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clinically significant non-protein nitrogens

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Question
Answer
What has the determination of npn in the blood been used to monitor?   renal function  
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Concentration of nitrogen containing compounds was measured spectrophotometrically by converting nitrogen to ammonia and subsequent reaction with what reagent to form what color?   Nessler's reagent. Yellow color.  
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NPN are the products of what types of catabolism?   protein and nucleic acid  
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NPN in highest concentration in blood?   Urea  
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Excretory product of protein and AA metabolism   Urea  
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Synthesized in liver from ammonia and CO2 from the deamination of AA in urea cycle?   Urea  
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Filtered by the glomerulus. 40% reabsorbed in tubules.   Urea  
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Amount of urea reabsorbed depends on what?   Urine flow rate and hydration  
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Conversion of BUN to urea   multiply by 2.14  
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How much nitrogen does 60 grams of urea contain?   28 grams nitrogen  
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Refers to elevated levels of urea in the blood   azotemia  
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Refers to high levels of urea accompanied by renal failure   uremia  
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Pre-renal causes of azotemia   Reduced blood flow->less urea filtered dehydration, shock, corticosteroid therapy, high protein diet, fever, CHF  
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Renal causes of azotemia   decreased renal function. glomerular-nephritis, tubular necrosis, chronic renal failure, intrinsic renal disease.  
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Post-renal causes of azotemia   urinary tract obstruction - stones, tumor, severe infection  
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Causes of decreased plasma urea   decreased protein intake, late pregnancy, infancy, severe liver disease  
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Tests used to differentiate the cause of azotemia   BUN and Creatinine  
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Normal BUN:Creatinine ratio   10:1 - 20:1  
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Pre-renal conditions have what levels of BUN and Creatinine. What ratio?   increased BUN, Normal Creatinine. High BUN:Creatinine ratio  
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Post-renal conditions have what levels of BUN and Creat? What ratio?   increased BUN, increased Creat. High ratio.  
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Conditions resulting in decreased BUN:Creat ratio   decreased urea production. decreased protein intake, tubular necrosis, liver disease  
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First step in enzymatic methods for urea   conversion of urea to ammonia in presence of urease  
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Glutamate dehydrogenase (GLDH) coupled reaction for urea   kinetic method that measures rate of disappearance of NADH at 340nm.  
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NH4 + 2-oxoglutarate + NADH -> Glutamate +H2O + NAD+   GLDH coupled enzymatic reaction for urea  
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NH3+ + pH indicator -> color change   indicator dye method for urea  
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Conductometric method for urea   conversion of unionized urea to NH4+ and HCO3- results in decreased conductivity. Specific and Rapid.  
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Advantages of conductometric method   specific and rapid. ammonia does not interfere.  
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NH4+ + 5NaOCl + 2phenol -(nitroprusside/OH-)-> indophenol + 5NaCl + 5H2O -indophenol colored.   Berthelot Method for urea  
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What are disadvantages to Berthelot method?   Non-specific. Sensitive to interference from endogenous ammonia.  
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Measurement of urea by condensation with diacetyl monoxime in the presence of a strong acid and an oxidizing agent.   Forms diazine which is yellow and is measured at 540 nm  
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Disadvantages of diacetyl monoxime method / advantages   toxic reagents, non-specific ammonia does not interfere.  
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o-Phthaldehyde and naphthylethylenediamine react with what to form a chromogen?   urea  
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Proposed definitive method for urea   isotope-dilution mass spectrometry  
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Reference method for urea   urease/GLDH coupled enzymatic reaction  
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What inhibits urease and should be avoided for specimen collection?   sodium citrate and sodium fluoride  
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Why should specimens for urea analysis be refrigerated?   Urea is susceptible to bacterial composition  
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Specimens for urea concentrations   plasma, serum, or urine  
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Reference ranges for BUN serum and urine   6-20 mg/dL and 12 - 20 g/day.  
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conversion of mg/dL to mmol/L   multiply by 0.36  
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Who has slightly higher levels of urea?   adults over 60 and males  
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Who has slightly lower urea levels?   pregnant women and children  
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Synthesized in the liver from arginine, glycine and methionine   creatine  
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What is creatine converted to after being transported to muscle and what is it good for? What enzyme involved?   phosphocreatine - high energy source for muscle building. Creatine Kinase  
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How much creatine is converted to creatinine daily?   2%  
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Waste product of muscle metabolism   creatinine  
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What is formed when creatine phosphate loses phosphoric acid or when creatine loses water.   creatinine  
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How is creatinine excreted?   filtered by glomerulus and excreted in urine  
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What is plasma creatinine conc. a function of?   muscle mass, creatine turnover, and renal function  
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Causes of increased plasma and urine creatine   muscular dystrophy, polio, hyperthyroidism, trauma  
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Is creatinine elevated in cases of increased creatine?   creatinine is normal  
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Are plasma creatine levels elevated in renal disease?   NO  
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Are serum creatinine levels affected by a high protein diet? Muscle mass?   NO. Yes, proportional to muscle mass.  
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Why is Creatinine is a reliable indicator of kidney function as it relates to glomerular function?   1) It's filtered completely through glomerulus and not reabsorbed by nephron tubule 2) It's an endogenous substance  
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measure of the ability of the glomerulus to filter blood   creatinine clearance  
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Equation for creatinine clearance   urine creat x urine vol / plasma creat x 1440  
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correction factor for body mass   x 1.73/A  
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plasma conc of creatinine is ______ to creatinine clearance   inversely proportional  
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Reference Ranges for Creatinine Clearance   Male: 97-137 mL/min per 1.73 m2 Female: 88-128 mL/min per 1.73 m2  
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What is the Jaffe reaction   based on principle that creatinine reacts with picrate in an alkaline soln to form an orange-red complex.  
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Disadvantages of Jaffe   nonspecific and subject to positive interference from acetoacetate, acetone, glucose, ascorbate,pyruvate, cephalosporins  
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Adsorbent for jaffe   Fuller's Earth or lloyds reagent  
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Dry slide method for creatinine   coupled enzymatic reaction using creatininase, creatinase, sarcosine oxidase, and peroxidase.  
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method for creatinine using reagent strip   3,5-dinitrobenzoic acid (DNBA) + creatinine = purple product  
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Interferences of dry slide method   some + due to lidocaine. Not affected by acetoacetate or cephalosporins.  
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Proposed reference method for creatinine   HPLC  
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Specimen collection for creatinine   plasma, serum, or urine. Avoid hemolysis and icterus (esp Jaffe), refrig urine and adjust to pH 7  
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Major product of purine metabolism   uric acid  
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where is uric acid synthesized and where excreted   liver adn kidney  
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what form is nearly all uric acid in plasma   monosodium urate ions  
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At what conc can urate in plasma form crystals and precipitate?   >6.4 mg/dL  
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Disease states associated with hyperuricemia   gout, increased metabolism of cell nuclei(chemotherapy), renal disease, hemolytic/megaloblastic anemia, Lesch-Nyhan syndrome, dietary intake of purines, G6PD def, toxemia of pregnancy and lactic acidosis  
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Clinical Significance of hypouricemia   secondary to liver disease ro defective tubular reabsorption (Fanconi's syndrome), chemo with 6-mercaptopurine or azathioprine. Overtreatment with allopurinol  
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uric acid is readily oxidized to what in the presence of what   allantoin in the presence of uricase  
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caraway method for urea   oxidation of uric acid with subsequent reduction of phosphotungstic acid to tungsten blue at 650-700 nm. Lacks specificity.  
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what provides alkaline pH for phosphotungstic method   sodium carbonate  
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spectrophotometric method for uric acid using uricase   measures decrease in absorbance at 293 nm as uric acid is converted to allantoin by uricase  
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interferences in simple uricase method   proteins reduce sensitivity. Neg interference from hemoglobin and xanthine  
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Coupled enzymatic reactions for uric acid   measure hydrogen peroxide produced as uric acid is converted to allantoin.  
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What catalyzes the indicator reaction in uric acid coupled reactions   peroxidase or catalase  
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Interference in peroxide coupled reactions   bilirubin and ascorbic acid destroy peroxide. minimize with potassium ferricyanide or ascorbate oxidase  
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Uric Acid Ref Ranges Male:   3.5 - 7.2 mg/dL  
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Uric Acid Female   2.6 - 6.0 mg/dL  
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uric acid child   2.0 - 5.5 mg/dL  
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uric acid 24hr urine   250 - 750 mg/dL  
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uric acid increase or decrease with age, pregnancy, children, premenopause   increase,increase,decrease,decrease  
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Specimen for uric acid   plasma, serum, urine. separate from RBCs ASAP  
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Falsley increase uric acid   salicylates and thiazides  
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Falsely decrease uric acid   hemolysis, icterus  
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Deamination product of AAs due to work of digestive and bacterial enzymes on proteins in GI Tract   Ammonia  
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Released from skeletal muscle in exercise   ammonia  
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what happens to ammonia in liver   consumed by parenchymal cells in production of urea  
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are plasma ammonia conc dependent on renal function   no  
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what are high levels of ammonia associated with   being neurotoxic and associated with encephalopathy  
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what is ammonia conc useful for   monitoring progress of severe clinical conditions  
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High levels of ammonia seen in   liver disease, Reye's syndrome (children treated with asprin for virus), Renal failure, inherited defects in urea cycle  
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all enzymes in urea cycle are deficient in inherited disorders except for   argininosuccinase  
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ion exchange method for ammonia   NH3 adsorbed onto Dowex 50 resin, eluted, and quantitated with Berthelot reagent  
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ISE method for ammonia   measures change in potential as pH of ammonium chloride soln as ammonia diffuses across membrane  
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Enzymatic method for ammonia   GLDH. measures decrease in absorbance at 340 nm as NADPH is reduced to NADP+  
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Adult ammonia ref range   11-35 umol/L plasma 10-107 mmol N/day  
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child ammonia range   40 - 80 umol/L (higher than adults)  
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Specimen for ammonia   heparin or EDTA. Place on ice immediately. Centrifuge at 0-4C within 20min and remove plasma. Assay ASAP or freeze at -20C  
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errors causing increased Plasma ammonia levels   smoking,ammonium salts, asparginase, barbituates, diuretics, ethanol, analgesics, hemolysis  
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errors causing decreased plasma ammonia   diphenhydramine, lactobacillus acidophilus, levodopa, antibiotics  
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what causes reduction of nitrates to nitrites in urine   gram negative bacteria  
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