The physical properties and chemical composition of urine are highly variable and are determined in large measure by the quantity and the type of food consumed. The specific gravity is the ratio of the density to that of water, and it is dependent on the number and weight of solute particles and on the temperature of the sample. The weight of solute particles is constituted mainly of urea (73%), chloride (5.4%), sodium (5.1%), potassium (2.4%), phosphate (2.0%), uric acid (1.7%), and sulfate (1.3%). Nevertheless, urine osmolality depends only on the number of solute particles. The renal production of maximally concentrated urine and formation of dilute urine may be reduced to two basic elements: (1) generation and maintenance of a renal medullary solute concentration hypertonic to plasma and (2) a mechanism for osmotic equilibration between the inner medulla and the collecting duct fluid. The interaction of the renal medullary countercurrent system, circulating levels of antidiuretic hormone, and thirst regulates water metabolism. Renin, aldosterone, prostaglandins, and kinins also play a role. Clinical estimation of the concentrating and diluting capacity can be performed by relatively simple provocative tests. However, urinary specific gravity after taking no fluids for 12 h overnight should be 1.025 or more, so that the second urine in the morning is a useful sample for screening purposes. Many preservation procedures affect specific gravity measurements. The concentration of solids (or water) in urine can be measured by weighing, hydrometer, refractometry, surface tension, osmolality, a reagent strip, or oscillations of a capillary tube. These measurements are interrelated, not identical. Urinary density measurement is useful to assess the disorders of water balance and to discriminate between prerenal azotemia and acute tubular necrosis. The water balance regulates the serum sodium concentration, therefore disorders are revealed by hypo- and hypernatremia. The disturbances are due to renal and nonrenal diseases, mainly liver, cardiovascular, intestinal, endocrine, and iatrogenic. Fluid management is an important topic of intensive care medicine. Moreover, the usefulness of specific gravity measurement of urine lies in interpreting other findings of urinalysis, both chemical and microscopical.
The structure and physical properties of human fibrinogen and fibrin are reviewed along with methods for the detection of products of their metabolism. Interactions of human fibrinogen with thrombin, factor XIII, plasminogen, glycoprotein IIb/IIIa, and other proteins are related to their relevance to thrombosis and hemostasis. To the extent information is available, the structural determinants of these interactions are delineated, and kinetic and thermodynamic parameters associated with the interactions are listed. Individual steps in the reaction pathway for the conversion of fibrinogen to cross-linked fibrin are characterized. The altered hemostatic properties of mutational variants of fibrinogen are related to their altered structure. The structures of the genes coding for the polypeptide chains of fibrinogen are discussed along with the current state of knowledge of the control and regulation of fibrinogen synthesis. Fibrinogen catabolism and fibrinolysis are also reviewed.
Autoimmune thyroiditis, most notably Hashimoto's thyroiditis, appears to be increasing in prevalence and is now more easily detected by sensitive laboratory tests and more invasive procedures such as fine needle aspiration. During the last decade, marked progress has been made in the understanding of these diseases. There is a greater awareness of the interaction between the humoral and cell-mediated arms of the immune system in autoimmune thyroiditis. Recent studies implicate a subpopulation of suppressor T lymphocytes which have an antigen-specific defect, resulting in their suboptimal interaction with the helper T lymphocytes and subsequent autoimmune manifestations. There is some evidence that thyroid epithelial cells which inappropriately express HLA-DR may enhance presentation of thyroid antigens to the immune system, possibly significant in the initiation or enhancement of the autoimmune response. The presence of various antithyroid autoantibodies allows the use of laboratory assays to confirm the clinical diagnosis and predict the results of treatment. There appears to be predisposing genetic factors in the development of autoimmune thyroiditis, with some geographical and racial differences. Environmental factors, most notably dietary intake of iodine, have also been implicated in the pathogenesis of Hashimoto's thyroiditis. Several animal models have been developed addressing such issues. Ongoing studies in the areas of postpartum thyroiditis and childhood thyroiditis are helpful in clarifying their relationship with Hashimoto's thyroiditis. Graves' disease and postpartum thyroiditis are being investigated as possible causes of postpartum depression. The association of Hashimoto's thyroiditis and carcinoma of the thyroid gland is still controversial, but its relationship with malignant lymphoma is now well accepted. Thus, although the pathogenesis of autoimmune thyroiditis remains elusive, there has been significant refinement of the clinical diagnosis, and immunological abnormalities of specific intrathyroidal lymphocytes have been identified. Hopefully, these new areas of knowledge will assist in the treatment of these diseases and in the prevention of the development of malignant lymphomas of the thyroid gland.
Human immunodeficiency virus (HIV) is a cytoplasmic retrovirus which is transmitted via body fluids, especially through blood products and sexual contact, and is the causative agent of the acquired immunodeficiency syndrome (AIDS). Only about 5 to 10% of the patients infected with HIV contract AIDS; the great majority of infected people either develop a less aggressive condition (AIDS-related complex) or appear healthy. All persons infected with HIV may transmit the virus. In order to protect the national blood supply and to help in diagnosis, tests have been developed to identify infected persons. These include viral isolation techniques, enzyme-linked immunosorbent assay (ELISA), immunofluorescent assay (IFA), radioimmune precipitation (RIP) assay, Western blot, and, most recently, antigen identification and gene probes. Although the sensitivity and specificity among these methods varies, all are susceptible to false-positive and/or false-negative results. In order to understand the reasons for fragility in methodologies, it is necessary to appreciate several basic concepts related to the biochemistry, biology, pathophysiology, and genetic characteristics of HIV and related viruses. The purpose of this review is to present the strengths and weaknesses of each method, with emphasis on peculiar viral characteristics that lead to methodological defects or efficacies.
With the advent of binding assays for vitamin B12 in blood, the Schilling test, which involves administration of radioactive B12 to a patient and subsequent urine collection for 24 to 48 h, fell into disuse in many laboratories. However, the test is still the only way to actually measure whether vitamin B12 is being absorbed through the terminal ilium. By administering radioactive vitamin B12, along with a preparation of intrinsic factor (IF), lack of functional IF may also be demonstrated. The Schilling test requires that attention be paid to a number of parameters, including the amount of radioactive vitamin B12 administered and the completeness of the urine collection. These factors, and others required for correct performance of the test, are discussed in this article.
During differentiation cells are known to change their biological behavior according to their genotype. This is thought to be accompanied by a modulation of cell surface determinants expressed on the outer cell membrane. Vice versa, cell surface molecules are suggested to mediate extracellular signals to the genome. Most of these molecules integrated in the cell membrane have been proven to be glycoconjugates. The carbohydrate moieties of these molecules can be detected by means of lectins that are characterized by their ability to react specifically with distinct terminal sugar sequences. Thus, lectins have been used as appropriate tools for studying the modulation of functionally important membrane-associated molecules during the differentiation of cells, in particular of B- and T-lymphocytes. Moreover, lectins have been proven to distinguish between differentiated cells and malignant cell clones, according to the hypothesis that transformed cells possess a glycoconjugate profile that corresponds to the stage of differentiation at which they are arrested. Since lectins, like monoclonal antibodies, make it possible to study functionally important molecules that are associated with differentiation and malignancy, they might be of value for diagnostic purposes and, moreover, for analyzing malignant transformation.
Although iron chelation therapy with deferoxamine (DFO) has changed life expectancy in thalassemic patients, compliance with the rigorous requirements of long-term subcutaneous DFO infusions is unsatisfactory. This problem underlines the current efforts for developing alternative, orally effective chelators to improve compliance and treatment results. For the patient with transfusional iron overload in whom results of DFO treatment are unsatisfactory, several orally effective agents are now available. The most important of the new generation of oral chelators are deferiprone and ICL670. Total iron excretion with deferiprone is less than with DFO, but deferiprone has a better ability to penetrate cell membranes and may have a better cardioprotective effect than DFO. Current studies of the clinical efficacy and tolerability of ICL670 indicate that at a single oral dose of 20 mg/kg daily, it may be as effective as parenteral DFO used at the standard dose of 40 mg/kg daily. Combined chelation treatment, employing a weak chelator that penetrates cells better, and a stronger chelator with efficient urinary excretion, may result in improved therapeutic effect through iron shuttling between the two compounds. The efficacy of combined chelation treatment is additive and offers an increased likelihood of success in patients previously failing DFO or deferiprone monotherapy.
The revision of the subcellular model of hormone action is described, with an incorporation of potential autocrine mechanisms. A general overview of available assay methodologies considers the major disadvantages of earlier methods and describes in detail the current methodologies (sucrose gradient analysis, dextran-coated charcoal assays, ER-EIA, ERICA). A major concern with clinical correlations of response to hormone receptor levels is the quality assurance of the multicentric programs. Results from national and international programs are considered. The clinical correlations are divided into four major categories: (1) the response to hormone deprivation (oophorectomy or adrenalectomy), (2) the development of specific agents which exploit receptor mechanisms (antiestrogens) or inhibit steroid biosynthesis (aminoglutehimide), (3) the rates of recurrence of tumors following mastectomy, and (4) the correlation of hormone receptors with current adjuvant therapies.
A considerable volume of literature has accumulated concerning the association of aluminum with Alzheimer's disease. The pathogenic mechanisms resulting in Alzheimer's disease remain unknown, but recent investigations have focused on cytoskeletal abnormalities as perhaps the key lesion in Alzheimer's disease and related neurological disorders. The diversity of neuronal functions that are dependent on cytoskeletal integrity suggests that subtle effects on polymerization, assembly, transcription, or processing of cytoskeletal elements may have significant and far-reaching neurological effects. That aluminum may participate in the development of neuropathological lesions characteristic of Alzheimer's disease is suggested by evidence that aluminum is a potent cytoskeletal toxin, produces cognitive deficits in laboratory animals, and can be detected within abnormal neurons isolated from brain tissue from Alzheimer's disease patients. In this review, a critical look will be taken at the enigmatic role aluminum has played in Alzheimer's disease research, the possibility of its pathogenicity, and its use as a tool for the investigation of cytoskeletal changes that may result in the biochemical and, ultimately, clinical manifestations of Alzheimer's disease.
Diarrheal disorders are the result of excessive fluid and electrolyte loss through the gastrointestinal tract. Many different underlying mechanisms are known to cause diarrhea. Fordtran suggested that in secretory diarrhea the osmolality of stool water should be accounted for by its electrolyte contents. Therefore, the osmotic gap between the measured osmolality and that estimated from electrolyte contents should be small. In osmotic diarrhea, due to the presence of the osmotic agent, there should be a greater gap between the measured and the estimated osmolalities. Osmotic gaps varying from 100 to 40 mOsm have been used arbitrarily in literatures to define the underlying pathogenesis. Because of the uncertainty, the usefulness of these measurements remains in question. In this article, methods used to measure stool osmolality and electrolyte contents are reviewed. Limitations of these measurements are discussed. Measurements derived from various diarrheal disorders revealed that the basic concepts put forward by Fordtran are corrected. However, we found that the osmotic gaps (measured osmolality - 2 [Na + K] in secretory diarrheal disorders are frequently negative numbers. In osmotic diarrhea, the osmotic gap (greater than 160 mOsm) is substantially greater than the figures used in the literature. In many diarrheal disorders the osmotic gap falls between the two extremes and the pathogenesis is multifactorial in origin. Under these circumstances, stool osmolality and electrolyte measurements provide little insight into the underlying mechanism causing the diarrhea. Furthermore, stool contains many biologically active organisms which can alter the stool osmolality. Unless these effects are appreciated, an inaccurate interpretation of these measurements may result.
We are presently at a point in human pineal research where we have recognized through melatonin assay the presence of pineal dysfunction in a variety of disease categories. Melatonin may now be quickly and accurately quantified in a range of body fluids, and our well-developed knowledge of the basic biochemistry and neuroanatomical connections of the pineal enables us to see at least how abnormalities in melatonin secretion occur, if not why. The reported increases in melatonin secretion in early malignancy with a reduction in secretion during the neoplastic process is interesting, as is the great decline in the elevated melatonin level of oncological patients following institution of chemotherapy. The correlations between estrogen receptor status of breast cancer and melatonin level, and between neoplastic status of the prostate and melatonin secretion, points to interesting differential diagnostic utilities of melatonin analysis in these conditions. Furthermore, an etiological involvement of melatonin in neoplasia is suggested by experiments which have demonstrated the capacity of melatonin to induce mitotic arrest, and to increase the affinity of mammary carcinoma estrogen receptors for their substrate. These are important observations among many others of direct relevance to research and treatment in oncology, and warrant much further investigation. Melatonin assay may also prove useful in the prognostic monitoring of patients treated for melatonin-secreting pineal tumors, and in such cases may form a logical part of follow-up investigation in the screening for metastatic complication. In psychiatry research, melatonin analysis has functioned as a tool by which alterations in pineal function within specific psychiatric diagnoses have been demonstrated and assessed. Its uses in the assessment of the effects of antidepressant drugs on central beta-receptor function, as a tool in the investigation of light-induced alterations in pineal function in manic-depressive individuals, and as a tool in the investigation of the putative pineal-adrenocortical functional interaction have produced the fundamental building blocks of modern research into the pineal and psychiatry. The experimental clinical utility of melatonin assay is not localized to oncology and psychiatry, and significant alterations in melatonin secretion have been reported in several other disease categories. Indeed, the demonstration of markedly elevated melatonin secretion in patients with spina bifida occulta might suggest that assay of melatonin in amniotic fluid could be useful as an experimental adjunct in the prenatal diagnosis of this condition.(ABSTRACT TRUNCATED AT 400 WORDS)
For several decades the standard blood preservative solution consisted of citrate, dextrose, and, later, phosphate (ACD and CPD). In 1978 a new solution containing adenine (CPDA-1) was introduced to permit extension of red cell shelf life from 21 to 35 days. The success of CPDA-1 and the high percentage of blood units processed into components (estimated 87% in 1983) have stimulated a burst of research and development activity to develop improved red cell preservation systems. Most of these systems have taken the form of "additive solutions" in which the blood is drawn in CPD or CP2D and processed into components. Then the packed red cells are stored by addition of a solution customized for their preservation. This review evaluates these additive solutions in detail. Innovative systems for pH control (buffers and resins) have also been explored. Our review concludes with discussion of the safety aspects of new preservative solutions and the methodological problems of evaluating these solutions.
Cyclosporine is an 11-amino acid cyclic peptide immunosuppressant that has revolutionized organ transplantation. Alone or in combination with prednisone and azathiaprine, it is preferred in hepatic, cardiac, and high-risk renal transplantation. Its unusual primary structure of hydrophobic, N-methylated amino acids results in a compact conformation in the crystal which changes to multiple conformations in hydrophilic solvents. The unusual structure produces unusual pharmacokinetic behavior which is still poorly understood. The metabolism occurs predominately in the liver and is affected by several drugs known to alter hepatic metabolism. At least ten metabolites have been identified but are inadequately characterized. The unique behavior of cyclosporine necessitates therapeutic drug monitoring (TDM) for individualization of therapy. Cyclosporine has been monitored in both whole blood and plasma by both RIA and HPLC with significantly different results for each combination. When cyclosporine is assayed by HPLC in a compulsive regimen of TDM, a correlation is observed between immunosuppression, toxicity, and concentration. To distinguish renal or hepatic toxicity from rejection, biopsies, clinical status, and blood concentrations of cyclosporine must be simultaneously analyzed. After extensive experimental and clinical study, cyclosporine remains an enigma with clear clinical benefit.
Radioisotope conjugated to monoclonal antibody products has been used for imaging tumors targeted by the antibody. As imaging progresses, new sets of procedural and technical questions arise. In this chapter, we discuss several current problems in imaging tumor with radiolabeled monoclonal antibody. These include (1) methods for selection of specific antibody and, once the particular antibody is selected, which fragment form is to be used; (2) imaging procedures: what are the optimum imaging parameters, such as optimum time for imaging after administration of tracer and considerations regarding background subtraction; and (3) noninvasive quantitative techniques: quantitation of localization of antibody indirectly from quantitative information in the images.
The salicylates are the most commonly used analgesic, antipyretic, and anti-inflammatory drugs. They are available in hundreds of preparations, many of which are over-the-counter medications. The easy access to large quantities of the drug and the widespread perception that the drug is harmless have contributed to salicylate intoxication becoming a serious and common problem, particularly among the pediatric and geriatric populations. Salicylate is still the major drug for the treatment of rheumatic diseases. The use of salicylate in high doses for the management of these patients requires close monitoring of serum salicylate levels because of the large interindividual variation in dose-serum level relationships and the narrowness of the therapeutic range. Thus, both for the management of patients intoxicated with salicylate and patients who are on high-dose salicylate therapy, the measurement of serum salicylate levels is an important clinical laboratory service. Recent research on the inhibitory effect of aspirin on platelet aggregation has led to the prophylactic use of aspirin in low doses as an antithrombotic drug. This new therapeutic use of aspirin can be aided by monitoring low serum levels of salicylate and perhaps aspirin itself. This article reviews the current state of the knowledge of the pharmacokinetics and clinical toxicology of salicylate, the clinical usefulness of salicylate measurement by the clinical laboratory, and recent development in the analytical technology for salicylate analysis.
The increasing frequency of cervical neoplasia among younger women and the increased invasiveness of these tumors has led to a considerable growth in research into this disease. Conventional methods (epidemiology, cytology, and immunology), while being extremely useful, also have significant limitations. Recent advances in techniques for the manipulation of DNA now make it possible to analyze tissues for the presence of viral genomes. This review introduces these techniques and describes their application to the search for herpes simplex virus and human papillomavirus sequences in cervical tissue. The significance of the findings both for the mechanism of transmission of the disease, and also the consequences for early detection and hence more successful treatment, are also discussed.
An instrument for laser microprobe mass analysis (LAMMA) has been available for applied research for several years. The value of this sensitive microanalytical technique has been demonstrated in various fields of science, including medicine and biology. The LAMMA instrument comprises two laser systems. The first is a weak continuous pilot laser that can be aimed at a region of interest of the sample, e.g., a histological microtome section, using a light microscope. The second is a powerful pulsed laser that can evaporate the selected area. The generated ions are then analyzed in a time-of-flight mass spectrometer. The advantages of LAMMA present new perspectives in biomedical research. Elements can be located and analyzed down to the parts per million-level with a 1 micron spatial resolution. Speciation and/or fingerprinting of (in)organic molecules may be obtained. However, the technique is destructive, the quality of the light-microscopic view is poor, and quantification is still under investigation. In this review, the analytical characteristics of LAMMA are evaluated, including instrumental features, detection sensitivity, ionization parameters, molecular speciation, and quantification. Applications of LAMMA in medicine are reviewed, covering the microanalysis of a wide range of physiological and toxic constituents (alkali metals, calcium, aluminum, titanium, fluorine and iodine, heavy metals, transition elements, and molecular compounds).
Thyroid function tests are one of the most common of endocrine laboratory investigations requested by general clinicians. The tests used therefore have to be efficient at identifying thyroid disease, monitoring treatment, and handling large numbers of tests. Recent advances in methodology have expanded both the range of in vitro thyroid function tests available and the techniques by which the well-established tests may be performed. This article reviews the methods and analytical and clinical performance of the routine tests currently available, concentrating particularly on the relatively new ones, and speculating on their role in strategies for the laboratory investigation of thyroid function.
Currently, advances in molecular technology involving recombinant DNA have led to dramatic breakthroughs in genetic diseases, cancer research, and identification of foreign DNA. Of particular interest is the impact these tools have made and will make on the clinical laboratory. We describe the techniques and their effects on clinical testing in the chemistry laboratory by using selected examples of available applications. Specific examples include carrier detections and prenatal diagnosis in cystic fibrosis and hemophilia, and sickle cell anemia.
Myoepithelial cells form an integral part of the secretory and ductular portion of most glands. They share a common origin with lumenal epithelial cells and influence proliferation and differentiation of developing terminal glandular buds by producing a scaffold of basement membrane proteins. Their contractile capacity, controlled by hormonal and neural mechanisms, plays an important role in propulsion of secretions. Furthermore, myoepithelial cells maintain glandular structural integrity and transport metabolites to secretory cells. The advent of modern immunochemistry made identification of specific myoepithelial cell markers possible which facilitated studies on their presence and behavior in disease processes. Although the significance of many myoepithelial alterations is speculative, some have proved valuable in determining the histogenesis of glandular lesions.
