Showing posts with label Clinical Chemistry. Show all posts
Showing posts with label Clinical Chemistry. Show all posts

05 February 2012

Primary amyloidosis Ppts latest 50 Published articles



Primary amyloidosis: Primary amyloidosis is a disorder in which abnormal proteins build up in tissues and organs. Clumps of the abnormal proteins are called amyloid deposits.

Primary amyloidosis can lead to conditions that include:
    Carpal tunnel syndrome
    Heart muscle damage (cardiomyopathy) leading to congestive heart failure
    Intestinal malabsorption
    Liver enlargement
    Kidney failure
    Nephrotic syndrome
    Neuropathy (nerves that do not work properly)
    Orthostatic hypotension (abnormal drop in blood pressure with standing)

Primary  AL Amyloidosis
by Matthew  Volk
http://www.med.unc.edu/medicine/web/12.1.08%20Volk.%20Amyloid,%20LCDD.ppt

Cardiac  Amyloidosis
by Ann Isaksen
https://medicine.med.unc.edu/education/internal-medicine-residency-program/files/ppt/11.10.09%20Isaksen%20cardiac%20amyloid.ppt

Primary  Amyloidosis  Case Presentation & Discussion
By Warren  Brenner
http://hematology.wustl.edu/conferences/presentations/Brenner20031017.ppt

Immune Disorders:  HLA and Disease Associations and Amyloidosis
by Nancy L. Jones, M.D.
http://cmspath.edu/rfc/lectures11-12/jones/hla/jones-hla_and_amyloidosis.ppt

Protein  Structure Determination, Protein Folding, Molecular Chaperones, Prions Alzyheimer’s
http://www.uh.edu/sibs/faculty/glegge/lecture_18.ppt

Computational  Method for Predicting Amyloidogenic Sequences
by Bill Welsh
http://dimacs.rutgers.edu/Workshops/Neurodegenerative/slides/welsh.ppt

Alphabet  Soup and Interstitial Lung Disease
by Leslie  Scheunemann
http://www.med.unc.edu/medicine/web/3.26.08%20ILD%20Scheunemann.ppt

Latest 50 Published articles:

30 September 2009

Coagulation Testing



Coagulation Testing
By:Diane Jette
BioMedica Diagnostics Inc.

Composition of Blood
* Formed Elements
o Erythrocytes (RBC)
o Leukocytes (WBC)
+ Neutrophils
+ Eosinophils
+ Basophils
+ Lymphocytes
+ Monocytes
o Thrombocytes (Platelets)
* Plasma
o 92% water
o 7 to 9 % of solutes are proteins
+ 55 to 60% Albumin, 15% Globulins, 4% Fibrinogen
o Non-protein nitrogen substance, Enzymes, Antibodies, Electrolytes, etc.
o Serum: No fibrinogen or Factors II, V and VIII

Hemostasis is the arrest of bleeding from an injured blood vessel
* Vasoconstriction and compression of injured vessels
* Platelets adhere to the site of injury and form a platelet plug
* Platelets release factors to augment vasoconstriction and initial vessel wall repair
* Platelets provide surface membrane sites and components for the formation of enzyme/cofactor complexes in blood coagulation reactions

Coagulation Reactions Lead to the Formation of a Blood Clot
* Two pathways: Intrinsic and Extrinsic - Coagulation Cascade
* Formation of a prothrombin activator - complex of Factor Xa, Factor Va and procoagulant phospholipid on surface of platelets.
* Prothrombin activator cleaves prothrombin into two fragments to give Thrombin.
* Thrombin cleaves small peptides from fibrinogen to form fibrin monomers that polymerize.
* Thrombin activates Factor XIII to cross-link the fibrin to form an insoluble clot.

Coagulation Cascade
* Intrinsic Pathway: (APTT)
o Factors VIII, IX, XI, and XII.
o Activated on surface of exposed endothelium.
o Complexes form on platelet phospholipids.
* Extrinsic Pathway: (PT)
o Factors IV, V, VII, X
o Activated by Tissue phospholipids (Tissue Factor or Tissue thromboplastin) released into blood as a result of tissue damage.
* Common Pathway (Thrombin Time)
o Factors I and II
o Leads to the formation of Fibrin Clot
o Thrombin time does not measure deficiencies in Intrinsic or Extrinsic pathway

The Role of Calcium
* Ca ions are needed for most of the reactions in the Coagulation Cascade
* Ca-chelating agents are used in vitro as anticoagulants (Citrate, EDTA, Oxalate)
* When Coagulation Factors are synthesized without Vitamin K they cannot bind Ca and lose enzymatic function

Regulatory Mechanisms
* Inhibition of Factor Activity
o Plasma protease inhibitors: anti-thrombin III (ATIII), *2-macroglobulin, *1 - antiprotease
o Heparin converts ATIII from a slow acting inhibitor to an instantaneous inhibitor of Thrombin, Factor Xa and Factor IXa
o Protein C and Protein S are serine proteases that cleave Factors VIII and Factor Va rendering them inactive

Fibrolysis
* Fibrin clot is degraded by protolytic enzymes and fragments dissolved in blood
* Process is catalyzed by Plasmin
* Plasminogen is converted to Plasmin
* Activation by tissue plasminogen activator (tPA) and urokinase
* Fibron degrades into large fragments X and Y then smaller fragments D and E

Regulation of Fibrolysis
* Plasminogen activator inhibitors (PAIs) and plasmin inhibitors slow the fibrolysis process
* tPA and urokinase have short half-lives and are rapidly cleared through the liver
* Unbound plasmin is instantaneously neutralized by 2-antiplasmin

Hereditary Coagulation Disorders
* Hemophilia A
o Factor VIII deficiency
o 80% of all Hemophilia cases
* Hemophilia B
o Factor IX deficiency
* Prolonged ATPP
o Recovered by dilution 1:1 with normal plasma
* Normal PT and Normal Bleeding Time
* Factor XI Deficiency
o 5 to 9% of European Jews
* 2-antiplasmin Deficiency

Acquired Coagulation Disorders
* Liver Disease
o Impaired clotting Factor synthesis
o Increased fibronolysis
o Thrombocytopenia
* Desseminated Intravascular Coagulation (DIC)
o Something enters the blood that activates factors
o Complication of obstetrics, infection, malignancy, shock, severe brain trauma
o Elevated PT, APTT, D-Dimer and other fibron degradation products

Circulating Anticoagulants
* Antibodies that neutralize clotting factor activity
* Factor VIII Anticoagulants
o Antibody
o Same profile as Hemophilia A
o Clotting time not restored by mixing with normal plasma
o Life-threatening condition

Lupus Anticoagulants
* Antibodies to phospholipid binding sites on clotting factors
* Prevent factors from accumulating on phospholipid surfaces
* Elevated APTT clotting times not corrected with mixing with normal plasma
* PT normal or slightly elevated.
* Non-specific depression of clotting factor activities (Factors VIII, IX, XI, XII)
* Test sensitivity increased by using diluted reagent
o Dilute ATPP reagent, Russell’s viper venom time, Kaolin time
o Clotting times corrected with the addition of phospholipids

Oral Anticoagulant Therapy
* Coumadin or Warfarin
* Inhibitor of Vitamin K dependant Factor synthesis
* Oral anticoagulant
* Dose regulated by therapeutic effect
* PT assay to measure INR
* INR range established for optimum therapeutic effect (typically 2.0 to 3.0)

Prothrombin Time: PT
* PT reagent contains Calcium ions and Thromboplastin from brain tissue (Rabbit).
* Thromboplastin (Tissue Factor) protein-lipid complex found in tissues outside blood vessels.
* Measures the function of the Extrinsic Pathway.
* Sensitive to Factors IV, V, VII, X.
* Provided as a lyophilized reagent.
* Used to monitor oral anticoagulant therapy (Warfarin / Coumadin).

PT Reagent Calibration
* Reagents are calibrated against standard PT reagent established by the WHO.
* ISI = International Sensitivity Index.
* ISI is assigned by the manufacturer for each lot of reagent using reference material traceable to WHO.
* The lower the ISI the more sensitive the Reagent
o ISI of 1.8 to 2.4 = Low sensitivity (North American Standard PT)
o ISI of 1.4 to 1.8 = Average sensitivity
o ISI 1.0 to 1.4 = High Sensitivity

PT: INR Values
* INR = International Normalised Ratio.
* MNP = Mean Normal Plasma.
* INR = (PT / MNP)ISI
* An INR of 1.0 means that the patient PT is normal.
* An INR greater then 1.0 means the clotting time is elevated.

INR Calculation
* Example 1
o MNP = 12.0 s
o ISI = 1.25
o Patient Plasma = 20 s
o INR = (20.0 / 12.0)1.25 = 1.9
* Example 2
o MNP = 12.0 s
o ISI = 1.85
o Patient Plasma = 17 s
o INR = (17.0 / 12.0)1.85 = 1.9
* Example 3
o MNP = 12.0 s
o ISI = 1.4
o Patient Plasma = 20 s
o INR = (20.0 / 12.0)1.4 = 2.0
* Example 4
o MNP = 12.0 s
o ISI = 2.0
o Patient Plasma = 20 s
o INR = (20.0 / 12.0)2.0 = 2.8

Expected PT Values

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Immunoassays



Immunoassays
By:Diane C. Jette, Ph.D.
BioMedica Diagnostics Inc.

Test Menu
* Infectious Diseases
* Cardiac Markers
* Cancer Screening
* Fertility / Hormones
* Drugs of Abuse
* Other Tests
o IgE
o Rh Factor
o Occult Blood
o TSH, T3, T4
o Blood Grouping

Test Formats Available
* Latex Agglutination
* DIPSTICK
* Card Format
* Cartridge Format
* ELISA Format with Plate Reader

Structure of IgG
(150,000 Daltons)
Antigen
Binding Site
Fab
Fc

Structure of IgM
(900,000 Daltons)
Binding Sites
Sandwich ELISA Procedure
* Antibody coated on plate.
* Incubate with sample containing antigen.
* Wash away unbound material.
* Incubate with antibody-enzyme conjugate.
* Wash away unbound conjugate.
* Add substrate.
* Enzyme acts on substrate and produces color change.
* Measure amount of color produced.
* Amount of antigen present is proportional to the amount of color produced.

ELISA: Sandwich Format
Sandwich Assay Results
Color Intensity (OD)
Antigen Concentration
Linear Range
ELISA: Inhibition Assay Procedure

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Urine Analysis



Urine Analysis
By:Diane C. Jette, Ph.D.
BioMedica Diagnostics Inc.


Reagent Test Strips
* Multiple test reagents on a ready to use test strip.
* Test results are read at different times after brief exposure to urine sample.
* The color on strip is compared to the reference color shown on test strip packaging.

Typical Test Strip Test
Glucose
Bilirubin
Ketone
Blood
Protein
Nitrite
Leukocytes
pH
Specific Gravity
Urobilinogen
Sensitivity_
* 4 to 7 mmol/L
* 7 to 14 mol/L
* 0.5 to 1.0 mmol/L (Acetoacetic acid)
* 150 to 620 g/L (Hemoglobin)
* 0.15 to 0.3 g/L (Albumin)
* 13 to 22 mol/L
* 5 to 15 cells/ L
* pH 5.0 to 8.5
* 1.000 to1.030
* 0.2 to 8 mol/L

Glucose
* Small amounts of glucose normally excreted by the kidney.
* Below sensitivity of the test.
* >6 mmol/L clinically significant.

Bilirubin

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04 May 2009

Evaluation of Laboratory Data in Nutrition Assessment



Evaluation of Laboratory Data in Nutrition Assessment
Presentation lecture by:Cinda S. Chima, MS, RD

Laboratory Data and the NCP

* Used in nutrition assessment (a clinical sign supporting nutrition diagnosis)
* Used in Monitoring and Evaluation of the patient response to nutritional intervention

Specimen Types

* Serum: the fluid from blood after blood cells and clot removed
* Plasma: fluid from blood centrifuged with anticoagulants
* Erythrocytes: red blood cells
* Leukocytes: white blood cells
* Other tissues: scrapings and biopsy samples
* Urine: random samples or timed collections
* Feces: random samples or timed collections
* Less common: saliva, nails, hair, sweat

Interpretation of Routine Medical Laboratory Tests

* Clinical Chemistry Panels
o Basic metabolic panel
o Comprehensive metabolic panel
* Complete blood count
* Urinalysis
* Hydration status

Clinical Chemistry Panels: Basic Metabolic Panel (BMP)

o Electrolytes: Na+, K+, Cl-, HCO3 or total CO2
o Glucose
o Creatinine
o BUN

Basic Metabolic Panel Charting Shorthand
Clinical Chemistry Panels: Comprehensive Metabolic Panel Includes
* BMP except CO2
* Albumin
* Serum enzymes (alkaline phosphatase, AST [SGOT], ALT [SGPT]
* Total bilirubin
* Total calcium
Phosphorus, total cholesterol and triglycerides often ordered with the CMP
Complete Blood Count (CBC)
* Red blood cells
* Hemoglobin concentration
* Hematocrit
* Mean cell volume (MCV)
* Mean cell hemoglobin (MCH)
* Mean cell hemoglobin concentration (MCHC)
* White blood cell count (WBC)
* Differential: indicates percentages of different kinds of WBC

Clinical Chemistry Panels: Urinalysis
Types of Assays

* Static assays: measures the actual level of the nutrient in the specimen (serum iron, white blood cell ascorbic acid)
* Functional Assays: measure a biochemical or physiological activity that depends on the nutrient of interest (serum ferritin, TIBC)
o (Functional assays are not always specific to the nutrient)
Assessment of Nutrient Pool
Assessment of Hydration Status

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Basic Clinical Chemistry Lab Testing



Clinical Laboratory Testing – Basic Clinical Chemistry
Presentation lecture by:Cecile Sanders, M.Ed., MT(ASCP),
CLS (NCA)

Purpose of Clinical Chemistry Tests

*Measure levels of substances found normally in human blood that have biological functions. Examples: Glucose, Calcium
*Detect or measure non-functional metabolites or waste products. Examples: Creatinine, Blood Urea Nitrogen (BUN)

Clinical Laboratory Testing - Basic Clinical Chemistry

*Detect or measure substances that indicate cell damage or disease. Examples: Liver enzymes, such as ALT, Cardiac enzymes, such as CK-MB
*Detect or measure drugs or toxic substances. Examples: Dilantin, Drugs of abuse screen
* Types of Specimens for Chemical Analysis
*Whole blood, serum or plasma. The most common specimen is serum, collected in a tube with no anticoagulant so that the blood will clot.
*Urine – often 24 hour collections
*Others – Cerebrospinal Spinal Fluid (CSF) and other fluids

* Collection and Handling of Blood Specimens for Chemical Analysis
*Blood collection tubes for obtaining serum - Serum Separator Tubes (SST) do not have an anticoagulant but do contain a gel substance which will form an interface between the clot and the serum when the blood specimen is centrifuged. These tubes are sometimes referred to as “Tiger Tops”.

*Blood collection tubes for obtaining plasma
*Patient preparation; time of collection; & effects of eating on chemistry analysis

*Some specimens are increased or decreased after eating (ex. Glucose, triglycerides), so it is important to know what the test and collection method call for. Specimens for these tests are usually collected in a fasting state.

*Sometimes serum or plasma appears lipemia (milky) after a patient has eaten a fatty meal. Lipemia affects most chemistry analyses. The blood must be recollected when the patient is fasting.


* Clinical Chemistry Tests
*Normal or Reference Values – range of values for a particular chemistry test from healthy individuals
*Chemistry Panel grouping – some tests are “bundled” according to the system or organ targeted. Examples: thyroid panel, liver panel, cardiac panel, kidney panel, basic metabolic panel, etc.

* Commonly Performed Chemistry Tests or Analytes
*Proteins – essential components of cells and body fluids. Some made by body, others acquired from diet. Provides information about state of hydration, nutrition and liver function, since most serum proteins are made in the liver.

*Electrolytes – sometimes called “lytes”
*Includes sodium (Na), potassium (K), chloride (Cl) and bicarbonate (HCO3-)
*Collectively these have a great effect on hydration, acid-base balance and osmotic pressure as well as pH and heart and muscle contraction
*Levels differ depending on if inside vs. outside cells
*Important in transport of substances into and out of cells

*Minerals
*Calcium
*Used in coagulation and muscle contraction
*99% is in skeleton and is not metabolically active
*Influenced by vitamin D, parathyroid hormone, estrogen and calcitonin
*Hypercalcemia – occurs in parathyroidism, bone malignancies, hormone disorders, excessive vitamin D, and acidosis; may cause kidney stones
*Hypocalcemia – can cause tetany; occurs in hypoparathyroidism, vitamin D deficiency, poor dietary absorption and kidney disease

*Phosphorus
*80% in bone and rest in energy compounds such as ATP
*Influenced by calcium and certain hormones
*Iron
*Essential for hemoglobin
*Deficiency results in anemia; may be caused by lack of iron in diet, poor absorption, poor release of stored iron or loss due to bleeding
*Increased in hemolytic anemia, increased iron intake or blocked synthesis of iron-containing compounds, such as in lead poisoning


*Kidney Function Tests

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