bluuebunny/crossref_metadata_2025_split huggingface dataset:
Estimation of serum sex hormone–binding globulin by five direct and two indirect methods
Authors: Brotherton Janet 
Published: March 2007

AbstractSerum sex hormone‐binding globulin (SHBG) has been estimated over the range 2–190 nM by six commercially available methods. The Farmos immunoradiometric assay (IRMA) and the Pharmacia‐LKB DELFIA fluoroimmunoassay, which use the same monoclonal antiserum, produced statistically identical but widely spread results. The Techland polyclonal radioimmunoassay produced similar results but was considerably less sensitive, and agreement with the other two direct methods was poor at low concentrations. These three direct methods gave slightly higher results than those obtained with the two indirect methods: the BioMerieux concanavalin‐A–binding method and, especially, the Serono ammonium sulphate precipitation method. The Diagnostic Products monoclonal IRMA gave results that were up to double the values obtained with the other five methods and is considered unsatisfactory. Monoclonal antibodies are now available for the direct estimation of SHBG, although an amino acid sequence has not yet been published for this protein, and there is still some controversy about the size of its structural units.
Isolation of human herpesvirus‐6 from clinical specimens using human fibroblast cultures
Authors: Luka Janos , Okano Motohiko , Thiele Geoffrey 
Published: March 2007

AbstractThe isolation and characterization of human herpesvirus‐6 (HHV‐6) has been hindered by the lack of cell lines useful for its rapid propagation. Recently, we have reported that the MRC‐5 cell line (human diploid lung fibroblasts) was susceptible for HHV‐6 infection. In this study, we report on the isolation of HHV‐6 from the peripheral blood or buffy coat of three chronic fatique syndrome patients, one post‐liver transplant patient, and one severe chronic active Epstein‐Barr virus syndrome patient using the MRC‐5 cell line. Additionally, it was observed by Southern blot hybridization studies that four of five isolates had different restriction enzyme fragment patterns than the isolate obtained from the National Institutes of Health with Eco RI. These data suggest the usefulness of the MRC‐5 cell line in the isolation and characterization of HHV‐6 from various patients.
Differentiation of myeloperoxidase and glandular peroxidase in biological fluids: Application to human saliva
Authors: Venezie Ronald D. , Jenzano Joyce W. , Lundblad Roger L. 
Published: March 2007

AbstractA modification of the assay system for peroxidase activity in human mixed saliva using 2,2′‐azinobis‐(‐ethylbenzthiazoline‐6‐sulphonic acid) (ABTS) as the substrate is reported. The modifications permit differentiation between glandular peroxidase (SPX) and myeloperoxidase (MPX) activity. In addition, factors endogenous to saliva such as thiocyanate did not adversely affect the modified assay system. The basic peroxidase assay utilized 1.0 mM ABTS and 0.1 mM H2O2 in 0.1 M sodium acetate, pH 4.7. The addition of 100 mM NaCl to the assay inhibited 90% of the activity of purified MPX but only 40% of the SPX activity in parotid saliva presumed to be free of MPX. Addition of Cl− to the assay had varied effects on the mixed saliva SPX activity (44–68% inhibition), suggesting that MPX is present in varying amounts in mixed saliva. Assay of mixed saliva from patients with different states of periodontal health indicated that the amount of Cl− inhibition is dependent on amount of inflammation and thus amount of MPX present. The results suggest that the modified assay yields a reliable estimate of relative SPX activity in mixed saliva.
Acetylcholine receptors from normal human muscle: Concentration, purification, and use in radioreceptor assays for autoantibodies
Authors: Wu James T. , Knight Joseph A. , Miya Terry G. , Bringhurst Cory 
Published: March 2007

AbstractWe measured the concentration of acetylcholine receptor (AChR) in various normal human muscles, obtained at autopsy, in order to find a more reliable and convenient source than the amputated leg muscle from diabetic patients for the isolation of AChR. We found that human calf muscle contains the highest concentration of AChR, approaching the concentration of receptor required for the radioreceptor assay for autoantibodies in myasthenia gravis (MG). Because the amount of contaminant proteins in the receptor preparations affect the sensitivity and precision of the assay, various chromatographic techniques were tested to improve the purity of the receptor preparation. We found that both G‐100 Sephadex and DEAE Sephacel chromatographies were effective in removing contaminant proteins. DEAE Sephacel chromatography is particularly useful because the procedure provides higher recovery of receptor. Against the same pool of autoantibodies from patients with MG, AChRs from diabetic and normal leg muscle exhibit similar affinities. We conclude that AChR from normal human calf muscle can be used in radioreceptor assays for measuring autoantibodies in patients with myasthenia gravis.
Correlation of histoplasma capsulatum polysaccharide antigen with the severity of infection in murine histoplasmosis
Authors: Williams Brian J. , Connolly‐Stringfield Patricia , Bartlett Marilyn , Durkin Michelle , Blair Robinette , Connolly Kathleen , Wheat L. Joseph , Garringer Todd , Tewari Ram P. 
Published: March 2007

AbstractWe sought to determine if Histoplasma capsulatum polysaccharide antigen (HPA) levels correlate with the extent of infection in murine of histoplasmosis. Separate groups of mice were inoculated intratracheally with varying numbers of H. capsulatum yeast cells. After 1 week, HPA levels and fungal burden (quantitative culture of lung and spleen and histopathologic stain of lung) were determined in lung and spleen, and HPA levels in serum. HPA levels, cultures and histopathological stain results of lung and spleen tissue showed a direct correlation with increasing inoculum size. HPA levels in serum also correlated with the size of inoculum. H. capsulatum antigen in lung correlated with silver stain scores of lung tissue, (R = 0.948, P <0.001) and with quantitative culture scores of lung, (R = 0.929, P <0.001). HPA levels in spleen tissue also correlated with spleen culture scores, (R = 0.724, P <0.001). These results indicate that determination of HPA level in serum and tissue may be a useful test in evaluating the severity of diseases as well as efficacy of antifungal therapy in histoplasmosis.
A novel igm‐indirect hemagglutination test for the serodiagnosis of acute toxoplasmosis
Authors: Yamamoto Yoshimi Imoto , Hoshino‐Shimizu Sumie , Camargo Mário E. 
Published: March 2007

AbstractA new indirect hemagglutination (M‐Toxo‐IHA) test was standardized for the serodiagnosis of acute toxoplasmosis. The reagent for this test consisted of stabilized human red cells coated with a Toxoplasma gondii heatstable alkaline‐solubilized extract which reacted predominantly with IgM antibodies found in serum samples from patients with a recent infection. Almost no reactivity was verified with serum samples from chronic or ancient toxoplasmosis. The reagent preparation was simple and obviated prior fractionation of parasite components reactive with IgM antibodies. Rheumatoid factor or high levels of IgG antibodies did not interfere in the M‐Toxo‐IHA test. A total of 358 serum samples was assessed for the evaluation of this test, comprising sera from patients with recent and ancient toxoplasmosis, from nonrelated diseases, as well as from clinically healthy individuals with no previous T. gondii infections. Serodiagnostic performance of the M‐Toxo‐IHA test in terms of sensitivity, specificity, efficiency, and positive and negative predictive values was verified, and the values obtained were 1.000, 0.985, 0.989, 0.952, and 1.000, respectively, in relation to IgM‐immunofluorescence or IgM‐capture ELISA. Six different batches of reagent, successively produced for the current study, provided reproducible results and were stable for at least 22 months at 4°C.
Immunoreactivity of nerve lipid antigens in leprosy
Authors: Vemuri Nalini , Mukherjee Rama 
Published: March 2007

AbstractNeural lipid antigens, namely, galactocerebroside and ganglioside, have been implicated in demyelinating diseases. We were interested in finding the role of these antigens in leprosy neuritis. The humoral immune response to these lipid antigens was quantitated by enzyme‐linked immunosorbent assay in sera from 91 leprosy patients and 18 normal individuals. Our data revealed the presence of antibodies to total nerve lipids (TNL) and galactocerebroside (GalC) and a significantly low level to ganglioside (Gg) in all the categories of leprosy. No antilipid antibodies were detected in normals. Anti‐TNL and anti‐GalC antibodies were highest in tuberculoid leprosy patients. Statistically significant positive correlation was observed between anti‐TNL and anti‐GalC antibodies in lepromatous borderline, tuberculoid, and neuritic patients.
Rapid and specific cyclosporine assay for the du pont aca® discrete clinical analyzer performed directly on whole blood
Authors: Hansen Joel B. , Lau H. Phillip , Janes Christopher J. , Miller Warren K. , Wiebe Donald A. 
Published: March 2007

AbstractA rapid, sensitive, and specific enzyme immunometric assay for cyclosporine in whole blood has been developed for the aca® analyzer, using a monoclonal antibody from Sandoz, Ltd. Between‐run CVs ranged from 6.5 to 7.6% for samples containing between 60 and 400 ng/mL cyclosporine. Sensitivity was better than 25 ng/mL in the assay, which has an effective upper range of greater than 600 ng/mL. Two correlation studies compared cyclosporine values from the Du Pont method to those determined by HPLC procedures in three hospital laboratories. The results from a total of 120 whole blood samples with CsA between 20 and 800 ng/mL showed excellent correlation between the methodologies. HPLC and Du Pont CsA values from 10 day serial studies also correlated well for samples from a kidney, kidney‐pancreas, heart, and two liver transplant patients. We conclude that the Du Pont CSA assay provides accurate and reproducible results in a convenient format in less than 30 minutes.
The phadiatop test allows adequate screening for atopy with a marked reduction in cost
Authors: Salkie Mark L. 
Published: March 2007

AbstractSera from 35 patients aged 5 years or younger and sera from 95 patients 6 years and older were assayed for both total lgE level, allergen‐specific lgE by RAST and either the Phadiatop Paediatric or Phadiatop procedures, depending on age. Good agreement was found allowing exclusion of specimens anticipated to be RAST negative with an expected reduction in RAST testing of up to 80% of the present workload, without significant exclusion of RAST positive specimens.
Measurement of human ventricular myosin light chain‐1 by monoclonal solid‐phase enzyme immunoassay in patients with acute myocardial infarction
Authors: Uji Yoshinori , Sugiuchi Hiroyuki , Okabe Hiroaki 
Published: March 2007

AbstractA monoclonal solid‐phase enzyme immunoassay has been developed for the detection of human serum ventricular myosin light chain‐1. Cross‐reactivity of this with human skeletal muscle myosin was observed, but the enzyme immunoassay with the sera of patients with acute myocardial infarction gave similar results with radioimmunoassay. The human ventricular myosin light chain‐1 levels in the healthy subjects were 0.2–6.6 ng/ml in males and 0.2–4.1 ng/ml in females. Withinrun and betweenrun precision (CVs) of the assays was on the order of 2.3–4.7% and 4.3–8.7, respectively. Sensitivity of the assay was 1.0 ng/ml, and working range was 5–100 ng/ml. In all patients with define acute myocardial infarction, serum ventricular myosin light chain‐1 levels increased three‐ to tenfold the upper reference range within 6 hr after the onset of chest pains. Two types of subtrend were discovered: its levels remain elevated for 3–4 days and its levels increased and then decreased 1–2 days after the initial rise but became elevated again for the next 4–7 days after the onset of chest pain, which is in contrast to the case with all conventionally used biochemical cardiac markers.
Evaluation of serum sialic acid and carcinoembryonic antigen for the detection of early‐stage colorectal cancer
Authors: Tautu Carmen , Alhadeff Jack A. , Pee David , Dunsmore Marlene , Prorok Joseph J. 
Published: March 2007

AbstractVarious expressions of elevated serum sialic acid (total sialic acid, TSA: lipid‐associated sialic acid, LASA; LASA/TSA; TSA normalized to total protein, TSA/TP) have been evaluated and compared with increased serum carcinoembryonic antigen (CEA) levels for the detection of early‐stage colorectal cancer. This evaluation was done blindly on a coded panel of 320 sera from staged colorectal cancer patients and controls provided by the Mayo Clinic‐National Cancer Institute Diagnostic Bank. Unlike the findings of a previous preliminary study (Tautu et al., JNCI 80:1333–1337, 1988), the ratio of LASA/TSA was not useful for detecting early‐stage (Dukes A and B) colorectal cancer. However, TSA and TSA/TP values were significantly elevated in each colorectal cancer subgroup compared with normal controls. TSA and TSA/TP values displayed a marginally better discriminatory power than CEA values in the case of Dukes A subgroup with respect to normal controls. CEA still appears to be the best single overall marker for discriminating between colorectal cancers and controls. However, multiple marker analysis using CEA and TSA (and related markers) appears to be more sensitive than CEA alone for detecting colorectal cancer.
Direct enzyme‐linked immunosorbent assay: A simple immunoassay using leishmania donovani promastigote for diagnosis of kala‐azar
Authors: Mukerji Krishna , Pal Abhijit , Naskar Kshudiram , Ghosh Dilip K. , Basu Debashish , Mallick K. K. 
Published: March 2007

AbstractFor immunodiagnosis of kala‐azar enzyme‐linked immunosorbent assay (ELISA) and immunofluorescence testing (IFAT) are commonly used. In IFAT, whole parasite antigen and in ELISA the soluble antigen have been used. Preparation of ELISA antigen has certain inherent difficulties. We have developed a simple, specific, and quantitative immunoassay, “direct ELISA” for diagnosis of kala‐azar. Intact formalinized promastigote suspension has been used to combine with the antibodies of the patient sera. The colour developed in the supernatant by the enzyme conjugate combined on the parasite surface was measured with a spectrophotometer. The test was able to detect kala‐azar‐specific antibodies at very high serum dilution and could discriminate between kala‐azar and the common diseases prevalent in Asia. The optical densities of the sera of different control groups were significantly low. The method has potential for use as a diagnostic tool in less well equipped laboratories.
New method for the determination of pancreatic amylase evaluated
Authors: Lorentz Klaus , Seiler Dieter , Nagel Dietmar , Naegele Ulrich , Koller Peter , Bais Renze , Bayer Peter M. , Keller Herbert , Puukka Matti , Ruostesuo Juhani , Puukka Raija , Rosalki Sidney B. , Foo Ying 
Published: March 2007

AbstractWe have carried out a multicenter evaluation of a new reagent carrier for Reflotron®, specific for pancreatic amylase where the salivary isoenzyme is inhibited by two specific monoclonal antibodies. This new procedure combines easy handling with low imprecision (median CV < 3%) in control material, serum, heparinized blood, and plasma and close correlation (r = 0.991 to 0.999) with established manual and automated methods. The same close correlation was found with values obtained from either venous or capillary finger‐stick blood. Salivary amylase up to 54 kU/L (37°C) was inhibited to about 97%. Endogenous interference by hemoglobin, bilirubin, triglycerides, cholesterol, or hematocrit was found to be negligible within a wide range of interferent concentrations. Out of a panel of 28 commonly used drugs it was shown that only two (ascorbic acid and paracetamol), and then only at toxic concentrations, caused a deviation in amylase activity of greater than 10%. From the results of this study we conclude that this new method is suitable for highly precise and accurate measurements of pancreatic amylase in emergency and routine laboratories.
Leukocyte membrane markers on cell populations defined by six flow cytometric bitmaps
Authors: Uz̆arević Branka , Batinić Drago , Petrovec̆ki Mladen , Marus̆ić Matko 
Published: March 2007

AbstractPeripheral blood samples from healthy adults, patients with transplanted bone marrow, and healthy children were analyzed for the proportions of cells positive for CD2, CD4, CD8, CD19, CD56, CDw65, and KiM8 cell membrane markers. Six cell gates were defined on granularity vs. cell size (L90°LSx‐FALS) flow cytometric display for each blood sample: large lymhocytic, small lymphocytic, monocytic mononuclear, granulocytic, and all‐cell gate, and the listed markers were determined in each of them. Considerable differences in marker‐positive cell proportions were found between healthy adults and children, and particularly between healthy adults and patients with transplanted bone marrow. Marker‐positive cell frequencies corresponded to biologic distribution of three leukocyte populations within the defined gates, in a general agreement with known specificities of the antibodies used. A considerable degree fo variations in the positions of gates drawn for different samples and the numbers of cells counted in them was observed. Still it appeared that all six gates, otherwise commonly used in flow cytometric analysis, could be precisely defined and yielded reproducible data. Large and small lymphocytic gates yielded very similar marker frequencies, revealing that in cases where three leukocyte populations were not clearly delineated on (90°LSxFALS) display, a smaller lymphocytic gate could be safely drawn in order to avoid contamination with monocytes.
Low frequency and low level of elevation of serum CA 72‐4 in human carcinomas in comparison with established tumor markers
Authors: Wu James T. , Carlisle Pam 
Published: March 2007

AbstractWe evaluated a new circulating tumor marker, CA 72‐4, by comparing its frequency of appearance and level of elevation with other established tumor markers in serial serum specimens from patients with various carcinomas. We found that CA 72‐4, though highly expressed and widely found in various tumor tissues, is present at low concentration and frequency in the serum. In breast, colon, ovarian, and pancreatic carcinomas, only 21%, 30.9%, 16%, and 26.5% of specimens, respectively, showed elevated CA 72‐4. When elevated, the level of elevation was also low, much lower than that of the dominant markers. Poor response of CA 72‐4 to therapy was especially noticeable in serial specimens. In most cases, the CA 72‐4 remained normal for the entire series while other markers remained at elevated levels. However, changes of the level of CA 72‐4 usually paralleled those of other markers but at a much lower concentration. Simultaneous measurement of CA 72‐4 and CA 19‐9 appears useful to differentiate colorectal from pancreatic carcinomas when they all contained elevated levels of CA 19‐9. There was a much higher ratio of CA 72‐4 to CA 19‐9 with colon than with pancreatic and other carcinomas (247 ± 524 vs. 4.7 ± 6.8).
Sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for (anti‐human T‐cell leukemia virus type I) IgG in serum using a synthetic peptide, Cys‐Env gp46(188‐224), as antigen
Authors: Kohno Takeyuki , Sakoda Iwane , Ishikawa Eiji 
Published: March 2007

AbstractA sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for (anti‐human T‐cell leukemia virus type I) IgG (anti‐HTLV‐I IgG) in serum using a synthetic peptide, Cys‐env gp46(188‐224) of HTLV‐I, is described. Anti‐HTLV‐I IgG in test serum, which had been incubated with excess of inactive β‐D‐galactosidase to eliminate interference by anti‐β‐D‐galactosidase antibodies, was reacted simultaneously with 2,4‐dinitrophenyl‐bovine serum albumin‐Cys‐env gp46 (188‐224) conjugate and Cys‐env gp46 (188‐224)‐β‐D‐galactosidase conjugate. The complex formed consisting of the three components was trapped onto polystyrene balls coated with affinity‐purified (anti‐2,4‐dinitrophenyl group) IgG. After washing to eliminate nonspecific IgG in the test serum and excess of the β‐D‐galactosidase conjugate, the complex was eluted from the polystyrene balls with ϵN‐2,4‐dinitrophenyl‐L‐lysine and transferred to polystyrene balls coated with affinity‐purified (anti‐human IgG‐γ‐chain) IgG. β‐D‐Galactosidase activity bound to the (anti‐human IgG‐γ‐chain) IgG‐coated polystyrene balls was assayed by fluorometry. This assay was more sensitive and useful than the immune complex transfer enzyme immunoassay using Cys‐Arg‐env gp46(188‐209) and other methods using HTLV‐I as antigen.
Tumor markers CA 19‐9 and CA 195 are also useful as markers for cystic fibrosis
Authors: Wu James T. , Olson June , Walker Kay 
Published: March 2007

AbstractWhen monoclonal kits are used we can no longer detect highly elevated serum concentrations of carcinoembryonic antigen in cystic fibrosis (CF) patients as we could earlier (Pediatr Res 10:235–236, 1975). Instead, we find increased concentrations of CA 19‐9 or CA 195 in the CF sera. The serum levels of CA 19‐9 not only reflect the pulmonary condition of CF patients but also respond well to antibiotic therapy. Several lines of evidence suggest that the elevated serum concentration of CA 19‐9 is derived from sputum and corresponds with the amount of sputum in the lung. Correlations between CA 19‐9 and CA 195 in random and serial specimens from both patients with CF and patients with pancreatic carcinoma suggest that all sera contain heterogeneous, Lewis blood group‐related epitopes and the proportions of various epitopes are different among individual patients. When monitored on multiple tumor markers, the pattern of CF is different from that of pancreatic carcinoma although both usually show elevated CA 19‐9. Our study indicates that both CA 19‐9 and CA 195 can be used as sensitive markers for the early detection of exacerbation in CF patients. © 1992 Wiley‐Liss, Inc.
Novel and ultrasensitive noncompetitive enzyme immunoassay (hetero‐two‐site complex transfer enzyme immunoassay) for α‐human atrial natriuretic peptide
Authors: Hashida Seiichi , Ishikawa Eiji 
Published: March 2007

AbstractA novel and ultrasensitive noncompetitive enzyme immunoassay (hetero‐two‐site complex transfer enzyme immunoassay) for α‐human atrial natriuretic peptide (α‐hANP) in plasma is described. α‐hANP was biotinylated using N‐hydroxysuccinimidobiotin and trapped onto an anti‐α‐hANP [6–28] IgG‐coated polystyrene ball. After washing, biotinylated α‐hANP was eluted from the polystyrene ball with HCI and was reacted with 2,4‐dinitrophenyl‐fluorescein‐bovine serum albumin‐disulfide‐rabbit anti‐α‐hANP [6–28] IgG conjugate. The complex formed was trapped onto (anti‐2,4‐dinitrophenyl group) IgG‐coated polystyrene balls and, after washing, reacted with avidin‐β‐D‐galactosidase conjugate. The polystyrene balls were washed, and the complex of the three components was eluted with ϵN‐2, 4‐dinitrophenyl‐L‐lysine and transferred to anti‐fluorescein IgG‐coated polystyrene balls. After washing, the complex was released from the polystyrene balls by reduction with 2‐mercaptoethylamine and transferred to (anti‐rabbit IgG) IgG‐coated polystyrene balls. β‐D‐Galactosidase activity bound to the last polystyrene balls was assayed by fluorometry. The detection limit of α‐hANP [1–28] was 3 fg (1 amol)/tube. Interference by plasma proteins was eliminated by separation of peptides from proteins using a molecular sieve. The assay range of plasma α‐hANP [1–28] was 0.04–120 ng/L, and plasma levels of hANP in healthy subjects (11–56 ng/L) were measured without concentration. © 1992 Wiley‐Liss, Inc.