Lecture 9 • Chemical Chemistry o Kidney Assays-renal  Blood urea nitrogen (BUN) measures the amount of urea nitrogen

, a waste product of protein metabolism, in the blood  Urea is formed by the liver and carried by the blood to the kidneys for excretion. Because urea is cleared from the blood stream by the kidneys, a test measuring how much urea nitrogen remains in the blood can be used as a test of renal function.  There are, however, many factors besides renal disease that can cause BUN alterations, including protein breakdown, hydration status, and liver failure.   Normal blood urea nitrogen in the dog is between 6 and 27 mg/dl Increased BUN • An increase in the BUN level is know as azotemia. An elevated BUN may be caused by: o Impaired renal function o Congestive heart failure as a result of poor renal perfusion o Dehydration o Shock o Hemorrhage into the gastrointestinal track o Excessive protein intake or protein catabolism • Diseased or damaged kidneys cause an elevated BUN because the

severe liver failure causes a reduction of urea in the blood. • Because urea is an end product of protein metabolism.  Decreased BUN • A decreased BUN may be seen in: o Liver failure o Malnutrition o Overhydration from prolonged intravenous fluids o Pregnancy (due to increased plasma volume) • Because urea is synthesized by the liver. • Extensive bleeding inot the gastrointestinal (GI) tract will also cause an elevated BUN because digested blood is a source of urea.kidneys are less able to clear urea from the bloodstream. • In conditions in which renal perfusion is decreased. such as high-protein tub feeding. • A patient who is severely dehydrated may also have a high BUN due to lack of fluid volume to excrete waste products. • Just as deyhydration may cause an elevated BUN. Because the BUN is affected by numerous non-renal .  Creatinine • An assessment of the BUN is used as a gross index of glomerular function. may also cause the BUN to increase. a diet high in protein. BUN levels rise. overhydration causes a decreased BUN. such as hypovolemic shock or congestive heart failure.

 The exocrine part of the pancreas secretes and enzyme-rich juice that contains enzymes necessary for digestion into the small intestine. which is roughly a measure of kidney function. • The normal serum creatinine in the dog is between 0.7 and 1. • Serum creatinine level is another way of determining kidney function. The amount produced is relatively stable in a given animal. held together in one stoma. Creatinine is a protein produced by muscle and released into the blood. If kidney function falls.factors. The three primary pancreatic enzymes are: • • • Trypsin Amylase Lipase . it is a line sensitive indicator of declining renal function than serum creatinine.6 mg/dl o Pancreatic Assays  The pancreas is actually two organs. the creatinine level will rise. one exocrine and the other endocrine. The endocrine part of the pancrease is involved with carbohydrate metabolism through secretion of insulin (which lowers blood glucose levels) and glucagon (which elevates blood glucose) into the blood. The creatinine level in the serum is therefore determined by the rate it is being removed.

typsin is present. • The Knox Test mixes feces with a gelatin (protein) solution.  Serum Trypsin-Like immunoreactivity (TLI) • • • Detects trypsinogen leakage into the blood (More accurate) Determines exocrine pancreatic insufficiency (EPI)-Low TLI values High TLI values indicate pancreatitis and feline IBD. If the gel comes off the film during mixing. • Trypsin activity is more readily detectable in feces than in blood. • The x-ray film test uses the gel on an undeveloped x-ray film to test for the presence or absence of trypsin. • Fecal samples should not be more than 1 day old as bacteria will proliferate and produce their own proteolytic enzymes which will give false positive results. Trypsin is normally found in the feces and its absence is abnormal. Exocrine – Trypsin • Trypsin (trypsinase) is a proteolytic enzyme that aids digestion by catalyzing the reaction that breaks down the proteins of ingested food. The x-ray film is placed in a slurry of feces and bicarbonate.  Cobalamine and Folate • Cobalamine and folate concentrations determine small intestinal . If tripsin is present in the feces it will digest the protein and prevent the solution form becoming a gel.

flair-ups of chronic pancreaitis. • Normal amylase in dogs is between 339 and 1007 IU/L  Exocrine – Lipase • Lipase’s function is to break down long-chain fatty acids of . intestines and pancreas) • The rise in blood amylase is not always proportional to the severity of pancreatitis. o Cobalamine(B12) absorbed in ileum o Folate(Folic acid) is also produced by bacteria • Cobalamine and folate are absorbed indifferent parts of the intestines. • Calcium-binding anticoagulants like EDTA should not be used if amylase determination will be run. (IBD) Low Folate levels indicate duodenum and jejunum problems Increased Folate levels indicate Small intestinal bacterial overgrowth.  Exocrine – Amylase • Amylase’s function is to break down starches and glycogen in sugars.function. (Levels also from liver. or obstrucion of the pancreatic ducts. • • • Low cobalamine levels indicate ileum problems. Increased levels of amylase appear in blood duing acute pancreatitis. Amylase requires the presence of calcium for its activity.

• • Normal lipase in dogs is between 267 and 1769 IU/L Spec cPL (Canine pancreas specific lipase) o Immunoassay test used to determine pancreatitis – 98% accuracy o Tests for specific pancreatic lipase o Does not measure liver and kidney lipase values as normal blood lipase values.  Endocrine – Glucose • The endocrine part of the pancreas is made up of small . o More specific lipase tests are performed to determine pancreatitis. and intestines) • Test methods are usually based on the hydrolysis of an olive oil emulation into fatty acids using the patients plasma. Increased levels of lipase appear in blood during episodes of pancreatitis. (Lipase also released from liver. Calcium-binding anticoagulants like EDTA should not be used if lipase determinations will be run. Lipase requires the presence of calcium for its activity.lipids. kidneys. because amylase and lipase levels increase in only 50-60% of all pancreatitis disease. • • Lipase is more sensitive for detecting pancreatitis than amylase.

it is best to use sodium fluoride as anticoagulant. • Type 1 Diabetes occurs due to immune system destruction of the . the blood glucose lowers. lowing the glucose value by about 10% per hour. • • Normal glucose level in a dog is between 79 and 127 mg/dl Problems with insulin causes high blood glucose and the disease Diabetes. • Glucose utilization depends on the amount of insulin and glucagon in the blood. As insulin levels increase. RBCs continue to metabolize glucose after collection. When insulin levels are decreased (as in Diabetes). blood glucose rises.microscopic clusters of cells called Islets of Langerhans. These islets features three main types of cells: o Alpha cells – secrete glucagon in response to low serum glucose o Beta cells – secrete insulin in response to high serum glucose o Dalta cells – secrete the hormone somatostain • The blood glucose level is used as a indicator of carbohydrate metabolism in the body and to indirectly measure endocrine function of the pancreas. • Serum and plasma must be separated from the erythocytes immediately after blood is collected. If samples cannot be separated immediately.

and anorexia. Type 2 diabetics animals may be producing no. but this insulin is failing to achieve its normal action. Blood glucose level is high (hyperglycemia) • Depending on the level of hyperglycemia. • Diabetes is generally characterized by increased thirst (polydipsia). or even excessive amounts of insulin. increase urination (polyuria). • Type 3 Diabetes occurs with some other disease or condition damage the pancreas and/or prevents bata islet cell function. • Type 2 Diabetes is characterized by obesity. normal amounts. but is rare in dogs. glucose may be detected in the urine (glycouria) • There are numerous secondary conditions that can occur from diabetes if it is not controlled including renal failure.insulin-producing (beta) cells of the pancreas. weakness. and deposition of a precipitate called amyloid into the pancreatic islets. loss of tissue sensitivity to insulin. Severe necrotizing pancreatitis and pancreatic tumors cause type 3 diabetes. liver failure. Patients with this form of diabetes often must be treated with insulin injections. too little. Type 2 diabetes is the most common form of diabetes in cats. There may be some genetic component to the cause of diabetes in dogs. This is the predominant form of primary diabetes in dogs. . but is rare in cats. fatigue.

They are produced by the liver and provide osmotic pressure for maintaining blood volume and blood pressure. persistent recumbency.and poor circulation to extremities (with possible amputation) • Depending on its severity.  Albumins account for most (about 60%) of the plasma proteins. myositis. or with corrective diet. CK is found in most abundance in striated muscle and therefore is extremely specific for muscle damage. albumin may be lost in the urine (proteinuria) . laceration. electric shock. vigorous exercise. it cannot determine which muscles are damaged or indicate the severity of muscle damage. globulins. diabetes may be treated with insulinreplacement therapy (insulin shots). They are also important in binding and transporting other substances in the blood. Decreased albumin (hypoalbuminemia) is commonly seen in diffuse where production is impaired. • Normal CK level in dogs is between 7 and 218 IU/L o Plasma Proteins  There are over 200 plasma proteins. • While specific to muscle. surgery. The three most important commonly assayed types of proteins are albumins. o Skeletal Muscle Assay  Creatine Kinase (CK) • • Previous name was creatine phopokinase (CPK) In all mammals. • Examples of muscle damage include intramuscular injections. and fibrinogen. In renal disease. and hypothermia. bruising.

dehydration. Acute inflammation often elevates fibrinogen. • Fibrinogen is often determined using the Heat Precipitation Fibrinogen test.  Fibrinogen. alpha-2. • Total proteins determinations are made in practice with the use of the Refractometer. all serum .5 mg/dL. Some types of globulins also produced by the liver. In general. and altered protein breakdown or excretion. alpha-1 and alpha-2 proteins levels increase in the presence of inflammation.  Serum Electrophoresis • The level of individual proteins (fractions) in serum can be determined in commercial laboratories. altered protein distribution. is an important clotting faction produced by the liver. The TP may be affected by altered hepatic synthesis. are produced by cells of the immune system. Most of these types transport lipids and fat-soluble vitamins in the blood. This method is based on the fact at a certain pH. • • The most common method is via serum electrophoresis. Globulins are roughly divided into alpha-1. known as antibodies. beta. or over-hydration. Others.5 and 7. and gamma globulins.  Total Proteins • Total proteins (TP) in a measure of all plasma proteins in the blood. which accounts for only about 4% of the total plasma proteins. including fibrinogen. Normal TP in dogs is between 5.

• Protein migrate in the following order: o Albumin o Alpha globulins (increased during inflammation) o Beta globulins o Gamma globulins  Most antibodies are gamma globulins therefore gamma globulins become ore abundant following infections or immunizations  If an antibody-secreting cell. and blood. tissue. it grows into a clone secreting single kind of antibody molecule. but the degree of that charge varies with each type of protein. fluid.proteins are negatively charged. called a plasma cell. The positive charge draws the proteins toward the positive pole at different rates depending on the degree of their negative charge. Examples are: o Sodium*** o Potassium*** . becomes cancerous.  Electrolyte Assays • Electrolytes are salts in the body that conduct electricity and are found in the body. • Serum proteins can be separated by placing serum on special paper or in a gel and applying an electrical charge. This type of cancer is called a plasma cell myeloma.

too little is called hypoKalemia. Proper balance is essential for muscle coordination. Normally. dehydration.1 mEq/L • Calcium . too little is called hypoNAtremia. o The kidneys regulate fluid absorption and excretion and maintain a narrow range of electrolyte fluctuation. Too much potassium is called hyperKalemia. and concentration. fluid absorption and excretion. nerve function. o Normal Na+ in dogs is between 138 and 152 mEq/L o Normal K+ in dogs is between 3. Too much sodium is called hyperNAtremia.4 and 5. heart function. and disease. caused by poor diet.o Calcium*** o Magnesium o Chloride o Bicarbonate • Sodium & Potassium o Sodium (Na+) is concentrated in the extracellular fluid between tissue cells and potassium (K+) is concentrated in the intracellular fluid within cells. results in an imbalance. medication. Too much or too little sodium or potassium. sodium and potassium are filtered and excreted in the urine and feces according to the body’s needs.

their constant secretion of parthormone leads to marked Ca+ removal form the bones (osteomalacia). cardiac muscle contractions. o Normal Ca in dogs is between 8. o If the parathyroids become cancerous. and frequent. PTH and Vit D causes Ca+ to come out of bone and enhance intestine absorption of Ca+ and phosphorus. easy fractures. o High blood Ca is called hypercalcemia. o The parathyroids are endocrine glands involved with calcium balance. hormone release and function of body cells. o Kidney damage can result in parathyroid hyperactivity (due .o Calcium (Ca) is primarily tied to bone density but it has many other functions. o Ca and phosphorus have an inverse relationship. Low blood Ca is called hypocalcemia.9 and 11. Osteomalacia characterized by soft and thin bones and teeth. Ca has major roles in nerve function. muscle contractions. They secrete parathormone (PTH) is response to low serum Ca+ and secrete less PTH with the Ca+ is normal or high. (PTH causes an increase in calcium).5 mg/dL o EDTA or oxalate anticoagulants should not be used when collecting blood for Ca assay because they bind with Ca and make it unavailable for assay.

muscle and digestive disease o Causes of Potassium deficiency as vomiting. This condition is called Renal Secondary Hyperparathyroidism. diarrhea and diuretic .to phosphorus retention) and thus also lead to osteomalacia. • Magnesium o Magnesium is needed for utilization of ATP o Primarily in bone and skeletal muscle  Only 1% in blood o Hypomagnesemia     • Hypokalemia – non resolving with supplementation Hypocalemia – hypoparathyroid disease Myopathy and neuropathy – critically ill Convulsions and cardiac arrest if severe Potassium-vomiting/diarrhea causes potassium problems o Potassium is an intracellular electrolytes o Potassium needed for normal kidney function  Sodium/Potassium pump • High sodium intake decreased potassium due to excretion. o Potassium has function in cardiac. skeletal.

o Hyperkalemia   Causes bradycardia Renal failure.o Hypokalemia  Symptoms – weakness. arrhythmias and walking abnormalities in cats. muscle cramps. FLUTD and Addison’s disease . lethargy.