Application of a human immortalized fibroblast cell line in laboratory diagnosis of autoimmune diseases

Many autoantibodies reacting with cellular and nuclear components have been described in sera of patients with autoimmune diseases. The most important diagnostic markers for those diseases are antinuclear antibodies (ANA). The first choice for laboratory diagnosis of autoimmune diseases is to use cultured monolayer cells as a nuclear substrate. Up to now the HEp-2 cell line derived from a human carcinoma of the larynx, appears to be the most sensitive and specific nuclear substrate. The cultured fibroblast monolayer cells have also been applied to detect the ANA, although the application was not recommended by one study. Thus to evaluate the applicability of our immortalized human fibroblast cell line (CCFS-1/KMC) as a nuclear substrate, commercial HEp-2 MBL monolayer cells was used as the standard substrate. The results of this report showed the applicability of the CCFS-1/KMC cell line as a nuclear substrate to detect the ANA of autoimmune diseases. The sensitivity of this fibroblast cell line was the same as both of the HEp-2 nuclear substrates (HEp-2 and HEp-2 MBL). The specificity of the CCFS-1/KMC cell line was similar to the HEp-2 substrate. Since the specificity of both of the above substrates were lower than the standard nuclear substrate HEp-2 MBL, therefore, if the specificity can be improved by changing the protocols of the substrate preparation, the CCFS-1/KMC cell line will be a good nuclear substrate for detecting the ANA of autoimmune diseases.     

Clinical and laboratory basis for the diagnosis and understanding of neonatal metabolic diseases

The evaluation, classification, and treatment of carpal instability continues to be one of the most controversial topics in hand surgery. One explanation is the lack of standardized findings and radiographic criteria. No good modality to date has been proposed to accurately aid in evaluating carpal instability. Wrist arthroscopy has become increasingly useful in the assessment of mechanical wrist pain. With this technology, specific patterns of injury can more accurately be identified by direct visualization. This will also aid in implementing a reproducible, less subjective means of describing wrist instability. The treatment can then be focused on management of the specific ligamentous injury responsible for the pattern of instability visualized. The following is an attempt to categorize these instability patterns as seen through arthroscopy and provide our proposed treatment options.     

Field validation of laboratory tests for clinical diagnosis of sheep-associated malignant catarrhal fever

Until recently, sheep-associated malignant catarrhal fever (SA-MCF) was diagnosed mainly on the basis of clinical presentation and histopathological changes. Using clinically diagnosed field cases, we have evaluated a seminested PCR and a competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA) and compared these assays in the diagnosis of SA-MCF in cattle with histopathology as a provisional "gold standard." Samples from 44 cattle with clinical signs suggestive of SA-MCF were examined by histopathology, PCR, and CI-ELISA. In addition, samples from healthy cattle were evaluated by PCR (n = 96) and CI-ELISA (n = 75). Based on histopathology, 38 of the 44 clinical cases were classified as SA-MCF positive, 3 were classified as inconclusive, and 3 were classified as SA-MCF negative. The sensitivity of PCR was 95 to 97%, whereas the specificity ranged between 94 and 100%. The CI-ELISA showed a sensitivity of 56 to 87% and a specificity between 91 and 100%. In the field, there is good correlation between the diagnoses of SA-MCF by histopathology, PCR, and CI-ELISA. These data also confirm the close association of ovine herpesvirus 2 with SA-MCF in Switzerland.     

Update on laboratory tests for the diagnosis of pulmonary disease in HIV-1-infected individuals

Various diagnostic tests, both specific and nonspecific, are available in the clinical laboratories for diagnosing human immunodeficiency virus-1 (HIV-1) infection and associated respiratory pathogens. Pneumocystis carinii pneumonia remains the most common pulmonary disease in HIV-1-infected individuals and there have been no significant advances in the laboratory diagnosis of the pathogen beyond the traditional microscopic examination of specimens. In contrast, the greatest revolution in laboratory diagnostic testing has been for mycobacteria, with major advances resulting in significant reduction in the time necessary for isolation and identification to the species level. The application of the polymerase chain reaction for the identification of a variety of pulmonary pathogens observed in HIV-1 infected individuals is discussed.     

Clinical significance of the tests used in the diagnosis of pancreatic diseases

Different methods available for investigating patients for pancreatic disease are discussed. They first include measurement of pancreatic enzymes in biological fluids. Basal amylase and/or lipase in blood are truly diagnostic in acute pancreatitis but their utility is low in chronic pancreatic diseases. Evocative tests have been performed to increase the sensitivity of blood enzyme measurement. The procedure is based on enzyme determination following administration of pancreozymin and secretin, and offers a valuable aid in diagnosis of chronic pancreatitis and cancer of the pancreas. They are capable of discerning pancreatic lesions but are not really discriminatory because similar changes are observed in both diseases. The measurement of urinary enzyme levels in patients with acute pancreatitis is a sensitive indicator of disease. The urinary amylase excretion rises to abnormal levels and persists at significant values for a longer period of time than the serum amylase in acute pancreatitis. The fractional urinary amylase escretion seems to be more sensitive than daily urinary measurement. The pancreatic exocrin function can be assessed by examining the duodenal contents after intravenous administration of pancreozymin and secretin. Different abnormal secretory patterns can be determinated. Total secretory deficiency is observed in patients with obstruction of excretory ducts by tumors of the head of the pancreas and in the end stage of chronic pancreatitis. Low volume with normal bicarbonate and enzyme concentration is another typical pattern seen in neoplastic obstruction of escretory ducts. In chronic pancreatitis the chief defect is the inability of the gland to secrete a juice with a high bicarbonate concentration; but in the advanced stage diminution of enzyme and volume is also evident. Diagnostic procedures for pancreatic diseases include digestion and absorption tests. The microscopic examination and chemical estimation of the fats in stool specimens in different conditions of intake are still important screening tests. Isotopic estimates of steatorrhea and distinction between labeled triolein and oleic acid absorption do not provide greater diagnostic discrimination than traditional procedures. 131I labeled proteins permit a good evaluation of a negative nitrogen balance. Sophisticated procedures to estimate exocrine pancreatic insufficiency are based on the study of endoluminal digestive processes at several times and different level of the small intestine. They permite esclusion of extrapancreatic factors interfering in digestion and absorption functions. The endocrin pancreatic function is evaluated by mean of oral tolerance test an radioimmunoassay of blood insulin. It is generally agreed that "diabetes" caused by insulin deficiency and digestion and absorption defects are the result of diffuse pancreatic destruction. Many methods are now available investigating patients with pancreatic disease but the single use of one of them is never satisfactory...     

A quality control program for the radioimmunoassay laboratory

The major problems in applying quality control to radioimmunoassay measurements are (a) nonlinearity of the dose-response curve, and (b) nonuniformity of the residual variance. A logit-log transformation of the dose-response variables combined with an iterative weighted regression analysis appears to overcome most of the difficulties. This technic is particularly helpful when applied to substandard runs where extraneous standard points tend to distort assay results. The authors describe a quality control program that involves recording control values on calendar and histogram formats, monitoring assay variables by charting, and comparing the computer-calculated slope with the graphic plot to reveal "outliers." This program is useful in guiding technologists to locate possible causes for "out-of-limits" runs.     

Autoimmune liver diseases: the relation to other autoimmune pathology

Nebraska Hospital Market Areas (HMAs) with significantly and consistently high admission rates for Ambulatory Care Sensitive Conditions (ACSC) were identified using the Blue Cross and Blue Shield of Nebraska (BCBSN) small area analysis data. Two-thirds of those hospitalized were surveyed by Wiese Research of Omaha, Nebraska to determine if the cause for the high admission rates was related to outpatient care quality. BCBSN members living in HMAs without high admission rates were also surveyed for comparison purposes. The survey revealed that satisfaction with outpatient care was high, and that access (geographic or financial) was not a problem for either the test or control group. The test group did delay (for a short time) obtaining outpatient care when compared to the control group. Most of the control group stated that they obtained outpatient care the day symptoms started. Although the majority of the test group obtained outpatient care within a week, they were not as prompt in obtaining outpatient care as the control group. The types of conditions studied (ACSCS) are conditions that can rapidly become severe, therefore prompt treatment is imperative. The main reason given for chronic delays of both groups were that the condition "suddenly became serious." There may be implications for providers to use more "compliance-gaining" strategies in problem area, and for insurers to offer contract endorsements which would encourage members to obtain prompt treatment and preventive healthcare.