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Oligoclonal bands and quantitation of IgG in cerebrospinal fluid as indicators of multiple sclerosis.

The authors evaluated two laboratory procedures in respect to usefulness in evaluating patients suspected of having multiple sclerosis (MS). They classified 185 patients as having MS (MS-positive) or as not having MS (MS-negative). Agarose electrophoretic patterns of cerebrospinal fluid (CSF) samples were inspected for the presence of oligoclonal bands. CSF immunoglobulin G (IgG) was quantitated and compared with total CSF protein for 175 patients. Oligoclonal bands were present in specimens from 77% of the MS-positive group; elevated percentages of IgG (18% or more of total CSF protein) were present in specimens from 20%. Oligoclonal bands were present in specimens from 0.6% of the MS-negative group; elevated percentages of IgG were found for 1.4% of the MS-negative group. The presence or absence of oligoclonal bands in agarose electrophoretic patterns is sensitive (77%) and very specific (99%) in respect to MS. CSF IgG quantitation using the authors' criteria does not add useful information.

Demyelinating Diseases↗

Fluorescent enzyme immunoassay for antibody to single- or double-stranded DNA.

Fluorescent enzyme immunoassays have been developed for antibody to either single-stranded (ssDNA) or double-stranded (dsDNA) DNA. Preliminary testing of several clinically distinct groups of persons is consistent with the suggestion that the former assay (anti-ssDNA antibody) has sufficient sensitivity and specificity to be used as a generalized test for the detection of possible systemic lupus erythematosus in a heterogeneous clinical population. Further evaluation may ultimately establish its utility in the diagnosis of other related disorders. The sensitivity and specificity of the anti-dsDNA antibody fluorescent enzyme immunoassay are compatible with its use as a confirmatory test in the laboratory diagnosis of systemic lupus erythematosus. Qualitative comparisons between both assays and the more classic laboratory procedures involved in the diagnosis and treatment of systemic lupus erythematosus were quite favorable. The enzyme immunoassay method appears to be acceptably reproducible, and rheumatoid factor has neither a suppressive nor an enhancing effect on test values.

Adult↗

The influences of sample distribution and age on reference intervals for adult males.

This investigation was designed to improve reference information and to evaluate the influences of sample distribution and age on the derived reference intervals. Specimens from 127 men were collected after a 12- to 14-hour fast and analyzed by 60 different laboratory procedures. Differences in the reference intervals derived, using three separate statistical methods, appeared to be unimportant clinically, but the percentile method was preferred because it required no assumptions concerning the frequency distribution. Relationships between age and analyte concentrations were examined by linear regression analysis, and the analytes were placed in one of three groups, according to the significance of this relationship: greatest significance (P less than or equal to 0.01), 18 analytes; intermediate significance (0.01 less than or equal to P less than or equal to 0.05), 12 analytes; and least significance (P greater than 0.05), 30 analytes. The age-related changes for each analyte were evaluated according to analytic variation, population dispersion, and clinical relevance. Age-dependent reference intervals for adult males are recommended for albumin, cholesterol, phosphorus, and sedimentation rate.

Adult↗

Alpha heavy-chain disease presenting as goiter.

A patient with alpha heavy-chain disease (alpha HCD) originally had presented with a goiter from an extramedullary plasmacytoma. Alpha HCD was diagnosed by both standard laboratory procedures and high-resolution two-dimensional gel electrophoresis. The fragment of the alpha chain had a molecular weight of 38,000 and consisted of seven major charge forms, with an isoelectric point range of 5.2-5.6. Since alpha HCD usually involves the gastrointestinal tract, the thyroidal involvement in this case is unusual.

Adult↗

Equality of the in vivo and in vitro oxygen-binding capacity of haemoglobin in patients with severe respiratory disease.

The in vivo and in vitro oxygen-binding capacity of haemoglobin was determined on 10 occasions in nine patients who required mechanical ventilation. The in vitro sample was tonometered with 97% oxygen for 10 min and then with air, while the in vivo sample was obtained after 20 min of lung ventilation with pure oxygen. Subsequent laboratory procedures were identical for both samples. The mean oxygen-binding capacity of haemoglobin in vitro and in vivo samples were almost equal (1.365 +2- 0.010 and 1,366 +/- 0.007 ml per g Hb). When the measured inactive fractions of haemoglobin (carboxy- and methaemoglobin) were taken into account, these values increased to 1.392 +/- 0.005 and 1.392 +/- 0.007 ml per g Hb respectively.

Aged↗

Evaluation of antibiotic usage: a comprehensive look at alternative approaches.

Current problems related to the use of antibiotics in the United States are summarized. In 1979, pharmaceutical manufacturers shipped +1.55 billion worth of anti-infective drugs. It is estimated that in approximately one-half of all cases that involve administration of antibiotics in the hospital, either the medical condition does not require antibiotic treatment, the most effective and least expensive drug is not chosen, or the correct dosage or duration of therapy is not prescribed. Much of the high cost of antibiotic therapy can be attributed to the use of expensive antibiotics of the cephalosporin and aminoglycoside groups and to the excessive duration of antibiotic prophylaxis in surgery. This review presents methods that assess the magnitude of the problem by audit and analyzes the corrective approaches that have been suggested. The major issues of concern related to the use of antibiotics are the complex series of considerations that lead physicians to prescribe antibiotics and the problem of patient expectation and compliance. Excessive usage of antibiotics must be viewed as part of the problem of overusage of all drugs and laboratory procedures.

Anti-Bacterial Agents↗

Histochemical demonstration of endothelial superoxide and hydrogen peroxide generation in ischaemic and reoxygenated rat tissues.

OBJECTIVE: The aims were to test and evaluate two novel and independent histochemical methods for detecting the initial postischaemic burst of superoxide and hydrogen peroxide in buffer perfused rat tissues during reflow after 60 min warm ischaemia. METHODS: The first is a high manganese/diaminobenzidine technique, in which superoxide oxidises Mn2+ to Mn3+, which in turn oxidises diaminobenzidine to form amber coloured polymers, observable by light microscopy. The second is a high iron/diaminobenzidine technique, in which hydrogen peroxide oxidises diethylenetriaminepenta-acetate chelated Fe2+ to form intermediate species, which in turn oxidise diaminobenzidine similarly to Mn3+. Various isolated organs of the rat were rendered ischaemic for 60 min, and reperfused with oxygen or air equilibrated buffers containing diaminobenzidine and either Mn2+ or Fe2+. Tissues were fixed by perfusion with Trump's solution and processed for light microscopy. RESULTS: Both manganese and iron methods consistently showed the appearance of reaction product on the luminal surfaces of arterial, capillary, and venular endothelial cells in lung, heart, and intestine of the rat during the first 2 to 3 min of reoxygenation after ischaemia. The histochemical reactions were nearly absent in non-manganese-treated and non-iron-treated controls. Superoxide dismutase strongly inhibited Mn2+/diaminobenzidine reaction product formation and catalase strongly inhibited Fe2+/diaminobenzidine reaction product formation, when tested in specially perfused lung preparations in which these specific antioxidant enzymes were concentrated. CONCLUSIONS: These histochemical techniques provide direct, visual evidence that a burst of reactive oxygen species is generated in postischaemic rat tissues. The Mn2+/diaminobenzidine and Fe2+/diaminobenzidine techniques permit investigation of the endothelium derived reactive oxygen by simple laboratory procedures available to almost any investigator at low marginal cost. The endothelial oxidants so revealed may be of pathophysiological significance in a variety of cardiovascular disorders.

Abdomen↗

Referrals for radiological examination of the large bowel. Pre-radiological examinations, tests and referral letters.

The aims of the present study were to assess the frequency of examinations or tests before referral for a barium enema, to identify predictors for the use of such examinations or tests and to evaluate the quality of the referral letters. 1852 patients completed a questionnaire on symptoms and examinations or tests performed before referral. Symptoms, examination procedures, laboratory data and reason for referral were registered in 1781 referral letters and were compared with the information given by the patients. Serum haemoglobin was measured in 76%, per-rectal examination in 45%, faecal occult blood tests in 37% and proctosigmoidoscopy in only 16% of the patients. High age was a significant predictor for all examinations, and patients with rectal bleeding and rectal pain were more often examined rectally, although this was omitted in several patients with such symptoms. All evaluated information was reported significantly more frequently by the patients than by the doctors in the referral letters. The very low figures for rectal examinations and the lack of completeness and adequacy of the information in the referral letters underline the need for efforts to change doctors' behaviour when investigating and referring patients with possible colorectal disease.

Adult↗

Validity of urinary biomarkers of exposure to tobacco smoke following prolonged storage.

BACKGROUND: The utility of using biomarkers of smoking in epidemiological studies depends not only on the validity and precision of the laboratory procedure but often on the long-term stability of the analytes of interest in stored biological samples. METHODS: We retrieved urine samples collected in 1976-1977 from women included in a cohort study in Utrecht and for whom information on smoking status was available. Creatinine and thiocyanate were measured in 1976-1977 on fresh samples. Cotinine and creatinine were analysed in 1988 on urine stored at -20 degrees C. RESULTS: Measurements of creatinine more than 10 years apart showed a correlation of 0.95 and equal means. Cotinine measurements made in 1988 allowed a clear separation of smokers and non-smokers (sensitivity 92%, specificity 100%), suggesting that concentrations retained their discriminant value even after 10 years of storage. CONCLUSION: These results emphasize the possibilities offered by long-term storage, under proper conditions, of biological samples for subsequent determination of analytes which may emerge as the study progresses.

Biomarkers↗

Improved leaf area index based biomass estimations for Zostera marina L.

The application of special scanning technologies in plant population studies makes it now possible to offer reliable indirect estimations of Leaf Area Index (LAI). This has stimulated the adaptation of related biomass assessment methods and has provided a way to simplify tedious laboratory procedures whilst avoiding destructive sampling. Particularly, above-ground biomass for Zostera marina L. has been expressed depending linearly on Leaf Area Index. Nevertheless, we demonstrate that this approach produces biased estimations. It is also shown that expressing leaf dry weight by means of an allometric function of length and width can eliminate bias. Furthermore, the dominant term of the associated power series expansion becomes the aforementioned linear representation in terms of Leaf Area Index. The consistency of the estimation methods derived from the allometric model was tested using data from a Z. marina meadow. Consequently, the improved method is expected to become a valuable tool for the reduction of the uncertainty associated with the estimation of above-ground biomass through the use of Leaf Area Index.

Biomass↗

Identification and preliminary antigenic analysis of swine influenza-like viruses isolated during an influenza outbreak at Fort Dix, New Jersey.

The sequence of events and the laboratory procedures that resulted in the identification of swine influenza-like viruses isolated during an influenza outbreak at Fort Dix, New Jersey in January and February of 1976 are described. Preliminary antigenic analysis suggested that the isolates from Fort Dix are closely related to a 1975 isolate of swine influenza virus and distinguishable from earlier swine influenza strains.

Antigens, Viral↗

Circulating immune complexes in experimental streptococcal endocarditis: a monitor of therapeutic efficacy.

An important problem in the management of infective endocarditis has been the delineation of laboratory procedures that are sensitive, reliable indicators of therapeutic efficacy. Because circulating, complement-containing immune complexes of the IgG type (CICs) have been demonstrated in most humans with infective endocarditis, serum CIC levels during the natural course of the infection and in response to penicillin therapy were studied in 42 rabbits with right-sided endocarditis due to Streptococcus salivarius. A significant rise in the level of CICs in both 21 control rabbits and 21 treated rabbits was observed after induction but before treatment of infective endocarditis (P less than 0.01). In the 17 successfully treated rabbits, CIC levels fell sharply during the first week of therapy and remained at preinduction levels thereafter (P less than 0.005). In contrast, CIC values did not change significantly either in control animals or in the four treated animals with refractory endocarditis, although in the latter animals, serum bactericidal titers remained less than or equal to 1:32. These findings suggest that serial measurements of CIC levels during antimicrobial therapy of infective endocarditis may aid in monitoring therapeutic efficacy.

Animals↗

Herpes simplex virus encephalitis: laboratory evaluations and their diagnostic significance.

Laboratory procedures were compared with brain biopsy findings in 113 biopsy-proven patients with herpes simplex virus (HSV) encephalitis and 93 biopsy-negative individuals. Examinations of brain tissue by histopathology, immunofluorescence, and electron microscopy demonstrated evidence of HSV infection in 56%, 70%, and 45% of proven cases and apparently false-positive results in 14%, 9% and 2% of those biopsy-negative. Serologic assessments revealed that HSV encephalitis occurred as both a primary (30%) and recurrent (70%) infection. Among patients with HSV encephalitis, 28% failed to seroconvert or seroboost within one month of the onset of disease. Titers of passive hemagglutinating and IgG immunofluorescent antibodies increased fourfold in the cerebrospinal fluid in 74% and 94%, respectively, of patients with proven disease. Similar percentages of patients had antibody ratios in serum and cerebrospinal fluid of less than 20 over the same interval. These data indicate the need for development of noninvasive diagnostic procedures.

Adolescent↗

Biomaterial-associated infection of gentamicin-loaded PMMA beads in orthopaedic revision surgery.

In two-stage orthopaedic revision surgery, high local levels of antibiotics are achieved after removal of an infected prosthesis through temporary implantation of gentamicin-loaded beads. However, despite their antibiotic release, these beads act as a biomaterial surface to which bacteria preferentially adhere, grow and potentially develop antibiotic resistance. Gentamicin-loaded beads were retrieved from 20 patients with prosthesis-related infections. Excised tissue samples were taken for routine culture, while beads were analysed in an extensive laboratory procedure. Extensive culture procedures indicated the presence of bacteria on gentamicin-loaded beads in 18 of the 20 patients involved, while 12 of these 18 patients were considered free of infection by routine culture. Nineteen of 28 bacterial strains isolated were gentamicin resistant and cultures from three patients yielded highly gentamicin-resistant sub-populations. It is concluded that routine culture of excised tissues in orthopaedic revision surgery is inadequate to ascertain full eradication of infection, especially as infecting, antibiotic-resistant bacteria preferentially adhere to and grow on gentamicin-loaded beads. Extensive examination of the bead surfaces is proposed as a more reliable indication that infection has been eradicated.

Anti-Bacterial Agents↗

Extensive human DNA contamination in extracts from ancient dog bones and teeth.

Ancient DNA (aDNA) sequences, especially those of human origin, are notoriously difficult to analyze due to molecular damage and exogenous DNA contamination. Relatively few systematic studies have focused on this problem. Here we investigate the extent and origin of human DNA contamination in the most frequently used sources for aDNA studies, that is, bones and teeth from museum collections. To distinguish contaminant DNA from authentic DNA we extracted DNA from dog (Canis familiaris) specimens. We monitored the presence of a 148-bp human-specific and a 152-bp dog-specific mitochondrial DNA (mtDNA) fragment in DNA extracts as well as in negative controls. The total number of human and dog template molecules were quantified using real-time polymerase chain reaction (PCR), and the sequences were characterized by amplicon cloning and sequencing. Although standard precautions to avoid contamination were taken, we found that all samples from the 29 dog specimens contained human DNA, often at levels exceeding the amount of authentic ancient dog DNA. The level of contaminating human DNA was also significantly higher in the dog extracts than in the negative controls, and an experimental setup indicated that this was not caused by the carrier effect. This suggests that the contaminating human DNA mainly originated from the dog bones rather than from laboratory procedures. When cloned, fragments within a contaminated PCR product generally displayed several different sequences, although one haplotype was often found in majority. This leads us to believe that recognized criteria for authenticating aDNA cannot separate contamination from ancient human DNA the way they are presently used.

Animals↗

HPtaa database-potential target genes for clinical diagnosis and immunotherapy of human carcinoma.

Tumor-associated antigens (TAAs) have been the most actively employed targets in the clinical diagnosis and treatment of human carcinoma, such as PSA in the diagnosis of prostate cancer and NY-ESO-1 in the immunotherapy of melanoma and other cancers. However, identification of TAAs has often been hampered by the complicated and laborsome laboratory procedures. In order to accelerate the process of tumor antigen discovery, and thereby improve diagnosis and treatment of human carcinoma, we have made an effort to establish a publicly available Human Potential Tumor Associated Antigen database (HPtaa) with potential TAAs identified by in silico computing (http://www.hptaa.org). Tumor specificity was chosen as the core of tumor antigen evaluation, together with other relevant clues. Various platforms of gene expression, including microarray, expressed sequence tag and SAGE data, were processed and integrated by several penalty algorithms. A total of 3518 potential TAAs have been included in the database, which is freely available to academic users. As far as we know, this database is the first one addressing human potential TAAs, and the first one integrating various kinds of expression platforms for one purpose.

Antigens, Neoplasm↗

Body habitus and coronary heart disease in men. A review with reference to methods of body habitus assessment.

Table 1 is a synopsis of the major findings from an extensive literature on the association between human body habitus and coronary heart disease. Whilst some studies have used quite sophisticated laboratory procedures to quantify body fat most have relied upon anthropometric measurements to determine some component of body habitus. Of these, body weight and height are the simplest measurements and are, therefore, well-suited to large-scale prospective studies. Height and weight are highly reproducible measurements, although in the short term, weight can have considerable physiological variation associated with gastric emptying and state of hydration. Less reliable measurements than height and weight are skinfolds and body circumferences, both of which have been used extensively in cross-sectional and prospective analyses. For skinfolds, both the inter and intra-observer variability is affected by the measurement technique, location of the skinfold site, the skinfold caliper used and skinfold compressibility. As measurement error has been shown to be a function of skinfold thickness, accurate and repeatable skinfold measurements are particularly difficult to make in the obese. In these subjects, it is not always possible to locate a specific anatomical bony landmark or to pull a parallel skinfold away from the underlying tissue. Furthermore, in the extremely obese it is sometimes possible for a skinfold to be thicker than the jaws of the currently available commercial calipers. Alternately, body circumferences are obtainable in all subjects and have greater reproducibility than skinfolds. They are, therefore, the preferred method in obese subjects. However, there is considerable work to be done to establish their association with body fatness. The evidence examined in this review suggests that body weight is a poor predictor of coronary heart disease. Some studies have reported no difference in the body weight of coronary heart disease patients compared to subjects free of the disease, others found the body weight of subjects with coronary heart disease to be slightly greater, and one found the body weight of cardiac patients to be less than controls. Height, however, is associated with coronary heart disease in prospective studies with long-term and shorter-term follow-up periods and case-control designs. Fetal, infant and childhood under-nutrition may link shorter adult height and susceptibility to cardiovascular disease. Many researchers have studied the relationship between overweight and coronary heart disease by using a surrogate measurement of body fatness such as relative weight or a weight-for-height index. In general, results produced by these studies suggest weight-for-height indices, particularly the often used body mass index, are not strong predictors of coronary heart disease. Indeed case-control designs have consistently failed to show a relationship between body mass index and coronary heart disease. Inconsistent results from prospective studies, however, are difficult to interpret. To further confuse the situation, the body mass index has been examined in relation to different coronary heart disease end-points and adjusted for different confounding variables. Explaining the inconsistent results on the basis of length of follow-up is also not straightforward. When follow-up periods exceed 20 years, and sample size is small, however, this closer association has not been found, even with a long follow-up period. Whilst some studies have found no association after 15, 13 and 12 years others have reported a relationship after 8.5, 10, 12, 10 and 7 years. The 22 year follow-up evidence from the Framingham Study shows the strongest 'independent' association between body mass index and coronary heart disease. (ABSTRACT TRUNCATED)

Adult↗

Electron-trapping probability in natural dosemeters as a function of irradiation temperature.

The electron-trapping probability in OSL traps as a function of irradiation temperature is investigated for sedimentary quartz and feldspar. A dependency was found for both minerals; this phenomenon could give rise to errors in dose estimation when the irradiation temperature used in laboratory procedures is different from that in the natural environment. No evidence was found for the existence of shallow trap saturation effects that could give rise to a dose-rate dependency of electron trapping.

Aluminum Silicates↗