Body Fluids

TBW%; M:60 F:50 %

ECF: 20% , 14 l = interstitial fluid + plasma
-Intravascular (blood plasma, lymph) - 5%
-Extravascular - Interstitial - 15% (ultrafiltrate of plasma - less proteins)
Major cation: Na´+ , Anions: Cl-, HCO3-
F= fluid from which cells take up O2 and nutrients and discharge metabolites into

ICF: 40% - 28L (2/3)
Major cations: K+, Mg+
anions: Protein, organic phosphates

ECF vs. ICF

Na+: EC; 142 IC: 9
K+: E; 5 , I:135
Cl-: E:103, I: 9
-Seperated by membranes
-Permeable to water, but not so much to solutes Na+, Cl- , conc. of these = effective osmolarity for each compartment
-ECF effective osmolarity increase, cell shrink (ICF decrease) + vv.

Interstital fluid vs plasma fluid
-Fluid moves across capillary membranes
-Freely permeable to all plasma substances except proteins
= plasma protein conc. determine effective osmolarity

ECV vs. ICV
-ECV; Enlarge with net gain of body fluid, (decrease with net loss)
-ICV: Varies with effective osmolarity of EC-compartment, (increase osmolarity -> decrease ICV)

Measurements:
Dilution method
-Triated water: Total body water
-Mannitol: ECF
-Evans blue: plasma volume
Conc. in plasma measured,
Volume = amount of substance present/ conc. in plasma

Blood volume: 7% of b.w.
-M: 7.7%,5.5l, 70ml/kg
-F: 6.5-7%

TBV= plasma (ECF) + cellular elements (ICF)
Plasma volume + Hematocrit
Blood volume = plasma vol. /1/Ht
SA: 2.1-3.8 L/m2

Composition:
-Plasma; ~55% ( 91% water, 7% proteins, 2%nutrients)
-Cellular elements: Ht (45%)
Normo/Hypo/Hyper-volemia
-Hypo: hemoconc. , hemodilution., dehydration
-Hyper: overhydration,
-Changes in RBC volume due to osmosis across cell membrane - isontonic/hypertonic/hypotonic


Blood volume: Decrease with age
-Newborn: 80 ml/kg
-15 years old: 70ml/kg

Gender: Males > women
Temperature: Cold weather - decreased volume
Pregnancy: 20-30% increase during early stages
Excercise: Releae of EP -> more RBC's
Posture: Standing - reduced
Altitude: Increase in high altitudes
Emotion: excitement - SY-stim. - splenic contraction

Measurement:
-Direct:Tapping out all the blood
-Indirect:
1) Determinating plasma volume:
-Indicator dye,dilution technique
PV = amount of dye injected - amount excreted / average conc. of dye in plasma
-Radioisotopes

2) Determination of RBC volume
-i) By hematocrit value
-Centrifugate,
TBV = 100 x amt. of plasma / 100 - PCV
ii) Radioistope method
-eg. Cr , injected, then measure radioactivty in sample

1) Specific gravity =relative density with respect to water
-Blood; 1056 ( 1052-1063) kg/m3
-Plasma; 1027
-RBC: 1090
Depends on:
-Ht , plasma proteins, water content of blood
Measurements ( indicate Hb content of blood)
-Direct; Pyconometers - blood vs. water equal volumes- ratio of wt. :> specific gravity
-Indirect
a) Hammar Schlags - Equalizing the 2 densities of miscible liquid to that of blood (chloroform, benzene)
b)
Philin van syke's Cu-sulphate method
-the specific gravity of blood is compared with solutions of known density

Viscosity of Blood
-4-5x higher than water
(plasma- x1.8)
n= property of a fluids that measures its internal resistance to flow
-Primarily determined by hematocrit
( Ht ~45% - Viscosity: 4-5)
-Anemia: decreased viscosity - decreased peripheral resistance
-Polycytemia: increased n -

> In large vessels - Ht changes cause greater changes in viscosity ( Ht has little effect on peripheral resistance)

Temperature & Viscosity
-Increase T- Increase n
~2% inc. n - for every C

Flow:
-Low flow allows molecules to stick together which may increase viscosity

>High viscosity blood coagulate more easily

Viscosity is also affected by:
-plasma composition ( significant inc. in Ig's)
-resistance of cells to deformation ( abnormally rigid RBC's - hereditary spherocytosis)

Measurement:
-Viscometer

Hematocrit:
-% of RBC's in whole blood
Ht value = volume of cellular elements of blood in relation who whole blood volume
-Depends mainly on RBC count & RBC volume
M: 0.44 +/- 0.05
F: 0.38 +/- 0.04
Newborns: 50-60%

Meausurement:
-Enables also evaluation of other things; whole blood volume, MCV..
-Centrifugation
i)Classical method: Venous blood - centrifuge- capillary
(multiply by correction factor 0.92)
ii)Microhematocritic set

Plasma
-Volume: 5% of b.w. ( 3.5 L)
Serum = clotting factors removed
Functions of plasma
-Transport substances
-clotting
-colloid osmotic P

Plasma osmolality: 280-295 msOsm/Kg H20
-Mainly due to Na+ & Cl-, HCO3-
Colloid osmotic P:

Plasma proteins:
-Osmotic P of plasma proteins > osmotic P of proteins of interstitium - oncotic P gradient ;
restraining fluid filtration + favours reabsoprtion of fluid into capillary

-Buffering - 15% of buffering capacity of blood (plasma)
-Special functions; Carriers, clotting,antibodies..

Plasma proteins; 60-80 g/dl
-Albumin- 1g in 100 ml plasma - osmotic effect 6 mmHg
-Globulin - 1 g in 100 ml - 1.5 mmHg
-Fibrinogen

> Hypoproteinemia: prolonged starvation,malabsoprtion..

RBC's
1)Composition/Structure
-Non nucleated, no organelles
-d: 7.2 um , th: 2.1 um vol: 85fl S: 130um2
(SA all RBC's 50x60 m - football field)
O: Bone marrow

-Water 70% , Dry substance 30%
-Membrane - lipid bilayer, flexible - cytoskeleton (spectrin, ankyrin, band 3 protein)
Fragility - increased: sperocytosis (abn. cytoskeleton)
decreased - microcytic anemia, sickle cell anemia
(small size, normal cytoskeleton)
-Stroma:
-Hb: -Ions ,-Enzymes
Functions;
-Carry O2
(HB some CO2)
Count:
M: 4.3-5.3 x 10^12 /l
F: 3.8 - 4.8 x 10^12/l
Newborn: 7-8 x 10^12/l

Hyperythrosis: Increased count
-Polycytemia
-Polyglobulia
-Excercise, altitudes,hemconc...

Erythrocytopenia - decreased count;
Physiological; Sucklings
Pathological: Anemia

Metabolism
-Glycolysis;
-Anerobic - 90-95% -> lactate
ATP used for: channels, phosphorylation, glycolysis
-2-3-BPG - regulate affinity
-NADH - Met-Hb-reductase - maintain heme as Fe2+

HMP: (5-10% of glucose)
NADPH - GpX

ESR:
-Blood in suspension - charges keep RBC dispersed
-Helmholtz- double - layer: RBC - , plasma proteins +
= Mutual repulsion
-Suspension stability - measured according to its reciprocal value = sedimentation

Factors influencing ESR
i)Plasma proteins; Quantive changes of ratio between plasma lipids
ii)RBC Count + Size
iii) Lipidemia - conc. of lipids
iv) Plasma pH

ESR Values;
M: 2-5 mm/1st h
F: 3-8 mm/1st h (less RBC, more fibrinogen)
-2nd hour value should not exceed 1st
M ESR = age/2 , F= age+ 10 / 2

AGE M F ( Newborns 0-2 mm/hr Children 3-13 mm/hr <50 < 15 mm/hr < 20 mm/hr

• 50

< 20 mm/hr < 30 mm/hr Indications
-severity of disease, monitoring ( patho. changes)
Increased ESR
-Physiological; elderly, pregnancy, menstruation
-Pathological: - inflammations, hepatic diseases, neoplastic processes, Proteinemias, leukemia, anemia
( PCV, viscosity low - reduced resistance to sediment.)

-Extremely high >80 mm/1st hour - sepsis, autoaggresive diseases, renal failure..

Decreased ESR
-polyglobulia ( viscosity- resistance to sedimentation high)
-allergic diseases
-sickle-cell anemia
-cardiac insufficiency

Measurement:
Fahreaus-Westergren
i) Classical
ii)Accelerated

HB:
Structure:Ferrous protophyrin (Fe2+)
(70% of body iron in Hb)
-4 polypeptide chains (HbA 2 a, 2 B)
Synthesis/catabolism:
-BM, Liver
900 g total, 0.3 destroyed/0.3 g synthesized /h
Catabolism: RES + liver

Hemolysis: Destruction of RBC's , release of free
-osmotic, physical,chemical, toxic,immunologic,hereditary
-Haptoglobin
Functions:
-O2 carrying
-Some Co2 carrying
-Buffering
Quantity:
-M: 135-170 g/l
F:120-160 g/l
Newborns: 190 g/l
Sucklings: 110 g/l
1 g Hb carries 1.34 ml O2- 1l blood ~200 mlO2
MCH: 29- 32 pentogram
MCHC: 32-35 g/dl
Derivatives:
Physiological
1) OxyHb: Hb with O2 bound to Fe2+ in heme
2) deoxyHb: without the O2
3)CarbaminoHb: CO2
Pathological
i)CarboxyHb: CO (displaces oxygen)
ii) MetHb: Fe2+ -> Fe3+ (drugs, oxidizing agents)
-Doesn't carry oxygen
Types:
Embryonic
i)Gower I ( 2 zeta , epsilon)
ii) Gower II ( 2 alpha, 2 epsilon)
iii) Portland (2 zeta, 2 gamma)
Fetal
2 alpha, 2 gamma - less avid binding to 2,3-BPG
(Facilitate movement of O2 from maternal -> fetus)
Adult
HbA (2 a, 2 B)
HbA2 - <2% - ( 2 alpha, 2 delta)
HbA1c - glycosylated Hb

Hb switching during development
Regulated mainly by oxygen availability
-Relative hypoxia (+) FHb production,
erytrhopoeting (+) globine production
Hemoglobinpathies:
-HbS- sickle cell anemia (Val -> Glu , B-chains)
Thalassemias:
-Alpha (gene deletion- Microcytic, hypochromic anemia)
-Beta - Various mutations -
Excess; B (super high O affinity) Excess A (precipitate)

Measurement:
a) Determination of Hb conc. in blood
-Spectrophotometry
b)Mean Hb content per RBC
~32 pg
-Hb conc. g/l blood / RBC count

c)Calculation of mean corpuscular Hb conc.
-0.3-0.35 (30-35%)
Hb conc. g/L blood / Ht x 1000

WBC
-leukocytes; true cells (nucleus, organelles, active met.)
Count: 4-9 x 10^9 /l blood
Newborns: 15-17 x 10^9/l blood
-Diurnal rythm (morning lower count)

>9 : Leukocytosis
Physiological:
i)Food intake, activity, pregancy..
ii) Pathological (inflammation, leukemia)


Leukocytopenia <4
-Physiological: Fasting, cold enviroment..
-Pathological: Depression of BM

Indications WBC count:
-Body's resistance to infection
-BM function

WBC count: Melanger /flask method
Differential WBC count: Stain, blood film,
NEUTROPHILS Eosinophils Basophils 56-64 % 1-3 % 0.5-1% 10-12 um in diameter 13-14 um in d 9-10 um 1-5 Nuclei;
Arneth,Hynek => age of nuclei
Young forms - unlobed Nucleus - 2 segments -Circulating
-Marginating
-Tissue Compartment
-Reserve (BM) -Red light granules Few dark blue granules
-Heparin
-Histamine -bacterial infection Inc. In allergic & parasitic diseases -infections
-hyperresponsiveness Staining: Pink Red Blue T1/2 ~ 6h Agranulocytes Lymphocytes, T,B,NK Monocytes (Histiocytes) -24-40% 3-8% 7-14 um 15-25 um -big round & dark blue nucleus
-Only narrow rim of cytoplasm around nucleus -Big kidney -like nucleus

Differential WBC Count:

PROPERTIES OF WBC 1) Ameboid motion-movement
in
tissues
2) Chemotaxis –
response
to
chemical
substances
3) Tigmotaxis –
ability
to
adhere
at
the
surface
of
foreign
bodies
4) Diapedesis –
ability
to
pass
through
the
capillary
wall
5)Phagocytosis –
destruction
of
foreign
particles
Before
phagocytosis:

• Opsonization: opsonins
  on
  the
  surface
  of
  the
  „target“
  (bacteria)
  

• adherence: at
  the
  surface
  by
  means
  of
  pseudopodia
  

PHAGOCYTOSIS ⇒⇒⇒⇒
phagosome,
phagolysosome,
digestion
–
enzymes
Capacity
to
phagocyte :
Microphages
–
Neu
(5-20),
Eo
(Ag-Ab) :Macrophages
–Mono
(100)
OTHER FUNCTIONS OF WBC -Granulocytes; When activated in response to chemical stimuli, their vesicle membranes fuse with plasma membrane
&granule contents are released (degranulation) Mononuclear leukocytes:
-Primary- antigen (foreign body) recognizing cells
-The sub-classes differentiated between different surface proteins that co-relate with their functions
- Neutrophils Eosinophils Basophils Immunite mechanism -Phagocytotic cells;
Migrate to areas of infection or tissue damage , engulf foreign bodies & destroy them
-Intiate respiratory burst (create oxygen radicals that destroy foreign material at site of infection) -Destroy parasites;
-granules= lysosomes With hydrolytic enzymes & cationic proteins which are toxic to worms -Allergic reactions, asthma -Hyperresponsiveness/hypersensitivity reactions; e.g. allergic reactions -Histamine: stimulate smooth m. cell contraction & increased vascular permeability -Enzymes: proteases, B-glucuronidases, lysophospholipase-
degrade microbial structures & assist in remodelins of new tissue Blood Coagulation -Fibrinolysis (profibrinolysin) -Heparin D B-Cells T-Cells NK Cells Monocytes -Secrete anti-bodies
in response to antigen binding -The sub-classes differentiated between different surface proteins that co-relate with their functions
-target virally infected & malignant cells for destruction -Precursors of tissue macrophages
-Macrophages enter inflammatory cites and phagcytose cellular debris &
microorganisms -Macrophages in spleen remove damaged RBC's from circulation

Hematopoesis
-Bone marrow
-In presence of appropiate hemopoetic signals hemapoetic stem cells; proliferate, differentiate, mature -> blood cells

Cytokines
-Developing progenitor cells in marrow grow nearby stromal cells ( FB's, endoth., adipocytes, Macs.)
-Form EXCM & Secrete growth factors
Hemapoetic GF's
-Inidvidual GF; (+) P,D,M of progenitor cells
+ may activate functions in mature cell
-Some GF's act on multiple growth lineages, others more specified targets
-HGF-receptors; cytokine superfamily of receptors
i)Binding of Ligand - JAK's-STAT
-Ras/Raf/MAP bind to phosp. CK-RC
-Binding often transient due to: SOCS
e.g
IL-1,, TNF, => GM-CSF
IL3 (M-CSF)

-Erythropoetin
-Stimuli: Hypoxia
O; kidneys - pertitubular capillaries

Inhibitory factors of erythropoesis:
-decreased levels of stimulatory factors