materi kul biokimia darah.blok 1.5. des 2014
TRANSCRIPT
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THE BLOODHAS MANY
FUNCTIONS
Biochemistry Department
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SPECIAL TOPICS
PLASMA PROTEINS ANDIMMUNO-GLOBULINS
RED AND WHITE BLOODCELL
HEMOSTASIS ANDTHROMBOSIS
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*Functions of Blood
Blood performs anumber of functions
dealing with:
Substance distributionRegulation of bloodlevels of particular
substancesBody protection
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*Disti!utionBloo
d
tra
nspor
ts:
Oxygen from the lungs andnutrients from the
digestive tract
Metabolic wastes from
cells to the lungs andkidneys for elimination
ormones from endocrine
glands to target organs
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*R"#ul$tion
Blood maintains:
!ppropriate body temperature byabsorbing and distributing heat
"ormal p in body tissues usingbuffer systems
!de#uate fluid volume in thecirculatory system
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*O%"%i"& of Blood Cicul$tion
Blood leaves theheart via arteries
that branchrepeatedly until
they becomecapillaries
Oxygen (O)* andnutrients diffuseacross capillarywalls and enter
tissues
'arbon dioxide('O)* and wastes
move from tissuesinto the blood
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*O%"%i"& of Blood Cicul$tion
Oxygen+deficient blood
leaves the capillaries andflows in veins to the heart
,his blood flows to thelungs where it releases
'O)and picks up O)
,he oxygen+rich bloodreturns to the heart
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*P'(sic$l C'$$ct"istics $nd )olu*"
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*Blood )olu*"
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*Co*+on"nts of W'ol" Blood
Figure 1
Withdraw blood and
place in tube1 2 Centrifuge
Plasma
(55% of whole blood)
Formed
elements
Buff coat!leu"occtes and platelets
(#1% of whole blood)
$rthroctes
(5% of whole blood)
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*Blood
Figure 10.12
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*Co*+osition of Blood
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*Co*+osition of Blood
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*Blood Pl$s*$
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*Pl$s*$
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*Ot'" co*+on"nts of +l$s*$
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*Fo*"d
"l"*"nts
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*Fo*"d El"*"nts
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*H"*o+oi"sis ,H"*$to+oi"sis
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*An Erythrocyte (RBC)
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* Erythrocyte Membrane Composition
http://www.ruf.rice.edu/~bioslabs/studies/sds-page/rbcmembrane.html
f l bi
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Structure of Hemoglobin
Red Blood Cells
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Red Blood Cells Found in higher concentration in ma
than in emale !la"ma
Biconca#e di"c"$ anucleate$e""entiall% no organelle"
&la"ma mem'rane contain" spectrina !rotein that gi#e" them (e)i'ilit%
Structural characteri"tic" contri'uteit" ga" tran"!ort unction
Biconca#e "ha!e that ha" a huge"urace area relati#e to #olume
Di"counting *ater content$er%throc%te" are more than +,-hemoglo'in
AT& i" generated anaero'icall%$ "the er%throc%te" do not con"umethe o)%genthe% tran"!ort. no
mitochondria/
Erythrocyte Function
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Erythrocyte Function
Er%throc%te" are dedicated to re"!irator% ga"tran"!ort
Hemoglo'in re#er"i'l% 'ind" *ith o)%gen and mo"to)%gen in the 'lood i" 'ound to hemoglo'in
Hb + O2 HbO
2
For*ard reaction occur" in the !ulmonar%ca!illarie"
0e#er"e reaction occur" in the "%"temic ca!illarie"
Hemoglo'in i" com!o"ed o the !rotein glo'in$ madeu! o t*o al!ha and t*o 'eta chain"$ each 'ound toa heme grou!
Each heme grou! 'ear" an atom o iron$ *hich can
Fate and De"truction o
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Fate and De"truction oEr%throc%te"
Hb Conservation and Recycling
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Hb Conservation and Recycling
Iron
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Erythropoiesis
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y p
A hemocytoblast transforms into a committed cell called the proerythro-
blast roerythroblasts develop into early erythroblasts !he developmental path"ay consists of three phases
hase # $ ribosome synthesis in early erythroblasts hase % $ hemoglobin accumulation in late erythroblasts and
normoblasts hase & $ e'ection of the nucleus from normoblasts "hen cell has
&() of the Hb it "ill ultimately have and formation of
reticulocytes Reticulocytes then become mature erythrocytes
Regulation and Reuirements for
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Regulation and Reuirements forErythropoiesis
Hormonal Control o
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Hormonal Control oEr%thro!oie"i"
Erythropoietin +echanism
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y p
Figure 1
&mbalance
'educes 2
leels
in blood
$rthropoietin
stimulates red bone
marrow
$nhanced
erthropoiesisincreases 'BC
count
*ormal blood o+gen leels ,timulus! -po+ia due todecreased 'BC count.
decreased aailabilit of 2to
blood. or increased tissue
demands for 2
&mbalance
,tart
/idne (and lier to a
smaller e+tent) releases
erthropoietin
&ncreases
20carring
abilit of blood
Dietar% 0e1uirement" o
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% 1Er%thro!oie"i"
H lbiG dG P d
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http://www.mun.ca/biology/desmid/brian/BIOL3530/DB_Ch09/fig9_24.jpg
Hemoglobin Genes and Gene Products
Hemoglobin Genes and Gene Products
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http://www.blackwellpublishing.com/korfgenetics/figure.asp?chap=13&fig=Fig13-1http://www.embryology.ch/anglais/qblood/blut03.html
g
HbF: 2and 2 HbA1: 2and 2
HbA2: 2and 2HbE: 2and 2
Umbilical vesicle
liver
spleen
Bone marrow
H lbiS Ch
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Hemoglobin Structure Changes
http://www.mfi.ku.dk/PPaulev/chapter8/images/8-3.jpg
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Depends on pH ([H+]), CO2, BPG (DPG), Temp
Factors Affecting Binding of O2
pH down
BPG or T up; right shift
pH up
BPG or T down; leftshift
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Erythrocyte metabolism
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Erythrocyte metabolism
Red Blood Cell Metabolism
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Red Blood Cell Metabolism
Metabolism These pathways are essential or o!ygen transport and
maintaining the physical characteristics o the RBC. $mbden0eerhof glcoltic pathwa
"enerates #0$ o energy needed by RBC%s
"lucose is metaboli&ed and generates two molecules o 'T( )energy*.
-e+ose monophosphate shunt Metaboli&es +,10$ o glucose.
-'(/ is end product
(rotects the RBC rom o!idatie inury.
Most common deect is deiciency o the en&yme glucose,,phosphate dehydrogenase (03P4)
3 the pathway is deicient4 intracellular o!idants can%t be neutrali&edand globin denatures then precipitates. The precipitates are reerredto as-ein6 bodies. )Must use supraital stain to isuali&e them.*
Red Blood Cell Metabolism
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Red Blood Cell Metabolism
ethemoglobin reductase pathwa Maintains iron in the errous )Fe2* state.
3n the absence o the en&yme )methemoglobin reductase*4
methemoglobin accumulates and it cannot carry o!ygen.
7eubering0'apaport shunt 'llows the RBC to regulate o!ygen transport during
conditions o hypo!ia or acid,base imbalance.
(ermits the accumulation o 245,(" which is essential
or maintaining normal o!ygen tension4 regulating
hemoglobin ainity
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E
rythrocyt
emetabo
lism
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E
rythrocyt
emetabo
lism
(entose phosphate pathway
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p p p y
'naerobic glycolysis and 245,B("
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g y y
245,B(" and hemoglobin,o!ygen binding
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eo!y/b ainity to bind 245,B(" is 100,oldhigher that that o 6!y/b
Binding o 245,B(" to /b decreases /b ainityto bind 627 right shit o the cure
2 mmol8l
9 mmol8l
mmol8l
/bsaturation
p62
tissues
lungs
:ungs
6nly a small decrease
o o!ygen loading
Tissues
;igniicant increase
o o!ygen unloading
/b o!ygen saturation and erythrocyte metabolism
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(hosphoructo
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lucose transporter!
integral membrane protein )12 membrane,spanning helices*
a channel or the glucose transport
insulin,independent transporter
Glycolysis in erythrocytes
1 ,ource of 89P :actate, the end product
Coer #0$ o energy re>uirement
2 enerate 2.:0bisphosphoglcerate (2.:0BP)
a maor reaction pathway or the consumption o glucose in erythrocytes the speciic binding o 245,B(" to deo!yhemoglobin decreases the o!ygen ainity o
hemoglobin and acilites o!ygen release in tissues
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2,3-bisphosphoglycerate
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8llosteric effector of haemoglobin!
binds to deo!yhaemoglobin )a central caity capable o binding 245,B("*
decreases haemoglobin?s 62ainity
Clinical aspects!
3n people with high,altitude adaptation or smo
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Glutathione$limination of -22and organic hdropero+ides
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2 2
1. Coactor or the glutathione pero!idase)remoes /262ormed inerythrocytes*
2. 3noled in ascorbic acid metabolism
5. (reents protein @;/ groups rom o!idi&ing and cross,lin
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"enerates -'(/ , reduction o glutathione )eliminates /262ormed in erythrocytes*
Clinical apect!
lucose030phosphate dehdrogenase deficienc
Causes hemolytic anemia )decreased production o -'(/ , reduced protection
against o!idatie stress , haemoglobin o!idation and /ein& bodies ormation4
membrane lipid pero!idation and hemolysis*
/emolytic crises are eocated by drugs )prima>uine4 sulphonamide antibiotics* and
oods )broad beans*
The most common en&yme deiciency disease in the world )100 million people*
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6!yhaemoglobin62
Superoxide dismutase
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/aemoglobin
MethaemoglobinMethaemoglobin reductase
;upero!ideSuperoxide dismutase
/262
Catalase
62A/
Glutathione peroxidase
/26
";/
";;"
Glutathione reductase
-'(/
-'(A(entose phosphate
pathway
";/,reduced orm ";;",o!idi&ed orm o glutathione
Haemoglobin autoxidation 5$ o the haemoglobin undergoes o!idation eery day a constant lu! o 62
D,
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2
-em 0 Fe2;0 2 -em 0 Fe:; 0 2
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a high concentration in erythrocytes
accelerates the dismutation 62D,to /262
H2!2 remo$e
1 Catalase
2 lutathione pero+idase
%& 'atalase
a erric haem group bound to the actie site
catalyses decomposition o /2
62
to water and o!ygen
2/262 2/26A62
2& Glutathione peroxidase
remoes / 6 by coupling its reduction to / 6 with o!idation o reduced glutathione
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remoes /262by coupling its reduction to /26 with o!idation o reduced glutathione)";/*
/262A2";/";;"A2/26
lutathione reductase reduces o!idi&ed glutathione bac< to reduced
";;"A-'(/A/A 2";/A-'(A
-'(/, the pentose phosphate pathway )glucose,,phosphate dehydrogenase*
Cooperation of glutathione pero+idase and catalase
The concentration o /262 is raised, catalase becomes more important )high =m or/262*
(ow-molecular mass antioxidants
= tocopherol (itamin $)
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=0tocopherol (itamin $)
(resent in the erythrocyte membrane
(reents lipid pero!idation )chain,brea
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R6; H molecules4 atoms or ions capable to react aidly with lipids )membranes*4
proteins4 nucleic acids and impair their structure and unctionRadicals R6; with an unpaired electron
3ncreased R6; production and8or decreased degradation H o!idatie stress
3mportant reactions, ;upero!ide generation 62A e
,7 62D,
, Fenton reaction Fe2AA /2 62 7 Fe5AA 6/ A 6/D,
, /aber,Ieiss reaction 62D,A /262 7 6/ A 6/
D,
, ;upero!iddismutase );6* 62D, A62
D, 7 /262 A 62, Catalase )C'T* 2 /262 72 /26 A 62, "lutathionpero!idase )"(J* 2 ";/ A R,6,6/ 7 ";;" A /26 A R6/
2 ";/ A /2627 ";;" A 2 /26
R6; in the erythrocytes
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/igh 62 content
' transition metal Fe )2A75A*4 contained in /b
/b autoo!idation
/emFe2AA 62 K )/em Fe2AL62K /emFe
5AL 62D,* K /emFe5AA K 62
D,
Regeneration o /b methemoglobinreductase
Erythrocytes are rich in antio!idant en&ymes ;64 "(J4 C'T =ey role o glutathione ),glutamyl,cysteinyl,glycin4 ";/*
Reaction o "(J
irect reaction with molecules modiied by R6;
";/ regeneration by glutathionreductase
";;" A -'(/ A /A7 2 ";/ A -'(A
(entose phosphate pathway is the source o -'(/ A /A
-'(/ and antio!idant protection
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-'(/ A /A is produced in a reaction cataly&ed by glucose,,
phosphatedehydrogenase )"(*
"( deiciency (robably the most re>uent en&ymopathy )L100 million people4 500 genetic
ariants o the en&yme*
Fre>uent in mediterranean countries4 tropical 'rica4 'sia
Results in hemolytic anemia4 worsened by some oods )broad beans* or drugswhich act as o!idants
/emolysis is caused by o!idatie impairment o membrane lipids )pero!idation*4proteins )o!idation o thiol groups to disulides*
Methemoglobinemia
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'bout 5$ o /b is conerted to met/b and bac< in 29 hours4 but
met/b concentration is normally ery low Causes o increased met/b concentration
3nherited deiciency o methemoglobinreductase 3ncreased susceptibility o abnormal /b ariants to autoo!idation )/bM4
/b; , sic
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The blue people of Troublesome Creek. Science 1982. lustrace !alt Spit"miller.
Erythrocyte ,isorders
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y y
Anemia .nsufficient Erythrocytes
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Anemia2 Decrea"ed Hemoglo'inContent
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Content
Anemia Abnormal Hemoglobin
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Anemia Abnormal Hemoglobin
olycythemia
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/hite Blood Cells
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Neutrophils 0polys12 +31s afterleaving bone marro"2 stay incirculation #4-#% hours then move into
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other tissues5 Become motile2phagocyti6e bacteria2 antigen-antibodycomple7es and other foreign matter5Secrete lyso6yme5 8ast #-% days5 9:-;:) of total /BC
Eosinophils5 8eave circulation and
enter tissues during inflammatoryresponse5 revalent in allergicreactions5 Release chemicals that helpdestroy tape"orms2 flu
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Responsible for antibody production5 Studiede7tensively "ith the immune system5 %4-%:) of
total
Basophils least common5 8eave circulation andmigrate through tissues2 play a role in bothinflammatory response and allergic reactions5roduce histamine and heparin5 45:-#) of total
/BC
Monocytes remain in circulation for & days2 leavecirculation and become macrophages5 hagocyticcells5 Can brea< do"n antigens and present them to
lymphocytes for recognition5 &-9) of total
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*Poduction of L"u.oc(t"s
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*Fo*$tion of L"u.oc(t"
*Fo*$tion of L"u.oc(t"s
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Figure 17
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*L"u.oc(t"s Disod"s/ L"u."*i$s
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*L"u."*i$
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latelets
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*Pl$t"l"ts
Genesis of Platelets
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Figure 17
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*Pl$t"l"t Plu# Fo*$tion
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*Co$#ul$tion
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*Co$#ul$tion
Figure 17.
*D"t$il"dE t f
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E%"nts ofCo$#ul$tion
Figure 17.
* Co$#ul$tion P'$s" 0/ T&o P$t'&$(s toPot'o*!in Acti%$to
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*Co$#ul$tion P'$s" 1/ P$t'&$( toT'o*!in
* Co$#ul$tion P'$s" 2/ Co**on P$t'&$(s to t'"Fi!in M"s'
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*Clot R"t$ction $nd R"+$i
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*F$ctos Li*itin# ClotGo&t' o Fo*$tion
In'i!ition of Clottin# F$ctos
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F$ctos P"%"ntin# Und"si$!l" Clottin#
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107/127
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108/127
* H"*ost$sis Disod"s/T'o*!o"*!ol(tic Conditions
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*P"%"ntion of Und"si$!l" Clots
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*H"*ost$sis Disod"s
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*H"*ost$sis Disod"s/ Bl""din# Disod"s
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*H"*ost$sis Disod"s/ Bl""din# Disod"s
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*H"*ost$sis Disod"s/ Bl""din# Disod"s
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*H"*ost$sis Disod"s/ Bl""din# Disod"s
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Blood =rouping
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AB> Blood =roups
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Agglutination Reaction
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!ransfusion
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Rh Blood =roup
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120/127
Erythroblastosis Fetalis
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,iagnostic Blood !ests
ype and Crossmatch: determination of ABO
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and Rh blood types. Red cells tested againstantibodies
Complete Blood Count
Red Blood Count: number of RBCs/ microliter ofblood
Hemoglobin Measurement: grams ofhemoglobin/!! m" of blood. #or a male$ %&'$female (&) g/!! m"
Hematocrit Measurement: percent of blood thatis RBCs
*hite Blood Cell Count: +$!!!&!$!!! /microliterof blood
Hematocrit
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Glycosylated haemoglobin Hb)%#
ormed by hemoglobinNs e!posure to high plasma leels o glucose non,en&ymatic glycolysation )glycation*, sugar bonding to a protein normal leel /b'1, +$ a buildup o /b'1, increased glucose concentration
the /b'1leel is proportional to aerage blood glucose concentration oer preious
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1 g g
weeuantities othese glycated hemoglobins are noted )patients monitoring*
Sugar C/6 A -/2C/2#rotein
Sugar C/ - C/2#rotein
Sugar C/2 -/ C/2 #rotein
Schiff base
Gl$cos$lated protein
%madori reaction
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*Su**$( of Fo*"dEl"*"nts
Table 1
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*Su**$( of Fo*"dEl"*"nts
Table 1