環境 (生理或病理) 產生可逆性改變
調整細胞的構造 (大小、數量、型態)及機能

➢ Hyperplasia 增生 ➢ Atrophy 萎縮
➢ Hypertrophy 肥大 ➢ Metaplasia 化生

Cellular Adaptations (Cause)

Injurious Stimulus Cellular Response

↑ demand, trophic stimulation
Hyperplasia, Hypertrophy
(e.g. growth factors, hormones)

↓ nutrients, stimulation Atrophy

Chronic irritation Metaplasia
(chemical or physical)
Hyperplasia 增生

• increased cell number 數量增加

• 細胞分裂，通常體積也增大
• Physiologic
• Hormonal (pregnancy breast)
• Compensatory (regenerate liver after resection)
• Pathologic
• Endometrial hyperplasia (子宮內膜增生)
• Benign prostatic hyperplasia, Skin warts
• fertile soil in which cancer may eventually arise
Hypertrophy 肥大
• increase in cell size
• non-dividing cells synthesis more structural components, not
cellular swelling
• heart and the skeletal muscles
• May occurred with hyperplasia

pregnancy normal

normal
pregnancy
 Atrophy 萎縮
• Decrease in cell size and number
• Physiologic:
• thyroglossal duct, menopause uterus 停經子宮

• Pathologic: denervation muscle

senile atrophy
reduced blood supply
• Cause Normal Atrophic
• Disuse, Denervation, Ischemia
• Inadequate nutrition (cachexia)
• Loss of endocrine stimulation
• Aging (senile atrophy)
• Pressure (tumor compression)

The degradation of cellular proteins mainly by ubiquitin-

proteasome pathway
Metaplasia 化生
• one adult cell type to another adult cell type
• Epithelial: columnar to squamous (most common)
• may induce malignant transformation
• Connective tissue: bone formation in muscle (myositis ossificans)
• Mechanisms: stem cells differentiate along a new pathway by signals generated by
cytokines, growth factors, and extracellular matrix components in the cell's environment.

squamous
metaplasia
 Hyperplasia
Atrophy
增生
萎縮

Hypertrophy
肥大
Metaplasia
化生

• Hypoxia 缺氧 (ischemia缺血 心衰竭 貧血)

• Physical agents (冷 熱 放射線)
Causes of cell injury
• Chemicals & drugs

• reduced oxygen; chemical; • Infectious agents (細菌 病毒)

microbial infection • Immunologic reactions (autoimmune diseases)
• Nutritional imbalances
• Genetic derangement
Sequences of biochemical and morphologic
changes in cell injury

Cell Injury & Death

Injurious Stimulus Cellular Response
Acute and self-limited Acute Reversible Injury
Progessive and severe Irreversible Injury➙Cell death
(including DNA damage) (Necrosis, Apoptosis)
Subcellular alterations in
Mild chronic injury
various organelles

Irreversibility
• Severe mitochondrial dysfunction (No oxidative phosphorylation and ATP)
• Profound membrane dysfunction
Reversible cell injury
• Cell swelling • Fatty change
• First manifestation of injury • Lipid vacuoles in the cytoplasm
• Failure of energy-dependent ion pumps • Mainly in hepatocytes and myocardial cells
• Hydropic change or vacuolar degeneration

Cellular change in Reversible injury

• cell swelling, organelles swelling→ Hydropic swelling (water volume )
• plasma membrane blebbing
• ribosomes detach from ER
• chromatin clumping
• laminated structures (myelin figures)
Renal tubule Reversible injury

surface blebs
eosinophilia of cytoplasm
swelling of occasional cells
Cellular change in Irreversible injury

• increased eosinophilia, loss of cytoplasmic RNA

• swelling↑, disrupt cell membranes

unable to maintain membrane integrity

• disruption of lysosomes
• amorphous density in swollen mitochondria
• profound nuclear changes
codensation (pyknosis)
fragmentation (karyorrhexis)
dissolution of the nucleus (karyolysis)
• laminated structures (myelin figures)
Mechanisms of cell injury
• response depends on the nature, duration, severity of injury
• consequences depend on the type, state, and adaptability of injured cell

(2)
(1) (3)
(4)

(1)Depletion of ATP
(1) & (2) Mitochondrial damage

↑permeability
(open MPT pores)
(2) Defects in membrane permeability

(3) Influx Ca 2+
extracellular
1.3 mmol

cytoplasmic Ca2+
<0.1 µmol

enzymes activated by calcium

(4) Free radicals
• chemicals with a single electron in outer orbit
• Reactive oxygen species (ROS)
• oxygen-derived free radical
• (O2- H2O2 OH. ONOO-)
• imbalance between free radical generating
& removing systems
→ oxidative stress
Remove free radicals
• Antioxidants
• Vitamins E and A, Ascorbic acid and Glutathione
• Iron and Copper
• Transferrin, Ferritin, Lactoferrin, Ceruloplasmin
• Enzymes
• Catalase, 2 H2O2 ➙ O2 + 2 H2O
• Superoxide dismutases (SOD), 2 O2- + 2 H ➙ H2O2 + O2
manganese-SOD, in mitochondria copper-zinc-SOD, in cytosol
• Glutathione peroxidase, H2O2 + 2 GSH ➙ GSSG + 2 H2O,
or 2 OH + 2 GSH ➙ GSSG + 2 H2O

Examples of cell injury

and necrosis
endoplasmic reticulum

ribonucleoprotein
Example 1: Ischemic cell injury
Example 2: Ischemia-reperfusion injury

• Reperfusion may recover reversible cell injury

• But new damaging during reperfusion

• Reoxygenation →↑oxygen free radicals from parenchymal, endothelial cells, leukocytes

• ischemia ↓antioxidant defense →↑radicals

• membrane injury → Intracellular Ca2+ overload

• cytokines and adhesion molecules by hypoxic cell & endothelium → Inflammation

• activation of the complement pathway

• Some IgM deposit in ischemic tissues,
• when reperfusion, complement proteins bind
Example 3: Chemical (toxic) injury

• Direct cytotoxic
(combine with some critical molecular component or cellular organelle)
• mercuric chloride poisoning,
binds membrane sulfhydryl, ↑permeability,
↓ ATPase-dependent transport
• Cyanide poisons mitochondrial cytochrome oxidase and blocks oxidative
phosphorylation.

• Toxic metabolites
• P-450 covert CCl4 conversion to free radical CCl3
• (CCl4 + e ➙ CCl3 + Cl-)
Morphology of necrosis(壞死) and apoptosis(凋亡)
Necrosis 壞死 vs Apoptosis 凋亡
• loss of membrane integrity
• enzymatic digestion of cells
• frequently a host reaction
• pathologic (Physiologic or pathologic)
• Geographic area of cell death (individual cell)
• apoptosis & necrosis sometimes coexist (share some common features and mechanisms)

Feature Necrosis Apoptosis

Cell size Enlarged (swelling) Reduced (shrinkage)
Nucleus Pyknosis → karyorrhexis → Fragmentation
karyolysis
Plasma membrane Disrupted Intact
Cellular contains Enzymatic digestion; leak out Intact; released in
apoptotic body
Adjacent inflammation Frequent No
Physioloic or pathologic pathologic Often physiologic
Cellular change in Necrosis

• Enlarged (swelling) cell size

• cytoplasmic blebs
• Pyknosis ➙ karyorrhexis ➙ karyolysis
• Enzymatic digestion
• Disrupted cell membrane
• leakage of cellular components
• Frequent adjacent inflammation

Na+ Ca2+ (10-3M vs 10-7M)

ATP

K+
Morphology of Necrosis
• increased eosinophilia
• loss RNA ↑denatured proteins in cytoplasm
• glassy homogeneous (loss of glycogen particles)
• vacuolated cytoplasm (enzymes digest organelles)
• initially clumped nuclei
• calcification (fatty acids → calcium soaps)

Normal kidney tubules Early (reversible) ischemic injury Necrosis (irreversible injury)
Patterns of tissue Necrosis 壞死
depend upon organ and circumstances

• Coagulative 凝固 necrosis
• preserved architecture of dead tissues

• Liquefactive 液化 necrosis
• digestion of dead cells → liquid

• Caseous 乾酪 necrosis

• Other necrosis
• Gangrenous 壞疽 necrosis
• Fat necrosis 脂肪壞死
• Fibrinoid necrosis 纖維蛋白樣
Coagulative necrosis凝固性壞死

wedge-shaped kidney infarct (yellow) normal kidney (N) and necrotic

cells in the infarct (I)
Coagulative necrosis
preservation of the basic outline
myocardial infarct eosinophilic anucleate fibers
Normal myocardium. Leukocytes in the interstitium
 Liquefactive necrosis液化性壞死 Liquefaction, no outline or structure
Ex: brain necrosis, pus

kidney fungal infection

abscess
(white cells and
cellular debris)

Caseous necrosis 乾酪性壞死

• distinctive form of coagulative necrosis • No cellular outlines, not disappear by lysis

• tuberculous infection • coagulated cells & amorphous granular debris
Gangrenous necrosis 壞疽
• not a distinctive pattern of cell death
• usually for limb, lost blood supply → coagulation necrosis
• If superimposed bacterial infection
→ liquefactive action (wet gangrene)

Fat necrosis
• not a specific pattern of necrosis.
• acute pancreatitis (lipase hydrolyzes triglyceride)
• fatty acids + Ca → chalky white (fat saponification)
Fibrinoid necrosis
• injured blood vessels
• accumulation of eosinophilic plasma proteins obscures
the underlying alterations in the vascular wall

Apoptosis 凋亡
• Cell program to die
• activate enzymes degrade nuclear DNA, nuclear, and cytoplasmic proteins
• cell membrane intact, but structure altered → phagocytosis
• rapidly cleared, before leak out its contents
• no inflammatory reaction
• Can be physiologic, adaptive, and pathologic
Physiologic Apoptosis

• Embryogenesis

• Hormone-dependent involution in the adult

– endometrial breakdown in menstrual cycle,
– Ovary after menopause, lactating breast after weaning
– prostatic atrophy after castration

• Cell deletion in proliferating cell populations

– intestinal crypt epithelia (maintain a constant number)

• Eliminate potentially harmful self-reactive lymphocytes

• Death of host cells served their useful purpose,

– neutrophils in acute inflammation
Pathologic Apoptosis
• Cell injury
– DNA damage by radiation, anticancer drugs, hypoxia
– Accumulation of misfolded proteins (ER stress)
• Viral diseases (viral hepatitis)
• duct obstruction induce atrophy of pancreas, parotid gland, and kidney
• Tumor cell death

Cellular change in Apoptosis

↓ Reduced (shrinkage) cell size
↓ chromatin condense, fragment
↓ cytoplasmic budding
↓ phagocytosis of the extruded apoptotic bodies
• Intact cell membrane; altered structure (orientation of lipids)
• Intact cell contents; may be released in apoptotic bodies
• No Adjacent inflammation
Morphology of Apoptosis
• single cells or small clusters of cells
• eosinophilic cytoplasm, small nuclear, dense chromatin
• quickly phagocytosed, no inflammatory reaction

Mechanisms of Apoptosis
• Initiation phase (activate caspases)
Inducers Extrinsic pathway: TNF and Fas ligand
Intrinsic : withdrawal of growth factors, hormones
injurious agents (radiation, toxin, free radical)
cytotoxic T cells (directly activate caspases)
Adapter proteins Cytochrome C, Initiator caspases
Regulators Bcl-2 family, inhibit or promote
Mechanisms of Apoptosis
• Execution phase
• (enzymes act to cause cell death)
Executioner caspases activate latent
cytoplasmic endonucleases and
proteases that degrade nuclear and
cytoskeletal proteins.
formation of apoptotic bodies, also express new ligands for binding and uptake by
phagocytic cells.

Mitochondrial (Intrinsic) pathway

• triggered by
• loss of survival signals (hormone, cytokine …)
• DNA damage
• accumulation of misfolded proteins (ER stress)
• Regulate mitochondrial permeability by
• Pro-apoptotic molecule (in mitochondrial membrane, BAX, BAK)
• Anti-apoptotic (protective) Bcl-2 family (BCL2, BCL-XL, MCL1)
• Sensors (BH3-only proteins: BAD, BIM, BID, Puma, Noxa)
• Release death inducers (cytochrome c) into cytoplasm →activate caspase
Intrinsic pathway

Apoptosis Inducing Factor

(apoptosis activating
factor-1)

(Anti-apoptosis)

Intrinsic pathway

When stress, lost Bcl-2 and/or Bcl-x and replaced by pro-

apoptotic members (Bak, Bax, and Bim) → ↑permeability
Death receptor (Extrinsic) pathway

• Induced by TNF Family of Receptors

• Fas (CD95) engaged by Fas ligand (FasL)
– elimination of self-reactive lymphocytes
– mutation in Fas or FasL → autoimmune diseases
– some cytotoxic T lymphocytes to kill virus-infected and tumor cells
• TNF
• bind type 1 TNF receptor (TNFR1) → adapter protein TRAD
(TNF receptor-associated death domain containing protein) binds to FADD-
→ caspase activation
• activate nuclear factor-κB (NF-κB) and stimulating degradation of the inhibitor
of NF-κB
Extrinsic pathway

Fas ligand
• Fas cross-linked by its ligand (FasL)
• ≧ 3 molecules of Fas come together
• form a binding site for FADD
• attached to death receptors
Fas-associated
death domain • binds pro-caspase-8
• Multiple pro-caspase-8 → active caspase-8
• caspase activation by
• This pathway inhibited by a protein (FLIP)
which binds to pro-caspase-8
Execution Phase
• by proteolytic Caspase
• Initiator (Intrinsic: caspase-9; Extrinsic: caspase-8 (-10)
• Executioner (caspase-3 and caspase-6)
– act on many cellular components
– cleave cytoskeletal and nuclear matrix proteins
Extrinsic
Intrinsic
Apoptosis in health and disease
• Protein misfolding
– unfolded protein response (↑chaperones, ↑degradation)
control by chaperones
misfolded will be ubiquitinated and proteolised
– ER stress: can’t cope the accumulation → apoptosis
– neurodegenerative (Alzheimer, Huntington, Parkinson) diseases
Apoptosis in health and disease

• Disorders associated with dysregulated apoptosis

• defective apoptosis and ↑ cell survival
• p53 mutation: cell fail to die, accumulate mutations → cancer
• mutated Fas or FasL: failure to eliminate dead cells (source of self-antigen)
→ autoimmune disorders

• increased apoptosis and excessive cell death

• loss of cells → neurodegenerative diseases,
• ischemic injury in myocardial infarction and stroke,
• death of virus-infected cells
 Necroptosis

• hybrid necrosis & apoptosis

loss of ATP, cell swelling, generate ROS, release
lysosomal enzymes, membrane rupture
“ caspase-independent ” programmed cell death
triggered by TNFR1, Fas, virus, genotoxic agent
recruits receptor associated kinase 1 & 3 (RIP1 & RIP3)

• physiologic & pathologic conditions

Formation bone growth plate
Cell death in steatohepatitis, acute pancreatitis,
reperfusion injury
Parkinson disease
necrosis morphology
host defense against CMV apoptosis mechanism
(encode caspase inhibitor)
Pyroptosis(inflammation & apoptosis)
• occurs in cells infected by microbes
• microbes recognized by cytoplasmic immune receptors
→ activate inflammasome (cytosolic danger-sensing protein complex)
• activate caspase-1
• active IL-1 (leukocyte recruitment & fever)
• caspase-1 & caspase-11
→ cell swelling
membrane rupture
inflammatory mediators
→ cell death
Intracellular Accumulations
3 categories:
• Normal cellular constituent (water, lipids, proteins, carbohydrates)
• Abnormal substance
• Exogenous
• Endogenous (abnormal metabolism)
• Pigment
 1. abnormal metabolism defects in packaging
& transport normal substance
→ fatty liver

2. Mutation abnormal endogenous substance

defects in folding, packaging, transport or
secretion
→ alpha 1-antitrypsin deficiency
Accumulation of
abnormal proteins

1. enzymes deficiency
→ storage diseases

2. inability to degrade phagocytosed

exogenous particles
→ hemosiderosis and carbon pigment
accumulation
Lipids

• triglycerides
• cholesterol/cholesterol esters
• Phospholipids
– myelin figures in necrotic cells
• lipids & carbohydrates complex → lysosomal storage diseases

• DDx water or polysaccharides (glycogen)

• Fat (frozen tissue sections, Sudan IV or Oil Red-O stains)
• Glycogen (periodic acid-Schiff (PAS) reaction)
Steatosis (Fatty Change)
• triglycerides accumulations in parenchymal cells
• Mainly in liver, also in heart muscle, kidney
– Alcoholic
alters mitochondrial & microsomal functions

– Nonalcoholic
➢ diabetes, obesity
➢ toxins (CCl4), malnutrition: ↓ apoproteins
➢ anoxia: fatty acid oxidation
➢ starvation: ↑fatty acid mobilization from peripheral stores
Cholesterol and Cholesterol Esters

• Atherosclerosis
• Xanthomas
– Intracellular cholesterol within macrophages → foamy cells
• Inflammation and necrosis
– Foamy macrophages (injured cell membrane)
• Cholesterolosis
– foamy macrophage in lamina propria of gallbladder
• Niemann-Pick disease, type C
– lysosomal storage disease
– mutated cholesterol trafficking enzyme

foam cells gallbladder cholesterolosis

Proteins
• Reabsorption droplets in proximal renal tubules
• Renal diseases, proteinuria, glomerular filtration
• → proximal tubule reabsorption into vesicles
• pink hyaline droplets in tubular cell
• Reversible process; can metabolized & disappear

Protein reabsorption droplets in the renal tubular epithelium

Proteins

• Russell body
– plasma cells excessive synthesis immunoglobulins
→ large distended ER
→ homogeneous eosinophilic inclusions

• α1-antitrypsin deficiency
– protein mutation, slow folding, aggregate in liver (pink globule)
– ↓circulating enzyme → emphysema
Defects in protein folding

• Ribosome arranged polypeptide into

– α helices or β sheets (function & transport)

• Chaperone
– aid proper folding & transport across the ER,
Golgi complex and beyond
– normally synthesized or induced by stress
(heat-shock proteins, hsp70, hsp90) & "rescue"
shock-stressed proteins from misfolding

• ubiquitin (heat-shock protein)

– mark abnormal folding protein
– (for proteasome complex degrade)
Chaperone protect unfolded protein from degradation, guide into organelle

Chaperone repair misfolded protein

apoptosis Repair failure, proteins are targeted for

degradation in the proteasome,
Diseases Caused by Misfolding of Proteins

Disease Affected Protein Pathogenesis

Cystic fibrosis Cystic fibrosis trans- Loss of CFTR leads to defects in
membrane conductance chloride transport
regulator (CFTR)
Familial LDL receptor Loss of LDL receptor leading to
hypercholesterolemia hypercholesterolemia
Tay-Sachs disease Hexosaminidase β subunit Lack of the lysosomal enzyme →
storage GM2 gangliosides in neurons
Alpha-1-antitrypsin α-1-antitrypsin Storage of nonfunctional protein in
deficiency hepatocytes causes apoptosis;
enzymatic activity↓ in lungs → elastic
fiber destruction → emphysema

Creutzfeld -Jacob Prions Abnormal folding of PrPsc causes

disease neuronal cell death
Alzheimer disease Aβ peptide Abnormal folding of Aβ peptides→
aggregation within neurons & apoptosis
Hyaline change

• Any material which homogeneous, glassy, pink appearance in H&E stain slides

• by a variety of alterations

• Intracellular hyaline
– reabsorption droplet, Russell body, Mallory body (alcoholic
hyaline in the liver)

• Extracellular hyalin
– Hyaline collagen in old scars
– hyaline arteriolosclerosis
plasma protein & basement membrane material
Glycogen
• abnormal glucose or glycogen metabolism
• clear vacuoles within the cytoplasm
• PAS reaction (rose-to-violet color), diastase digestion
• In proximal convoluted tubules, liver, heart
• glycogen storage diseases (glycogenoses)
– macrophage enzymatic defects to break down glycogen
– accumulation with secondary injury and cell death
Pigment

• colored substances

• Exogenous Pigments
 anthracosis carbon or coal dust
 Tattooing (pigments in dermal macrophages)

• Endogenous Pigments
 Lipofuscin (lipochrome, aging pigment)
 Melanin (melanocyte, brown-black pigment)
 Hemosiderin (hemoglobin-derived, yellow brown)
Lipofuscin (aging pigment)
• insoluble brown pigment
• lipid & phospholipid polymers complex with protein
• from lipid peroxidation of subcellular membranes

• finely granular, usually perinuclear location

– aging liver and heart, severe malnutrition, cachexia

• not injurious to the cell or its functions

Lipofuscin granules in cardiac myocyte

Melanin

• endogenous brown-black pigment

tyrosinase
tyrosine oxidation dihydroxyphenyalanine

• in Melanocyte
Hemosiderin
• hemoglobin-derived
• golden yellow-to-brown granular pigment
• Iron carried by transferrins stored in cell with apoferritin → ferritin micelles
• ↑iron, ferritin → hemosiderin granules

• a few hemosiderin in normal phagocytes

• (bone marrow, spleen, and liver)

Hemosiderin Prussian blue reaction

potassium ferrocyanide (colorless)
Yellow-brown granular iron ferric ferrocyanide (blue)
pigment
Pathologic Calcification

• abnormal calcium salts deposition

• together with a few iron, magnesium, other salts

• Two forms
Dystrophic calcification
 deposition occurs locally in dying tissues
 normal serum calcium, normal calcium metabolism
Metastatic calcification
 deposition in normal tissues
 Hypercalcemia, disturbance in calcium metabolism
Dystrophic calcification

• in Necrotic areas
– coagulative, caseous or liquefactive
– atheromas of advanced atherosclerosis
– aging or damaged heart valves

• fine, white granules or clumps

• In H&E basophilic, amorphous granular
• Intracellular, Extracellular or both
• heterotopic bone (ossification)
• psammoma bodies
– lamellated configurations
Metastatic Calcification

• Hypercalcemia
• ↑parathyroid hormone
• destruction of bone tissue (myeloma, immobilize)
• vitamin D-related disorders (vitamin D intoxication)
• renal failure (phosphate→2nd hyperparathyroidism)

• In normal tissues
• or accentuates dystrophic calcification
• noncrystalline deposits, or hydroxyapatite crystals
• Usually, cause no clinical dysfunction
• Massive renal deposits (nephrocalcinosis)
• → renal damage
Cellular Aging

• progressive ↓cellular function & viability

• accumulation of sublethal injury
↓ proliferative capacity
↓ cell life span

Decrease in human physiologic

capacities as a function of age
Mechanisms of cellular aging
• Genetic and environmental factors combine to cause aging

• Accumulation of DNA damage (defective DNA repair mechanisms)

• Replicative senescence (↓telomere, ↓cells divide capacity)
• Defective protein homeostasis (damaged protein, ↓protein)
impaired chaperone and proteasome function
• Nutrient sensing (calorie restriction, ↑sirtuins counteracts aging)
growth factors (IGF, mTOR) promote aging
Cellular Senescence

• Cells have limited replication capacity

children > older people > Werner syndrome
(premature aging, defective DNA helicase, repair)
After a fixed number of divisions
→ nondividing state (cellular senescence)

• Telomere attrition

• Activation of tumor suppressor genes

• ↑ CDKN2A (p16) levels with age
• decline in self-renewal potential
Telomeres

• short DNA repeated TTAGGG

• at ends of chromosomes
• ensuring complete replication
• protect chromosomal termini

• somatic cells replicate

→ telomeres shortened
→ cell cycle arrest
Telomerase

• RNA-protein complex
• RNA template
• maintain telomere length
• Telomerase activity
repressed by regulatory proteins
germ cell > stem cell
> somatic tissues (~0)
• Cancer cells
• telomerase reactivated
• telomeres not shortened
Nutrient sensing system

• Caloric restriction increases longevity

• ↓ IGF-1 signaling, ↑ sirtuins

• Insulin & insulin-like growth factor 1 (IGF-1)

• promote anabolic state, cell growth & replication
• downstream targets: AKT and mTOR

• Sirtuins (histone deacetylase)

• deacetylate and activate DNA repair enzymes
• adapt to stress (food deprivation, DNA damage)
• ↓ metabolic activity, ↓ apoptosis, ↑ protein folding
• ↓ harmful effects of oxygen free radicals
• ↑ insulin sensitivity & glucose metabolism