biochem ospe nmu

blood specimens are 3 types

1- whole blood

2- plasma

3- serum

whole blood includes ?

anticoagulant added ?

includes all cellular components (RBCs, WBCs, platelets) and fluid components

yes, it includes anticoagulant

plasma is

anticoagulant ?

the liquid portion of blood without blood cells

yes it includes anticoagulant

serum is

anticoagulant ?

the liquid portion of blood (plasma) without the clotting factors

no its without anticoagulant

what is this specimen ?

does it includes anticoagulant

whole blood

yes it includes anti coagulant

what is this specimen ?

does it includes anticoagulant

blood plasma sapecimen

it includes anticoagulant

what is this specimen ?

does it includes anticoagulant

this is serum specimen

no it doesnt include anticoagulant

2 types of collecting tubes

without anticoagulant

with coagulant

3 types of blood sample

capillary

arterial

venous

when is capillary blood simple is required

required when only few drops are needed

when is arterial blood simple is required

used to measure arterial blood gases like oxygen , co2 , Ph

when is the venuous blood sample is required

its the specimen of choice for most routine laboratory

3 types if venipuncture methods

needle and syringe

butterfly needle

evacuation tube system

2 venipuncture sites

1- the cubital fossa in 3 veins (basilic veins, median cubital vein,,cephalic vein )

2-wrist and dorsum of the hands are also commonly used

9 things needed for needle and syringe venipuncture

gloves

syringes

needles

gauze sponges

tourniquet

evacuated collecting tubes

alcohol wipes

bandages

sharp container (safety box)

sample transport is characterized by 4

1. Rapid delivery to the lab

2. Seal the tube cap & Pack the specimen in an insulated, leakproof container

3. Specimens that must be cooled (e.g., ammonia) in crushed ice. 4. Wrap specimens that require protection from light (e.g., bilirubin)

what specimen must be cooled

ammonia samples

what samples requires being wrapped to be protected from light

bilirubin sample

repeating thaw and freezing what should i do

nooooooooooooooo

this is unacceptable

tips for specimen storage

1. Store specimens in an appropriate temperature

2. Repeat thaw and freezing of samples is unacceptable

3. Keep all specimen away from bright light

mention a qualitative

method

semi-quantitative

quantitative method

for glucose measurement in urine

fehling sol.

urine strip

colorimetry

principle of benedict and fehling test veeeeeeeeeeeeeeeeeery important and discuss it below

Glucose + 2Cu⁺² +2H₂O → Gluconic acid + Cu₂O +4H⁺

reducing sugars (have free aldehyde or ketone functional groups) can reduce cupric ions Cu⁺² (blue) in benedict reagent to cuprous ions Cu⁺ (RED)

benedict/fehling test disadvantage

not specific for glucose

what substance is highl specific for glucose used in its measurement

glucose oxidase

glucose oxidase method reaction is ?

its a double sequential enzymatic reaction in the presence of an indicator that changes in color in the presence of glucose

active reagents in urine glucose dipstick 2?

glucose oxidase & peroxidase

when does glucose appear in urine

glucose appears in urine when blood glucose levels are more than 180mg/dl

what is the renal threshold for glucose

when glucose level in the blood is more than 180mg/dl

what are the 2 catigories of causes of glucosuria

hyper glycemic glucosuria

normoglycemic glucosuria

hyperglycemic glucosuria is sub classified into 3

diabetes mellitus

increased anti insulin hormones

stress glucosuria

what are anti insulin hormones

thyroxine , growth hormone , adrenaline

normoglycemic glucosuria is classified into 2 types

renal glucosuria due to kidney disease

glucosuria in pregnancy (low renal threshold)

Low renal threshold معناها انه مش شرط السكر يبدا يظهر في الurine عند

180mg/dl

لا بيظهر قبلها كمان

what is the principle of protein identification in a sample using the biuret method

in the reaction cupric (Cu+2) complexes with N atoms involved in peptide bonds of protein

so violet color appears and its intensity is proportional to the number of peptide bonds which reflects protein concentration

من الاخر ال Cu+2

بيمسك في النيتروجين اللي في الامينو اسيدس الاحماض الامينيه

ولما بيتركبو علي بعض بيعمل لون بنفسجي

what are plasma proteins

they are proteins present in the blood plasma

normal range for total plasma proteins

6-8gm/ dl

types of plasma proteins include 3

1-albumin (50%-60%) normal range of plasma albumin is (3.5-5g/dl)

2-globulin (alpha,beta,gamma)

3-fibrinogen

how are plasma proteins measured

they are measured by colorimeter

how does colorimeter work

عشاان تفهمه بس

it measures the absorption of light

how does colorimeter work while measuring plasma proteins

the colorimeter measures the absorption of light so the intensity of the violet color is proportional to the concentration of total plasma proteins

albumin in urine relation to kidney health

• A healthy kidney doesn’t let albumin pass into the urine.

• A damaged kidney lets some albumin pass into the urine.

is there any protein in urine for a normal healthy individual

normally small amounts of protein are present in urine

amount of total urinary protein and total albumin in urine

normally total urinary protein excretion is less than 150mg per day

and less than 20mg/day of albumin

what does abnormal albumin levels in urine reveal

albumin in urine can reveal a developing renal pathology at an early stage

protein in urine can be detected by 4

1-urinary test strips

2-the biuret test

3-heat coagulation test

4-colorimeter and immunochemical method

protein in urine other name ?

scientific name يعني

proteinuria

active reagent in urine strips (dipsticks) for protein

tetra-bromophenol blue

فاكر بتاع ال

agarose gel electrophorises هو هو

what does urine strip color change to when proteins are present in urine

greeeeeeeeeeeeeen

urine strip sensitivity to albumin and globulins

urine stripes are highly senitive to albumin and less sensitive to globulins

lipoprotein structure formed from 2 parts

lipid part which contains 3

phospholipid

cholesterol

and triglycerids

non lipid part which contains apoprotein

2 methods for separation of plasma lipoproteins

1-electrophoresis

2-ultracentrifugation

migration of lipoproteins in an electric field depends on 2?

and we are talking about which method of separation of plasma lipoproteins ?

1- amount of -ve charge

2- molecular weight

electrophoresis

separation of plasma lipoproteins using ultracentrifugation depends on ?

ratio between protein and lipid (density)

ترتيب ال Lipoproteins

after separation using ultracentrifuugation

from up to down

chylomicron

vldl

ldl

hdl

albumins and FA

ترتيب ال Lipoproteins

after separation using electrophoresis

from cathode to anode

chylomicron

ldl

vldl

hdl

الاختلاف هنا بين ال

ldl و vldl

functional plasma enzymes

conc in plasma relative to tissues ?

function known ?

substrates presence ?

site of synthesis?

effect of disease ?

examples ?

higher conc in plasma than tissues

have known functions

their substrates are always present in blood

liver

decrease in liver disease

ex: clotting factors (prothrombin)

lipoprotein lipase

pseudo choline esterase

non functional plasma enzymes

conc in plasma relative to tissues ?

function known ?

substrates presence ?

site of synthesis?

effect of disease ?

examples ?

present in plasma in v.low conc in comparison to tissues

no known functions

their substrates are absent from the blood

different organs like heart brain liver and skeletal muscles

different enzymes increase in different organ diseases

ex: amylase & lipase

CK&LDH

ALT&AST
alkaline phosphatase & acid phosphatase

sources of non-functional plasma enzymes 4

1-increase the rate of enzyme synthesis

2-obstruction of normal pathway

3-increase permeability of cell membrane

4-cell damage with the release of its enzyme content

say an example on each of the following sources of non func plasma enzymes

1-increase the rate of enzyme synthesis

2-obstruction of normal pathway

3-increase permeability of cell membrane

4-cell damage with the release of its enzyme content

1- bilirubin increases the rate of synthesis of alkaline phosphatase in obstructive liver diseases

2- obstruction of bile ducts increases alkaline phosphatase

3-as in tissue hypoxia

4-cell damage with the release of its enzyme content into the blood like myocardial infarction and viral hepatitis

medical importance of measuring the non functional plasma enzymes 2

diagnosis of diseases (detection of diseased organ)

prognosis and follow up the disease before and after treatment

سلايد رقم 37 دا مش عارف اعمل فيه اي انا مش هكتب الاختصارات دي ل ايه بس هي منطقيه يعني

لوجالبلك الكلمه كامله هتفهم انها الاختصار ان شاء الله لو مش هتمشي معاك ذاكر النقطه دي من البي دي اف بقا

examples of medically important non functional plasma enzymes 8

amylase & lipase

CK&LDH

ALT&AST
alkaline phosphatase & acid phosphatase

amylase and lipase increase in

pancreatic diseases

CK increases in

hheart brain and skeletal muscle diseases

LDH increases in

liver blood and heart diseases

ALT increases in

liver diseases

AST increases in

liver heart muscles diseases

acid phosphatase increase in ?

cancer prostate

alkaline phosphatase increase in ?

increase in obstructive liver disease bone disease and hyperparathyroidism

iodine reaction with starch gives color ?

deep blue color

iodine reaction with maltose gives color ?

no color produce

اوعي تتغفل ;)

enzymes activity in normal room temp and if cooled to temp like 2 C or heated until like 100 C

in normal temp works fine

if heated or cooled enzyme wont work

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