Blood Volume1

7/18/2019 Blood Volume1

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Plasma Volume + Packed Cell Volume = Total Blood Volume

Blood = 45% Formed Elements (Cells) + 55% Liquid Elements (Plasma)

!Normal neutrophil• 2-5 lobes

! Hypersegmented neutrophil

• More than 5 lobes

• Pernicious Anemia

BLOOD VOLUME! Total amount of blood in the circulation

! Normal blood volume

" Male: approx. 5.8 L

" Female: approx. 4.5 L

! Definition of terms:

" Normovolemia

# Normal volume of blood in the body

" Hypovolemia

# Decrease in the volume of the blood

" Hypervolimia

# Excessive volume of blood in the body

" Oligemia

# Total reduction of blood volume

# A condition in which the total volume of

the blood is reduced

FACTORS RELATED TO DECREASE ( ) BLOOD VOLUME

! Loss of whole blood

" Hemorrhage

! Loss of RBC

" Spherocytosis

# An auto-hemolytic anemia

! Loss of plasma

" Burned patients, Dehydration

! Loss of body water

" Decrease consumption of water

FACTORS RELATED TO INCREASE (

) BLOOD VOLUME! During blood transfusion ! Intravenous fluid

! Due to increase fluid intake

FORMULA:

! Plasma volume should comprise 55% of the total blood

volume ! Packed cell volume should be round 45% of the total blood

volume

" Total reduction of blood volume ! Packed Cell Volume

" Measured by counting with use of radioactive

materials

# Radioisotopes – for the determination

of Packed Cell Volume (PCV)

# Examples:

- Cr-51, P-32, Radio Iron- Inject Radioisotopes: attach to

RBC ! Plasma Volume

" Measured by intravenous injection of foreign

substances of known amount and concentration

that will allow the measurement of the body fluid" Could be radio active material (radioisotope iodine

131 and with the use of dyes)! Examples of Dyes:

" Evan’s Blue

" Congo Red

" Labeled Human Serum Albumin

NORMAL MATURATION OF CELLS3 Principles:

1) Cytoplasmic Differentiation! Several Factors:

" Loss of Basophilia

# Associated with color blue

# Nucleus

- Acidic part of the cell

- Methylene Blue# Cytoplasm

- Basophilic part of the

cell- Eosin

" Cytoplasmic Granules

# Increase in granules, the

more mature

" Elaboration of hemoglobin

# Increase in hemoglobin, the

more mature

2) Nuclear Maturation! Subdivisions:

a) Structure & Cytochemistry

" Presence of round/oval

nucleus# Young cell-compact

" Large nucleus to cytoplasm

ratio

# The larger the

nucleus, the moreimmature

# The larger thecytoplasm, the more

mature

" Nuclear chromatin

# Rich in RNA

# The more nuclear

chromatin, theyounger the cell

" Decrease in the number of

nucleoli# Found on the most

immature stage

# The more lobulated,the more mature

b) Changes in the shape of nucleus

" Lobulation of nucleus

# The more lobulated,

the more mature

3) Reduction of cell size! The smaller the cell, the more mature! The larger the cell, the more immature



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HEME SYNTHESIS

! First Step

" Condensation of succinyl co-enzyme A + glycine in

the presence of pyroxidal phosphate and deltaaminolevulinic acid synthesis will react to co-enzyme A

and glycine to produce delta aminolevulinic aciddehydrogenase

" Synthesis of nucleus

# Spleen – graveyard of RBC

RBC (120 days)"

Hemoglobin# $

heme globin (reabsorbed protein) # $

Iron (Fe) Porphyrin

HEMOGLOBIN! Red coloring pigment of the blood

! Main component of RBC

! 35% of the total RBC

! Sometimes called respiratory pigment because of its capacity to

transport O2 and CO2

! A gram of hemoglobin can carry approximately 1.34mL of

Oxygen and 3.47mg of Iron

! The adult RBC mass of 600grams hemoglobin can carryapproximately 800mL of Oxygen

! Women need 400-800µg of iron supplement

! Conjugated CHON (Protein) that is made up of 1 globin molecule ofheme groups and the globin is divided into 4 polypeptide chains

! Each chain is attached to 1 heme group

1 Globin Molecule

! Polypeptide chain

" Made up of amino acid

! In normal adult hemoglobin, it is made up of2 alpha and 2 beta chain

" Alpha chain

# Made up of 141 amino acid # First Amino Acid – Valine

# Last Amino Acid – Arginine

" Beta chain

# Made up of 146 amino acid

# First Amino Acid – Valine

# Last Amino Acid – Histidine

! The rest of the globin chain contains 146 amino acids

FACTORS AFFECTING THE HEMOGLOBIN AFFINITY ( ATTACHMENT)WITH OXYGEN

I. Body Temperature! An increase in the body temperature causes the

hemoglobin to release oxygen more readily! * % In Body temperature = % in Oxygen released

" Shift to the right

# % In the release of Oxygen &release of uptake in Carbondioxide

II. Blood pH/Bohr Effect! The relationship of the blood pH and oxygen affinity

is what we call the Bohr effect which tells us anincrease in pH favors the uptake of Oxygen

! * % pH = Alkaline

! * $ Blood pH = % uptake of Oxygen

" Shift to the left

# % in the uptake of Oxygen &

release of Carbon dioxide

III. 2,3 Diphosphoglycerate (DPG)

! An increase amount of DPG level will cause a

diminish oxygen affinity" Shift to the right

# % DPG = $ Oxygen (release)

affinity

IV. Carbon Dioxide! An increase of Carbon dioxide will diminish the

affinity of Oxygen

" Shift to the right

V. Presence of HbF (Fetal Hemoglobin)! An increase of HbF in the fetus will cause increase

oxygenation of the fetal blood

" Shift to the left

" *For adults: Shift to the right (Not Normal)

VI. Presence of Hemoglobin Variance! The number of normal hemoglobin balance will show

either shift to the left or right! Different types of Hemoglobin

" HbM & HbO – Right

" HbA2 & HbA – Left

VII. Decrease affinity of oxygen and increase carbon dioxide

! Haldane Effect

" $ Affinity = Right

$ Body Temperature – shift to the LEFT

%HbF adult – shift to the RIGHTHaldane Effect – shift to the RIGHT% Oxygen uptake – shift to the LEFT

$2, 3 DPG – shift to the LEFT

Heme Heme

Heme Heme



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CLASSIFICATION OF HEMOGLOBIN

A. Normal Hemoglobin! Capable of transporting blood gases particularly

Oxygen & Carbon dioxide

" According to Function

# Oxyhemoglobin- Arterial blood

- Bright red- Oxygen

# Reduced Hemoglobin

- Venous blood

- Dark red- Carbon dioxide

! Hemoglobin in Adult ( post-neonatal )

" HbA

# Constitutes majority of adult Hg

# 96-98% Hg

# 2 ! & 2 " chains

" HbA2

# Normal adult Hg

# 1.5 – 3% total types of Hg

# 2 ! & 2 # (delta) chains

" HbF

# Major hg in fetus (90-95% hg)

# 0.5 – 1% hg

# 2 ! & 2 $ (gamma) chains

! Embryonic Hemoglobin (Pregnant )

" Gower 1

# 2 % (epsilon) & 2 & (zeta) chains

" Gower 2

# 2 ! & 2 % chains

" Hemoglobin Portland

# 2 & (zeta) & 2 gamma chains

B. Abnormal/Non-functional Hemoglobin! Formed due primarily to a defect in the

polypeptide chain of hemoglobin molecule! Category:

1. Carboxyl Hemoglobin (HbCO)

" Reacts with Carbon monoxide in

which the affinity of CO 218xgreater than Oxygen at 37°C

" Do not transport oxygen that may

lead to hypoxia & finally death of

the patient

" Produces cherry red blood color

" Concentration to non-smokers:

0 – 2.3%

" Smokers: 2.1 – 4.2%

" Test for HbCO

# Katayamas Testa. 10 mL Distilled H2O

b. + 5 drops of bloodfrom patients/ controlblood

c. + 5 drops ofAmmonium sulfide

d. + Acidify with acetic

acid

! if NORMAL BLOOD : Dirty greenish, Brown

! if HbCO (+): Rose red color

! Other Test for HbCO:

r Sanderman Dethionite Test

r Palmer’s Test

2. Sulfhemoglobin

" Blood with combined sulfide

" Benign condition

" With significant effect of

cyanosis

" Cause & effect of drug

" Mauve lavender color

3. Met Hemoglobin" A.k.a.

# Hemiglobin

# Oxidized Hemoglobin

# Ferri Hemoglobin

" Reversible reaction

" Normal individual

approximately 1% ofmethylene blue

" Characteristic color:

Chocolate brown

VARIANCE OF HEMOGLOBIN! Hemoglobin S

" Causes sickling of RBC

" Confined to blacks

" Homozygous state causes sickle cell anemia

" Heterozygous state causes/shows sickle cell

fomite

! Heterozygous – carrier parents

! If both parents HbS(+) = Sickle Cell Anemia

! HbS + HbA = Sickle cell Trait

! HgS + HgS = Sickle cell Anemia

! HgS + HgA = Sincle cell shape (carrier)

! Hemoglobin C

" Found also among blacks, rare in whites

" Appears also in homozygous & heterozygous

state

" If present, it demonstrate target cell

! Target cell

# Condensation of Hg at thecenter/periphery

# A.k.a.

# Bull’s eye cell

# Codocyte

# Leptocyte

# Mexican hat cell

# Yersinia enterocolitica

! Hemoglobin D

" Most common for HbD Purijab & HbD Los Angeles

" Both alpha & beta chain abnormally have beenreported

" Shows no sickling of RBC

% Happy Hormones:& Serotonin& Adrenaline& Epinephrine

[ Blood pH becomes basic

“Humble yourselves, therefore, under God’s mighty hand, that he may lift youup in due time. Cast all your anxiety on him because he cares for you.”

- 1 Peter 5:6,7



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HEMOGLOBINOMETRY! Screening test associated with anemia

FACTORS ASSOCIATED WITH INCREASED OF HEMOGLOBIN

! Hyperchromia

# %Increase intensity in the color of hemoglobin

! Polycythemia

# %Increase in RBC

! High altitude# Depletion of oxygen

# Hypoxia & will trigger the release of erythropoietin

in kidney & Bone marrow & RBC! Dehydration

# Loss of fluid within the body

! Heart disease

# Patient with heart problem will have %

hemoglobin pigmentation because of the

compensation of erythropoietin

FACTORS IN DECREASE HEMOGLOBIN CONCENTRATION

! Pregnancy

# Competition for hemoglobin with the fetus

# Ferric sulfide/iron supplements needed ^! Anemia

! Women# Females naturally have low hemoglobin

because of menstruation

# Males naturally have high hemoglobin because

of testosterone (which is associated withmaturation/production of RBCs)

CAUSES OF LOW HEMOGLOBIN COUNTS

! Normal low hemoglobin counts

" It is quite common for women to experience low

hemoglobin counts during pregnancy.

" Some people might also experience this as a

natural way of life – it is simply how their bodyworks.

" Low counts in these cases shouldn’t be alarming.

! Conditions and diseases that cause fewer than normalred blood cells

" Some conditions can cause lower numbers of red

blood cells which can lead to a low hemoglobin

count

" Anemia, cancer, cirrhosis (scarring/ tumor in the liver |

Kirros means yellow), kidney disease and leadpoisoning are just a few reasons.

! Conditions and diseases that destroy red blood cells

" Sometimes your body cant produce red blood cells

fast enough.

" Conditions like Sickle cell anemia,

vasculitis(Inflammation of blood vessels) and an enlargedspleen can all destroy red blood cells quickly

which leads to low hemoglobin.

! Blood loss

" Losing large amounts of blood can easily lead to

low hemoglobin.

" This can also be a warning sign of internal blood

loss.

! Iron deficiency

" Iron is required to create hemoglobin, so if your

body needs more iron, your hemoglobin counts

are probably low.

" In fact, this is the most common cause of

anemia

! Vitamin deficiency

" Vitamins such as B12 or foliate help your bodycreate red blood cells. If you aren’t getting enoughyour hemoglobin levels can drop

! Blood disorders

" Some conditions, such as some cancers, can lead

to low hemoglobin. These blood disorders meanthat your bone marrow isn’t producing red bloodcells fast enough.

" Aplastic anemia

# Bone marrow disorder

# Incapable of releasing RBC

HOW TO INCREASE HEMOGLOBIN COUNTS! Fruits and vegetables

" Look for fruits of all kinds and vegetables that arebursting with leafy green color.

" Cabbage, peas, beans, lettuce, broccoli and

tomatoes are good places to start.

" You can opt for all kinds of fruits, such as

grapefruits, oranges, bananas, raspberries, kiwiand mango

! Vitamin C (Ascorbic acid)

" This is a vitamin that helps your body absorb iron,

which is essential to hemoglobin production.

" Eat foods that contain Vitamin C and take a

supplement that helps your body get even more ofit.

! Other foods

" The staples of a healthy diet can keep yourhemoglobin high.

" Nuts of all kinds, fish and lean meats, poultry,

eggs, organ meats and even fortified cereals canhelp.

! Herbs

" There are many herbs that up your hemoglobin

counts,

" Including fenugreek seeds, rosemary, basil,

thyme, sage, nettle leaf and yellow dock root,among others.



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THINGS TO AVOID

! Iron blockers

" Anything that blocks iron absorption is bad.

" Avoid things like coffee, foods with high levels offiber, tea, and some antacids and medications

! Foods with oxalic acid

" This can prevent absorption, and should be

avoided." Spinach is the biggest culprit among foods with

oxalic acid

" Don’t combine leafy green with fruits because it causesvery high oxalic acid

! Foods with gluten

" Wheat products, such as breads or pasta can

cause trouble for those who have gluten allergies.

" But they can also lead to problems with iron, and

they can lead to low hemoglobin.

" Reduce your intake of foods that contain gluten

" White pasta/white bread inhibit proper absorption

SYMPTOMS OF LOW HEMOGLOBIN COUNTS

! General symptoms" In general, the signs of low hemoglobin include

dizziness, headache, fatigue and a generalfeeling or tiredness. It might be tough to

concentrate. You can experience an irregularheartbeat or notice the skin, gums or nail bedsare very pale in color .

" In addition, you might experience shortness of

breath, palpitations and chest pain with more

severe cases of low hemoglobin.

! Rare symptoms

" Other symptoms that might occur but are relatively

rare include excessive sweating, vomiting andpersistent heartburn(because of too much acidity or very

low/decrease of hemoglobin).

" You might experience swelling in your arms orlegs, and could even notice bloody stool when you

go to the bathroom

! Symptoms in children

" The symptoms in children can be more severe, aslow hemoglobin can lead to long-termconsequences. Look for paleness, a rapid heartbeat, a lowered ability to concentrate, poor

neurological development and disturbedbehavior patterns

" High hemoglobin in children are NORMAL

METHODS OF HEMOGLOBIN DETERMINATIONI. Colorimetry

II. Specific gravity methodIII. GasometricIV. Chemical method

I. COLORIMETRY! 2 types

1) Direct visual colorimetrya) A tallguist method

# Measures hemoglobin using an

absorbent pad.

# The color is match using a standard

b) Dare’s method

# The blood is drawn through capillaryaction between 2 glass plates

c) Acid hematin

# Hemoglobin in a known quantity of

blood in the presence of acid such as

0.1 N HCl is converted to acidhematin. Which is brown coloredsolution diluted with distilled water

drop by drop until it matches thestandard solution

C.1 Sahli – hellige methodC.2 Sahli – hayden methodC.3 Sahli – adam’ s method

C.4 Osgood-haskin methodC.5 New comer

d) Alkaline hematin

# Same principle except we use an

alkaline solution produces alkalinehematin

D.1 Wu methodD.2 Clegg and king method

Equivalent hemoglobin

14-18gms

/dL Male

12-16gms

/dL Female

14-26gms

/dL Children



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SPECIFIC GRAVITY = DENSITY OF SUBSTANCEDENSITY OF WATER

2) Photoelectric colorimetrya) Oxyhemoglobin method

# Measures only normal hemoglobin

# Reagent:

• 0.007N NH4OH or 0.1%

NaCo3 solution

# Wavelength: 540nm

b) Cyanment hemoglobin# Also known as ferriHbCN

# Considered as the most accuratemethod of hemoglobin determination

since It measures all Hb except theSulfhemoglobin

# Reagents:

• Drabkins reagent

KCN2 (Potassium cyanide)

K3Fe(CN)6 (potassium ferricyanide)

NaHCO3 (Sodium bicarbonate)

Distilled water

# Principle: blood is diluted in a solution K3Fe(CN)6, KCN,

K3Fe(CN)6 oxidizes hemoglobin to hemoglobin. Thegemiglobin will further oxidize the KCN to form

cyanmethemoglobin and that cyanmethemoglobin has a

borad absorption max 540.

II. SPECIFIC GRAVITY METHOD! Ratio of the weight of a volume of the blood to the

ratio of the weight to the same volume of water atroom temperature.

!

! CuSO4 (Copper sulfate) METHOD

# Commonly used in blood banks

# Easily acquire fast

# May start 1.055 in counting

# Approximation of donors

# Heavy CUSO4, the blood will sink & vice versa

# Sink if:' >12.5 g/dL (can donate blood)

# Suspended blood if:

' <12.5 g/dL (rejected)

III. GASOMETRIC METHOD! Base on ability of hemoglobin to carry 1.34mL of

oxygen/gram of hemoglobin.! Also called indirect method! Disadvantage:

# Abnormal hemoglobin may not be measure

# Ex.

' vanSlyke,' Haldane' smith

IV. CHEMICAL METHOD! Another indirect method based on the principle that gram

of hemoglobin can carry 3.47 mg of Fe (Iron) ! Ex.

# Wongs

# kennedy’s method

ABNORMAL HEMOGLOBIN1) Several test for abnormal hb pigments:

a. Naked eye testb. Dilution test

c. Alkali spot plate testd. Katayamas test (Important, nakakalason sa dugo)e. Hand spectroscopic testf. Spectrophotometric method

“Be strong and of a good courage, fear not, nor be afraid of them: for the LORD thy God, he it is that doth go with thee; he will not

fail thee, nor forsake thee.”

- Deuteronomy 31:6

Blood Volume1

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