Clinical Chemistry-2 Lecture Prelims PDF

CLINICAL CHEMISTRY-2 LECTURE PRELIMS TOPIC 1. ENZYMES PT. 1 Lecturer/s: Mr. Christian Villahermosa, RMT, MSMT August 10, 2023 ENZYMES...

CLINICAL CHEMISTRY-2 LECTURE PRELIMS TOPIC 1. ENZYMES PT. 1 Lecturer/s: Mr. Christian Villahermosa, RMT, MSMT August 10, 2023 ENZYMES ENZYMOLOGY Biologic catalysts ○ Hasten chemical reactions Study of enzymes Are not consumed during the reactions We have to look for: Do not undergo a chemical change after the ○ Activity of Enzymes reactions Different reactions they mediate Significance in diagnosis, Why do doctors include enzyme tests as part of lab tests possible diseases the doctor can for their patients? explore if a certain enzyme is Enzymes operate as biological catalysts that hasten or increased speed up chemical reactions. ○ Chemical reactions they catalyze They facilitate/mediate the fruition of certain biochemical ○ Clinical uses reactions. Enzymes make us thrive on this planet. Enzymes do not undergo change. HOW AN ENZYME INTERACTS INSIDE THE BODY MAIN ACTION OF ENZYMES Catalyze/interact with substrate to mediate/facilitate chemical reactions. Enzymes are found in the cells and tissues. They are present only in the circulation at a limited level (low level) The bulk of enzymatic concentrations are present in the cells/tissues where they facilitate chemical reactions If they are present at increased levels in the blood, we can point to a problematic organ. EXAMPLE… - ALT (alanine transaminase) is found in the cells of the liver. - It is found in low concentration/circulation but once it increases in circulation, it will be detected by the Pentra machine. - The doctor assumes there is something wrong with the liver of the patient. - Enzyme + substrate (object of choice) → basic requirement TESTING FOR ENZYMES - Enzyme alone/ substrate alone → no reaction will is NOT DIAGNOSTIC for a certain disease. occur Purpose: For the doctors to be alerted as to which body part, - Enzyme + substrate = enzyme-substrate complex cell/tissue has a problem. which produces a product and the enzyme - This helps the doctor on which area to focus. - During this reaction, the enzyme will not be - “Narrowing down” of the possible disease consumed and the form will not be changed - Disease is Non-specific - The intact enzyme bound to the substrate is still the There are enzymes found in so many body parts, rendering same enzyme that was expelled after products were enzyme tests for them as impractical. formed. It retains the same appearance before binding. EXAMPLE → LDH Role: For the substrate to turn into product. (Helper) LDH is found in the RBCs, many tissues, in the Only the substrate underwent change. The enzyme heart, etc., deeming it expensive and unnecessary. remained intact. (Michaelis-Menten Theory) [see below] Made by Miguel Astronomo and Daniel Budo 1  Linkage Catalyze one type Chymotrypsin reaction for a catalyzes the specific type of hydrolysis of peptide bond bonds (all types of protein) [not all, but If you have this they share the peptide specific type of bond] bond, it will bind to you. ACTIVE SITE Comparison of energy required for a chemical reaction to occur with an enzyme vs. without an enzyme Even if there is no enzyme, we can expect products to form because this is the basic rule (substrate will become product.) If there is NO ENZYME, products will still be formed, but at a longer time, and with the utilization of very high levels of energy. (Lock and Key Model, Emil Fisher in 1894) Binding of enzyme to a substrate to form a complex Y axis = Energy usage which will yield a product X axis = Time required to produce a product Enzyme = as is, no change, no consumption Red line refers to the process if there is no ACTIVE SITE ⇒ where the substrate binds to an enzyme presence enzyme ○ A lot of energy is needed, and the ○ Since there is a need to bind for the production time is slower compared to blue formation of complex and yielding of line product ○ Our body will choose the ‘with enzyme’ process – every energy we produce is ALLOSTERIC SITE precious. ○ Enzymes facilitate faster reactions The Allosteric site is where the activators and the because the alternative may take too long, inhibitors bind (not the substrate) even months. Activators promote the binding of the enzyme to Enzymes help break down triglycerides into simpler forms. the substrate. Inhibitors prevent the binding of the enzyme to the ENZYME SPECIFICITY substrate Enzymes may recognize and catalyze: Since BOTH of them are needed, they will bind to - a single substrate the enzyme in the allosteric site (different from the - a group of substrates that have specific active site) functional groups - A group of substrates that have a ALLOSTERIC INHIBITOR particular type of bond TYPES OF ENZYME SPECIFICITY TYPE REACTION TYPE EXAMPLE Absolute Catalyze one type of Urease catalyzes only reaction for a single the hydrolysis of urea substrate Group Catalyze one type of Hexokinase adds a reaction for all phosphate group to similar substrates hexoses Looks for a specific functional group An allosteric inhibitor (red) binds to the allosteric site to (amino group, prevent the enzyme from binding the substrate carboxyl group, ester, etc. made into Inhibitors prevent, so they make sure the substrate substrate) doesn’t bind to the active site. Made by Miguel Astronomo and Daniel Budo 2  ○ The shape of the enzyme is changed so they could not bind. NOTE! The names of some enzymes do not end in -ase. ○ Examples: pepsin, trypsin ○ Angiotensin-converting enzyme CLASSIFICATION OF ENZYMES IN ORDER 1. Oxidoreductases 2. Transferases 3. Hydrolases 4. Lyases 5. Isomerases 6. Ligases An allosteric activator (green) makes the enzyme better at ENZYMATIC REACTIONS binding the substrate Can occur in both ways. There is a forward and reverse reaction, thats why the arrow points both ways. TAKE NOTE! The substrate and the active site are not the same, 1. OXIDOREDUCTASES so they could not bind All enzymes in this class can perform oxidation and reduction What’s needed for them to bind is for the activator forward and reverse. to bind to the allosteric site Oxidation and reduction ○ Oxidation - removal of H ion Or it could be any electron, but in Do we have inhibitors in the body? If enzymes are needed this case this is appropriate for for chemical reactions, why is it that inhibitors exist? Clinical Chemistry Yes. There are chemical reactions that their ○ Reduction - acceptance of H ion products can worsen certain conditions. Gout / increased uric acid – xanthine oxidase will - E.C. 1.1.1.27 : L-lactate NAD+ Oxidoreductase produce uric acid, and normally the body will get - Lactate Dehydrogenase (removal of rid of it, uric acid is continuously produced and hydrogen from lactate) excreted. - This enzyme has a lactate ○ For patients with gout, we have to do substrate. something to prevent the production of uric acid. Allopurinol is an inhibitor of xanthine oxidase. Same with inflammation, as anti-inflammatory drugs act as inhibitors. Inflammation is created by the enzyme cyclooxygenase which produces prostaglandin. 1. In the forward reaction, the substrate is lactate NOMENCLATURE OF ENZYMES 2. As Lactate dehydrogenase reacts, it becomes pyruvate as hydrogen is converted (left lactate) 1. Substrate + -ase (not true to all) 3. This is an oxidation reaction (removal of H ion) a. Lipid = lipase a. We base it on the substrate b. Ester = esterase c. Protein = protease LOOKING AT IT IN REVERSE… Pyruvate is the substrate, and H binds back to 2. Reaction it catalyzes pyruvate to become lactate a. Oxidation = oxidase This is now a reduction reaction. b. Reduction = reductase c. Hydrolysis = hydrolase 2. TRANSFERASES d. Dehydrogenase = remove hydrogen atoms Transfer of functional groups other than hydrogen e. Decarboxylase = remove carboxyl groups from one substrate to another 3. Enzyme Commission Nomenclature (E.C.) - E.C. 2.6.1.1 : L-Aspartate 2-Oxaloglutarate - E.C. 1.1.1.21 Aminotransferase 1st digit = class - Aspartate Aminotransferase (transfers 2nd digit = subclass amino group) 3rd and 4th = serial number Made by Miguel Astronomo and Daniel Budo 3  - Aspartate that has NH2 is transferred by 5. ISOMERASES AST to a-ketoglutarate that has no NH2. Rearrange the functional groups within a molecule - NH2 is being transferred. and catalyze the conversion of one isomer into - NH2 of aspartate is being transferred to another a-ketoglutarate. - As soon as a-ketoglutarate has NH2, it - Phosphoglycerate mutase becomes glutamate. - Aspartate, when removed with NH2, becomes oxaloacetate. O-P=O-O is attached to the 3rd carbon 3. HYDROLASES Now it is attached to the 2nd carbon. Therefore it is Hydrolysis of various bonds named 2-Phosphoglycerate Addition of water to a bond resulting in bond breakage 6. LIGASES ○ Loss of substrate Catalyze the joining of two large molecules by - E.C. 3.1.1.3 : Triacylglycerol lipase forming a new chemical bond Accompanied by an ATP-ADP interconversion 1. Lipase with water will break down TAG (triacylglycerol.) 2. It will become Diacylglyercol with 1 Free Fatty Acid 3= 2+1 4. LYASES Catalyze the removal of groups from Substrates without hydrolysis or oxidation; the product contains double bonds or a ring TERMS It breaks down a big molecule, WITH NO WATER INVOLVED. Substrate ○ Enzyme is strained ○ Acted upon by the enzyme ○ Product should always contain either a ○ Specific double-bond or a ring. Isoenzyme ○ Different form, but with the same action Cofactor ○ Non-protein molecule ○ Helper of enzyme / promote enzymatic attachment to the substrate Some enzymes require cofactor, some do not Apoenzyme ○ Polypeptide portion ○ Inactive enzyme ○ Enzyme that requires a cofactor BUT has not met a cofactor Holoenzyme ○ Apoenzyme + Coenzyme ○ Enzyme that has met a cofactor Made by Miguel Astronomo and Daniel Budo 4  ○ Only if it becomes a holoenzyme can it bind with the substrate. - Some enzymes require cofactors - Without cofactors: inactive apoenzyme - With cofactors: active holoenzyme COFACTORS TWO TYPES Activators or Coenzymes (Bishop) ○ Activators → inorganic substances Does NOT contain carbon ○ Coenzymes → organic substances Contains carbon Other literature divides cofactors based on their binding tightness. Since there is NO COFACTOR YET, the cofactor in Prosthetic groups: Tightly bound place will have to be COPPER first. ○ Metal ions (activators) or organic This is an inorganic substance. → Activator molecules ○ Change to the configuration of the COFACTOR enzyme/link substrate to the enzyme/ Prosthetic groups coenzyme - Heme ○ Copper is a member - Flavin adenine dinucleotide (FAD) ○ Activators are members Coenzymes Coenyzmes: loosely bound - Flavin mononucleotide (FMN) ○ Organic molecules - Thiamine pyrophosphate (TPP) ○ “Second substrate” for enzyme reactions - Nicotinamide adenine dinucleotide (NAD) - Lipoic acid Metal ions - Zn2+ - Cu2+ - Mg2+ PROENZYME a.k.a Zymogen Role: enzyme precursor / immature enzyme Not all enzymes have proenzymes – there are enzymes where once they are created, they are good to go. There are certain enzymes, most especially digestive enzymes, they express as a proenzyme EXAMPLE: trypsinogen and trypsin Trypsinogen is the proenzyme, whereas trypsin is the active enzyme. Some enzymes require a coenzyme to facilitate the reaction. The apoenzyme binds the coenzyme and then the Why is it that some enzymes need to undergo proenzyme substrate. before it has to become active form? The coenzyme is a part of the catalytic domain and will either In Trypsinogen, it is produced by the pancreas. donate or accept functional groups, allowing the reaction to They are proteases (digest proteins) occur. If the pancreas will produce trypsin right ahead, that Once the product is formed, both the product and the is grounds for death – trypsin is a protease. If coenzyme are released. pancreas will produce trypsin right away, it will digest the pancreas, leading to pancreatitis and killing the patient. For safety, if the substrate protein is not present yet, they will exist in their proenzyme form, which will not do anything because IT HAS NO ACTIVE SITE. Trypsinogen will exit the pancreas and go to the intestine, where proteins are present. Trypsinogen will become trypsin will become proteins. Only in this case, ONLY TRYPSIN IS THE PROTEASE. Made by Miguel Astronomo and Daniel Budo 5  ENZYME KINETICS MICHAELIS-MENTEN THEORY E + S → E-S → P + E Vmax Y axis - velocity of the enzymatic reaction X axis - substrate concentration Low substrate, more substrate - Our enzymes are limited. - Enzymes are fixed in number, substrates are not. In other words, if the substrate count is low, there is an enzyme available to bind to them, that’s why velocity is going upwards because enzyme and substrate is available. - There will come a time where it will just stop (Vmax) (maximum velocity) because all enzymes are saturated. All enzymes have been bound to a substrate at this moment. - They could not accommodate more substrate. - This explains the slow digestion. Made by Miguel Astronomo and Daniel Budo 6  CLINICAL CHEMISTRY-2 - LECTURE PRELIMS TOPIC 2. ENZYMES PT. 2 / CLINICAL SIGNIFICANCE Lecturer/s: Mr. Christian John Villahermosa, RMT, MSMT August 17, 2023 LINEWEAVER-BURK PLOT 1. SUBSTRATE CONCENTRATION First-Order Reactions ○ Are those which proceed at a rate exactly proportional to the concentration of ONE REACTANT ○ In enzymes, when the reaction rate is directly proportional to the substrate concentration Second-Order Reactions ○ Are those in which the rate is proportional to the product of the concentration of TWO REACTANTS or THE SQUARE OF THE CONCENTRATION OF ONE REACTANT Similarities between 1ST and 2ND order is that the 𝐾𝑚 relationship between SUBSTRATE conc. and VELOCITY. is 1 1 1 𝑉 = 𝑉𝑚𝑎𝑥 [𝑆] + 𝑉𝑚𝑎𝑥 DIRECTLY PROPORTIONAL. (S = V | ↑S ↑V) - Meaning, the MORE substrates you add, the A double-reciprocal plot of the Michaelis-Menten FASTER is the velocity. constant which yields a straight line - Which is true because if there are substrates, there are products nga form. Difference between Michaelis-Menten Plot & Zero-Order Reactions Lineweaver-Burk Plot ○ When the substrate concentration is high Presents the same thing as with the enough to saturate all available enzymes Michaelis-Menten Plot (MM Plot), the difference Na saturate na lahat ng enzymes between the 2 is that the lineweaver-burk plot (LB thus it attained → Maximum Plot) presents a STRAIGHT LINE compared to the reaction velocity (Vmax) HYPERBOLA of Michaelis-Menten’s plot. ○ A higher concentration of substrate will no Another difference is the unit of X and Y axis of the longer result in increased reaction rate lineweaver-burk plot is the RECIPROCAL of ↑S = No change in V Michaelis-Menten which are the Velocity and Substrate concentration 2. ENZYME CONCENTRATION ○ Velocity makita na sa Y axis; Substrate makita na sa X axis. The HIGHER the enzyme level, the FASTER the reaction will proceed. The point of Lineweaver-Burk Plot is to illustrate in another graph form how the enzyme to substrate reaction occurs. Once the reaction has reached its Vmax it cannot anymore produce any reaction but when enzyme is added the reaction But the main graph that we are going to use is the MM Plot, will proceed once again eliminating Vmax. because in LB Plot, we CANNOT appreciate the reaction ↑E = ↑V happening because the line is just continuous, unlike in MM Plot where we can appreciate the reaction or the relationship 3. pH between velocity and the substrate concentration due to the pH + 7.0 - 8.0 (most enzymes) curve that it forms. ○ ALP → Has a pH: 9 Changes in pH may DENATURE the enzyme. FACTORS THAT INFLUENCE ENZYMATIC REACTIONS ○ Denature = Destroyed Each enzyme operates within a specific pH range These factors either INCREASE or DECREASE the rate of reaction Made by Miguel Astronomo and Daniel Budo 1 How to ensure desired pH in reagents? 5. COFACTORS (Activators or Coenzymes) - The Reagent box contains BUFFERS. It ensures Non-protein entities that must bind to particular desired pH is followed; NO MORE MANUAL enzymes before a reaction occurs measurement. ○ Activators: Metallic or Nonmetallic ○ Coenzymes: Nucleotide phosphate and 4. TEMPERATURE Vitamins 37° C is the OPTIMAL temp for enzymes (Body Temp) 6. ACTIVATORS Denaturation @ 40-50°C (HOT) Proper substrate binding Inactivation @ 4-10°C (COLD) Link substrate to the enzyme or coenzyme ○ Will become INERT Undergo oxidation or reduction ○ The enzyme you refrigerated for it to inactivate is the same enzyme you will be getting if you restore it back to its optimal 7. INHIBITORS temp Interfere with enzyme reactions ○ Preservation of blood samples in Clinical 3 types of Inhibition Laboratories is REFRIGERATION. ○ Competitive Assay temperatures: 25, 30, or 37°C ○ Noncompetitive ○ Uncompetitive 1. Competitive Inhibition: The inhibitor will bind to the ACTIVE SITE. - So since the active site is occupied by this inhibitor, the substrate cannot bind to the active site where it usually binds. 2. Noncompetitive Inhibition: The inhibitor will bind to the ALLOSTERIC SITE. - The allosteric site of the enzyme is occupied by the inhibitor changing now the FORM or the SHAPE of the active site making it unable for the substrate to bind to the active site. 3. Uncompetitive Inhibition: The inhibitor will bind to the ENZYME-SUBSTRATE COMPLEX. - This inhibitor is considered smart because it waits for the Enzyme and Substrate to bind first before it binds to the complex which inhibits the reaction process. reductase ing agent IF WE WANT ENZYMES TO PRODUCE A PRODUCT, WHY DO WE ALLOW INHIBITORS TO EXIST? Pargyline Monoamine Antihypertensive oxidase agent Examples of Enzyme Inhibitors with Therapeutic Applications Penicillin Transpeptidase Antibacterial agent Inhibitor Target Enzyme Effect or Application HMG-CoA = 3-hydroxy-3-methylglutaryl coenzyme A. Allopurinol Xanthine oxidase Treatment of Gout is the accumulation of URIC ACID. gout Uric acid came from the breakdown of PURINE. In order for the purine to become a uric acid it Aspirin Cyclooxygenase Antiinflammatory needs the help of XANTHINE OXIDASE. agent Thymidylate Antineoplastic So if we eat food that is rich in purine our xanthine oxidase 5-Fluorouracil will now convert it into uric acid for it to be urinated. synthetase agent Lovastatin HMG-CoA Cholesterol-lower Too much accumulation of uric acid will now be deposited into our joints and will lead to gout. Made by Miguel Astronomo and Daniel Budo 2  In patients with gout, uric acid is avoided and the Increase in PRODUCT concentration. drug ALLOPURINOL inhibits Xanthine oxidase thus ○ How could this be related to enzyme uric acid production is LOWERED. (Decrease only; activity? not eliminated totally) Because if there is no enzyme, there is no product produced. This is now why inhibitors are allowed to exist. The more product is formed the more enzymes are present. LOCK AND KEY THEORY Decrease in SUBSTRATE concentration ○ How could this be related to enzyme activity? Because if the substrate is low, meaning to say that there is an enzyme that converts the substrates to products. The more na ning baba nag level sa substrates the higher is the concentration of the enzyme. Decrease or increase in COENZYME concentration ○ There are coenzymes that can be ALTERED, DISAPPEAR, UNAFFECTED or MULTIPLIED when they interfere in an enzymatic reaction. The STRUCTURE of the substrate is ○ What is measured here is how many COMPLEMENTARY to the size and appearance of coenzymes are altered, disappeared, the active site. unaffected or multiplied. So when they interact they will just merge as one ○ There is NO FIXED measurement. perfectly. An increase in the concentration of the altered coenzyme According to one journal, this theory is too perfect, too good to be true. They believe that our body is not THIS PERFECT. INDUCED FIT THEORY Measuring enzymes directly is difficult. ENZYMATIC ASSAYS 1. Coupled- Enzyme Assay Coupling the activity of the enzyme being tested to another, more easily detectable, enzyme ○ Measured here is the activity of ANOTHER ENZYME. ○ This is because enzymes are There is a CLASH between the structure of the difficult to catch or measure substrate and the active site of the enzyme. directly, so we need to examine If a substrate comes close to its target enzyme, the another thing, in order to active site will CHANGE ITS STRUCTURE to represent our TARGET ENZYME. accommodate the substrate Coupled-Enzyme meaning you are using The fact the substrate is CLOSE TO THE ACTIVE MANY ENZYMES. SITE, is enough to change the structure of the Indirect measurement active site to accommodate the substrate. Example of coupled assay reaction: The enzyme Lysozyme helps kill bacteria by binding to the polysaccharide coating of the bacteria. The fit is not perfect, so the shape of the active site changes to fit the polysaccharide substrate. This change of shape of the active site is called INDUCED FIT. By initiating this “induced fit”. the enzyme breaks the Hexokinase (HK) = Auxiliary Enzyme polysaccharide, ultimately helping kill the bacteria Glucose-6-phosphate dehydrogenase (G-6-PD) = Indicator Enzyme MEASUREMENT OF ENZYME ACTIVITY In this type of reaction (CK Enzyme), what is measured here In the laboratory we do not measure the enzymes directly, is how many Oxidized NADP+ is formed into reduced but we measure their ACTIVITY; By measuring their activity NADPH. we will now come up with their concentration. Made by Miguel Astronomo and Daniel Budo 3  This information is true because, if you can produce But in the laboratory, we do not test for elastase, trypsin, 1 NADPH that is equivalent to 1 CK. pepsin and others. We ONLY test for AMYLASE and LIPASE ○ 1:1 Ratio to diagnose ACUTE PANCREATITIS. 1 NADPH will not be formed without CK ENDOCRINE AND EXOCRINE FUNCTION OF THE 2. Fixed - Time Method (End point) PANCREAS Reactants are combined Endocrine = HORMONES from Pancreatic Islets. Reaction proceeds for a designated time ○ Exits to the Bloodstream. Reaction is stopped Exocrine = Mainly ENZYMES from Acinar cells. Measurement is made of the amount of ○ Exits through the Pancreatic duct. reaction that has occurred. AMYLASE (AMS) We are given with only 1 RESULT; meaning the E.C 3.2.1.1 – 1,4-D-Glucan Glucanohydrolase measurement will only come at the END Breakdown of starch and glycogen to monosaccharides 3. Continuous - Monitoring Method (Kinetic Assay) ○ Polysaccharide → Monosaccharide. Multiple measurements of absorbance Activators: CALCIUM and CHLORIDE change In order for us to know if there are fluctuations in the absorbance while the machine is assessing your sample. ○ Increasing in absorbance ○ Decreasing in absorbance CALCULATION OF ENZYME ACTIVITY INTERNATIONAL UNIT (IU) Amount of enzyme that will catalyze the reaction of 1 MICROMOLE OF SUBSTRATE PER MINUTE Amylase follows the LINKAGE specificity; they are bound to under specified conditions react with structures with α-1,4-glycosidic bonds. if a structure is present which has that type of bond, INTERNATIONAL UNIT PER LITER (IU/L) amylase will react. Enzyme concentration Used MOSTLY in the hospitals in the laboratory. KATAL UNIT (mole/s): SI unit Amount of enzyme that will catalyze the reaction of 1 MOLE OF SUBSTRATE PER SECOND under specified conditions. ○ 1.0 IU = 16.7 nkat PANCREATIC ENZYMES The type of amylase that is present in humans is ALPHA (α) These enzymes are: Beta (β) is found in plants and microorganisms AMYLASE This picture, the red arrows shows the way alpha LIPASE amylase breaks down a polysaccharide in a RANDOM manner. as long as there is an α-1,4-glycosidic bond TISSUE SOURCES OF AMS SALIVARY GLANDS ○ Salivary AMS or PTYALIN ACINAR CELLS OF THE PANCREAS ○ Pancreatic AMS Amylase is also found in other organs but their presence in that organ is negligible. The fact also that the amylase has 2 major sources makes the amylase NON-SPECIFIC. If the patient has increased amylase concentration then you can narrow it down to 2 organs. The pancreas does not only produce amylase and lipase, it also produces many other enzymes such as, elastase, trypsin, pepsin. PROPERTIES OF AMS Made by Miguel Astronomo and Daniel Budo 4  MW: 50,000 – 55,000 ○ P3 is usually what's checked when we run Readily filtered by the glomerulus due to its small a serum sample to check its amylase level molecular weight. content ○ Once inside the glomerulus it is NOT HIGH P3 is indicative of having REABSORBED by the tubules. acute pancreatitis ○ Thus we can detect amylase in URINE. These subtypes will only be measured through The first amylase enzyme ELECTROPHORESIS. discovered was from a urine ○ because our machines can only detect the sample. GENERAL amylase; not specific. MOUTH – initial digestion of starch by salivary AMS ○ Inactivated in the stomach DIAGNOSTIC SIGNIFICANCE SMALL INTESTINE – final digestion by pancreatic Acute Pancreatitis AMS ○ Rise: 5-8 Hrs. after the onset of an attach If the inflammation of the ISOENZYMES OF AMS pancreas started at 6pm, later P-type isoamylase 11pm pa mag start ug rise ang ○ Pancreas level sa amylase ○ P3: Predominant in AP ○ Peak: 24 Hrs. S-type isoamylase (None pancreas) ○ Normalize: 3-5 days ○ Salivary gland Salivary Gland Lesions ○ Lungs (levels not significant) ○ Parotitis – disease in parotid glands ○ Fallopian tubes (levels not significant) (Mumps) Intra-abdominal diseases Each type of isoamylase has 3 subtypes: ○ Negligible S – 1, 2, 3 P – 1, 2, 3 WHAT IS ACUTE PANCREATITIS? DUCT OBSTRUCTION Acute pancreatitis has 3 possible causes: There is something that OBSTRUCTS the 1. Duct Obstruction pancreatic duct. 2. Acinar Cell Injury ○ Possible cause of obstruction is 3. Defective Intracellular Transport GALLSTONES AND PARASITES. because the gallbladder is near Remember that in our pancreas we have proenzymes which the pancreas. acts as a safety net for the pancreas, since proenzymes are ○ The obstruction caused by the gallstone inactivated protease, no digestion in itself will happen. causes IMPAIRED BLOOD FLOW. ○ Thus no oxygen can pass through But if the proenzymes are activated WITHIN the pancreas ○ No oxygen = NECROSIS then that would lead to some serious trouble and would ○ Will also lead to the PREMATURE cause the pancreas to be digested by its own enzyme. ACTIVATION of the enzymes. Made by Miguel Astronomo and Daniel Budo 5  Pancreas produces Amylase, LIPASE (LPS) Lipase and Protease. E.C. 3.1.1.3 – Triacylglycerol Acylhydrolase Which attacks CHO, FATS and Hydrolyzes the ESTER LINKAGE OF FATS to PROTEINS. produce alcohols and fatty acids. It can destroy the pancreas if activated prematurely ALCOHOLISM ○ Everytime you drink, THICKENED PROTEIN BLOBS will start to deposit in your pancreatic duct. ○ Until such time, it will totally obstruct the pancreatic duct. ○ Alcohol is also TOXIC to the Acinar cells. ○ That is why chronic alcohol ingestion is bad for the pancreas. Lipase is bound to react with (Target) ESTER 1 (R1) and ACINAR CELL INJURY ESTER 3 (R3) Can be caused by: Alcohol, Drugs, Trauma, Ischemia and Viruses. TISSUE SOURCES OF LPS Which leads to the accidental activation of the Primarily in the PANCREAS enzymes in the pancreas. Also present in liver, stomach, small intestine, white blood cells, fat cells and milk. DEFECTIVE INTRACELLULAR TRANSPORT Our tongue has lipase and it is called LINGUA Inside the acinar cells there is a pathway for the LIPASE. enzymes that NEEDS to become proenzyme and enzymes that DOES NOT need to become a PROPERTIES OF LPS proenzyme. MW: 45,000 For example trypsinogen. This enzyme should have Enzymatic activity is specific for the fatty acid a different pathway from other enzymes released by residues at positions 1 and 3 of the TAG molecules the pancreas such as the amylase and lipase. Substrate must be an emulsion for activity to occur If trypsinogen goes to the pathway where amylase Reaction rate is accelerated by the presence of and lipase goes then, trypsinogen will be activated COLIPASE and BILE SALTS into trypsin. ○ Both of them are needed for the reaction of This is problematic since the trypsinogen was lipase to occur. activated within the pancreas itself and will now ○ The substrate must be an EMULSION for lead to the digestion of its own cells. the activity to occur. Lipase could not act on a lipid if it ALCOHOLISM AND PANCREATITIS is not emulsified (emulsified by Chronic alcohol ingestion results in the: bile). Secretion of protein-rich pancreatic fluid that leads Bile is formed in the LIVER and to the deposition of thickened protein plugs and stored in the GALLBLADDER. obstruction of small pancreatic ducts Transient increase in the pancreatic exocrine DIAGNOSTIC SIGNIFICANCE secretion and contraction of the sphincter of Oddi Direct toxicity in the acinar cells Almost exclusive to the diagnosis of ACUTE PANCREATITIS ○ Rise: 4-8 hours after onset of an attack MACROAMYLASEMIA ○ Peak: 24 hours This is the most common type of Hyperamylasemia ○ Persist for about 8-14 days ○ ↑Amylase yet essentially NORMAL > 14 days rise suggests POOR Persistent increase in serum amylase is seen PROGNOSIS for pancreatic cyst without clinical symptoms Can be due to Cancer ○ In these patients, the amylase binds to the (one of the deadliest) ANTIBODIES or IMMUNOGLOBULINS. Found also in other intra-abdominal conditions ○ AMS molecules combine with IgG or IgA. ○ Duodenal ulcers, perforated peptic ulcers, ○ This molecules combined together will now intestinal obstruction, acute cholecystitis become a big structure Normal in conditions with SALIVARY GLAND ○ Which cannot be filtered out by the involvement glomerulus. ○ Patient has increased amylase yet normal ○ So it will GO BACK to the blood lipase; possible issue is in the salivary circulation. gland. ○ Thus, having increased AMS yet the ○ If this was Acute pancreatitis bothe patient feels normal. amylase and lipase should be high. Made by Miguel Astronomo and Daniel Budo 6  CLINICAL CHEMISTRY-2 - LECTURE PRELIMS TOPIC 3. LIVER ENZYMES / CLINICAL SIGNIFICANCE Lecturer/s: Mr. Christian John Villahermosa, RMT, MSMT August 28, 2023 ASPARTATE AMINOTRANSFERASE Vit. B6 is acquired through diet, thus a patient serum should contain enough Vit. B6 in order for the reaction to occur. ENZYME 2.6.1.1 CLASSIFICATION Why do these enzymes exist? SYSTEMATIC L-Aspartate: 2-oxoglutarate Functions in the synthesis and degradation of NAME Aminotransferase amino acids. ○ 1 amino acid is broken down and 1 amino FUNCTION AND Transfer of an amino group acid is born. PRODUCT between aspartate and alpha-keto ○ One amino acid has to go and 1 amino acids to form oxaloacetate and acid has to be produced because if you glutamate compare the two, GLUTAMATE is more FORMER NAME Serum glutamic-oxaloacetic important. transaminase (SGOT or GOT) ○ Because glutamate is a NEUROTRANSMITTER COFACTOR Pyridoxal phosphate (Vitamin B6) What is the purpose of the keto acid formed from the enzymatic reaction? Ketoacids formed are ultimately oxidized by the tricarboxylic acid. ○ They are sent to the TCA cycle (Tricarboxylic Acid Cycle) to become an energy source. ISOENZYMES FRACTIONS Both enzymatic reactions (AST & ALT) involve 1 Inside the hepatocyte, AST has a higher AMINO ACID and 1 KETOACID. concentration than ALT. ○ In the forward reaction: Involves the giving ○ Cell cytoplasm (ASTc) up of the amino acid group of L-Aspartate Predominant form in serum to 2-Oxoglutarate ○ Mitochondria (ASTm) ○ In doing so, glutamate and oxaloacetate Increased in disorders producing are formed CELLULAR NECROSIS 2-Oxoglutarate → L-Glutamate (Amino acid) Location of the Hepatocellular Enzymes L-Aspartate → Oxaloacetate (Keto acid). MODERN NAME is derived from the amino acid substrate of the forward reaction (In this case: Aspartate) FORMER NAME is derived from the products of the forward reaction. Case from Henry’s A patient with Cirrhosis (scarring of the liver) both AST and ALT should be INCREASED. Now when an ALCOHOLIC patient with CIRRHOSIS undergoes testing both the enzymes are at normal ranges. The reason behind the normal test results of an alcoholic w/ cirrhosis patient is that TOO MUCH ALCOHOL INTAKE can ELIMINATE Pyridoxal Phosphate (Vit. B6). The major diagnostic hepatocellular enzymes are ○ Without Vit. B6 → AST/ALT reaction will located at various sites in the hepatocyte, giving not occur Made by Miguel Astronomo and Daniel Budo 1  rise to different patterns of enzyme release with ALP was NOT REQUESTED because if the hepatocyte is different causes of injury. damaged at one end (hepatocyte facing the bloodstream) the Alanine aminotransferase (ALT) and the other hepatocyte facing the canaliculi end REMAINS cytoplasmic isoenzyme of aspartate INTACT. aminotransferase (ASTC) are found primarily in - Remember that AST and ALT are found in the the cytosol. cytoplasm and when the viral infection destroys the ○ With membrane injury, as in viral or hepatocyte, it will cause leakage. chemically in-duced hepatitis, these - Ni gawas na nuon si ALT and AST enzymes are released and enter the (Specifically ASTC; not so much ASTm) sinusoids, raising plasma AST and ALT - Causing now increased AST and ALT in our serum. activities. Mitochondrial aspartate aminotransferase ASTc is leaked along with ALT but not ASTm (ASTm) is released primarily with mitochondrial injury caused by ethanol, as in alcoholic hepatitis. Hepatobiliary (ALP) Alkaline phosphatase (ALP) and y-glutamyl ○ ALP is what is tested in this disorder transferase (GGT) are found primarily on the ○ Common example: GALLSTONE or canalicular surface of the hepatocyte. OBSTRUCTION sa bile duct ○ Bile acids accumulate in cholestasis and ○ If there is an obstruction in the bile duct, dissolve membrane fragments, releasing the bile cannot pass through therefore it bound enzymes into plasma. will go back. GGT is also found in the microsomes, represented ○ Which now damages the hepatocytes, as pink rings in the figure; microsomal specially the part of the hepatocyte facing enzyme-inducing drugs, such as phenobarbital and the canaliculi. Dilantin (phenytoin), can also increase CGT synthesis and raise plasma GGT activity. The location of the enzyme can be a clue of what disorder a person might have. All 3 of them are LIVER ENZYMES but AST is found in: Cytosol and Mitochondria they have different directions and diagnosis. ALT is found in: Cytosol ALP is found in: One part only (hepatocyte facing the DIAGNOSTIC SIGNIFICANCE canaliculi) Limited mainly to the evaluation of hepatocellular disorders and skeletal muscle involvement In diagnosing AMI it is not useful because a rise in AST does not automatically refer to AMI. Remember in AMI time is of the essence; it has to be diagnosed immediately ○ ACUTE MYOCARDIAL INFARCTION Rise: 6-8 hours Peak: 24 hours Normalizes within 5 DAYS Not useful in the One end of the hepatocyte is near the canaliculi, the diagnosis of AMI other is near the blood vessel. ○ Pulmonary embolism ALP is seen in the area of the hepatocyte closest to ○ Congestive heart failure the bile. ○ Acute hepatocellular Disorders Viral hepatitis: 100 times upper LIVER DISORDERS limit of Normal (ULN) Hepatocellular (ALT & AST) Ex. 30-60, in this case ○ Diseases that damage the hepatocyte the ULN is 60, directly. 100 times of 60 that is e.g. Hepatitis and Cirrhosis the usual value of the ○ Hepatitis B is a viral infection and can be patient if the patient has acquired through sex or contaminated Acute hepatocellular blood products Disorder. ○ Which will go through our blood circulation Liver Cirrhosis: 4 times ULN and will travel all throughout the body. ○ Muscular Dystrophies: 4-8 times ULN ○ Once they reach the liver, from the blood ○ Inflammatory conditions vessel, the viral infection will target the hepatocyte facing the bloodstream. ○ So what is damaged is the entire hepatocyte.That's why in hepatocellular disorders what is requested is AST and ALT alone. Made by Miguel Astronomo and Daniel Budo 2  ALANINE AMINOTRANSFERASE 4. Kidney 5. Pancreas 5. Pancreas 6. Spleen ENZYME 2.6.1.2 6. Erythrocytes 7. Erythrocytes CLASSIFICATION Intracellular level: 7000 Intracellular level: 3000 SYSTEMATIC L-Alanine: 2-oxoglutarate times more than plasma times more than plasma NAME Aminotransferase AST is not used as a marker of a heart problem because it is FUNCTION AND Transfer of an amino group found in SO MANY OTHER tissue sources. PRODUCT between alanine and alpha- keto - We have TROPONIN I for cardiac tissues. acids to form glutamate and pyruvate PHOSPHATASES FORMER NAME serum glutamic-pyruvic transaminase (SGPT or GPT) Alkaline Phosphatase Acid Phosphatase (ALP) (ACP) COFACTOR Pyridoxal phosphate (Vit. B6) Tissue Sources: Tissue Sources 1. Liver - focus 1. Prostate (richest a. Sinusoidal and source) bile canalicular 2. Bone membranes 3. Liver 2. Bone - focus 4. Spleen a. Osteoblasts 5. Kidney 6. RBC 3. Intestine 7. Platelets 4. Spleen The same chemical reaction with the AST, the only 5. Placenta difference is the substrate used and the product 6. Kidney produced. ○ Instead of L-Aspartate on the amino group ALKALINE PHOSPHATASE (ALP) substrate we now use L-Alanine which will yield Pyruvate ENZYME 3.1.3.1 2-Oxoglutarate → L-Glutamate (Amino acid) CLASSIFICATION L-Alanine → Pyruvate (Keto acid) SYSTEMATIC Orthophosphoric Monoester NAME Phosphohydrolase (Alkaline DIAGNOSTIC SIGNIFICANCE Optimum) Confined mainly to the evaluation of hepatic disorders (hepatocellular disorders): ALT elevation Optimum pH 9.0 -10.0 is frequently higher than AST due to its LONGER HALF-LIFE in the blood FUNCTION AND Catalyze the hydrolysis of various ○ 47 hours (ALT) vs. 17 hours (ASTc) PRODUCT phosphomonoesters at a certain ○ HALF-LIFE is the time needed by the pH enzyme to half its level in the blood. Liberate inorganic phosphate Note: in acute hepatocellular injury, initially AST > from an organic phosphate ester ALT but ALT > AST within 24-48 hours (except in with the concomitant production of ALCOHOL-INDUCED HEPATOCYTE INJURY an alcohol which damages the mitochondria; ○ Na release si mitochondrial AST tungod COFACTOR Magnesium and Zinc kay na damage through alcohol. ○ Mitochondrial AST has a half-life of 87 hours; ○ De Ritis ratio: 3-4:1 Compares the value of AST and ALT (Respectively) TISSUE SOURCES Aspartate Alanine Aspartate (ALT) 4 MAJOR ISOENZYMES: Aminotransferase (AST) Placental ALP Liver ALP 1. Cardiac tissue - 1. Liver - highest Bone ALP highest 2. Kidney Intestinal ALP 2. Liver 3. Heart ○ Common in patients w/ Blood Type B or O 3. Skeletal Muscles 4. Skeletal muscle and are SECRETORS Made by Miguel Astronomo and Daniel Budo 3  Are individuals that their blood type can be determined on their body fluids. e.g. tears, saliva, breast milk, semen. 80% Secretors In persons with Type A blood type their Intestinal ALP is bound to RBCs. ACID PHOSPHATASE (ALP) ENZYME 3.1.3.2 CLASSIFICATION SYSTEMATIC Orthophosphoric Monoester NAME Phosphohydrolase (Acid Optimum) Optimum pH 5.0 FUNCTION AND Catalyze the hydrolysis of various PRODUCT phosphomonoesters at a certain pH Liberate inorganic phosphate from an organic phosphate ester with the concomitant production of an alcohol COFACTOR Magnesium and Zinc Made by Miguel Astronomo and Daniel Budo 4  CLINICAL CHEMISTRY-2 - LECTURE PRELIMS TOPIC 4. / Phosphatases and Its Clinical Significance Lecturer/s: Mr. Christian John Villahermosa, RMT, MSMT August 17, 2023 Are individuals that their blood PHOSPHATASES type can be determined on their body fluids. e.g. tears, saliva, breast Alkaline Phosphatase Acid Phosphatase (ALP) (ACP) milk, semen. 80% Secretors Tissue Sources: Tissue Sources In persons with Type A blood type 1. Liver - focus 1. Prostate (richest their Intestinal ALP is bound to a. Sinusoidal and source) RBCs. bile canalicular 2. Bone membranes 3. Liver The most commonly tested isoenzyme of ALP are LIVER 2. Bone - focus 4. Spleen and BONE a. Osteoblasts 5. Kidney 6. RBC because if we look into placental and intestinal 3. Intestine 7. Platelets diseases, there are other tests specific for them. 4. Spleen Application of ALP testing in the lab is purely in 5. Placenta bone and liver disorders 6. Kidney ○ e.g if a person has high ALP supposedly it should be differentiated into which ALKALINE PHOSPHATASE (ALP) isoenzyme Because if ALP is high you will not know ENZYME 3.1.3.1 which organ or tissue has a disease if its CLASSIFICATION isoenzyme is not differentiated SYSTEMATIC Orthophosphoric Monoester NAME Phosphohydrolase (Alkaline In medical practice (In other contries)… Optimum) ALP is tested, not to have an idea of what disease a patient might have, but it is used for Optimum pH 9.0 -10.0 CONFIRMATION that the patient has this type of disease. FUNCTION AND Catalyze the hydrolysis of various Examination is done first → After Enzyme is tested PRODUCT phosphomonoesters at a certain pH Methods used to differentiate the isoenzymes and other Liberate inorganic phosphate multiple forms of ALP from an organic phosphate ester Electrophoresis with the concomitant production of an alcohol Stability to denaturation by heat or chemicals Response to the presence of selected inhibitors COFACTOR Magnesium and Zinc Affinity for specific lectins Immunochemical characteristics Isoenzymes (Electrophoresis) LIVER → migrates fastest BONE PLACENTAL INTESTINAL → Migrates slowest Liver ALP: migrates the fastest and most anodal; It 4 MAJOR ISOENZYMES: has 2 Classifications which points to different Placental ALP disorders which vary in severity Liver ALP ○ Major liver fraction: usually in the EARLY Bone ALP STAGES of hepatobiliary conditions Intestinal ALP Disease is only in the bile duct ○ Common in patients w/ Blood Type B or O and the liver and are SECRETORS Made by Miguel Astronomo and Daniel Budo 1  It suggests that the patient has a 3M Urea - Inhibits bone ALP hepatobiliary condition. Since it is in its early stages we Abnormal Isoenzymes (Abnormal Fractions associated with can easily manage the disease Neoplasms): Ectopic production of an enzyme by malignant itself. tissues; NOT PRESENT in normal individuals only in patients ○ Fast liver fraction (Alpha1 liver): Found in with a certain type of cancer. metastatic carcinoma of the liver and hepatobiliary diseases (valuable indicator This isoenzymes are produced by cancer cells of obstructive liver disease) The cancer started somewhere CARCINO PLACENTAL ALKALINE PHOSPHATASE else 1. Regan Isoenzyme Bone ALP: Osteoblast activity 2. Nagao Isoenzyme Placental ALP 3. Kasahara Isoenzyme Intestinal ALP - Found in all of the following cancers: Hepatocellular/ gastric/ maxillary/ Isoenzymes (Heat Stability) pulmonary/ bladder carcinoma ALP activity is measured BEFORE AND AFTER heating the serum at 56°C for 10 min. Similarities to the placental isoenzyme PLACENTAL → Most heat stable 3% - 15% in cancer patients Produced by CANCER CELLS INTESTINAL LIVER Carcino are found in cancer patients only and most of their BONE → Most heat labile properties is SIMILAR with PLACENTAL ALP Placental ALP: Most heat stable (resist heat Not all cancer patients produce this enzymes; only denaturation at 65°C for 30 min) 3-15% Intestinal ALP This isoenzymes are NOT USED TO DIAGNOSE Liver ALP CANCER due to its small numbers Bone ALP ABNORMAL ALP ISOENZYMES When you heat a patient serum you can target the BONE REGAN ISOENZYME and LIVER only, since these isoenzymes are the most tested Detected in various carcinomas in the laboratory, the other 2 are not so much regarded. ○ Increased in ovarian (highest incidence) Both are heat sensitive yet heating can still be used and gynecological cancers (better used to detect the presence of the last 2 isoenzymes as therapy monitor) under heat stability ○ Can be used to determine the EFFECTIVENESS OF THERAPY HOW TO KNOW IF THE MEASURED ACTIVITY IS FOR Ex. For a cancer patient after a BONE ALP OR LIVER ALP therapy it should go down, but if it If the activity is < 20% of the before level that is doesn’t it indicates that the BONE (Most heat sensitive) therapy is not working for the ○ Na denature ang bone ALP after heating patient. If the activity is >20% of the before level that is MOST HEAT STABLE OF ALL ALP: resists LIVER denaturation (65°C for 30 min) ○ PRESUMED only to be liver because there ○ It assumes the properties of PLACENTAL are other isoenzymes above the liver ALP in heating which are much stable. Inhibited by phenylalanine and migrates like bone ALP Isoenzymes (Chemical Inhibition) ○ In electrophoresis BONE ALP is its Using chemical inhibition, it is Impossible to differentiate the equivalent when regan isoenzymes four fractions; It is DIFFICULT to measure ALL FOUR. migrate. There is no chemical that can detect all four NAGAO ISOENZYME It has the same properties with Regan except on the Phenylalanine - Inhibits intestinal ALP and SOURCE OF CANCER and CHEMICAL INHIBITION placental ALP to a much greater extent than liver Identical to the Regan Fraction (considered as a and bone ALP variant of Regan) Levamisole - Inhibits bone ALP and liver ALP Inhibited by both Phenylalanine and L-leucine (Henry) Detected in metastatic carcinoma of pleural ○ you will not know if the patient has bone surfaces and adenocarcinoma of the pancreas related disease or liver related disease and bile duct since both of them are inhibited and ○ The source of adenocarcinoma is the through that we cannot measure them. epithelial cells Made by Miguel Astronomo and Daniel Budo 2  DIAGNOSTIC SIGNIFICANCE (ALP) ○ Cysts and carcinoma Hepatobiliary Disorders ○ Parasitic infection ○ Biliary Tract Obstruction: 4-10 times ULN Intrahepatic (in the liver) due to increased synthesis of the enzyme ○ Viral hepatitis induced by CHOLESTASIS (Translocation ○ Cholestasis in pregnancy (due to estrogen) of ALP to the sinusoidal surface) RARE in the general population Since the bile flow was blocked, yet COMMON in most pregnant the bile will go back to where it woman came from Increased ESTROGEN → SLOW until the pressure exerted by the bile flow bile on the hepatocyte will damage the hepatocyte itself May lead to CHOLANGITIS (Inflammation on the bile duct; Will now then cause the ALP on side effect of cholestasis) the canalicular surface to Normally our bile contains its own antibody (IgG), translocate in the sinusoidal which helps our body fight off micro bacterial or surface parasitic infection that could potentially enter the Hepatocellular Disorders bile flow and damage the liver. ○ Hepatitis and cirrhosis (usually less than 3 In cases of blockage, the antibody from the normal times ULN) flow of bile is no longer to be found; thus, the normal flora found in our small intestine could go up the duct and cause infection. Which now leads to cholangitis which could cause SEPSIS (if bacterial infection will spread) Cholestasis is most of the time ASYMPTOMATIC; it will only show its signs and symptoms if something obstructed already the bile duct. Hallmarks: Charcot’s triad ○ (1) Fever, (2) Right upper quadrant pain, (3) Jaundice Reynold’s pentad ○ (1) Fever, (2) Right upper quadrant pain, (3) Jaundice, (4) Shock (low blood volume), (5) Altered mental state Tokyo Guidelines are also used BONE DISORDERS Paget Disease (osteitis deformans; increased ALP) ○ Gain in bone mass (disordered/ unsound In cases of hepatobiliary disorders, ALP has a new bone) higher concentration compared in cases of ○ Normally our bones are REMODELED hepatocellular disorders ANNUALLY (10%). This is due to the fact that most ALP are found in ○ In paget disease, remodelling is MORE the canalicular end of the hepatocyte frequent yet the quality of remodelled bone is of NOT GOOD quality. Most of the time,tests for ALP in cases of hepatobiliary osteomalacia and rickets disorders. ○ The difference between the two is that osteomalacia occurs in ADULTS while Any blockage on the bile ducts could lead to rickets occurs in CHILDREN. CHOLESTASIS ○ Due to the LACK OF VITAMIN D ○ Bile cannot pass through; hampers the Deformed bones flow of bile through the bile duct. Hyperparathyroidism Osteogenic carcinoma CHOLESTASIS Healing bone fractures Failure of bile excretion or transport During periods of physiologic bone growth Extrahepatic (outside liver) ○ Gallstones (cholelithiasis and NORMAL PREGNANCY choledocholithiasis) Increased ALP; due to the increase of placental Lith = Bato/Stones ALP Cholelithiasis → blockage of the Can be detected between weeks 16 and 20 and Cystic duct persists until the onset of labor Choledocholithiasis → blockage of the Common bile duct Made by Miguel Astronomo and Daniel Budo 3  May be elevated in hypertension, preeclampsia, Useful in investigation of rape: Vaginal washings threatened abortion are examined for seminal fluid ACP activity, which can persist for up to 4 days GENETIC ABNORMALITY ○ Presumptive evidence of rape: ACP activity > 50 U/L = POSITIVE Decreased ALP is observed in HYPOPHOSPHATASIA (An ○ If ACP is detected there is semen inborn error of metabolism) contamination because the semen went - Defect in the gene that encodes for ALP through the prostate gland isoenzymes (Kidney, liver, bone) Increased ACP ACID PHOSPHATASE (ACP) Bone disease ○ Associated with OSTEOCLASTS ENZYME 3.1.3.2 Paget’s disease CLASSIFICATION ○ because it includes osteoclasts also Breast cancer with bone metastases SYSTEMATIC Orthophosphoric Monoester ○ Spread through the bone NAME Phosphohydrolase (Acid Optimum) Gaucher disease Optimum pH 5.0 ○ Cause by a deficiency of the enzyme GLUCOCEREBROSIDASE leading to the FUNCTION AND Catalyze the hydrolysis of various buildup of fatty substances in certain PRODUCT phosphomonoesters at a certain organs pH ○ it hydrolyzes glycolipids; within the cell there is a glycolipid that it should degrade. Liberate inorganic phosphate from an organic phosphate ester with the concomitant production of Thrombocytopenia an alcohol Increased ACP Resulting from excessive platelet destruction from COFACTOR Magnesium and Zinc