Chemistry

Define diluent, solute, solvent, solution, molar solution. Solve the following: - A buffer is made by adding 2 parts of solution "A" to 5 parts of solution "B". How much of each solution is needed to make 70 mL of the buffer? How would you prepare 500ml of 0.85% saline? Is this w/w or w/v solution? 4.25ml of saline to 495.75ml of DI water. w/v. How would you prepare 500ml of 10% bleach solution? Is this a w/w or w/v solution? 50ml bleach to 450ml water 1:10 dilution Write directions for preparing one liter of a 2 m (mol/L) solution of NaOH.

- 70/7= 10 - 2x10= 20 - 5x10= 50 - So, you would take 20 mL of solution A and 50 mL of solution B to get 70 ml buffer solution. - C1V1=C2V2 - 1 MXV1= 100 mL X 0.1 M = V1= 100 mL x 0.1 M / 1 M = V1= 10 mL - The 10 mL of the 1 M HCl is added to 90 mL of H2O to make the 100 mL of 0.1 M HCl solution. - 500 mL = 5 x 0.85% g = 4.25 g - Use the weight out of 4.25 g of saline to add it to the 4.95.75 mL of DI water to the W/V. Diluent: a substance used to dilute something. Solute: a dissolved substance especially a component of a solution present in smaller amount than the solvent. Solvent: A solvent is a substance that dissolves a solute, resulting in a solution. Solution: is a special type of homogeneous mixture composed of two or more substances. In such a mixture, a solute is a substance dissolved in another substance, known as a solvent. Molar Solution: is another standard expression of solution concentration.

State the importance and value of measuring glycosylated hemoglobin levels compared to a fasting glucose level.

A blood test can measure the amount of glycosylated hemoglobin in the blood. The glycosylated hemoglobin test shows what a person's average blood glucose level was for the 2 to 3 months before the test. This can help determine how well a person's diabetes is being controlled over time. A fasting blood glucose test can be useful to see how well the body is able to manage blood sugar levels in the absence of food. When we do not eat for several hours, the body will release glucose into the blood via the liver and, following this, the body's insulin should help to stabilize blood glucose levels.

Draw and define the following pipet types: Ostwald folin, mohr, serological, and volumetric. Include marking interpretation, etched rings, and correct reason for use.

A pipette is a piece of volumetric glassware used to transfer quantitatively a desired volume of solution from one container to another. Pipettes are calibrated at a specified temperature (usually 68°F [20°C] or 77°F [25°C]) either to contain (TC) or to deliver (TD) the stated volume indicated by the etched/painted markings on the pipette side. Pipettes that are marked TD generally deliver the desired volume with free drainage; whereas in the case of pipettes marked TC the last drop must be blown out or washed out with an appropriate solvent. Ostwald Folin: Used for more viscous liquids, similar to a volumetric pipet in that there is a bulge at the bottom. However, the bulge is more rounded than that of a volumetric pipet. The Ostwald folin is a TC blow out pipet. Mohr: Has a maximum and minimum volume. They are particularly tricky as you can deliver too much volume and ruin your measurement. TD self-draining. Serological: Has a maximum and minimum volume. They are particularly tricky as you can deliver too much volume and ruin your measurement. TC blow out. Volumetric: Similar to the Ostwald folin in shape in that there is a bulge in center. The bulge in a volumetric pipet is more barrel shaped than that of the more bulbous Ostwald folin pipet. TD self-draining.

State the significance of an increased sweat chloride value, delineate procedure used for obtaining a result.

A sweat test measures the amount of chloride, a part of salt, in Sweat. It is used to diagnose cystic fibrosis (CF). People with CF have a high level of chloride in their sweat. CF is a disease that causes mucus build-up in the lungs and other organs. In the first part of the test, a colorless, odorless chemical (pilocarpine) and a little electrical stimulation is applied to a small area of the arm or leg to encourage the sweat glands to produce sweat. A person may feel tingling in the area, or a feeling of warmth. This part of the test lasts about five minutes. To understand what the sweat test results mean, a chloride level of: Less than or equal to 29 mmol/L = CF is unlikely regardless of age. Between 30 - 59 mmol/L = CF is possible and additional testing is needed. Greater than or equal to 60 mmol/L = CF is likely to be diagnosed.

Relate the following analytes with the disease state associated with an abnormal value: ACP ALT Copper pro-BNP ALP Indirect bilirubin Ammonia lactic acid CK Direct bilirubin Creatinine Vitamin B12 Amylase Uric acid Creatine 25-hydroxy Vitamin D

ACP- Elevated in prostate cancer ALP - elevated in liver 7& bone diseases. Elevated in hepatitis, cirrhosis CK - Elevated with AMI, muscular dystrophy. • Amylase- Elevated in acute pancreatitis, other abdominal diseases, mumps. ALT and SGPT: Primarily liver, hepatitis x100; Elevated in liver disease, Hepatitis Indirect bilirubin - Elevated in prehepatic, post hepatic and some types of hepatic jaundice Direct bilirubin - Elevated in liver disease, obstructive jaundice. Uric acid - Elevated in gout, renal failure, ketoacidosis, lactate excess, high nucleoprotein diet, leukemia, lymphoma, polycythemia. Decreased in renal tubular defects an after administration of ACTH. Copper - Deficiency in malnutrition, malabsorption, chronic diarrhea, hyperalimentation, observed in premature infants. Elevated in smoking, inflammatory conditions, pregnancy and estrogen Ammonia - Elevated in liver disease, hepatic coma, renal failure and Reyes syndrome • Creatinine - Elevated in kidney disease Creatine - Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), long-term memory deficits, Alzheimer's disease, and stroke. Pro-BNP - renal failure Lactic acid - signs of decreased O2 in tissue Vitamin B12 - megaloblastic anemia (pernicious anemia) 25-hydroxy Vitamin D - HIV disease progression

Explain how water and sodium concentrations are regulated by ADH and aldosterone levels, state the relationship between aldosterone and ADH and list diseases associated with abnormal aldosterone and ADH levels.

Aldosterone Aldosterone regulates the balance of salt and water in the body and can trigger the release of ADH Elevated Cushing's Disease Decreased Addison's Disease ADH (vasopressin) is secretion if stimulated Produced in Hypothalamus (HT) Stored and secreted from the posterior pituitary gland Target tissue is the kidney ADH controls the water balance Diabetes insipidus (too little ADH); symptoms look like diabetes mellitus

Relate elevated or decreased aldosterone levels to primary Cushing's or Addison's disease. Be able to draw the adrenal cortex hormone axis.

Aldosterone Elevated Cushing's Disease Decreased Addison's Disease Aldosterone regulates the balance of lytes and can trigger the release of ADH ADH (vasopressin) is secretion is stimulated o Produced in Hypothalamus (HT) Stored and secreted from the posterior pituitary gland Target tissue is the kidney ADH controls our water balance Diabetes insipidus (too little ADH); the symptoms looks like diabetes mellitus

Classify, amikacin, amitriptyline, carbamazepine, digoxin, disopyramide, gentamicin, lidocaine, lithium, phenobarbitol, nortriptyline, phenytoin, primidone, procainamide, NAPA, quinidine, theophylline, tobramycin, valproic acid, and vancomycin, into therapeutic drug types (cardiac, antiepileptic, etc).

Analgesic (Pain): (Acetaminophen)-Tylonel and (Salicylic Acid)-Aspirin. Antiasmatic (Asthma): (Theophylline)-Aminophylline and Caffeine. Anticonvulsant (Epilepsy): (Carbamazepine)-Tegretol, (Ethosuximide)-Zartonin, Primidone (Mysoline) and Valproic Acid (Depekene). Antibiotic (Bacterial Infection): Gentamicin (Garamycin), Tobramycin (Nebcin), Amikacin (Amkin), Chloramphenicol (Chlormycetin), and Vancomycin (Vancocin). Cardiacs (Antiarrhythmic): Disopyramide (Norpace), Lidocaine (Xylocaine), Procainamide/NAPA (Pronestyl), Digoxin (Lanoxin), and Quinidine (Kinidin). Anti-Psychotic (Depression or Schizophrenia): Amitriptyline (Elavil), Nortriptyline (Aventyl), Imipramine (Tofranil), Doxepin (Sinequan), and Lithium.

Define anion gap. Calculate anion gap for the following values Na=145mmol/L, K=4.2 mmol/L, Cl=100mmol/L, HCO3= 28 mEq/L. Is the Anion gap normal?

Anion gap - Difference between unmeasured anions and unmeasured cations. Normal range (Na++K+ )-(Cl-+HCO3 - ) is 10-20 or NA+ -(CI-+HCO3 - ) is 7-16 145-(100+28)=17 (145+4.2)-(100+28)=21.2. The anion gap is abnormal.

Identify bands seen on serum protein electrophoresis gels and if abnormal state the disease state that may be present. List migration order of bands from anode to cathode and from cathode to anode.

Anode (+) Albumin, α1, α2, β, γ (-) Cathode Albumin Decreased in Hepatic disease, Impaired digestion, Nephrotic syndrome, and Burns Increased in rare except in shock α1 Increased in Inflammation and Spina Bifida α2 Decreased in Liver disease Increased in Nephrotic syndrome, SLE, and RA β Increased in Inflammation and Lipid disease Γ Decreased in α-Gammaglobulinemia and Hypogammaglobulinemia Increased in Multiple myeloma, Waldenstroms, MGUS, Chronic infection.

Define azotemia; uremia. State the normal BUN:creatinine ratio. List reasons for an increased ratio. Name methods used to quantify BUN in serum.

Azotemia: is a biochemical abnormality, defined as elevation, or buildup of, nitrogenous products (BUN-usually ranging 7 to 21 mg/dL), creatinine in the blood, and other secondary waste products within the body. Uremia: is a clinical syndrome marked by elevated concentrations of urea in the blood and associated with fluid, electrolyte, and hormone imbalances and metabolic abnormalities, which develop in parallel with deterioration of renal function. Normal BUN: 10-20 BUN/Creatine ratio. A high BUN value may be caused by a high-protein diet, Addison's disease, tissue damage (such as from severe burns), or from bleeding in the gastrointestinal tract. High BUN-to-creatinine ratios occur with sudden (acute) kidney problems, which may be caused by shock or severe dehydration.

Identify the disease correlation(s) associated with abnormal urea, creatinine, creatine, uric acid and ammonia serum levels. Discuss dietary effects for each analyte. BUN/urea:

BUN/urea Pre-renal Heart failure Shock Dehydration Decreased blood volume High protein diet Renal Nephro-Glomerulonephritis Renal failure Tubular necrosis Nephrotic syndrome Post-renal Calculi, stones Renal obstructions Creatinine Blood levels reflect muscle mass and turnover of creatine; elevates with renal disease Least affected by diet of NPNs Creatine Synthesized in liver Elevates with muscle disease Converted to phosphocreatine in muscle (high energy source) When energy is needed Creatinine is formed and enters plasma Uric acid Increased: Gout Ammonia Breakdown product from proteins Levels not dependent on renal function

Discuss compensation and buffer systems responsible for maintaining arterial blood pH.

Bicarbonate Buffer System (20:1 ratio bicarbonate to carbonic acid) Lungs (pCO2) Metabolic (Bicarbonate or Total CO2) Carbonic acid ↔ Bicarbonate + H+ • Chloride Shift Chloride and HCO3- ions can move between the ICF (inside the RBC) and ECF (plasma) o They trade places to maintain charge balance Phosphate Buffer System ( Excretion) o Phosphates are found in the plasma and RBC o They are active in the renal system for excreting H ions Hemoglobin Buffer System o H ions can bind to hemoglobin to control pH The buffer systems functioning in blood plasma include plasma proteins, phosphate, and bicarbonate and carbonic acid buffers. The kidneys help control acid-base balance by excreting hydrogen ions and generating bicarbonate that helps maintain blood plasma pH within a normal range.

State the principle and list the major reagents used in the following methods of analysis: Biuret Jaffe Jendrassik-Grof

Biuret: is the total protein using alkaline copper solution causing a reaction with peptide bonds for a color change of purple. Thus, causing a color intensity from the concentration. Jaffe: is the creatinine with alkaline picric making creatinine picrate. This can lead to problems with picric acid. Jendrassik-Grof: is the bilirubin. You take the patients serum and mix it with sodium acetate, diazotized sulfanilic acid and ascorbic acid and alkaline tartrate causing a color change of blue. 600nm.

Define and discuss CEA, alpha fetoprotein, hCG as related to type of tumor marker. Include advantages and disadvantages of use as a tumor marker.

CEA (Carcinoembryonic Antigen) Liver and Colon tumor markers Increased in smokers Alpha Fetoprotein Ovarian tumor markers o Oncofetal markers are present in fetal development and then disappear hCG Testicular tumor markers • Advantages As tumor gets worse it metastasizes, the marker levels increase Can predict treatment staging Aid in a predictor of disease outcome Successful treatment Disadvantages There are no ideal tumor markers to date

State the possible clinical significance of elevated CK, AST, ALP, ALT, ACP, GGT, amylase, lipase, and LD levels in serum based on textbook and MLAB 2220 information.

CK • "Creatine Kinase" Indicates general disease state of muscle tissue or brain. Isoenzymes are more specific for the tissue affected. AMI (CK-MB) Duchenne muscular dystrophy (50-100 x ULN) o Stroke; cerebral damage (CK-BB) o Muscle disease and damage (CK-MM) • AST (SGOT) Aspartate Cardiac, liver and skeletal muscle Indicative of; AMI, congestive heart failure, pulmonary embolism, acute hepatocellular disorders; skeletal muscular disease • ALP Obstructive jaundice ▪ Post hepatic disease • ALT (SGPT) Alanine Liver Acute hepatic disorders (usually higher than AST) • ACP Bone, prostate, (kidneys, RBC, liver, spleen) • GGT Liver, Kidney , prostate, pancreas and brain Often to occult alcoholics • Amylase Pancreas and salivary glands Acute pancreatitis and mumps indicator • Lipase Pancreas (stomach and SI) Acute pancreatitis, cholecystitis, intestinal obstructions • LD o AMI 1>2 Liver disease (viral hepatitis, cirrhosis) Anemias ▪ Lymphoblastic leukemia, Muscle disease and damage

Discuss CK and LD isoenzymes, myoglobin, troponin, sensitive CRP and BNP, including source and importance in assessing a myocardial infarct. Include the significance of a "flipped" LD pattern; and list the timeline for each analyte increase after a myocardial infarct.

CK: is found in brain (BB), heart (MB), and skeletal muscle (MM). CK will rise within 3-6 hours of MI o Peaks at 24hrs o Back to normal within 3-4 days. CK-MB will rise above 6%. LD: are Isoenzymes of HHHH (Heart and RBC) LD1: HHHM (Kidney) LD2: HHMM (Lungs) LD3: HHMM (Muscle and Liver) LD4: MMMM (Muscle and Liver) LD5: Will rise in 12-24hrs of MI o Peak in 3-4 days and back to normal within 7-10 days LD1 > LD2 Myoglobin: is the heme protein found only in skeletal and cardiac muscle and will rise in 3-4hrs on MI. Troponin: has 3 complex of proteins that bind to the thin filaments of striated muscle (cardiac and skeletal). These levels rise within 4 hours following an MI o Remain elevated for 10-14 days. CRP: is in the blood of patients with diverse inflammatory diseases but is undetectable in healthy patients; Synthesized in the liver o Elevated in MI. BNP is used in specific for cardiac failure and elevated in MI

Compare chylomicrons, VLDL, LDL, and HDL as to composition, size, density and other nomenclature.

Chylomicrons: Floats on top and causes the triglycerides to be less dense. VLDL: is before beta, can be formed in the live and carries triglycerides to the tissue, density is 1.006-1.94. LDL: present in beta, made in VLDL and is carries from Cholesterol to tissue. Known as BAD Cholesterol. Density is 1.063-1.063. HDL: present in alpha, from the liver and intestines, Cholesterol in the tissue is takes to the liver to be excreted. Known as the Good Cholesterol. Density is 1.063-1.21.

State which areas of the serum protein electrophoresis patterns exhibit abnormalities for the following: Cirrhosis Acute inflammation Nephrotic syndrome Chronic Infection Alpha 1 antitrypsin deficiency monoclonal gammopathy Plasma polyclonal gammopathy

Cirrhosis: is elevated in polyclonal in gamma with the beta gamma bridging. Nephrotic syndrome: has decreased albumin and has an increase in alpha 2. Alpha 1 antitrypsin: has a decreased in alpha 1. Plasma: has fibrinogen, extra bands between beta and gamma. Acute inflammation: as elevated levels in alpha 1, and alpha 2. Chronic infection: has elevated levels in alpha 1, alpha 2 and gamma. Monoclonal gammopathy: elevation in 1 immunoglobulin causing a M spike and decrease in other fractions. Polyclonal gammopathy: has a defused elevation in gamma.

Describe continuous flow, centrifugal, and discrete analysis by definition and give examples of each analyzer.

Continuous Flow- liquids (reagents, diluents, and sample) are pumped through a system of continuous tubing. Samples are introduced in a sequential manner, following each other through the same network. A series of air bubbles at regular interval serve as separating and cleaning media • Ex: DXC,DXI • Centrifugal Analysis- analytical techniques that uses the force generated by centrifugation to transfer and then contain liquids in separate cuvettes for measurement at the perimeter of spinning rotor • Ex: DXC,DXI • Discrete Analysis- approach to automated analysis in which each sample and accompanying reagents are in a separate container. They have the capability of running multiple tests one sample at a time, or multiple samples one test at a time • Ex: DXC, DXI

State the generic and brand names for digoxin, phenytoin, carbamazepine, theophylline, and valproic acid.

Digoxin: brand or generic name is Lanoxin, Phenytoin: brand or generic name is Dilantin Carbamazepine: brand or generic name is Tegretol Theophylline: brand or generic name is Aminophylline Valproic Acid: brand or generic name is Depekene

List the NTR (normal therapeutic ranges) for digoxin, phenytoin, carbamazepine, theophylline, and valproic acid.

Digoxin: normal range is 0.5-2.0 ng/mL Phenytoin: normal range is 10-20 mcg/mL Carbamazepine: normal range is 40-10 ug/mL Theophylline: normal range is 10-20 mcg/mL Valproic Acid: normal range is 50-100 mcg/mL

Describe what is meant bycircadian/diurnal and give examples of analytes that are diurnal/circadian in nature.

Diurnal rhythms. - a circadian rhythm that is synchronized with the day/night cycle. Ultradian rhythms. - biological rhythms (e.g. feeding cycles) with a period much shorter (i.e., frequency much higher) than that of a circadian rhythm. There are many examples of circadian rhythms, such as the sleep-wake cycle, the body-temperature cycle, and the cycles in which a number of hormones are secreted. Infradian rhythms have a period of more than 24 hours. The menstrual cycle in women and the hibernation cycle in bears are two good examples.

Describe types and the use of tumor markers in the laboratory and their advantages and disadvantages. (Include alpha fetoprotein, CEA, hCG, PSA, CA125, CA 15-3, CA27-29, CA19-9)

Enzyme: tumor markers are PSA, ACP (prostate) and ALP (bone) Hormones: tumor marker are B HCG (testicular) and ACTH (lung). Oncofetal: tumor markers are CEA (liver and colon) and AFP (ovarian). CA19-9 (pancreatic) CA125 (ovarian) CA1-15-3 (breast) BRCA-1 and 2 (breast and ovarian)

Explain the relationship between total iron binding capacity and serum iron. Define ferritin, transferrin and % saturation. List conditions indicated by low ferritin levels and high ferritin levels.

Ferritin - a spherical protein shell composed of 24 subunits with a molecular mass of 500 kDa. The protein can bind up to 4,000 iron molecules, making it a large potential source of iron. Ferritin is decreased in iron deficiency anemia and increased in iron overload an HH. Transferring Saturation %- percent of transferrin molecules that have iron bound. A ratio of serum iron (actual iron in the serum) and serum transferrin or TIBC (potential quantity of iron that can be bound). Serum iron/TIBC=% saturation. Transferrin elevated in iron deficiency anemia. Transferrin decreased in iron-overdose, hemochromatosis, chronic infection, malignancy.

Describe classical and current methodologies available for glucose determination, including principle, reagents, limitations, and names of the methods.

Folin Wu, Nelson somogi and Benedicts; Clinitest urine Measures all redusing sugars Molybdenum blue-Cu++ and glucose react to form red color when copper is reduced. Reacts with sugars (not sucrose or polysaccharides). • Orthotoluidine Measures all aldose sugars Schiffs base reaction resulting in a green color. Reacts with aldose and hexose sugars Hexokinase Measures all sugars with 6 carbons Reacts with hexose sugars. Uses NADPH+ at 340nm GOD Specific for Beta D-glucose Uric acid, Bilirubin, and Vitamin C can interfere with test causing elevated values Glucose + oxygen → hydrogen peroxide + gluconic acid o Hydrogen peroxide + dye chromagen → color absorbance change Glycated hemoglobin (glycosalated) Measures HgbA1c, is an indicator of glucose values for a 8-10wk span

Recognize total T4, free T4, T3U and TSH values, seen in primary hypo- and hyperthyroidism versus pregnancy and kidney disease.

Free T3 and T4 not affected by TBG; pregnancy have increased tbg levels and liver disease or renal disease have decreased levels Hypothyroidism Total T4 ↓ Free T4 ↓ T3U ↓ TSH ↑ Hyperthyroidism Total T4 ↑ Free T4 ↑ T3U ↑ TSH ↓ Pregnancy Total T4 ↑ Free T4 Not affected T3U ↓ TSH ↓ Kidney Disease Total T4 ↓ Free T4 Not affected T3U ↑ TSH ↓

State the techniques that may be used to perform serum or urine osmometry.

Freezing Point Depression has sample at -7C. The freezing point of a solution is less than the freezing point of the pure solvent. This means that a solution must be cooled to a lower temperature than the pure solvent for freezing to occur. The sample attains equilibrium through a series of freezes and thaws and the drop in freezing point from 0 is related to osmolality or particle content. Farther the drop larger the number of particles present or higher the osmolarity. Vapor Pressure Depression is when a sample is placed on filter disk in sealed chamber. Dew points of these sample is measured. The drop in dew point is related to the vapor point pressure. The farther the drop the greater the osmolality or more particles present. Vapor pressure lowering is a colligative property of solutions. The vapor pressure of a pure solvent is greater than the vapor pressure of a solution containing a nonvolatile liquid. This lowered vapor pressure leads to boiling point elevation. When something is volatile is able to evaporate very quickly. Ranges: Serum Normal: 275-300 m0m/kg Urine Normal: 300-900 m0m/kg

Differentiate GC, HPLC, TLC, ion exchange, and column chromatography as to stationary phase, mobile phase, additional components, and principle.

Gas Chromatography Substances are volatized and partitioned and related to retention time Mobile phas is N2, He or Ar Stationary phase is inert packing; diatomaceous earths; polymers and beads Samples cannot be reused; oven, detectors and readouts; blood alcohols; confirmatory methods HPLC (High performance liquid chromatography) Solvent is pumped through column and retention time is related to compound Mobile phase is organic solvents Stationary phase is Inert packing; silica gel, C18 Therapeutic drug analysis; detectors and readouts; confirmatory methods TLC (Thin layer chromatography) Solvent by capillary action moves up the stationary phase and partition occurs Mobile phase is organic solvents Stationary phase is silica, cellulose, alumina, detrains Uses Rf calculations Screening Test Ion Exchange Cations or anions either attach by charge to resin or are eluted Mobile phase is liquids of differing pH and ionic concentration and composition Stationary phase is cation and anion resins Hg A1C Column Chromatography Large particles excluded by small pores Mobile phase is organic solvents Stationary phase is Beads; dextran, polyacrylamide, agarose Research

Differentiate GC, HPLC, TLC, ion exchange, and column chromatography as to stationary phase, mobile phase, additional components, and principle.

Gas Chromatography: is a substance that volatized and partitioned and related to retention time. The mobile phase is N2, He or Ar. The stationary phase is inert packing, polymers, and beads. The samples cannot be reused. HPLC (High performance liquid chromatography): is a solvent that is pumped through column and retention time is related to compound. The mobile phase is organic solvents, and the stationary phase is inert packing with silica gel, known as C18. TLC (Thin layer chromatography): is the solvent by capillary action moves up the stationary phase and partition occurs. The mobile phase is organic solvents, and the stationary phase is silica. Screening Test is used. Ion Exchange: is cations or anions that are attached by charge to resin. The mobile phase is liquids of differing pH and ionic concentration and composition and the stationary phase is cation and anion resins. Known as Hg A1C. Column Chromatography: are large particles excluded by small pores. The mobile phase is organic solvents, and the stationary phase is beads.

Define gluconeogenesis, glycolysis, glycogenesis, and glycogenolysis

Gluconeogenesis: a metabolic pathway that results in the generation of glucose from certain non-carbohydrate carbon substrates. Glycolysis: the metabolic pathway that converts glucose C₆H₁₂O₆, into pyruvate. Glycogenesis: the process of glycogen synthesis, in which glucose molecules are added to chains of glycogen for storage. Glycogenolysis: the breakdown of glycogen to glucose-1-phosphate and glycogen.

Discuss how glucose, protein and chloride CSF values change in a case of bacterial meningitis.

Glucose and Chloride: decreased Protein: increased

Differentiate HDN, acute hepatitis, cirrhosis, and biliary obstruction when looking at abnormal laboratory data.

HDN Excess levels of unconjugated bilirubin Total levels rarely above 5mg/dL No free bilirubin in urine Excess urobilinogen in feces and urine Acute Hepatitis Greatly increased ALT and AST Other liver enzymes increased Cirrhosis Greatly decreased albumin ALT and AST slightly elevated Elevated GGT if alcoholic liver Biliary Obstruction Increased Total and direct bilirubin Increased urine bilirubin Decreased fecal and urine urobilinogen Greatly elevated ALP

State the historical clinical significance of measuring prealbumin.

Historically, albumin levels have been used to determine the nutritional status of a patient, but they are relatively insensitive to changes in nutrition. Albumin has a half-life of 20 days. Serum albumin is affected by the patient's hydration and renal function. Typically it takes 14 days to return to normal when the pool has been depleted.

List and describe the function of the hormones secreted by the following endocrine glands: 33.1-Hypothalamus (CRH, TRH, PIF, GRH) 33.2-Anterior pituitary (LH, FSH, GH, TSH, PL, ACTH) 33.3- Posterior pituitary (oxytocin, ADH) 33.4- Thyroid (T3, T4, calcitonin) 33.5- Parathyroid (PTH) 33.6- Adrenal gland (aldostrone, cortisol, adrenaline, catacholamines) 33.7- Pancreas (insulin, glucagon) 33.8- Reproductive glands (androgens, testosterone, estrogens, progesterone) 33.9- GI tract (gastrin) 33.10- Pineal gland (melatonin)

Hypothalamus (CRH, TRH, PIF, GRH) CRH (Corticotrophin Releasing Hormone) Target gland/tissue is the Anterior Pituitary Stimulates the secretion of ACTH TRH (Thyotropin Releasing Hormone) Target gland/tissue is the Anterior Pituitary Causes release of TSH and PRL (prolactin) PIF (Prolactin Inhibiting Factor) Regulator on prolactin production GnRH (Gonadotropin Releasing Hormone) Target gland/tissue is the Anterior Pituitary Causes the releases of LH and FSH Anterior pituitary (LH, FSH, GH, TSH, PL, ACTH) LH (Luteinizing Hormone) Target gland/tissue is the Ovaries and Testies Responsible for the ovulation and progesterone production and testosterone production FSH (Follicle Stimulating Hormone) Target gland/tissue is the Ovaries and Testies Follicular growth and estradiol secretion Spermatogenesis GH (Growth Hormone) Target gland/tissue is the Whole body/ Peripheral tissues Responsible for growth TSH (Thyroid Stimulating Hormone) Target gland/tissue is the Thyroid Causes the releases of T3 and T4 PL (Prolactin) Target gland/tissue is the mammary glands Production of milk ACTH (Adrenocorticotropic Hormone) Target gland/tissue is the Adrenal Cortex Responsible for the release of cortisol and some aldosterone 33.3- Posterior pituitary (oxytocin, ADH) Oxytocin Target gland/tissue is the Smooth Muscle contractions Pictocin is given to promote uterine contractions ADH (Antidiuretic Hormone) Target gland/tissue is the Kidneys and ACTH release Synthesized in the hypothalamus Stored and secreted from the posterior pituitary Directly controls water balance Disease hypo ADH- Diabetes insipidus Thyroid (T3, T4, calcitonin) T3 and T4 Target gland/tissue is all tissues with physiological effect of metabolism Iodine oxidation Calcitonin Target gland/tissue is the Bones Inhibits PTH and resorption Parathyroid (PTH) PTH (Parathyroid Hormone) Target gland/tissue is the Bones and Kidneys Promotes Calcium reabsorption from kidney, Bone resorption, and Vitamin D secretion Adrenal gland (aldostrone, cortisol, adrenaline, catecholamines) Aldosterone Target gland/tissue is the Kidneys Mineral corticosteroid Physiological effect Sodium reabsorption Cortisol Target gland/tissue is the Whole Body Glucocorticosteroid Physiologic effect increases lipolysis and gluconeogenesis Adrenaline (Epinephrine) Target gland/tissue is the Heart, Blood vessels, Smooth muscle, Eye, Metabolic actions The increase in epinephrine stimulates the heart, raises blood pressure, constricts small blood vessels, releases sugar stored in the liver, and relaxes certain involuntary muscles while it contracts others. These changes in the body prepare it for fight or flight; meaning the body is more alert, physically stronger, and has greater energy Catecholamines Target gland/tissue is the Liver, Muscle, Adipose Fight, Flight, and Fright Increased blood pressure and catecholamines Pancreas (insulin, glucagon) Insulin Target gland/tissue is all of the Tissues Beta cells isles of Langerhans Decreases plasma glucose Glucagon Target gland/tissue is the Liver Alpha cells isles of Langerhans Increases plasma glucose Reproductive glands (androgens, testosterone, estrogens, progesterone) Androgens Produced in the Testes Testosterone Target gland/tissue is the Whole Body Produced in the Testes Estrogens Target gland/tissue is the Uterus Produced in the Ovaries Progesterone Prepares uterus for pregnancy Produced in the Ovaries GI tract (gastrin) Gastrin Target gland/tissue is the Stomach Increases secretion of HCl Pineal gland (melatonin) Melatonin Target gland/tissue is the Brain and Smooth Muscle Neurotransmitter Serotonin Target gland/tissue is the Hypothalamus and Anterior Pituitary Suppresses GH and GnRH release

State the function of insulin, glucagon, epinephrine, ACTH, glucocorticoids, and thyroid hormones in plasma glucose regulation.

Insulin Allows glucose to enter tissues and leave blood stream Comes from B cells in islets of Langerhans Glucagon Insulin antagonist, alpha cells Epinephrine Insulin antagonist, cortisol: up blood glucose levels, adrenal gland ACTH Release of cortisol and some aldosterone Glucocorticoids Ex. Cortisol physiologic effect increases lipolysis and gluconeogenesis "Up blood sugar" Thyroid TSH is an insulin antagonist

Relate L/S ratio to fetal maturity. Describe the L/S ratio test and foam stability test.

L/S Ratio >2 indicates maturity Lecithin increases throughout pregnancy and allows lungs to inflate. Spingomyelin stays at a constant level As ratio goes to two and beyond fetal lungs are deemed to be mature and a C-section or delivery can be accomplished with little risk to the fetus Foam Stability test Shake the tube and look for foam The more foam, the more mature the lungs

Explain the effect, if any, moderate hemolysis and /or lipemia in serum would have on the chemical analysis of the following: potassium, LDH, CK, amylase, thyroxine, glucose, calcium.

LDH: hemolysis can lead to false elevation in LDH levels. CK: hemolysis can lead to false elevation. Amylase: hemolysis and lipemia have no effect. But opiates will cause false elevation of Amylase. Thyroxine: hemolysis and lipemia significantly elevate fT4 measurements. Glucose: hemolysis can falsely elevate glucose and lipemia can falsely decrease glucose levels. Calcium: hemolysis can falsely elevate glucose and lipemia can falsely decrease calcium levels.

Calculate LDL levels and cardiac risk given HDL, total cholesterol and triglyceride value.

LDL= CH-(TG/5)-HDL

State the significance and appropriate methods for measuring Lithium levels.

Lithium is a drug approved by the U.S. Food and Drug Administration (FDA) as an antipsychotic for the treatment of bipolar disorder. Occasionally, it can be used in other disorders in combination with an antidepressant that does not adequately treat a depression. This test measures the amount of lithium in the blood. Bipolar disorder can affect both adults and children. It is a mental condition marked by alternating periods of depression and mania. These periods may be as short as a few days or weeks or if months or years. During an episode of depression, you may feel sad, hopeless, worthless, and lose interest in daily activities. You may be fatigued but have trouble sleeping, experience weight loss or gain, have difficulty concentrating, and have thoughts of suicide. During a manic episode, you may be euphoric, irritable, have high energy and grandiose ideas, use poor judgment, and participate in risky behaviors. Sometimes you will have mixed episodes with aspects of both mania and depression. Lithium levels are monitored on a regular basis because lithium has a narrow therapeutic index. This means there is relatively little space between therapeutic and toxic levels. Too little drug, and the medication will not be effective. Too much drug and symptoms of lithium toxicity may develop.

Differentiate metabolic and respiratory acidosis and alkalosis using arterial pH, pCO2, and bicarbonate or total CO2 values.

Metabolic Acidosis pH pCO2 HCO3 - Primary Decrease Normal Decrease Partial Compensation Decrease Decrease Decrease Compensation Normal Decrease Decrease Respiratory Acidosis pH pCO2 HCO3 - Primary Decrease Increase Normal Partial Compensation Decrease Increase Increase Compensation Normal Increase Increase Metabolic Alkalosis pH pCO2 HCO3 - Primary Increase Normal Increase Partial Compensation Increase Increase Increase Compensation Normal Increase Increase Respiratory Alkalosis pH pCO2 HCO3 - Primary Increase Decrease Normal Partial Compensation Increase Decrease Decrease Compensation Normal Decrease Decrease

List causes of primary metabolic and respiratory, acidosis and alkalosis.

Metabolic acidosis has three main root causes: increased acid production, loss of bicarbonate, and a reduced ability of the kidneys to excrete excess acids. Metabolic alkalosis is primary increase in bicarbonate (HCO3−) with or without compensatory increase in carbon dioxide partial pressure (Pco2); pH may be high or nearly normal. Common causes include prolonged vomiting, hypovolemia, diuretic use, and hypokalemia. Respiratory acidosis involves a decrease in respiratory rate and/or volume (hypoventilation). Common causes include impaired respiratory drive (eg, due to toxins, CNS disease), and airflow obstruction (eg, due to asthma, COPD [chronic obstructive pulmonary disease], sleep apnea, airway edema). Respiratory alkalosis involves an increase in respiratory rate and/or volume (hyperventilation). Hyperventilation occurs most often as a response to hypoxia, metabolic acidosis, increased metabolic demands (eg, fever), pain, or anxiety.

Describe the possible clinical significance associated with abnormal Mg, Ca, Cu, P, and Fe values. List hormonal controls to maintain calcium levels in the blood. State the most common methods to quantitate the following minerals: Calcium, Iron, Phosphorous, and Magnesium. State the most common methods to quantitate the following minerals: Calcium, Iron, Phosphorous, and Magnesium.

Mg (Magnesium) Increase: Overloading on Antacids (MOM) Decrease: Diuretics or digoxin Paget's disease Alcoholism Renal stones Heart failure Ca (Calcium) Increase: Multiple myeloma Excess vitamin D Hyperparathyroidism Paget's disease Decrease: Acute pancreatitis Vitamin D deficiency Hypoparathyroidism GI blockages Malnutrition and Lack of sunlight Hormone: PTH (parathyroid hormone) Released when Ca is low stimulates bone resorption Increases reabsorption of Ca in kidney and gut Stimulates Vitamin D production Cu (Copper) Increase: IUDs Copper overdose Fungicides Kayser-Fleischer ring Decrease: Microcytic, hypochromic anemia Wilson's disease Menke's syndrome P (Phosphate) Increase: Renal failure Hypoparathyroidism Excess growth hormone Excess Vitamin D Decrease: Hyperparathyroidism Alcohol withdraw Vitamin D deficiency Excess antacids Fe (Iron) Increase: Hemochromatosis Decrease: Iron Deficiency Anemia Spectrophotometer

List current and classical methods used to determine Na, K, Cl, urea/BUN, creatinine, and total protein values.

Na (Sodium) Flame photometer Atomic absorption spectroscopy ISE Modified pH electrode for Na K (Potassium) Flame photometer Atomic absorption spectroscopy ISE Valinomycin membrane for K Cl (Chloride) Colorimetry ISE Coulometry; amperometric; titration Ag+ + Cl- → AgCl Titration time proportional to Chloride concentration BUN/urea o NPN + Nessler's reagent → yellow color Creatinine o Urea → 2NH4 + + HCO3 - (urease catalyzed) NH4 ++ nitroprusside = color (Berthelot reaction) NH4 ++2-oxoglutarate NADH → glutamate + NAD++H+ (glutamate dehydrogenase catalyzed) Jaffe reaction Total Protein Biuret Assay Albumin + globulin + Total protein

State the normal reference range for serum Na, K, Cl, BUN/urea, creatinine, albumin, bilirubin, cholesterol, glucose, and total protein serum levels.

Na (Sodium): 135-145 mmol/L K (Potassium): 3.4-5.0 mmol/L Cl (Chloride): 98-106 mmol/L of serum BUN/urea: 6-20 mg/dL Creatinine: Males: 0.6-1.2 mg/dL Females: 0.5-1.1 mg/dL Albumin: 3.5-5.5 g/dL ~60% of Total Protein (TP) Bilirubin: 0.2-1.0 mg/dL Cholesterol: < 140-200 mg/dL Glucose: 80-120 mg/dL Total Protein (TP): 6.5-8.3 g/dL

List the current classification system for determining normal, prediabetic, and diabetic individuals; using the American Diabetes Association Guidelines and oral glucose tolerance, fasting glucose and glycated hemoglobin testing. State the causes, differentiating characteristics, and treatment of the following diabetic conditions: Type 1, Type 2, and gestational diabetes

Normal value: <140 mg/dL Prediabetic: > or equal to 126 mg/dL Diabetic: > or equal to 200 mg/dL and show symptoms of diabetes Fasting Glucose: > of equal to 126 mg/dL Oral Glucose Tolerance Test: fast for at least 8 hours. Fasting is > or equal to 92 mg/dL. 1 Hour is > or equal to 180 mg/dL 2 Hours is > or equal to 153 mg/dL 75 g of glucose loaded Glycated Hemoglobin: 4-6 is normal, 6.5 or higher diabetes mellitus is indicated

Calculate serum protein values using total protein and % fraction from electrophoresis results.

Percentage of fraction % x total protein .08 x 7.4 = .592 g/dL

Define accuracy and precision.

Precision- How close the measured values are to each other. Accuracy- How close a measured value is to the actual (true) value. Mean - Sum of all numbers / total numbers Median - middle number in a data set Mode - Value that appears most often in a data set. Standard deviation - Square root of (Sum of X-mean2 / n-1) Coefficient of variation - SD / Mean X100 (5% or less is good) Confidence interval - 95% confidence: + 2SD

Discuss methodologies to assay bilirubin include principle, test name, interferents reagents, and purpose of reagents.

Principle: Traditional methods of measuring bilirubin are based on the reaction of bilirubin with a diazo reagent to form the colored compound azobilirubin. The diazo reaction can be accelerated by the addition of various chemicals. For example, Malloy-Evelyn3 used methanol, Jendrassik-Gróf4 used caffeine, and Walters-Gerarde5 used dimethyl sulfoxide (DMSO). Modifications of these methods included the addition of surfactants as solubilizing agents.6 Total (conjugated and unconjugated) bilirubin couples with a diazo reagent in the presence of a surfactant to form azobilirubin. The diazo reaction is accelerated by the addition of surfactant as a solubilizing agent. The increase in absorbance at 548 nm due to azobilirubin is directly proportional to the total bilirubin concentration. Methodology: Diazonium Salt. Test name: Total Bilirubin assay on the ARCHITECT c Systems™ and the AEROSET System. Interferents: Hemoglobin solutions at the above concentrations were prepared by addition of hemolysate to solutions of human serum albumin. Intralipid solutions at the above concentrations were prepared by addition of Intralipid to solutions of human serum albumin. Reagents: Surfactants 4.51% HCl 8.204 g/L 2, 4-dichloroaniline 0.81 g/L HCI 5.563 g/L Sodium Nitrite 0.345 g/L Surfactant 2.00% Purpose of reagents: To calculate total Bilirubin.

List two methods used for total serum protein analysis and state their principles.

SPE: Solid-phase extraction (SPE) is an extractive technique by which compounds that are dissolved or suspended in a liquid mixture are separated from other compounds in the mixture according to their physical and chemical properties. Biuret: The biuret test, also known as Piotrowski's test, is a chemical test used for detecting the presence of peptide bonds. The test is named so because it also gives a positive reaction to the peptide-like bonds in the biuret molecule. In this assay, the copper (II) binds with nitrogen present in the peptides of proteins.

Discuss the principles of laboratory tests used to measure salicylate, acetaminophen, ethanol, and barbituate levels (screening and confirmatory) and rules for proper collection.

Salicylate: Salicylic acid, the major metabolite of ASA, can be detected easily in urine using simple chemical spot tests such as ferric chloride or Trinder's reagent. In forensic cases, urine is often not available and the rapid detection of salicylate in whole hemolyzed blood can be difficult. Must draw 6 hours after ingestion to get the peak. Treatment for overdose is forced like alkaline diuresis. Acetaminophen: The test for acetaminophen is used to measure the level of drug in the blood to establish a diagnosis of overdose, to assess the risk of liver damage, and to help decide on the need for treatment. Prompt diagnosis and treatment are important for a positive outcome. Ethanol: The ethanol test, commonly known as alcohol test, is used for both medical and legal purposes. Samples and results for each use are usually collected and tested separately. Medical: medical testing is used to determine the level of ethanol in the blood to effectively treat the intoxicated person's symptoms. Barbituate levels: When a blood sample from a vein is needed, a vein in your arm is usually selected. A tourniquet (large rubber strap) may be secured above the vein. The skin over the vein will be cleaned, and a needle will be inserted. You will be asked to hold very still while your blood is collected. Blood will be collected into one or more tubes, and the tourniquet will be removed. When enough blood has been collected, the healthcare worker will take the needle out.

Draw an example of a Levy Jennings chart; define shift, trend, and outliers.

Shift: six or more data points fall around a new mean. Trend: An uninterrupted rise or decline from the mean for 6 or more consecutive data points. Outliers: The data points value is greater than +2SD or less than -2SD.

3. For each of the following instruments, list the major parts comprising the instrument: Spectrophotometer flame photometer Nephelometry Mass Spectrometry Potentiometry Chromatography

Spectrophotometer: light source, entrance slit, Monochromator, exit slit, sample cuvette, phototube, photo multiplier (PM), A/D, display. Flame photometer - A flame (burner), nebulizer (aspirator), photo detector. Nephelometry: light source, collimating optics, sample cell, collection optics including light scattering optics, detector optical filter and detector. Mass spectrometry: sample inlet, ionization source, vacuum pumps, mass analyzer, ionization detection = data. Potentiometry: indicator electrode, reference electrode, potential measuring device. Chromatography: sorbent coated glass, or plastic plate, closed container, solvent (TLC) (HPLC)Solvent, pump, injection port, column, detector, recorder. (GC) Gas, injection port columns, oven, detector, recorder.

Define steady state, trough and peak, half-life, and NTR as related to therapeutic drug monitoring.

Steady State: is the therapeutic range. Trough: is the point of where the concentration is low that is prior to next dosage. Peak: top level or max of administered drugs. Half-life: The time required to reach steady state depends on the elimination half-life of the drug, defined as the time required for the serum drug concentration to decrease by 50%. The half-life is itself determined by the metabolism and excretion rates of the drug. NTR: The drug can become ineffective of toxic if the patient levels are outside the therapeutic range.

List the disease states associated with abnormally high or low levels for the following hormones: 41.1- T4 41.2- Growth hormone 41.3- Cortisol 41.4- Prolactin 41.5- Parathyroid hormone 41.6- Gastrin 41.7- hCG 41.8- FSH 41.9- LH 41.10- Estradiol

T4 • High: Hyperthyroidisim, Grave's Disease Low: Hypothyroidisim, Hashimoto Growth hormone High: Gigantism, Acromegaly Low: Dwarfism, Hypopituitarism Cortisol High: Cushing's Disease Low: Addison's Disease Prolactin High: Headaches, Visual disturbances, Galactorrhea, Infertility Low: No clinical importance Parathyroid hormone High: Hypocalcemia, Hyperparathyroidism Low: Hypoparathyroidsm, Hypercalcemia Gastrin Zollinger Ellison Disease hCG High: Multiple pregnancies, HcG releasing tumors, Testicular cancer, Hydatidiform mole, Pancreatic tumors Low: Ectopic pregnancies, Spontaneous abortion FSH High: above 15, the success rate goes down to less than 5% and there is an increased chance of chromosomal abnormality. Elevated FSH levels also indicate diminished ovarian reserve which means there are fewer follicles or eggs and these are often of questionable quality. Low: ovaries are not making enough eggs. Pituitary gland is not working correctly. Hypothalamus deficiency LH High: In natural born woman, means menopause Low: pituitary disorder, anorexia, malnutrition, or are under stress. Estradiol: High: Estradiol may be suppressing FSH, problem with your ovarian reserve which could lead to infertility. Low: polycystic ovary syndrome (PCOS) or hypopituitarism, eating disorders, extreme endurance exercising and after menopause.

Calculate patient results when performing a standard or serial dilution given a concentration and patient and diluent volumes.

The patient result is Dilution X result. Dilution factor is solute / solute + solvent.

Explain the theory of electrophoresis.

The theory of electrophoresis is that it turns out that in fact the electrophoretic mobility of a molecule depends on its charge to mass ratio. Two different sized molecules with the same charge to mass ratio should run with the same mobility in a uniform electric field and a perfect world.

Differentiate total, conjugated (direct) and unconjugated (indirect) bilirubin, and list disease states associated with their elevation. Discuss formation, symptoms, abnormal lab data and test results for hepatic, pre-hepatic and post-hepatic disease.

Total Bilirubin = direct + indirect Indicative of liver disease Unconjugated (indirect) Bilirubin Directly from heme (RBC) destruction Insoluble, albumin carrier Elevated in pre-hepatic disease Conjugated (direct) Bilirubin Unconjugated bilirubin taken to liver Converted to conjugated by UDPGT (uridyldiphosphate glucuronyltransferase) Water soluble Secreted in to bile and intestines excreted into urine and stool and blood in the form of urobilinogen Pre Hepatic Formation: Excess levels of unconjugated bilirubin are presented to liver Symptoms: HDN, Hemolytic Anemia Abnormal Labs and Test Results: Liver enzymes not increased, Increase in unconjugated form, Excess bilirubin in urine and feces Hepatic Formation: Impaired cellular uptake or liver damage Symptoms: Coagulation problems not correctable with Vitamin K injections Abnormal Labs and Test Results: Increased total, and direct bilirubin, Increased urine bilirubin, urobilinogen, urobilin, Increase in liver enzymes Diseases Acute Hepatitis: Greatly increased ALT, AST, Other liver enzymes increased Cirrhosis: Greatly decreased albumin, Slightly elevated ALT & AST, Elevated GGT Reye's Syndrome: Abnormal ammonia and liver enzymes, Bilirubin is normal Liver Tumors: Mirrors Cirrhosis. Organic or Therapeutic Drug Toxicity: Acetaminophen Gilbert Disease and Crigler Najjar Syndrome: Cellular uptake impairment or enzyme deficiency, unconjugated bilirubin levels rise Post Hepatic Formation: Bilirubin can be conjugated and leave liver, but is blocked Symptoms: Coagulation problems corrected with Vitamin K injections, Clay colored stools Abnormal Labs and Test Results: Increased total and direct bilirubin, Increased urine bilirubin, Decreased fecal and urine urobilinogen, Greatly elevated ALP Diseases *Bile stones and bile duct blockages, Cholestasis

Calculate uncorrected creatinine clearance in mL/min given serum and urine creatinine values, collection time, and volume collected. (Write formula)

Uncorrected creatinine clearance= U Creatinine x vol. (ml) / P Creatinine x time (min) 24 hour volume (ml/day) / 1440 min/day = V in ml/min

State the significance of elevated gastrin and xylose levels.

While elevated gastrin may indicate tumors in your pancreas or duodenum, it can also be caused by other conditions. For example, gastrin may also be elevated if your stomach isn't making acid, or you're taking acid-reducing medications, such as proton pump inhibitors. With the xylose absorption testing procedure, high blood and urine levels of xylose are normal. They indicate good xylose absorption by the intestines. This suggests that the tested person's symptoms are likely due to another cause, such as pancreatic insufficiency or bile insufficiency.

Define zero order kinetics, first order kinetics, and substrate exhaustion. List factors which affect enzyme reactions.

Zero Order Kinetics: is a chemical reaction wherein the rate does not vary with the increase or decrease in the concentration of the reactants. First order kinetics: Reaction rate is directly proportional to substrate concentration. Substrate Exhaustion: The enzyme has bonded to all the present reactants.