Amniotic Fluid and HCG Lecture Lesson 10 PDF

ANALYSIS OF URINE AND BODY FLUIDS (LECTURE) - Urine regulation: - It is a clear, colorless, and sometimes yellowish o High urine = high fetal swallowing fluid which is found in pregnant women...

ANALYSIS OF URINE AND BODY FLUIDS (LECTURE) - Urine regulation: - It is a clear, colorless, and sometimes yellowish o High urine = high fetal swallowing fluid which is found in pregnant women o Lung fluid adds lung surfactant to the - It is usually produced during the first 12 days amniotic fluid following the conception o Phospholipids: one of the compositions - Purpose: surround the growing fetus in the of amniotic fluid that could assess the uterus lung maturity - Problems in the amount of amniotic fluid inside - Increased amniotic fluid peak at 800 to 1200 mL the uterus will lead to complication in the third trimester is the result of fetal urine - It is made up of fetal urine and fetal cells - During the first trimester of pregnancy, the Abnormal Amniotic Fluid Volume amniotic fluid is mainly made up of maternal Polyhydramnios (>1200 mL) plasma o Increased amniotic fluid volume - It is an ultrafiltrate of plasma o Causes: Decreased fetal swallowing of urine and neural tube defects STRUCTURE OF THE PLACENTA o Acute Polyhydramnios – associated with fetal edema, hydroxy fetalis, or fetal heart failure o Chronic Polyhydramnios – associated with fetal disorders (ex. poor fetal swallowing, toxemia during pregnancy, and mother has diabetes) Oligohydramnios (2.0 mg/dL Provide cushion for the fetus Allow fetal movement QUADRUPLE SCREENING TEST Stabilizes fetal temperature exposure - Done prior to performing Amniocentesis Infection control (IgG could pass through the - Check the hormones in the blood of the mother placenta coming from the mother) - Performed during the second trimester (14th up Proper lung development to 22nd week of gestation) Exchanges water and chemicals among the fluid, - It is used to evaluate whether the pregnancy has fetus, and maternal circulation increased chance of certain conditions such as Muscle and bone development down syndrome or neural tube defects Lubrication (to prevent webbing) Four Pregnancy Hormones Alpha-fetoprotein (AFP) – protein made by the AMNIOTIC FLUID VOLUME developing baby - During the first trimester, approximately 35 mL o Check for neural tube defects (NTD) or of amniotic fluid is derived primarily from the trisomy 21 (down syndrome) maternal circulation or maternal plasma Human Chorionic Gonadotropin (hCG) – - As the fetus grows after the first trimester of hormone made by the placenta pregnancy, the major contributor of the amniotic Unconjugated estriol (UE3) – hormone made by fluid will be the fetal urine and lung fluid the placenta and the liver of the baby o Volume: 200 to 500mL per day Inhibin A – hormone made by the placenta  ANALYSIS OF URINE AND BODY FLUIDS (LECTURE) AMNIOCENTESIS Test for Hemolytic Disease of the Newborn: - It is an invasive procedure because the amniotic o Detect the presence of bilirubin fluid is obtained by needle aspiration into the o Protect from light amniotic sac - Two procedures: AMNIOTIC FLUID vs. MATERNAL URINE ANALYTE AMNIOTIC FLUID MATERNAL URINE o Transabdominal amniocentesis Less Reliable (preferred) Protein + – o Vaginal amniocentesis (may cause Glucose + – leakage of amniotic fluid and More Reliable contamination of bacteria) Urea 300 mg/dL Creatinine < 3.5 mg/dL > 10 mg/dL - It is guided by an ultrasound - It is done by OB-GYN and trained doctors AMNIOTIC FLUID MATERNAL URINE - Maximum of 30 mL collected in sterile syringes The watery and typically - Discard first 2 to 3 mL for contamination The fluid, which surrounds the yellowish fluid, one of the chief - Protect specimens from light for bilirubin fetus within the amnion means of eliminating excess analysis for HDN at all times: amber tubes or water and salt black plastic tube covers Occurs inside the amniotic Produced by kidney fluid Watery hopefully, clear but Normal urine is either colorless sometimes yellow, green or or pale yellow in color with white specks Has a strong "fish-like" odor Odorless due to bacterial growth after urination Contains nutrients including Contains mainly water, proteins, carbohydrates, and inorganic salts, proteins, lipids, hormones, immune hormones, and a wide range of system cells, and the urine of metabolites baby Normal level is about one Normal range of urine is 0.6 to quart by 36 weeks after 2.6 L per person per day pregnancy Serves as a cushioning liquid to Responsible for excreting the fetus and facilitates the nitrogenous wastes, excess exchange of nutrients and salts, and other water-soluble water between mother and chemicals out of the body baby A true amniotic fluid leak The flow can be controlled by cannot be controlled squeezing muscles FERN TEST - Detects ruptured amniotic membranes - It is also used to diagnose early pregnancy - Procedure: o Vaginal Fluid → Slide (Air Dry for at least 5 to 7 minutes) → visualize under LPF o (+) Fern-like Crystal (Amniotic Fluid) SPECIMEN COLLECTION Method of Collection: Amniocentesis with ultrasound Up to 30 mL is collected in sterile AMNIOTIC FLUID COLOR Color Clinical Significance syringe Colorless Normal 2nd Trimester Amniocentesis: Assess genetic Traumatic Tap, trauma, defects or chromosomal abnormalities (Ex. Blood-Streaked intraamniotic hemorrhage Trisomy 21 and Down’s Syndrome) Yellow HDNF (Bilirubin) 3rd Trimester Amniocentesis: Fetal Lung Dark-Green Meconium Maturity and Fetal Hemolytic Disease (first fetal bowel movement) Dark Red Brown Fetal Death SPECIMEN HANDLING DIFFERENT TESTS FOR DISORDERS Test for Fetal Lung Maturity o Place on ice (deliver) Test for HDNF o Refrigerated or frozen - Hemolytic Disease of the Fetus & Newborn o Filtration: prevents loss of phospholipids - Also known as Optical Density or OD 450 Test for Cytogenetic Studies - To detect whether there is any hemolytic disease o Room temperature or body temperature - Absorbance of Amniotic Fluid  ANALYSIS OF URINE AND BODY FLUIDS (LECTURE) o Normal: Increased at 365nm and Test for Fetal Lung Maturity decreased at 550 nm - The production of lung fluid containing o HDN: increased at 450 nm (bilirubin) phospholipids can be used to assess or evaluate - Common cause: Rh incompatibility whether the fetal lung is matured and ready for - Results are plotted on a Liley Graph delivery o Zone I: nonaffected/ mildly affected - Most common complication of early delivery is fetus respiratory distress syndrome (RDS) o Zone 2: Moderately affected fetus - Lack of lung surfactant, which keeps the alveoli (requires close monitoring) open during inhaling and exhaling o Zone 3: Severely affected fetus (requires - Surfactant decreases the surface tension on the intervention) alveoli so they can inflate more easily - Usually checked during the 3rd trimester - Many laboratory tests are available for FLM - Interferences include: cells, mecomium, debris, and hemoglobin Lecithin/Sphingomyelin (L/S Ratio) - Reference Method - Lecithin: for alveolar stability o primary component of lung surfactant o high during 3rd trimester (35th week) - Sphingomyelin: serves as a control for the rise of lecithin o Produced at a constant rate o Start to increase during the 26th week - Mature fetal lungs: Ratio of > 2.0 - Cannot be done on a specimen contaminated by blood or meconium - It requires thin-layer chromatography Amniostat-FLM - Immunologic test for Phosphatidyl Glycerol - Uses antisera against Phosphatidyl Glycerol (not Test for Neural Tube Defects a major component of lung surfactant) - The neural tube forms the early brain and spine o The production of phosphatidyl glycerol - Alpha-fetoprotein (AFP) produced by the fetal is parallel to lecithin liver prior to 18 weeks’ gestation - Alternative for L/S ratio - Increased levels in maternal blood or amniotic - Not affected by blood or meconium fluid indicate possible anencephaly or spinal - Production of PG is delayed among diabetic bifida mother - Increased levels are found when skin fails to close over neural tissue Foam Stability Test (Foam or Shake Test) - Measure maternal blood first, then amniotic - Bedside testing fluid - Amniotic Fluid + 95% Ethanol → Shake 15 secs → - Alpha-fetoprotein could also reach the maternal Stand 15 mins circulation - Mature fetal lungs: (+) Foam/Bubbles - Diseases: o Ethanol is an anti-foaming agent o Spina bifida – the spine and the spinal o Continuous formation of bubbles in the cord do not form properly solution indicates sufficient amount of o Anencephaly – serious birth defect in lung surfactant which the baby is born without the parts - Cannot be used with contaminated amniotic of the brain and skull fluid - Screening Test: AFP o Increased in neural tube defects Microviscosity o Decreased in Down Syndrome - The presence of phospholipids decreases - Confirmatory Test: Acetylcholinesterase microviscosity (enzyme that hydrolyzes a neurotransmitter in - Measured by Fluorescence Polarization the body, acetylcholine) Lamellar Body Count - Lamellar Bodies (a.k.a. Type II pneumocytes) o Found in the lungs as dust cells o Responsible for production of alveolar surfactants o Spherical in shape and contains granules inside o Storage form of lung surfactant - Alternative method to detect the presence of phospholipids - adequate FLM: >32,000/uL lamellar body count - hematology analyzer is used for counting  ANALYSIS OF URINE AND BODY FLUIDS (LECTURE) OD 650 nm - Specimen: 1st Morning Urine, random urine, - Increased Lamellar = Increased OD (Absorbance) serum, or plasma - An OD of ≥ 0.150 is equivalent to: - Responsible for rescuing and maintaining corpus o L/S ratio of ≥ 2.0 luteum, which produces the hormone o (+) Phosphatidylglycerol progesterone. - Lamellar bodies absorbance: 650nm o It also produces estradiol and inhibit-A hormones Lamellar Bodies - Secreted immediately on implantation (6-8 days - Approximately 90% phospholipid and 10% after conception) protein - Doubling does not occur in ectopic pregnancy - Secreted by the type II pneumocytes of the fetal - HCG decreases to nondetectable 2 weeks after lung to the alveolar space at about 24 weeks of delivery gestation - Increase in amniotic concentration from 50,000 METHODS IN MEASURING HCG ACTIVITY to 200,000/mL by the end of the third trimester Immunologic test - OD of 150 at 650 nm correlates with L/S ratio of o Test kits are used 2.0 and the presence of PG o Direct or Agglutination Test (common) - Lamellar bodies are red in platelet channel when o Indirect or Agglutination Inhibition/ using hematology analyzer Hemagglutination Inhibition Radioimmunoassay: competitive binding assay Radio Receptor Assay Immunometric Assay Bioassays: uses test animal and done in vivo HCG Bioassays Animal Mode of Test Positive Result Test For Fetal Age Used Injection Formation of ≥ 2.0 mg/dL creatinine = 36 weeks Immature hemorrhagic Ascheim- female Subcutaneous follicles and Zondek Test for Fetal Well-Being and Maturity mice Corpus Lutea Test Normal Values Significance (enlarged ovary) Change of OD at Hyperemic Uterus Bilirubin scan HDN Mature 450 nm (>0.25) and Corpora virgin Marginal ear Neural Tube Friedman hemorrhagia Alpha-Fetoprotein < 2.0 female vein Defects (small spots in rabbit Fetal Lung ovary) L/S Ratio ≥ 2.0 Female Maturity Fetal Lung toad South Oogenesis Amniostat-fetal Maturity/ Hogben African Lymph sac (Extrusion of Positive Clawed eggs) Lung Maturity Phosphatidyl Glycerol 7 Frog Foam Stability Fetal Lung Spermatogenesis ≥ 47 Galli- Male Frog Index Maturity Subcutaneous (presence of Mainini Male Toad Microviscosity Fetal Lung sperm in urine) ≥55 mg Ovarian (FLM-TDx) Maturity Immature Optical Density Fetal Lung Frank- hyperemia ≥0.150 Female Subcutaneous 650nm Maturity Berman (enlarged red Rats Lamellar Body Fetal Lung ovary) ≥ 32,000/mL Ovarian Count Maturity Female Hyperemia Kupperman Intraperitoneal Rats (enlarged red ovary) - It is a hormone that is being produced by the cytotrophoblast cells in the placenta of pregnant women as a recognition of pregnancy - It is detected using pregnancy test - Male who are positive with pregnancy test have a possibility of having seminoma or testicular cancer - HCG is important to thicken the uterine lining to support the growing embryo and to tell the body to stop menstruation - There is a high HCG level after conception and it will continue to rise about 10 weeks of pregnancy - Peaks during 1st trimester of pregnancy (Increased blood, urine, amniotic fluid) - Composed of 2 subunits (dimer): o Alpha: HCG, LH, FSH, TSH o Beta: unique for HCG