Ferrokinetics

A. Iron metabolism

Excess Fe deposited in lysosomal membrane as Hemosiderin (pseudocrystalline complex) under LM as Prussian Blue Staining Dec. transferrin: 1)Collagen disease 4) RA

Fe - most abundant trace element - Stored in:

1)

Hepatocytes (parenchymal cells)

2) RE cells 3) BM

2)

Long stanging infection 5)chronic disease-erythropoietin

3)Malignancies (tumor, CA) Sideroblasts nucleated RBC w/ stored ferritin Siderocytes mature RBC w/ ferritin deficiency Fe stores are depleted; defects in Fe

- TIC = 3-5 g in adults; ave: 4g/ 4000 mg 65% Hb 2,500

mg in RBC

- 1 mg required for each ml of RBC produced - 20-25 mg erythropoiesis (95% - recycled Fe salvaged from
normal RBC turn over & Hb catabolism)

5) Iron

metabolism if there is:

- 1 mg/day (5% of Fe TO) newly absorbed balance Fe loss

by fecal & urinary excetion

- Remaining body Fe (1/3 of TIC) stored in

liver, spleen & BM

or carried in myoglobin & coenzymes of cytochrome electron transport protein (respiratory)

- Cytochrome hemoprotein; resp. for electron transport - Myoglobin has 1 heme pigment, bind w/ O2 tightly; muscle
protein

1. Inappropriate oral intake Fe content in food - Infants too long on milk diet - Native pop. w/ marginal & poor diets - Older people w/ limited food intake (tea & toast) 2. Insufficient or defective absorption from intestine
transport = dec. transferrin (Rheumatic arthritis) mense) (inflammatory

after

GIT operations (gastrectomy) or in chronic malabsorption states

3. Inefficient transport, storage or utilization of Fe - deficient

conditions

- Very low O2 conc release O2 by myoglobin

- hematin pigment w/ Fe; myo muscle - Storage iron:

4. Abnormal loss of Fe loss of circulating RBC (hemorrhage / Males: pathologic blood loss prime suspect (hemorrhoids occult carcinomas of bowel)

a. b.

Ferritin Fe + apoferritin (intracellular acceptor Hemosiderin apoferritin is unavailable

protein)

B. Iron absorption Regulated by intestines just enough Fe to cover losses w/o excessive absorption Normal dietary intake = 10-20 mg/day (1 mg absorbed) Dietary Fe Fe+3 (Ferric) or Fe+2 (Ferrous) only absorbed Source of Fe+2 (heme) liver Source of Fe+3 (non-heme) eggs & ampalaya (veggies) Reduction by acid pH of stomach & reducing subst. in duodenum & jejunum, sometimes in stomach Inc. Fe absorption: 1)Vit. C 2)AA 3)sugars

Loss of Fe in men & menopausal women hemorrhoid & bleeding neoplasm Accompanying factors in Fe depletion: 1)Malaria 3) Kala-azar (L. donovani) 2)Trypanosomiasis 4) Other intestinal parasites Abnormal loss of Fe:

Proper nutrition adequate Fe for body

1) 2)
IDA

Mens
hemolysis)

3) gastritis due to salicylates (MIDOL) 4)Hemosiderinuria(intravascular

Esophagealvarices

1) 3) 4) a.

Distorted appetite (PICA) (crunchy food) Pain Koilonychia (spooning)

2) Fatigued

6) Iron overload 1. Increased absorption Primary hemochromatosis genetically determined metabolic disorder

Dec. Fe absorption: 1)Antibiotics 2)Antacids 3)Oxalates 4) Phytates (cereal & veggie fiber) 5) Tannins from tea 6) Phosphates

- Fe deposited outside of phagocytes - Treatment: remove Fe thru phlebotomy

2 yrs)

(500 ml/weekly for in stores of Fe

once absorbed intestinal mucosal cells oxidize it ferric Fe temporarily stored as ferritin unabsorbed in bowel excreted severe ID inc. absorption to 30% to compensate for depletion Fe requirement pregnant & lactating women, children up to 2 yrs

b.

Hemosiderosis

moderate

deposited

- 1 in mononuclear phagocytosis (monocyte & macrophage)

c. Iron poisoning

1) 2)

Dietary Bantu, Africa cooking utensils, H2O & food Medicinal prolonged administration of medicine Fe

4) Iron transport & storage mucosal cells (intes) blood

bound to specific Fe transport protein (Transferrin) plasma beta-globulin synthesized from liver transferrin attach to receptors of developing RBC membrane release free Fe into RBC (incorporate into heme in mitochondria) 10-20% of T Body Iron stored as ferritin

- 30 mg/kg toxic - 200 mg/kg + fatal 3)

Transfusional Fe deposited in RE cells rather than in hepatic 2. Inc. RBC destruction

3. Ineffective erythropoiesis thalassemia, sideroblastic anemia

Hemoglobin

1. Transcription pre-mRNA production

Formation in developing RBC in BM (except fetal life) 1 week formation occupy 1/3 of RBC (2/3 H2O)

2. 4. 1.

Processing final mRNA formation Transfer tRNA collect AA from cytop. to

3. Translation mRNA leaves nucleus for ribosome in cytoplasm appropriate ribosome site Normal Hb variants: - identified by electrophoresis (also abnormal) Embryonic Hb (1st 3 months after conception)

Components:

1) 2) 3) 4) 1. 2.

4 molecules of nitrogenous subs. Protoporphyrin IX 4 iron atoms in Fe+2 state (heme w/ porphyrin) Globin protein component 2 sets of 2 different polypeptide 1 2,3 diphosphoglycerate (2,3-DPG) sometime resident in Produced in anaerobic glycolytic (Emden-Meyerhof) pathway generates energy for RBC Hb binds 2,3-DPG O2 affinity dec. Plasma level of 2,3-DPG dec Hb 2,3-DPG released Hb O2 affinity Adequate tissue oxygenation need adequate 2,3-DPG

chains center

a)

Portland b) Gower I c) Gower II

th

2. Fetal Hb (4 month of embryonic devt. birth) 80 Nb, <1 A 3. Adult Hb (1yr +) a. HbA, or HbA (22) 97% of Hb 20 Nb, 97A b. HbA2 (22) 2% of Hb - <0.5Nb, 2.5A
Hb derivatives: - Physiologic Hb OxyHb & reduced Hb readily converted to series of cmpds. By acids, alkalies, Oxidizing & reducing subs, heat & other agents

encourage Hb to release O2 to tissues Function: Carries CO2 reduced Hb Carries O2 oxiHb (1g Hb = 1.34 ml O2) O2 combining capacity of blood dir. Proportional to Hb, not RBC

1. 2. -

Hemiglobin or Methemoglobin (Hi) Fe+2 Fe+3 inability of

Hi to combine reversibly w/ O2 - Normal: 1.5% of Hb is Hi If - cyanosis & functional anemia Sulfhemoglobin (SHb) mix of oxidized, partially denatured Hb Hb oxidation S incorporated to heme rings green - Reduced back to Hb by erythrocyte enzyme sys. w/c form during oxidative hemolysis hemochrome Further oxi denaturation & pption of Hb as Heinz bodies Cant transport O2, but combine w/ CO2

Determination used to: 1. Screen for disease assoc. w/ anemia 2. Determine severity of anemia 3. Follow response to treatment for anemia

Physiologic factors w/c affect Hb concentration: 1. Altitude

2. 3. 1. 2. 3. -

Age (Nb = 14-20 g/dL 1 yr adults old) Sex (female = 12-16 g/dL; male = 14-18 g/dL)

4. Pregnancy dilutional anemia Pathologic factors w/c affect Hb conc: Anemia () Pulmonary disease Dehydration ( value) 5. Hemorrhage 6. Leukemia () 7. Polycythemia ()

Carboxysulfhemoglobin - Cant be reduced to Hb, remains in cells until they break down - In patients having treatment w/ sulfonamides or aromatic amine drugs (phenacetin, acetanilid) * Hb S hemoglobin (sickle)

3.

Carboxyhemoglobin (HbCO) endogenous CO during heme

degradation bilirubin 0.5 (CO) in blood & in hemolytic anemia - Hb combines w/ CO 210x greater than affinity for O2 (even if conc. In air is extremely low [0.102 to 0.04%)

4. Destruction of BM Location of Hb during Function & Degradation: In developing RBC (prorubricyte after RBC is released in circulation) RBC destruction remnants captured by phagocytic cells of RES (mostly spleen & liver, also BM) Iron & globin chain AA recycled for Hb synthesis Biliverdin degraded & excreted

A. 1. 2. 3.

Build up typical symptoms of toxic appear >40% saturated blood acute CO poisoning death

- Cant bind O2 Hemoglobinometry measure conc. Of Hb in blood circulation Specific gravity method drops of blood fall into 16 small bottles Drop falls in few sec. - sp.gr. than CuSO4 Drop rises in few sec. - sp.gr. than CuSO4 Drop of blood suspended for 15 sec then fall same sp. Gr as w/ CuSO4 soln of inc. Sp. Gr (results):

Heme production & structure requires formation of protoporphyrin IX & Fe Heme synthesis steps enzymatically directed; in erythroid precursors - Remnants of heme production: Free Erythrocyte Protoporphyrin (FEP) excess porphyrin in mitochondria complexed to Zn

CuSO4 Bottle labe;s: Hb values Adv: fast, simple, inexpensive Disadv: inaccurate Used by blood banks screening test for blood donors Female 1.053 (normal in soln.) = 12.5 g% Hb - abnormal globulin Oxygen method Hb combines w/ & liberates fixed quantity of

- Fe is diminished Measured in lab for diagnosis of disorders ( - Pb poisoning, IDA)

Male 1.055 = 13.5 g% Hb B. Gasometric methods

Ferritin aggregates in cytoplasm; storage Fe not used in heme syn.

a.

Globin chain production simple, nonconjugated proteins w/ AA only - (alpha), (Beta), (gamma), (delta), (epsilon), (zeta?) Greek

O2 blood hemolyzed w/ Saponin O2 collected & measured in Van Slyke apparatus b. CO method C. Colorimetric method

4 phases: (like protein syn)

a.

Direct

matching

methods

(Tallqvists

proc.,

Dares

proc.,

Spencers proc) - Blood color compared w/ colored standards w/ known Hb quantities - Adv: fast, simple, convenient; Spencers proc is accurate - Used at bedside by physician

b.

Acid hematin method (Sahli-Hellige & Haden Hausser) Hb

acid hematin (brown-colored soln. formed by adding 1/10 N HCl) compare w/ standard - Venous blood (EDTA or double oxalate) or capillary blood

c.

Alkali Hematin Methods blood + dil. NaOH boiled Hb

blue-green soln (alkali hematin) measure w/ colorimeter or compared w/ standard - Adv: accurate - Disadv: not accurately measure Hb of infant

d.

Oxyhemoglobin method blood + dil. NaCO3 or NH4OH Hb

OxyHb depth of resulting color measured w/ photoelectric colorimeter - Adv: fast, accurate

e. -

Disadv: Cu traces in diluting fluid can convert Hb MetHb value Cyanmethemoglobin method blood + FerriCN Hb Fe+2

Fe+3 form metHb + KCN stable pigment cyanmetHb - Conv. Time = 3 mins Absorbance dir. Prop. to amt. of Hb present (SHb not meas. Completely) Drabkins soln. (orig) NaHCO3 (1g) KCN (0.05 g) K3Fe(CN)6 (0.20g) Dist. H2O (1,000 ml) (new) anhydrous KH3PO4 KCN K. ferricyanide non-ionic detergent (Sterox S.E) or Triton x100 - Vol. of O2 corrected for temp. & pressure; Hb determined by: Vol. of O2 per 100 ml = gms. Of Hb per 100 ml 1.34 - Abnormal:

1. 2. 3. 4.

Multiple myeloma & Waldenstroms macroglobulinemia Lipemic lipids turbid serum/plasma

- globulin false add 0.1 g K2CO3 to 1L test soln. absorbance add 0.02 ml patients plasma to 6ml of cyanmetHb. HbC, HbS, Hbn abno turbidity (5 cyanmetHB: 5 Turbidity - WBC (+30,000) bacterial infection H2O); 1:2 dilution centrifuge cyanmetHb & read supernatant - Adv: none - Disad: time consuming, technique expensive equipment, inaccurate - Formerly used to calibrate instruments for determining Hb conc.

D. -

Chemical methods based on fact: Hb has fixed Fe quantity

(0.34%) - 1 g or 1000 mg Hb = 3.4 mg Fe Method of Wong blood + conc. H2SO4 w/ K persulfate Fe detached from Hb proteins ppt. w/ Tungstic filter off Fe contect of filtrate determined by colorimeter & Hb value calculated by: mg Fe/100 ml = gms. Hb per 100 ml 3.4

Adv: accurate; Disadv: time consuming, involve technique

- Used to calibrate instruments to determine Hb conc.