What is first order kinetics? in an enzymatic reaction, as more substrate is consumed, the reaction rate declines and becomes dependent on substrate concentration; this is a phrase of first-order dependence on substrate concentration when the rate of the reaction is proportional to the concentration of substrate or when the enzyme concentration is fixed and the substrate concentration is varied. What is kinetics? In an enzymatic reaction, when the reaction rate is constant at maximum value, the reaction rate depends only on enzyme concentration and is independent of substrate concentration; the rate of reaction is proportional to the zero power of the substrate concentration. What is a constant for a given enzyme acting under given conditions; the experimentally determined substrate concentration at which the enzymatic reaction velocity equals ½ of the maximum velocity of the enzymatic reaction? Michaelis-Menten constant (Km) What is a plot of the reciprocal of the velocity of an enzyme catalyzed reaction (ordinate; against the reciprocal of the substrate concentration (abscissa; x-axis)? Lineweaver-Burk plot What shows maximum activity in vitro in the pH range of enzymes in plasma? pH from 7 to 8 What are the activity values observed in pH as low and high? Low as 1.5 (pepsin) High as 10.5 (ALP) What can be controlled by the use of buffer solutions? Pronounced effects of pH on enzyme reactions. When does enzymes inactivation and denaturation occurs? Occurs at temperature of 60 degrees Celsius to 70 degrees Celsius. What is the accepted measuring temperature for enzymes in the clinical laboratory? 37 degrees Celsius What is a substance that reduces the rate of an enzymatic reaction and is classified as reversible or irreversible? Inhibitor What is a structural analog of the substrate that combines with the free enzyme and competes with the normal substrate for binding at the active site? Competitive inhibitor What is structurally different from the substrate, it binds at a site on the enzyme molecule that is different from the site? Noncompetitive inhibitor
Mobile robots are being utilized for what laboratory service? Mobile robots are being used for specimen transport within hospitals. They are loaded with a schematic blue print of the hospital and can be programmed to go from area to area to collect and transport specimens to the proper lab. Having instrumentation that is capable of sampling directly from a primary tube or centrifuging and aliquoting the plasma or serum into a secondary tube on its own will reduce what and increase what? Reduce-Specimen evaporation, turn around time, risk of disease transmission, specimen contamination Increase-efficiency Three processes of removing protein and other interferents are? Dialysis Column chromatography Filtration In immunochemistry, instrumentation is capable of separating bound and free fractions via which methods? •coated beads •coated •microtiter plates •magnetic and nonmagnetic microparticles Define carryover in regards to specimen transport during analytical testing The transport of a quantity of analyte or reagent from one specimen reaction into and contaminating a subsequent one. What sample introduction system utilizes a peristaltic pump? Continuous-flow system What sample introduction system utilizes pipets and is highly automated. Discrete systems Instrumentation must have what capability to store reagents? Refrigeration Instrumentation must have this to identify patients and tests to be performed on each sample. Bar code scanner Rate of transport of reaction mixture through the system to the measurement station affects what? Timing of the reaction on-time events on reagent addition (or activation) relative to measurement will affect what? Timing of reaction What are the five main means of mixing reactants in instrumentation? Forceful dispensing magnetic stirring vigorous lateral displacement a rotating paddle the use of ultrasonic energy Some temperature control and efficient heat transfer methods seen in instrumentation are… Air baths water baths contact with warm plates
Lymphomas or chronic lymphocytic leukemias are examples of ___. Lymphoid tumors Where do lymphoid tumors arise? From less mature stages in B-lymphocyte development What is a malignant proliferation of plasmacytoid cells producing IgM? Waldenstroms Macroglobulinemia What causes hyperviscosity that may require treatment by plasma exchange? IgM and IgM paraproteins What is the rare disorder in which a free Ig heavy chain is produced? Heavy Chain disease What is the most common form of Heavy Chain disease? Alpha chain disease What is chain disease associated with? The heavy chain of IgA What can cause malabsorption in alpha chain disease? Intestinal infiltration with clonal lymphocytes What are serum proteins or protein complexes that precipitate at a temperature lower than normal core body temperature? Cryoglobulins What is the result of precipitation in tissues? Vasculitis and ischemic injury to peripheral tissues of the lower temperature How do you lower a patient's cryoglobulin concentrations? Patient must be kept in a warm environment What do type I cryoglobulins consist of? Monoclonal immunoglobulins, often IgM How many days are serum usually refrigerated for cryoglobulin analysis? 7 days What is the next step after 7 days? Centrifugation What happens when cryoglobulins that precipitated in the cold are warmed back to 37C? They should redissolve What is a pathological process in which proteins are deposited in tissues as aggregates (amyloid), and form b-sheet structures? Amyloidosis What can result from amyloidosis? Kidney failure, heart failure, and peripheral neuropathies What are stable structures resistant to proteolysis and normal clearance mechanisms? Aggregates Aggregates accumulate over time. TRUE What happens when there is increased production of specific proteins, genetic variants of proteins, or fragments of proteins? Formation of amyloid Where are the tissues obtained in order to diagnose amyloidosis? From fat aspirates, rectal biopsy specimens, or biopsy specimens of affected tissues What can the b-sheet structure of the amyloid be stained with? Congo red dye What occurs as the result of increased concentrations of proteins including... AL amyloidosis
What are three clinical significances of LD3 when elevated? Pulmonary edema Pulmonary infarction Pancreatitis What are two clinical significances of LD4 and LD5 when elevated? Liver disease Skeletal muscle disease The LD isoenzyme pattern in renal disease is very similar to a normal pattern except for the higher _____________? Absolute values Define Diagnostic Window. The interval of time after an episode of injury during which plasma concentrations of the marker are increased, thereby demonstrating the occurrence of injury When does CK activity increase? 4-6 hours after the symptoms When does CK activity peak? 12-24 hours When does CK activity return to normal? 48-72 hours When does AST activity increase? 6-8 hours after the symptoms When does AST activity peak? 24-46 hours When does AST activity return to normal? 4th of 5th day When does LD activity increase? 8-12 hours after the symptoms When does LD activity peak? 48-60 hours How long does LD activity remain elevated? 10-14 days What does AMI stand for? Acute Myocardial Infarction What is a myoglobin? A heme protein that can bind oxygen reversibly found in cardiac and striated muscles How long does myoglobin take to increase after infarction? 1-3 hours How long does myoglobin take to double its initial volume after infarction? 1-2 When does myoglobin peak? 5-12 hours When does myoglobin return to normal? 18-30 hours What is myoglobin used in conjunction with to assess AMI? Troponin and CK-MB What is Troponin? A group of three proteins that function in muscle contraction by binding to the thin filaments of cardiac and skeletal muscles What are the 3 types of Troponin? Troponin T (TnT) Troponin I (Tn I) Troponin C (TnC) When does Cardiac troponin I (cTn I) increase after an AMI? 4-8 hours How long does Cardiac troponin I (cTn I) remain elevated after an AMI? 3-5 days When is a serial quantification of Cardiac troponin I (cTn I) necessary after an AMI? 3-6 hours 6-9 hours 12-24 hours interval
is Cardiac troponin T (cTnT) found in the body? cTnT is found in regenerating and diseased skeletal muscle expressed in small quantities What is GAMMA GLUTAMYL TRANSFERASE (GGT)? GGT is an transferase that transfers a terminal gamma-glutamyl from one compound to an acceptor compound (transpeptidation) What is an example of tissues containing significant amounts of GGT (descending order of activity per gram of tissue) ? Kidney Liver Pancreas Intestines All of the above E How is GGT activity measured? Szasz Method Nonhemolyzed serum Both A and B C How is GGT activity stored? Stable for at least one month at 4°C and for one year at -20°C. List the Clinical Significance of GGT: primarily used as a marker for the diagnosis of liver related disorders (intrahepatic or post hepatic obstruction detection of alcoholism and monitoring of alcohol intake urine contains significant amount of GGT elevated also in serum of patients receiving anticonvulsant drugs such as phenytoin and phenobarbital. All of the E Between male and female organism, who shows the higher GGT activities? Male GGT values are lowest in adolescents and rise steadily throughout life: True False A What does it usually mean when GGT and ALP enzymes are elevated and when GGT is normal but ALP is high? If both enzymes are elevated, liver disorder is suspected; If GGT is normal, and ALP is high, suggestive of bone disease What is Glutamyl Transferase (GGT)? Is an transferase What does GGT do? Transfers a terminal gamma-glutamyl from one compound to an acceptor compound (transpeptidation).
What do you call the enzyme synthesized by the liver that hydrolyzes the esters of choline? CHOLINESTERASE (CHS) What do you call an enzyme that is also known as “pseudo-cholinesterase”? CHOLINESTERASE (CHS) What does Acetylcholinesterase use as a substrate? Acetlycholine Where is Acetylcholinesterase found in the body? RBC, Spleen Gray matter, nerve endings A and B Stomach and Heart C. Acetylcholinesterase is the false cholinesterase or WBC cholinesterase. Evaluate this statement. True or False False Psuedocholinesterase uses what kind of substrate? Butyryl esters (butyrylcholine) is Psudeocholinesterase found in the body? serum, liver pancreas, gray matter A and B D. A and B Pusedocholinesterase has other reference names. Which of these is false. serum cholinesterase (SChE) or acylcholine acylhydrolase butyrylcholinesterase, choline esterase II acetylcholinesterase D. Manometric techniques measure liberated ____________ from the formation of acetic acid. Carbon Dioxide Carbon Monoxide Phosphate Dioxin Methane Gas A Electrometric techniques determine enzyme activity by measuring the ____ decrease resulting in the liberation of acetic acid. Salt Base pH Aerosol C
CA 125 is also used to detect ______ or _____ disease in patients who have undergone therapy and would be considered for second-look procedures residual or recurrent CA 125 is also elevated in non-_____ carcinoma including pancreatic, lung, breast, colorectal, gastrointestinal tumors and useful in determining the prognosis of patients with endometrial carinoma ovarian CA 125 is determined by what? determined by immunoradiometric essay CA 19-9 (sialylated Lexa) a marker for gastrointestinal cancer and is used primarily in patients with _____ _____? pancreatic carcinoma CA 19-9 has been approved by the FDA for quantitative measurement in serum and as an aid in monitoring _____ _____ patient pancreatic cancer Monoclonal immunoglobulin has been used as a marker for multiple ______? myeloma Monoclonal paraproteins appear as sharp bands in the _____ region of serum electrophoretic patterns. Globulin Bence Jones proteins is what? a free monoclonal immunoglobulin light chain in the urine What type of cancer is detected through cytoscopy or cytology of shed cells using tumor markers like NMP22, complement factor-H (CFH), fibronectin, telomerase, cytokeratins and survivin. Bladder Cancer Bladder cancer is measured what type of technique? fluorescence in situ hybridization (FISH) What type of proteins have functions that have been associated with regulating key reactions in the nucleus, such as DNA replication and RNA synthesis Nuclear Matrix Protein (NMP22) What detects the antigen human complement factor-H related protein, which is a variant of human complement factor-H Bladder Tumor Associated analytes Bladder Tumor Associated analytes functions in the alternative complement pathway by interacting with complement factor C3b to prevent what? cell lysis _______ is a cell surface glycoprotein express on mesothelial and mesothelin fragments, found in the circulation of patients with mesothelial tumor. Mesothelin
What three organs is Creatinine synthesized in? Kidneys Liver Pancreas True or False: Creatinine is present in all body fluids and secretions. True What is Creatinine a marker of? Glomerular Filtration Rate (GFR) What reaction is used in Creatinine? reaction The Jaffe reaction of Creatinine has a reaction with what? Picrate ion Creatinine analytical methods tend to lack what? Specificity Name two enzymes of Creatinine Creatininase creatinase What are four analytical methodology factors that are used in Creatinine? Enzymatic methods (Fig. 21-1) Hydroxide Wavelength Temperature “Concentration” What type of state is Urea? Azotemic state Urea is an underestimation of what? Glomerular filtration rate Urea is an outdated indicator of what function? Kidney What is a chemical method of Urea? Fearon reaction What is an enzymatic method of Urea based on? Preliminary hydrolysis of urea with urease What three types of Urea enzymatic methods approached is used? Spectrophotometric Dry Chemistry Systems Specificity What does a spectrophotometric approach consist of? Berthelot method and enzymatic What does the biochemistry and physiology of Uric Acid focus on? Urate pool Renal handling of uric acid A clinical significance of Uric Acid that involves, 20-plus inherited disorders of what metabolism? Purine metabolism Uric Acid is used as a measurement in management of what? Gout What does Uric Acid help determine on during Hypouricemia? Link to underlying conditions Other causes Name two analytical methodology of Uric Acid? Phosphotungstic acid (PTA) Uricase Which is more specific, PTA or Uricase? Uricase What does HPLC-based methods focus on during Uric Acid? Ion-exchange or reversed-phase What does reference values of Uric Acid indicate? Gout Purine-containing and purine-free diets What are Carbohydrates ? •Aldehyde or ketone derivatives of polyhydroxy (more than one -OH