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Medical Biochemistry

Medical Biochemistry: Principles and Experiments

John F. Van Pilsum
Robert J. Roon
Copyright Date: 1986
Edition: NED - New edition
Pages: 120
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  • Book Info
    Medical Biochemistry
    Book Description:

    Medical Biochemistry was first published in 1986. A good knowledge of biochemical analysis is essential for today’s health-care practitioners, who, with their patients, face a widening array of laboratory tests to aid in diagnosis. The requisite biochemical methods and principles are best understood if medical students perform their own experiments, yet most currently available laboratory manuals are intended for general biochemistry courses and lack the clinical orientation that could make them useful in a medical context. John Van Pilsum and Robert Roon have designed this laboratory manual specifically to introduce first-year medical students to clinical methods in biochemistry and to help them understand basic biochemical principles as they are applied top medical practice. Each chapter in Medical Biochemistry is devoted to a basic set of related problems and includes, along with laboratory procedures, a clear and readable introduction, a list of selected references, and questions. All of the experiments call for procedures that are used routinely in most clinical laboratories. The areas covered include: electrophoresis of blood proteins, enzymes as diagnostic indicators, lactate dehydrogenase isozymes, the determination of glucose, blood lipids, experiments with nucleic acids, inheritable diseases and genetic engineering, the use of radioisotopes in clinical biochemistry, glycosylated hemoglobin, steroid hormone formation, immunoelectrophoresis of serum proteins, radioimmunoassay of thyroxine, serum electrolytes and carbon dioxide, and the lecithin-sphingomyelin ratio of amniotic fluid. The contributors, besides Van Pilsum and Roon, include: Marilyn H. Koenst, John D. Lipscomb, James B. Howard, Esther F. Freier, Ivan D. Frantz, Denise M. McGuire, Howard C. Towle, Dennis M. Livingston, Ronald D. Edstrom, Robert P. Changler, Frank Ungar, Maureen A. Scaglia, James F. Koerner, and Charles W. Carr.

    eISBN: 978-0-8166-5569-4
    Subjects: Health Sciences

Table of Contents

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  1. Front Matter
    (pp. i-iv)
  2. Table of Contents
    (pp. v-v)
  3. Preface
    (pp. vi-vi)
  4. Acknowledgments
    (pp. vii-viii)
  5. Safety Precautions in the Laboratory
    (pp. ix-x)
  6. Venipuncture and Processing of Blood Samples
    (pp. xi-2)
  7. 1 Electrophoresis of Blood Proteins
    (pp. 3-15)
    John F. Van Pilsum, Robert J. Roon and Marilyn H. Koenst

    Human blood contains hundreds of individual proteins. The quantitation of many of these proteins (or groups of proteins) in the plasma is used as a diagnostic aid by physicians. Electrophoresis makes possible the separation and quantitation of six groups of plasma proteins; immunoelectrophoresis permits the identification of 15 to 20 plasma proteins.

    Electrophoresisis the movement of a charged particle or electrolyte in a solution under the influence of an electric field. The particles are in a solvent that is supported by an inert and homogenous stabilizing medium such as a paper or a gel. The movement of the proteins...

  8. 2 Enzymes As Diagnostic Indicators
    (pp. 16-24)
    John D. Lipscomb and James B. Howard

    The clinical application of enzymology was made possible by advances over the past 50 years in our understanding of cellular metabolism and enzyme kinetics. Enzymes are currently used in the clinical laboratory in three ways:

    1.They are reagentsemployed in specific methods for measuring the concentrations of metabolites, such as glucose, in the blood.

    2. The measurement of theactivity of enzymesin the blood can serve in the diagnosis and management of certain diseases.

    3. Thecharacteristics of the enzymesin the blood can be used to identify the organ or tissue involved in the disease state.

    During the...

  9. 3 Tissue Distribution of Lactate Dehydrogenase Isozymes
    (pp. 25-30)
    John D. Lipscomb

    Many enzymes are composed of multiple subunits that are identical in some cases and nonidentical in others. A few enzymes are active with either identical or nonidentical subunits as long as the final quaternary complex has the appropriate total number of subunits. This interchangeability of subunits leads to a family of enzymes within a single organism which catalyzes the same reaction. These isoenzymes, commonly called isozymes, have distinct differences in their physical properties (molecular weight, isoelectric point, denaturation temperature) as well as their catalytic properties (Km, turnover number, pH optimum). However, since the subunits usually arise from genes produced by...

  10. 4 Determination of Glucose in Serum and Urine
    (pp. 31-36)
    Esther F. Freier and John F. Van Pilsum

    Glucose is utilized in one or more ways (as a metabolic fuel, in the synthesis of other metabolic fuels, in the synthesis of cellular components, etc.) byalltissues in the body. The measurement of levels of glucose in the blood (and urine) is helpful in the diagnoses of a large variety of diseases and is the most widely performed clinical biochemical analysis. Many different methods for determining glucose levels are currently being used by hospital clinical laboratories. Thespecificitiesof these methods forglucosevary greatly, as do the principles of the various procedures. You, as future physicians, should be...

  11. 5 Enzymatic Analysis of Blood Lipids
    (pp. 37-46)
    Ivan D. Frantz and John F. Van Pilsum

    The lipids found in human blood are insoluble in water and must be solubilized and transported as complexes with proteins. The free fatty acids, which are added to the blood from adipose tissue, are transported as complexes with the serum albumin. The phospholipids, cholesterol (both free and esterified), and triglycerides are transported as protein-containing complexes called lipoproteins. The amounts of lipoproteins in the blood are a function of the nutritional state of the individual and have also been implicated in the etiology of certain disease states such as atherosclerosis. Alterations in the amounts of the various lipoproteins in blood are...

  12. 6 The Use of Recombinant DNA in the Detection of Genetic Abnormalities
    (pp. 47-53)
    Denise M. McGuire, Howard C. Towle and Dennis M. Livingston

    The structure and function of nucleic acids have in recent years assumed an increasingly important and exciting role in biomedical research. Through the techniques known as gene cloning, medically important products such as human insulin and growth hormone are now being produced in recombinant bacteria for treatment of patients. In addition, these advances are providing new ways to test DNA from individuals for genetic abnormalities and a better understanding of the molecular nature of such genetic disorders.

    DNA, or deoxyribonucleic acid, is the molecular store of genetic information. This information is sufficient to encode the sequence of every protein that...

  13. 7 Inheritable Diseases and Genetic Engineering
    (pp. 54-57)
    Dennis M. Livingston

    An expanding field of concern for the physician is the study of inheritable diseases and the genetic predisposition of individuals to contracting diseases. The purpose of this laboratory session is to describe: the genetics of inheritable diseases; how a genetic lesion results in a recognizable disease: and how health scientists can “treat” inheritable diseases.

    All human cells except the gametes contain two of each of 23chromosomes.One copy of each chromosome pair is inherited from the ovum and the other from the sperm. Which of the two copies of each pair is packaged into the gamete is a matter...

  14. 8 Use of Radioisotopes in Clinical Biochemistry
    (pp. 58-65)
    Ronald D. Edstrom and Robert P. Chandler

    Biochemical processes such as hormone-cell interactions, gene replication, and protein synthesis take place in dilute solutions or at low concentrations at membrane surfaces. These low concentrations require extraordinarily sensitive methods of detection and measurement. The high sensitivity of instruments used in detecting radioactivity permits the performance of experiments that would fail if ordinary chemical methods were used. Chemical methods of analysis are generally sensitive to a few nanomoles at best (10-9mole). By using radioisotopes, molecules can be detected in the femto- (10-15) to picomole (10-12) range.

    Many metabolic pathways have been elucidated with the aid of14C...

  15. 9 Determination of Glycosylated Hemoglobin
    (pp. 66-69)
    Marilyn H. Koenst and Ronald D. Edstrom

    Approximately 90% of normal adult human hemoglobin is the α2β2tetramer called hemoglobin A. Four to 7% of hemoglobin A is present as glycosylated hemoglobin, which contains covalently-bound glucose and can be distinguished from hemoglobin A by various methods. In persons with high blood-glucose levels the amount of the glycosylated forms of hemoglobin also will be high, and thus the measurement of glycosylated hemoglobin is of clinical importance. In this experiment we will measure the relative amounts of glycosylated hemoglobin in blood from normal and diabetic individuals.

    The hemoglobin of normal persons contains a few percent of molecules that have...

  16. 10 Biosynthesis of Adrenal Steroid Hormones
    (pp. 70-76)
    Frank Ungar

    The adrenal cortex converts cholesterol to biologically active steroid hormones. More than 30 steroid compounds have been isolated from adrenal tissue. The analysis of adrenal steroids in blood and urine is used in the diagnosis of a variety of disease states, e.g., Cushings disease, Addison’s disease, hypopituitarism, adrenal adenoma, diuretic abuse, cardiac failure. The purpose of today’s experiment is to demonstrate the formation of the adrenal steroids by an extract of bovine adrenal glands and the inhibition of their formation by the drug Metyrapone. This drug is used in the clinic to interfere with adrenal steroid formation and thus test...

  17. 11 Immunoelectrophoresis of Serum Proteins
    (pp. 77-86)
    Maureen A. Scaglia and James F. Koerner

    Only five protein fractions in human serum can be identified by the technique of simple electrophoresis. However, with the technique of immunoelectrophoresis, 15-20 proteins are identifiable. The serum proteins are separated first by electrophoresis in agar gel. Immunological differentiation of the separated proteins is achieved by adding specific antisera to a groove in the agar plate beside the separated protein fractions. Proteins and antisera diffuse through the agar and within 12 to 18 hours form precipitin bands. Immunoelectrophoresis is essential in the diagnosis of diseases of the immunoglobulin system.

    The separation of three serum proteins by electrophoresis, followed by their...

  18. 12 Radioimmunoassay of Thyroxine
    (pp. 87-91)
    Frank Ungar and John F. Van Pilsum

    Many compounds such as hormones, vitamins, and drugs are present in blood and tissues at such low concentrations that their quantification by chemical means, visible or ultraviolet spectroscopy, or even by biological assays is difficult. The levels of these compounds in tissues and biological fluids are of great diagnostic value, and the concentrations of a large number of these compounds can be ascertainedonlyby radioimmunoassays (RIA) in which levels as low as 10-12g (picogram) to 10-15g (femtogram) may be detected. Some examples of substances that are measured by radioimmunoassays are:

    pituitary hormones hematologic compounds

    adrenocortiotropin erythropoietin

    follicle stimulating ferritin...

  19. 13 Clinical Analysis of Serum Electrolytes
    (pp. 92-97)
    Charles W. Carr, Robert J. Roon and John F. Van Pilsum

    One of the major medical advances in this century has been the understanding and the management of fluid and electrolyte balance. Many pathological conditions in the human result in changes in the electrolyte composition of the extracellular fluid. Alterations in these components are often lethal and thus rapid procedures are required for the analysis of body fluids and remedial treatment of the imbalance. Procedures are presently available for the rapid determination of Na+, K+, Ca²+, Cl-, pCO2, pO2, pH, and bicarbonate.

    The concentration of the following electrolytes are routinely determined in the clinical laboratory for the purpose of diagnosis...

  20. 14 Lecithin/Sphingomyelin Ratio of Amniotic Fluid
    (pp. 98-101)
    Maureen A. Scaglia and John F. Van Pilsum

    A leading cause of death in premature babies is respiratory distress syndrome (RDS). RDS is caused by lung immaturity due to a lack of lung surfactant, a major component of which is a phospholipid called lecithin. Fluid containing lung lecithin flows from the lungs of the fetus into the amniotic fluid. By determining the amount of lecithin relative to the amount of sphingomyelin in amniotic fluid, an estimate of lung maturity can be made. Determination of the lecithin/sphingomyelin ratio involves extraction and precipitation of the phospholids, their separation by thin-layer chromatography, and comparison of sample spots with those of the...

  21. Appendix: Reagents
    (pp. 102-106)
  22. Back Matter
    (pp. 107-107)