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Cell Injury and Cell Death

Taught by Dr. Lewis
1. How can a smooth surface change to a rough surface see the pics in his slides
a. Addition of material, examples:
i. Leakage of proteins from inflamed vessels in fibrous pericarditis
1. lassical finding in acute inflammation is leakage of proteins
b. !emoval of material, examples:
i. "carring of tissue after necrosis
1. #ith necrosis have loss of tissue, what is left behind scars
a. $f the loss of tissue is in a cone shape or v shape the
underl%ing cause is ascular in nature
b. #hen a vessel is occluded, the tissue that is furthest awa%
from that vessel &'( still supplied b% that vessel is what
suffers the most
c. Addition and removal of material, examples:
i. irrhosis of the liver has two components
1. !emoval of tissue through necrosis and scarring
!. !egeneration of liver tissue in response to damage
a. Appear as nodules on the surface rough
b. )ewl% formed *nodules+ tend to be poorl%,non functional
isolated from blood suppl% due to the scarring
". $nduced b% alcohol and#or hepatitis
-. #hat is disease
a. .ur bodies response to in/ur% not the in/ur% itself
b. #hat is treated is not the in/ur%, but the response
i. 0.g. anti histamines, ice on an ankle sprain
c. #hat is often most damaging is the response, not the in/ur%
i. 0.g. gram negative sepsis
ellular $n/ur%
1. an be either:
a. !eversible
i. !ecover%
ii. Adaptation repeated exposures to an in/urious agent, cells adapt
b. $rreversible
i. )ecrosis alwa%s pathological
ii. Apoptosis ph%siological or pathological
2. ritical needs of a cell
a. 3embrane integrit%
b. 0nerg% prodution
c. "%nthesis of structural components
4. (%pes of in/ur%
a. H%poxia
i. Low ox%gen, not anoxia 5no ox%gen6
ii. 7lobal h%poxia
1. Lack of ox%gen throughout the entire bod%, caused b%:
a. Lack of air
i. 8ue to drowning or displacement of o- b% other
gasses 5nitrogen6
b. )ot being able to take the air in
i. "uffocation 5blocking 9-6
ii. "trangulation 5around carotids blocking blood to
brain6 fastest death
1
iii. :lugged pipes 5can;t get air to lungs6
c. $nabilit% to transport ox%gen
i. Anemia
ii. . poisoning
1. &inds to hemoglobin with high affinit%,
displacing 9- and also inhibits c%tochrome
oxidase so can;t make A(:
!. :atient is h%poxic but not c%anotic will be
cherr% red
". (reatment options: h%perbaric chamber with
ver% high p9- 5displace .-6 or transfusion
5if not have enough time6
iii. %anide poisoning
1. 'ncoupler, no A(:
!. Also inhibits c%tochrome oxidase
i. :ump failure
. 8ecreased blood volume 5bleeding6
i. $ncreased vascular space shock, vasodilation,
decreased pressure, less blood flow 5anaph%lactic
shock6
iii. Local h%poxia
1. 0.g. heart attack
iv. 3a/or effect of h%poxia
1. #a% less A(: production due to anaerobic gl%col%sis
!. - vs 1< atp,molecule of glucose 5need that substrate6
". heart and brain are ver% sensitive to h%poxia and ischemia 5which is a
cause of h%poxia6 anaerobiosis 5metabolic processes occurring is not
an option for us to live on
b. $schemia
i. $mpaired deliver% of blood 5less ox%gen and other metabolites too6
ii. )ot h%poxia
1. Lack of 9- so switch to anaerobic gl%col%sis in h%poxia
!. Atp will drop and plateau, go on with anaerobic gl%col%sis
iii. $n ischemia lack 9- and also lack other metabolites 5glucose6 and lose abilit% to
remove metabolites 5lactate6
1. Atp will drop continuall%, can;t get substrate in for anaerobic gl%col%sis
iv. auses
1. Arterial and venous occlusion 5once venous and arterial pressures are
e=ual, flow > 96
c. .x%gen related in/ur%
i. !eperfusion in/ur%
1. .cclude vessel, cells die, leak metabolites, reperfuse, blood and ox%gen
rush in, more damage mediated now b% free radicals
ii. #hat is a free radical
1. 3olecule with one unpaired electron when reacts with other
molecules it turns them into free radicals 5chain reactions6
iii. - mechanisms for generating free radicals
1. adding electrons to ox%gen
!. c%tochrome p249 metabolism
iv. 1 important ox%gen radicals, in bold
1. ox%gen can form supero$ide ion % &!'
a. not reactive but the rate limiting step so important
b. froms in mitochondria during oxidative phosphor%lation
!. two superoxides dismutate to form (!)!
". H-9- itself is not reactive but if it interacts with ?ous metals 5-@@
forms6 it forms hydro$yl radical *&('+ in ,enton cycle
-
Anow this
a. .HB is Cer% reactive and causes the damage
v. H-9- in neutrophils
1. ombines with l in presence of myelo pero$idase en-yme to from
bleach 5h%pochlorus acid6 to kill invading microbes
vi. Denton reaction c%cle
1. !eview the above steps in development of .HB
!. "uperoxide ion 59-B6 is dangerous for two reasons
a. $s the rate limiting step in generation of .HB 5b% forming
H-9-6
b. .-B !educes oxidiEed ?ic metals back into ?ous metals
i. .nl% ?ous metals can react with H-9- to form
h%drox%l radical and keeps fenton c%cle going
c. 9-B is generated all over the place but the bod% controls it to
prevent this c%cle )!' is what the body tries to control
most
". )ever need to add more iron, ox%gen and superoxide keep the c%cle
going
.. (he role of vitamin 5ascorbate6
a. $n normal situation it is a free radical scavenger 5good6
b. At sites of in/ur%, it can keep the fenton c%cle going b%
reducing ferric iron 51@6 into ferrous iron 5-@6
vii. Lipid peroxidation ox%gen related membrane damage
1. &(' acts at methylene groups in unsaturated fatt% acids 5the% flank
dbl bonds6
!. "ets up a C0!F reactive chain reaction
". .HB extracts an electron from fatt% acid, forms fatty acid radical that
reacts with ox%gen to from lipid pero$ide, goes on and oxidiEes other
fatt% acids and damages the membrane further
.. (his entire process is prevented b% the presence of itamin / *a
tocopherol+
a. Aliphatic h%drophobic tail allows it to sit in cell membrane,
causes chain termination of oxidation of fatt% acids
b. 8eficienc% makes %ou look old bc of all the damage
0. Citamin can increase production of Citamin 0
1
viii. .xidant defenses
1. Against superoxide 5.-B6
a. 1upero$ide dismutase
i. Actuall% speeds up the reaction and forms H-9-
less damaging than superoxide which can reduce
ferric iron back to ferrous -@ iron
ii. $t is located ever%where
iii. Lou gherigs disease mutation of "8 in brain that
increases its activit%, too much H-.-
!. Against H-.-
a. $n peroxisomes onl%
i. Catalase
1. High turnover rate
!. (akes H-9- and makes water
". .nl% protects against damage from
metabolism of odd chain fatt% acids 5which
makes H-.-6 and nothing else wh%, bc it
is onl% located in peroxisomes
b. 0ver%where else
i. 2lutathione pero$idase333
1. 4educed glutathione scavenges, accepts
electrons, forms water from H-9-
!. !e=uires glutathione reductase and )A8:H
from ::: to remain in reduced form
". .thers
a. 'ric acid
i. :urine metabolite that acts as a scavenger in plasma
ii. (oo much > gout
b. 3etal binding proteins 5lactoferrin, ceruloplasmin6
i. :revent D!00 ?ous metals from reacting with H-9-
b% binding them
ii. 8eficiencies cause hemochromatosis 5too much free
iron6 and #ilson;s disease 5copper6
c. Cit0
d. Cit
d. :h%sical $n/ur%
i. 0lectricit%, trauma, temperature
ii. !adiation damage i.e. ioniEing radiation
1. "plits water and forms h%drox%l radicals 5ma/or damage mech6
!. $schemic tissue will be resistant 5wh%, bc less ox%gen there to begin
with so less damage6
". 8osage of radiation is important 5remember, targeting rapidl% dividing
cells6
a. 199 units targets the marrow
i. mess up the immune s%stem and blood cells
ii. get bleeding and infections
iii. death within 1 weeks
b. 1999 units gets deeper, targets gut epithelium, latenc% is one
week and death in - weeks
c. -999 units targets most resistant organs
i. e.g. brain
ii. radiation in/ur% and pt is found in coma know the
brain is involved had to be a large dose
iii. death within 1 da%
2
e. hemical in/ur%
i. toxins and poisons free radical mediated damage
ii. carcinogens not man% human cancers are induced b% carcinogens
iii. sensitiEers hapten mediated damage
1. alter our bod% so our immune s%stem attacks us
!. e.g. cheap /ewelr% t%pe 2 H"!, metals leak out of the /ewelr% and alter
membrane proteins which are subse=uentl% attacked
iv. chem. $n/ur% can be classified as direct or indirect
1. direct c%anide 5c%tochrome oxidase inhibitor and uncoupling agent6
!. indirect drug,p249 damage
a. parent compounds themselves are not damaging but are
metaboliEed b% enE%mes in our bod% forming damaging
products damage will be specific to locations with enE%me
availabilit%
b. l2 > arbontetrachloride dr% cleaning industr%
i. 3etaboliEed b% :249 in the lier to form l1 which
in turn induces lipid peroxidation kills liver
c. Acetaminophen also metaboliEed b% p249 in the lier
i. :roduces intermediate ben-o5uinone
1. )ormall%, benEo=uinone is neutraliEed b%
glutathione, but during poisoning the
glutathione becomes depleted and so the
benEo=uinone accumulates and leads to liver
necrosis: treat with )BAcet%lc%stein
!. get protein thiol o$idation occurs
a. Cysteine residues *with 1(
groups+ in proteins can be
o$idi-ed6 resulting in the
formation of disulfide *1''1+
bonds
b. This results in conformational
changes in proteins6 loss of
en-yme actiity6 and protein
crosslinking
c. a A(:ase pumps have thiol
groups in active site, become
inactivated6 Ca leaks out of /4
and through the plasma
membrane increasing c%tosolic a
B disregulated calcium will activate
proteases, lipases, endonucleases,
etc.. And %ou can literall% eat the
cell alive
4eersible injury 5recover% or adaptation6 /ust talked about all the wa%s that damage can be inflicted, if
%ou don;t damage the cell too much the in/ur% will be reversible
1. 3orpholog% associated with reversible in/ur%
a. ell swelling see slide G2 and G4
i. #h% do cells swell two reasons
1. 0arl% on, start to produce less A(:
a. )a pump doesn;t work no more, intracellular )a increases and
water follows sodium
-. Later, tissue osmolalit% increases due to catabolism in in/ured cell
a. ells breaks down into small pieces, ups osmotic pressure in
tissue, water is drawn in
4
ii. #hat do the look like
1. ells will increase in siEe and staining will be less dense 5water dilutes
c%toplasm6
iii. Aidne% proximal tubules are ver% sensitive to swelling 5A()6 Dirst pic
b. 3itochondrial swelling
c. 1mall dense bodies in mitochondria 5big > irreversible6
d. 3embrane blebbing
i. 8ue to c%toskeletal damage
e. $ntracellular accumulations
i. Lipid accumulates ver% =uickl%, particularl% hepatoc%tes 5weekend in st martin6
ii. 3echanisms
1. $mpaired lipoprotein s%nthesis
a. 8ue to alcohol or protein malnutrition 5can; make apo b1996
-. 8ecreased fatt% acid oxidation
a. H%poxia
i. liver is site of fat metabolism
ii. need ox%gen for the process
iii. no ox%gen, so liver accumulates fat
1. $ncreased liberation of fat from peripheral stores
a. "tarvation can be starving but have a big bell%
iii. #hat does it look likeH
1. Fellow because fat is %ellow
<
-. &igger holding more fat
1. 3icroscopicall% see the fat cells 5slides G4 and G<6
iv. "top drinking and it all goes awa% reversible in/ur%, don;t stop could get
irrhosis 5irreversible6
-. (he adaptations to reversible in/ur% see slides 1<1 onwards for the histo pics
a. H%pertroph%
i. $ncrease in the siEe of a tissue or organ bc of an increase in the si-e of
indiidual cells
1. ells grow bc of increase amount of proteins and siEe of organelles
ii. 0xamples
1. 'terus during pregnanc%
-. Heart bc of h%pertension or exercise
a. Heart weight should be related to height
b. 0xercise muscle la%er increases a little and heart gains the
ability to dilate and thus increases stroke volume resulting in
a lower resting heart rate increase in the silhouette of the
heart but not much of an increase in weight
c. H%pertension muscle la%er become massive but ventricles
decrease in siEe smaller stroke volume 5compensated for b%
elevated resting heart rate6
i. ells in the subendocardial Eone are most vulnerable
to ischemia
d. .ther changes in a h%pertensive h%pertrophied heart
i. Health% heart A)D onl% in the atria, h%pertro heart
A)D in the ventricles and atria 5an attempt to lower
the load b% pissing out salt and water opposite of
rennin angiotensin s%stem6
1. )ote that A)D is expressed in the ventricles
of the fetal heart so this is an example of
*re expression of fetal genes+
ii. 'nhealth% heart expresses multiple growth factors
1. Dos, Iun, 07!D, T2,'B, $7DB1
-. All increase siEe and number of organelles
and sarcolemma in cells and, as a result, the
siEe of the cells themselves
1. $n other cells the same growth factors can
cause h%perplasia, but in heart onl%
h%pertroph% results
iii. Health% heart expresses adult contractile proteins
5alpha m%osin6, unhealth% heart reBexpresses fetal
contractile proteins 5beta m%osin6
1. Heart has an increased work load so seeks a
more energ% efficient contraction
-. &eta m%osin has less A(:ase activit% so it is
more efficient 5less A(:,contraction6
1. Bm%osin can;t adapt to higher pressures as
fast as Bm%osin could cause damage
down the road if pressure increases
G
iii. auses
1. 3echanical stimuli heart and skeletal muscle
-. 7rowth factor stimulation pubert%
1. $ncreased functional demand B unilateral nephrectom% 5one kidne%
grows to compensate for the loss of the other6
b. H%perplasia
i. $ncrease in the siEe of a tissue or organ bc of an increase in the number of cells
ii. 0xamples
1. Lactating breast
-. :rostate bc of endocrine d%sfunction 5over stimulation from
testosterone metabolites B 8H(6
iii. auses
1. 7rowth factor stimulation
-. 0ndometrial proliferation
1. allus formation
2. 0r%throid h%perplasia in h%poxia 5at elevation6
4. Cirall% induced H:C loves the s=uameous epithelium 5warts6
iv. :artial hepatectom%
1. !emove part of the liver and the remaining lobes will undergo
h%perplasia to reBestablish liver mass
c. Atroph%
i. 8ecrease in the siEe and function of a tissue or organ bc of a decrease in the
si-e6 function 78D number of cells % not associated with cell death
ii. auses
1. 8isuse 5voluntar% or neuronal6
-. 8ecreased blood suppl%
1. :oor nutrition
2. #ithdrawl of growth factors and endocrine stimulation
4. Aging and pressure aging causes atroph% of the brain
iii. 3echanisms
1. $ncreased catabolism
-. 'bi=uitin,proteosome or l%sosomal degradation
iv. 0xample of the ischemic kidne% 7old blat kidne%
1. Also suspect h%pertension in this patient
a. #hen kidne% is not getting blood rennin angiotensin s%stem is
activated, )a is retained, &: rises
d. 3etaplasia
i. 4eplacement of one differentiated cell type by another
1. )ot the actual cells changing, one line of cells is replaced b% the second
line which is derived from stem cells 5stem cells are the ones that are
altered6
ii. auses
1. hronic irriataion sites of high exposure to carcinogen and toxin
a. "=uamous metaplasia in the resp tract in smokers
i. "=uamous defends against toxins, replaces columnar
cells 5but lose ciliar% function6
b. olumnar gastric metaplasia of distal oesophagus in gerd
5barrets oesophagus6 columnar replaces s=uamous,
columnar is protective against the gastric acid bc it has a
mucous la%er
iii. A double edged sword
1. (he new epithelium is in some wa% protective, but %ou lose normal
function and end up at a ver% high risk of developing cancer
J
e. 8%splasia > 8$".!7A)$K08 8$DD0!0)($A(08 0LL" 5(!$::L0 86
i. 7bnormal#disorgani-ed growth of cells in a tissue *changes in si-e6 shape
and organi-ation+
ii. cells lose two thing uniformit% and architectural organiEation
iii. Definite precursor to cancer
iv. !eason we do pap smears
1. Drom basement membrane up should be progenitor, differentiating,
then s=uamous
-. )ormall%, surface cells should be pavement like cells with small nuclei
5s=uamous epithelium6
1. $n d%splasia have less mature cells showing up near the surface
f. Anaplasia
i. Complete loss of morphological differentiation
ii. (his $" cancer not a precursor but straight up cancer
iii. $n displasia %ou see differentiated cells and %ou know where the% came from
iv. Here the cells look nothing like their parents and have no idea where the% came
from
g. )eoplasia
i. 7utonomous new growth
ii. Drom benign fibrinoids to malignant carcinoma
iii. !emove the stimulus and the cells will not stop growing 5autonomous growth6
iv. Cs h%perplasia which is growth of normal cells that can be stopped if the
stimulus is removed
Irreersible injury # cell death 5necrosis or apoptosis6
1. )ecrosis
a. 4 *subsets+ slides J1 B L4
i. Coagulatie
1. "tandard
!. :reservation of outline of the cell for some da%s
a. 0nE%mes are all denatured b% acidosis and so proteol%sis is
inhibited
b. "tatus of the outline 5how intact is it6 can give %ou an estimate
of how long the cell has been dead for
". 'nderl%ing cause often ischemia , h%poxia in all tissues except brain
L
.. #hat does a stain look like B morpholog%
a. )uclear alterations slides 191 194
i. :%knosis
1. Cer% good indicator of cell death
!. Also present in apoptosis however so not
enough on its own to sa% necrosis is
occuring
". "hrinkage and darkening of nuclei
ii. Aar%ohexis
1. "mall dark nuclei begins to break apart
!. ell is definitely dead
iii. Aar%ol%sis
1. )ucleus disappears
b. H%pereosinophelia
i. 0xposure of eosin binding sites due to alterations in
tertiar% structure of proteins
c. Large mitochondrial densities 5small in reversible in/ur%6
0. ells eventuall% degraded b% proteol%tic enE%mes in #&;s
ii. Li5uefactie
1. :roduced b% neutrophils and macrophages in )" and the lungs
!. )o scarring, dead cells are completel% digested and %ou get formation
of a cavit% 5often filled with pus > abscess6
a. Allows the brain to maintain spatial arrangement critical for
normal function
b. Also, scarring can induce seiEures so should be avoided
". Bacterial and fungal infection or hypo$ic cell death in )" induce
damage b% enE%mes released from white cells 5stroke6
.. Ae% is the presence of inflammator% cells 5neutrophils and macs6
iii. Caseous
1. ream cheese necrosis
!. .nl% from tb infection and some s%stemic fungi
". Induced by # *underlying cause is+ granulomateous inflammation
.. #hat does a stain look like
a. $nflammator% border filled with amorphous granular debris
5tissue architecture is lost unlike coagulative6
i. ,at
1. .ften associated with acute pancreatitis or trauma to fatty tissue
5female in car accident s=uishes her breast and it later melts awa%
!. :ancreatic enE%mes 5lipases+ released and deregulated, eat tissue in the
peritoneal cavit% alive
". (issue will have chalk% white areas
a. Dat saponification
i. !eleased fatt% acids react with calcium and are
deposited making it chalk% looking
. fibrinoid
1. .nl% in small blood vessels
!. $nduced b% malignant hypertension6 asculitis6 immune mediated
damage
a. H%pertension elevated pressure pushes proteins into the wall
b. Autoimmune compliment activated on antigen antibod%
complexes fixed to the vessel wall
i. 2angrene
1. "ubset of coagulative
!. .ccurs in the feet and fingers of diabetics
19
". $nitial coagulative necrotic site is exposed to a bacterial infection,
subse=uent li=uefactive necrosis induces gangrene
b. 3echanisms
i. A(: depletion
ii. Low :H
iii. .x%gen radicals
i. $ncreased membrane permeabilit%
1. $s not the cause of swelling, cause of swelling is ionic inbalance
. $rreversible mitochondrial damage
i. Influ$ of calcium into cytosol % most important
1. $nterferes with signal transduction pathwa%s
!. all can be induced b% protein thiol oxidation 5occurred during
reversible stage6
".
-. Apoptosis
a. Cs necrosis
i. &iggest difference is control genes are involved here, in necrosis the cell loses
control and that;s wh% its inner constituents are exposed to 00nv
ii. 3embrane bound apoptitic bodies are phagoc%tosed no neutrophils, no
inflammation
iii. #ont affect an entire organ, simpl% removes single cells
1. $n necrosis get transient or permanent loss of function and permanent
scarring
i. 8)A fragmentation is not random, occurs at nucleosomes producing a ladder
pattern on gel electrophoresis
1. Cs necrosis where fragmentation is random, producing a smear on 70
. ). HA)0 of recover%, an% sign > cell death
1. Cs necrosis where nuclear chromatin marginates earl% during the
reversible stages of in/ur%
i. 2et e$ternali-ation of phophatidilserine
1. "ignal for apoptosis
!. &inding site for antibodies best way to diagnose#detect apoptosis
ii. ell permeabilit% labeling with 8)A d%es is not an option
1. &c in apoptosis the membrane is kept intact
!. $n necrosis the membrane is busted so permeabilit% 8)A d%es can get
in
b. #hen and where does it occur
i. :h%siological states
1. $n senescent cells
11
!. 8uring embr%ologic development
". $n immune mediated death
.. $n hormonal and c%tokine withdrawl
a. e.g. menstruation 5don;t want necrosis6
b. prostate after castration
c. lactating breast after weaning
0. immune related
a. withdrawl of $L- results in apoptosis of ( cells
9. ioniEing radiation
a. radiation damages 8)A, cell undergoes apoptosis to prevent
replication of damaged 8)A and cancerous growth
:. steroids , glucocorticoids
a. know that the% are antiBinflammator%
b. are so bc induce apoptosis of l%mphoc%tes
;. viral infections
a. apop is a defense mech via (L and granE%me insertion
b. adeno and pox prevent apoptosis
c. 82 death in A$8" pts ma% be induced b% apoptosis
ii. :athological states
1. Autoimmune diseases , t%pe 2 H"!;s
a. "elf reactive l%mphoc%tes should be removed during
maturation via apoptosis
!. )eoplasia
a. :41 induces apoptosis
b. Lose p41, no more apoptosis, neoplasia
c. 3orphological give awa% slide 119
i. Cell shrinkage 5protein cleavage and cross linking6
1. )ote that p%knosis occurs in both necrosis and apoptosis so look for
the cell siEe give awa%
ii. :resence of macrophages for apoptosis
d. !ole of stress
i. $nduces both apoptosis and necrosis
ii. 8egree of stress is ke%: light 5apop6 vs heav% 5necrosis6
iii. 0.g. a burn 5necrosis at the center, apop around the peripher%6
1. Areas greater than 21 > necrosis, less than > apoptosis
e. 3echanisms of Apoptosis
i. 3ultiple initiating pathwa%s that are tightl% controlled leading to a common
executioner pathwa% that prepares the cellular component for phagoc%tosis b%
macrophages
ii. $ntro to the pathwa%s
1. 0xtrinsic
a. ()D, DA",DA"L
b. $nitiating caspases J and 19
!. $ntrinsic pathwa%
a. 3itochondrial damage
b. &L-, &AM, :41, Apaf
c. $nitiating caspases L 5dna damage6 and 1- 5misfolded proteins6
". ommon executioner pathwa%
a. 0xecutioner caspases 1 and <
iii. Apoptosis in nematodes 5. 0legans6
1. ed 1 @ 2 are absolutel% re=uired for death
a. Are present in all cells and are controlled
!. ed L inhibits apoptosis
i. $mportant pla%ers in humans
1. &clB-
1-
a. Iust like ed L it inhibits apoptosis
b. .verexpressed in & cell l%mphomas translocation to $g7
c. !elative, &clBML acts the same wa%
d. :revent mitochondrial leakage of c%t c and bind to Apaf1
!. &ax
a. (runcated &cl- 5/ust missing &H2 section of &L- gene6 that
promotes apoptosis
b. &ax is upregulated by p0"
c. the balance
i. bax regulates bcl- b% forming heterodimers with it
ii. if %ou have more bax > apoptosis, all about balance
". ApafB1
a. Iust like ed2 it is also necessar% for apoptosis
b. 7long with cyt'c it actiates initiator caspase <
c. &cl - prevents this step
i. $nhibits release of %tBc from mito
ii. $n some species binds and inhibits Apaf1
.. aspases
a. Iust like ed1, necessar% for the process
b. %stein proteases that
i. Inactiate inhibitors of apoptosis
1. &cl- famil%
!. )ucleases responsible for 8)A
fragmentation
ii. cleae proteins at aspartic acid residues
1. causes cross linking and cell shrinkage
iii. deregulate proteins by separating regulatory and
catalytic subunits
i. inactiate inhibitors of apoptosis
. nuclease actiation
1. nuclear shrinking and dna fragmentation
c. "%nthesiEed as inactive precursors and then activated b%
proteol%tic cleavage via:
i. leavage from upstream caspases
ii. $nduced proximit%
iii. HoloenE%me formation
i. =hat steps follow 7ctiation of Caspases>
1. Dirst we see protein cleaage that is ver%
controlled and is followed up b% ?rotein
cross linkage so the protein degraded can be
packaged properl% into vesicles. (his is then
followed b% Cell shrinkage and 8uclease
actiation. #e see the 8uclear shrinkage
and D87 fragmentation into apoptotic
bodies.
f. (he 0xtrinsic :athwa%
i. 8ormal control of cell growth and survival
ii. (rail ligands

tumor necrosis factor'related apoptosis'inducing ligands
5()D, DasBL6 BN binding to trail receptors with homologous c%toplasmic death
domains BN receptors aggregate BN binding to adaptor proteins w death domains
5(!A88, DA886 BN induced proximit% and autocatal%tic activation of caspase
J BN apoptosis
1. )ote when DasBL binds onl% use DA88
!. #hen ()D binds both (!A88 and DA88 bind together
iii. (here are actuall% two ()D receptors
11
1. ()D!1 binding leads to apoptosis
!. ()D!- binding leads to increased gene expression and survival
a. Adaptor proteins activate protein kinases and )DBkb and
induce gene exp.
b. $s how ()D can help in would healing
i. ontrol
1. 8eco% receptors gobble up trail ligands
a. Have no death domains and so don;t bind adaptor proteins and
continue the process
!. BDlip
a. 7obble up procaspase J
". As in an% other control mech, these pla%ers are constantl% present in
small amounts and prevent accidental activation of apoptosis in a full
ph%siological response an% (!A$L ligands will blow past the deco%
receptors all about balance
. "ide note DA"BL
1. #hen an immune cell 5t cell,b cell6 becomes activated in the immune
response it upregulates DA"r
!. #hen the /ob is done, regulator% ( cells with DA"L on their surface
bind to the receptor and induce apoptosis in the immune cell
g. (he $ntrinsic :athwa%
i. Injury induced apoptosis
ii. $nitiated in 8)A damage, mitochondrial damage, protein damage and (L
mediated killing
iii. :la%ers see above for details
1. 3itochondria leaks
a. %t c with Apaf1 activates procaspaseL
b. "mac and 8iablo inactivate inhibitors or apoptosis 5$A:"6
i. $A:" > inhibitors of apoptosis 5e.g. survivin6
1. )ormall% inhibit activation of executioner
caspase 1
!. :41, &ax and &cl-
". :41 upregualtes &ax BN bax forms dimers with &cl- BN allows %tBc to
leak out of the mitochondria along with smac and 8iablo BN c%tBc with
apaf1 activate procaspase L BN smac and 8iablo consume inhibitors of
apoptosis allowing for activation of executioner caspase 1
i. 3ore on p41
1. Has two functions
a. Arrests growth if 8)A is damaged, induces 8)A repair
enE%mes 7A8824
b. $f the damage is too much induces apoptosis upregulate
Ba$
!. "o it acts as a natural checkpoint that preents the replication of
damaged D87
". Cer% important in radiation which induces 8)A damage
a. !adiation actuall% increases p41 protein 5without
transcription6 in normal cells and tumor cells
b. $f someone is p41 negative, can;t give them radiation therap%
.. )ot needed in steroid induced apoptosis of l%mphs
0. Anock out mutations in p41
a. ?rone to lymphomas
. :rocaspase L vs 1-
1. L is activated if there is 8)A damage
!. 1- is activated if there is protein damage
a. proteins are also ver% tightl% controlled
b. chaperones 5Heat "chock proteins6 ensure proper folding
12
c. if proteins are damaged have ubi=uitin and proteosome
induced degradation
d. if damage is too great activate procaspase 1- and get apoptosis
i. CTL mediated damage is also considered intrinsic
1. (L uses perforin to punch a hole in the membrane BN inserts
granE%me & BN granE%me & skips all initiator caspases and activates
executioner caspase 1
h. Applications of apoptosis
i. &L- antisense m!)A in phase - trials
1. &L- inhibits apoptosis
!. Antisense will bind sense BN make less &L- BN induce apoptosis BN
kill cancer
". #ould be helpful in p41 negative individuals 5can;t form bax6
ii. !ecombinant trails have been shown to target cancer cells
1. Administration of actual ()D or DA"BL was toxic
!. !eco;s are helpful
a. 7oing through the extrinsic pathwa%
b. 8on;t need to interact with p41 and &cl- 5good for b cell
l%mphoma6
iii. aspase inhibitors
1. #ill help in diseases caused b% excess cell death
ellular accumulations
1. Alcoholism sublethal changes 5all reversible if %ou stop drinking6 all of the following are
diagnositic of sublethal changes to the lier, not of alcoholism &'( in the western world
alcohol is a cause of sublethal changes to the liver
a. ,atty change % the earliest signal of lier stress
b. Alcoholic H%aline a sign of more severe stress
i. Aka 3allor% bod%
ii. )ot 199O diagnostic of alcohol induced damage but in the '" it usuall% is a
sign of an alcoholic liver
c. 3egamitochondria
i. )ormall%, should not be able to see mitochondria with a light microscope.
ii. Again, in '", usuall% associated with alcohol
d. 3itochondrial cr%stals small densities
e. :roliferation of 0! also occurs with some drugs
-. 7l%cogen accumulation associated with :ompe;s disease
a. an;t metabolise gl%cogen so it accumulates
b. #ill not reall% stain normall%, show;s up as clear areas in the cell
1. :rotein accumulation, causes:
a. :roteinurea and resorption in proximal tubules
i. (here are lots of diseases where %ou will put protein into the urine when the
glomeruli is damaged
ii. Also, the filtration of glomerulus is not perfect so %ou want to resorb whatever
protein got out the proximal tubules will activel% resorb this protein
iii. "omeone with proteinuria will reall% tax the proximal tubules and %ou can see
protein staining in the tubules
b. 0xcess secretor% proteins !ussel bodies in plasma cells
i. #hen plasma cells are producing lots of $g, the 0! grows and houses
eosinophilic protein inclusions known as !ussel &odies
c. 8efects in folding and protein transport
i. Alpha1 antitr%psin
1. 3utations slow folding of the protein
-. :rotein accumulates in the 0! of the liver
1. !esultant low plasma levels of the protein cause emph%sema
14
ii. %stic fibrosis
1. .ne of the proteins for the chloride channel does not separate from
chaperone
iii. DH
1. $mproper folding of receptors
2. 3ineral and pigment deposition
a. Anthracotic pigment
i. #e all have it in our lungs from living in an urban environment
1. "o smokers do not have an% blacker a set of lungs then us
a. Lots of variation in rates of clearance so can;t differentiate
between smokers and non smokers
-. A trul% black black lung is diagnostic of coal miners disease
ii. arbon deposition is the cause of the blackness
4. Hemosiderin
a. $ron deposition
b. 3a% see iron in macrophages due to excess degradation of !&;s
i. ).( 199O diagnostic of heart failure but can suggest heart failure
1. $n the bone marrow macrophages store lots of iron 5a normal thing6
c. an occur in the liver too hemosiderrhosis
i. Happens in hereditar% hemochromatosis
ii. Liver turns into a rust% color bc of iron oxidation
<. :olariEable foreign bodies
a. 0.g. talc 5put into crack, in/ected into the bod%, appears in vessels bc it does not dissolve6
G. Doreign bod% giant cells
a. 3acrophages surround foreign bodies in an attempt to digest
b. Happens in ruptured silicon breast implants
J. Lipofuscin hard to distinguish from hemosiderin
a. $nsoluble, brownish %ellow, intracellular
b. Accumulates with age in non dividing,slowl% dividing cells
i. )eurons, cardiac m%osites, hepatoc%tes
c. Lipid and protein complexes derived from peroxidation of pol%unsaturated fatt% acids in
subcellular membranes
L. 8%strophic calcification ver% important occurs under normal calcium levels
a. alcium accumulations at sites of chronic in/ur% or cell death e.g. atherosclerosis ,
hardening of the arteries or the valves of the heart
19. 3etastatic calcification
a. :roblem with calcium homeostasis so calcium accumulates in the bod%
b. H%perparath%roidism , renal issues , iatrogenic 5calium supplements6
i. alcium deposits in the lung and other tissues
Aging
1.
1<
B one of the things that decreases with age is population
doubling level
B occurs because telomerase activit% decreases with age
and so the abilit% to replicate the 8)A of a cell
eventuall% is done
B germ cells and stem cells maintain high levels of
telomerase
B cancer cells activate telomerase to replicate
indefinitel%
-. other factors affecting longevit%
a. caloric restriction diets ma% promote longevit%
b. weight AL.)0 does not cause problems it is the secondar% complications associated
with heav% weight that cause disease
c. free radical generation and clearance
d. disease that damage 8)A
i. werners and 8)A helicase
ii. Ataxia (elanactasia and 8)A breaks
1G