bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antibodies to Sm, RNP and SSB detected by solid‐phase ELISAs using recombinant antigens: A comparison study with counter immunoeloctrophoresis and immunoblotting
Authors: Delpech Annie , Gilbert Daniéle , Daliphard Sylvie , Le Loet Xavier , Godin Michel , Tron Francois 
Published: March 2007

AbstractSera from 64 patients with systemic lupus erythematosus (SLE) were tested for the presence of anti‐Sm, anti‐RNP, and anti‐SSB antibodies using commercially available solidphase enzyme‐linked immunosorbent assays (ELISAs) and recombinant nuclear proteins as substrates. The results were compared to those obtained with counterimmunoelectrophoresis (CIE) and immunoblotting (IBT) using a rabbit thymus extract (RTE) as the substrate.The ELSAs detected antibodies to Sm, RNP, and SSB in, respectively, 25%, 36%, and 15% of the SLE sera. Neither IBT‐positive/ ELISA‐negative nor CIE‐positive/ELISA‐negative sera were found, regardless of the specificity considered, suggesting that ELISAs using recombinant nuclear antigens are highly sensitive.Discrepancies were observed between the results obtained with these different techniques. In addition to sera positive by both IBT and ELISA but negative by CIE, a substantial number of sera had ELISA‐detectable anti‐Sm, anti‐RNP, and anti‐SSB antibodies which failed to react with the corresponding polypeptides by IBT. The reasons for ELISA/IBT discrepancies were explored; however, no single explanation was found. Instead, a higher sensitivity of the ELISA to detect antibodies directed against certain polypeptides, the possible inability of IBT using RTE as the substrate to detect antibodies reacting with conformational antigenic determinants, and false‐positive reactions in the ELISAs were suggested.Thus, it is still advisable to perform both IB and ELISAs simultaneously in well‐defined autoimmune diseases to further analyze the potential advantages of ELISAs using recombinant antigens. © 1993 Wiley‐Liss, Inc.
Rapid colorimetric detection of epidermal growth factor receptor mRNA by in situ hybridization
Authors: Iezzoni Julia C. , Kang Jin‐Han , Bucana Corazon D. , Reed Jon A. , Brigati David J. 
Published: March 2007

AbstractWe describe a rapid, formamide‐free, random oligomer‐enhanced in situ hybridization method in formalin‐fixed, paraffin‐embedded tissue sections using a biotinylated oligonucleotide probe for colorimetric detection of the mRNA transcript of the epidermal growth factor receptor (EGFR) gene, a putative protooncogene. Transitional cell carcinomas (TCC) of the urinary bladder and oral squamous cell carcinomas (SCC) were examined. A431, a human squamous carcinoma cell line that overexpresses EGFR mRNA, and mature skeletal muscle, Known not to express EGFR, served as control tissues. A biotinylated poly‐T oligonucleotide probe was used to evaluate the preservation of mRNA in the formalin‐fixed, paraffin‐embedded tissues. The EGFR mRNA and poly‐T oligonucleotide probes were constructed with a target‐specific, 5′ region and a 3′, non‐hybridizing, biotinylated tail. Random sequence oligomers nine bases long added to the probe cocktail eliminated the need for formamide and sheared carrier DNA in the method. The assay produced strong, specific staining for EGFR mRNA in all cases of TCC and SCC, and in the A431 tumors, but not in skeletal muscle. The assay was completed in approximately 90 minutes. This method may have widespread utility for rapid and specific detection of other mRNA sequences. © 1993 Wiley‐Liss, Inc.
Detection of antineutrophil autoantibodies by flow cytometry: Use of unfixed neutrophils as antigenic targets
Authors: Maher George M. , Hartman Kip R. 
Published: March 2007

AbstractAntineutrophil antibodies may be found in the sera of patients with chronic neutropenia as well as in the sera of a variety of patients with neutropenia and associated autoimmune or infectious disorders. We evaluated an immunofluorescent flow cytometric technique for the measurement of antineutrophil antibodies in serum. Sera from patients with suspected immune neutropenia were studied and compared with a group of sera from normal healthy individuals, as well as with sera from patients with rheumatoid arthritis and systemic lupus erythematosus. Of 159 patients with suspected immune neutropenia and a variety of associated clinical disorders, 59 (37%) were found to have evidence for enhanced binding of IgG to normal target neutrophils, interpreted as positive for antineutrophil antibodies. Whereas 0/37 nonneutropenic patients with typical RA had positive results, 51/244 (21%) of sera from nonneutropenic patients with SLE or other collagen vascular disorders showed enhanced IgG binding to neutrophils. Living neutrophils were used to study the effects of cellular activation, and increased antibody binding was observed with certain sera that contained IgG directed against activation‐dependent antigens. We found that, under controlled conditions, flow cytometry can be reliably used to detect antineutrophil autoantibodies, with unfixed, living neutrophils as antigenic targets. © 1993 Wiley‐Liss, Inc.
Enzyme immunoassay and characterization of pancreatic stone proteins in human urine
Authors: Tatemichi Noriyuki , Takahashi Chikako , Hayakawa Shinobu , Hayakawa Tetsuo , Shibata Tokimune , Kitagawa Motoji , Sobajima Hiroshi , Nakae Yasuyuki 
Published: March 2007

AbstractAn enzyme immunoassay of pancreatic stone protein (PSP) in human urine was developed. Mean analytical recovery of pure PSP‐S2‐5 added to urine was 102.3% (SD 5.9%), and the precision of the assay was 2.0‐2.7% within an assay and 2.5‐2.9% between assays. In healthy volunteers (age 20‐55 years), the mean value of the PSP concentration, expressed as ratios to urine creatinine, was 129 ± 88 (mean ± SD) μg/g without any differences for sex. Urine PSP correlated with urine N‐acetylglucosaminidase (NAG) (r = 0.354). The molecular forms of immunoreactive PSP in urine were characterized by using cation exchange chromatography (Mono S), SDS‐PAGE, N‐terminal sequence, and enzyme immunoassay analysis. The urine PSP, eluted at the position corresponding to PSP‐S2‐5 on cation exchange chromatography, was converted to PSP‐S1 by trypsin digestion. The difference in mobility on SDSPAGE between urine PSP and PSP‐S2‐5 seems to be due to a glycosylated undecapeptide (N‐terminal 1‐11). The proposed method offers a sensitive, specific, and reproducible tool for laboratory analysis of human urine PSP levels. © 1993 Wiley‐Liss, Inc.
Reticulocyte count with maturation fractions in pancytopenic evaluation by a fully automated counter
Authors: Lin Chung K. , Hsu Hui C. , Chau Wing K. , Jiang Mei L. , Chiu Chang F. 
Published: March 2007

AbstractUsing a fully automated reticulocyte counter, the roles of the reticulocyte count with maturation in pancytopenia were evaluated. Different groups of pancytopenia including aplastic anemia, infiltrative marrow disorder, hypersplenism, and megaloblastic anemia were recruited. All patients had bone marrow examinations for morphological diagnosis and reticulocyte evaluation using an automated counter. The roles of these parameters were then analyzed statistically in the differential evaluation among these conditions. The following subjects were studied: 292 normal subjects, 67 cases of aplastic anemia, 69 cases of marrow infiltration by different malignancies, 35 cases of hypersplenism, and 13 cases of megaloblastic anemia. The results showed that the absolute retlculocyte counts were lowest in the groups of aplastic anemia and megaloblastic anemia and highest in hypersplenism. Both showed significant differences from the infiltrative groups. The maturation fractions were most immature in the group of marrow infiltration and are significantly different from the other groups. It was concluded that the highest absolute reticulocyte count (>0.09 1012/L) obtained in pancytopenic patients suggests it to be a case of hypersplenism. The lowest counts ( < 0.03 1012/L) with lowest immature fractions ( < 10%) suggest the groups of aplastic or megaloblastic anemias. The highest immature fractions ( >30%) with a nearly normal reticulocyte count favor the group of marrow infiltration. © 1993 Wiley‐Liss, Inc.
Human thyroid peroxidase: Autoantibody recognition depends on the natural conformation
Authors: Berthold Heike , Steffens Ute , Northemann Wolfgang 
Published: March 2007

AbstractThyroid peroxidase (TPO) purified from human thyroid glands was used to study the correlation between natural conformation and its ability for binding of autoantibodies in sera from patients with autoimmune lymphocytic thyroiditis (Hashimoto's thyroiditis). Treatment with heat (60°C), urea, sodium dodecylsulfate (SDS), and dithiothreitol (DTT) decreased the autoantigenicity of TPO on average by 73.3%, 74.1%, 86.3%, and 91.8% measured by enzyme‐linked immunosorbent assay (ELISA), respectively. These data demonstrate, that the binding of the majority of autoantibodies to human TPO depends on its natural conformation and modification. © 1993 Wiley‐Liss, Inc.
Quantitation of hepatitis B virus DNA in serum by ammonium sulfate precipitation and molecular hybridization
Authors: Wen Long T. , Henneberger Michael , Nguyen Nicole , McPherson Richard A. 
Published: March 2007

AbstractWe developed a method to quantitate hepatitis B virus (HBV) DNA in serum by ammonium sulfate fractionation and DNA hybridization. Serum samples were precipitated with 45% saturated ammonium sulfate, resuspended in buffer, and spotted on a nylon membrane. Following denaturation in alkali, HBV DNA sequences on the membrane were detected by hybridization with a 32P‐labeled DNA probe of the entire HBV genome. Bound radioactivity was measured with liquid scintillation counting. Ammonium sulfate fractionation of positive samples increased assay sensitivity by 10–30‐fold compared to no treatment. Sensitivity for detection of cloned HBV DNA added to negative serum was 0.2 pg. Recovery of cloned HBV DNA added to negative serum before fractionation was equivalent to direct spotting of DNA onto the membrane in the absence of serum. This method enhanced HBV DNA recovery from serum into small volumes, thereby expanding the potential analytic range of spot hybridization assays. © 1994 Wiley‐Liss, Inc.
Evaluation of bioassay for toxicity of ciguateric fish and associated toxins
Authors: Hokama Y. , Shirai J. L. , Islam M. A. 
Published: March 2007

AbstractEvaluation of the mouse toxicity assay symptom of hind leg paralysis (HLP) with mouse death by statistical analysis is presented in this study. The fishes assessed were herbivores including Ctenochaetus strigosus (kole), Ctenochaetus hawaiiensis, Acanthurus sandvicensis (manini), and Mugil cephalus (mullet); and the carnivores, Cephalopholis argus (roi) and Cheilinus rhodochrous (po'ou). The latter can also be considered an omnivore. The extracts of both herbivore and carnivore species appeared to be most toxic when HLP occurred in the mice. Ninety‐three percent of the mice with HLP died, whereas when no HLP (NHLP) occurred, only 51% of the mice died. Carnivore flesh extracts (po'ou and roi) were least toxic with one death out of a total 22 mice. The unidentified toxin associated with HLP appears to differ in biological properties from that of ciguatoxin(s) in that it was not found in the flesh tissues of carnivores. Further chemical studies of this toxin(s) is being addressed presently. © 1994 Wiley‐Liss, Inc.
Mapping of antigenic epitopes within the recombinant human preproinsulin related to insulin‐dependent diabetes mellitus
Authors: Walter Michael , Berg Heike , Northemann Wolfgang 
Published: March 2007

AbstractThe human insulin domains, signal peptide, B‐chain, C‐peptide, and A‐chain, were highly expressed in Escherichia coli as recombinant proteins N‐terminally fused to glutathione‐S‐transferase and a histidinehexapeptide. The recombinant proteins were purified from insoluble cell fraction by affinity chromatography using metal chelating matrix, which was charged with Ni+2 iones. ELISA screening for autoantibodies directed to preproinsulin were performed with sera from patients with recently diagnosed insulindependent diabetes mellitus in order to localize the antigenic epitopes within the human preproinsulin. Of the patients, 14% had developed autoantibodies that recognized either the recombinant C‐peptide or the signal peptide. No reaction was observed with the A‐chain or B‐chain. © 1994 Wiley‐Liss, Inc.
Comparison of two Cobas Core enzyme immunoassays with other test systems for the detection of cytomegalovirus‐specific IgG and IgM antibodies
Authors: Steinmann J. , Weigel T. 
Published: March 2007

AbstractThe new Cobas Core CMV IGM and IgG enzyme immunoassays (Roche Diagnostic Systems, Basle, Switzerland) were evaluated for their ability to detect cytomegalovirus (CMV)‐specific IgM and IgG antibodies. Therefore, both were compared with some other commercially available and already established serological tests used in the laboratory diagnosis of CMV infection. These included the Abbott IMx CMV IgM and IgG assays, the Abbott CMV‐M EIA, the Gull CMV IgM and IgG immunofluorescence tests, the medac CMV‐IgM‐ELA, and the Enzygnost anti‐cytomegalovirus assay (Behringwerke). A total of 572 serum samples of various categories were examined and the results showed high concordances between all methods, ranging from 84.5% to 94.9%. In a follow‐up on renal transplant patients, the times of first detection of seroconversions were compared. Since a high overall agreement between the Cobas Core CMV IgM and IgG enzyme immunoassays and the other test systems were observed, these new assays represent useful and reliable tools for clinical CMV diagnosis. © 1994 Wiley‐Liss, Inc.
Comparison of techniques for minimizing interference of bilirubin on serum creatinine determined by the kinetic Jaffé reaction
Authors: Lolekha Porntip H. , Sritong Noppmats 
Published: March 2007

AbstractThis study compared the effect of sodium dodecyl‐sulfate, potassium ferricyanide, and preincubation technique to continuous‐flow analysis and prior deproteinization to correct the negative interference of bilirubin on serum creatinine found by the kinetic Jaffé reaction. Bilirubin increased to 684 μmol/L did not interfere with serum creatinine measured by the methods incorporated with dialysis or deproteinization. Trichloroacetic acid was the best protein precipitant. The reagent incorporated with sodium dodecyl sulfate was more appropriate to minimize bilirubin interference than reagent containing potassium ferricyanide. An increase in potassium ferricyanide concentration resulted in false positive creatinine values. Incorporation of both SDS and potassium ferricyanide in the reagent did not help in minimizing the bilirubin interference over use of each chemical alone. The 10 minutes of preincubation of the sample with alkaline buffer incorporating with either SDS or potassium ferricyanide before starting the Jaffé reaction was the appropriate way to overcome unconjugated bilirubin interference at a level of 342.0 μmol/L. However, the technique did not work uniformly with icteric patient sera containing conjugated, unconjugated, and delta bilirubin. This is a challenging problem that remains to be solved by the clinical chemist. © 1994 Wiley‐Liss, Inc.
Contribution of the host to test results in assays of staphylococcus epidermidis
Authors: Herndon Betty , Dall Lawrence , Suvarna Padma 
Published: March 2007

AbstractFrom two human populations (one pediatric and one adult), clinically diagnosed with Staphylococcus epidermidis (S. epidermidis) sepsis of similar severity, bacteria were isolated from pre‐antibiotic blood samples and evaluated for virulence. The LD50 of the bacteria in a mouse model was performed, with evaluation of animals dying acutely following intravenous S. epidermidis administration. More simple assays of virulence were also performed, including bacterial adherence to a fibrin clot and carbohydrate specific lectin binding. The eight pediatric‐host S. epidermidis isolates required a significantly larger dose to produce lethality in dosed animals (LD50) when compared to the 20 adult‐host S. epidermidis isolates. The fibrin clot assay, a test that has corroborated bacterial virulence in endocarditis models, did not differentiate the groups: all but one of the 28 isolates were well above the adherence seen with the ATCC control, suggesting endocarditis‐producing potential. Glycocalyx (slime) from eight of the more virulent isolates showed reactivity with a glucose‐specific biotinylated lectin which was lacking in other isolates. Necropsy of mice dying at 12 hr showed S. epidermidis strain differences in specific organ effects.Overall, this study demonstrates the utility of the LD50 to provide a highly sensitive quantification of bacterial virulence. Necropsy of test animals dying acutely has showed an apparent organ tropism of some of these isolates which are usually considered harmless commensals.©1995 wiley‐Liss, inc.
Colorimetrical rate assay for urinary dipeptidyl peptidase iv (dppiv) activity using a new substrate
Authors: Shibuya‐Saruta Hiroko , Sugiyama Masami , Kasahara Yasushi 
Published: March 2007

AbstractWe synthesized a new substrate glycyl‐L‐proline 3,5‐dibromo‐4‐hydroxyanilide (Gly‐Pro‐DBAP), for dipeptidyl peptidase IV (DPPIV). Its hydrolysis by DPPIV resulted in the formation of a chromophore, 2,6‐dibromo‐phenol‐indo‐p‐xylenol, and its maximal absorption wavelength (600 nm) was longer than that of p‐nitroaniline (415 nm) released from conventional substrate, glycyl‐L‐proline pnitroanilide (Gly‐Pro‐pNA). We also established the rate assay for urinary DPPIV activity using Gly‐Pro‐DBAP. The optimum pH was between 8.5 and 9.0. The apparent Km was 1.1 mmol/1. The detectable range was 2.5–350 U/I. No changes in blank values occured throughout the enzyme reaction in the optimum pH. Its value was also much lower than Gly‐Pro‐pNA. CVs for within‐run and between‐run were 1.1% (n=10) and 3.0% (n=10), respectively. Among tested peptidases, only DPPIV could hydrolyze Gly‐Pro‐DBAP. Among the protease inhibitors, only two, diprotin‐A and phenylmethylsulfonyl fluoride (PMSA), could inhibit DPPIV activity. The present method did not interfere with urinary ingredients such as hemoglobin. The correlation between the present (y) and conventional (x) methods is presented by the equation y=1.121×+0.096 (r=0.993). Thus the present method provides practical advantages over the conventional method for routine laboratory use.©1995 wiley‐Liss, inc.
Expression of matrix metalloproteinase matrilysin (mmp‐7) was induced by activated ki‐ras via ap‐1 activation in sw1417 colon cancer cells
Authors: Yamamoto Hiroyuki , Itoh Fumio , Senota Akinori , Adachi Yasushi , Yoshimoto Mitsuru , Endoh Takao , Hinoda Yuji , Yachi Akira , Imai Kohzoh 
Published: March 2007

AbstractThe matrix metalloproteinase matrilysin (MMP‐7) is a member of the matrix metallo‐proteinase gene family, which is believed to play an important role in tumor invasion and metastasis. We have previously found that matrilysin mRNA is specifically expressed in colorectal cancers and adenomas and that its message is localized in the tumor cells themselves. We examined the effects of activated Ki‐ras oncogene on the expression of matrilysin in colon cancer cells. We showed that both mRNA and the enzymatic activity of matrilysin were induced by the introduction of activated Ki‐ras into SW1417 colon cancer cells. To understand the mechanisms regulating this induction, we analyzed alterations of AP‐1 activity induced by activated Ki‐ras, using the chloramphenicol acetyltransferase assay. AP‐1 activity in SW1417 cells expressing activated Ki‐ras was higher than that in control cells. The gel‐shift assay also showed higher levels of AP‐1 binding protein in SW1417 cells expressing activated Ki‐ras than those in control cells. Our results suggest that activated Ki‐ras may play a role in inducing expression of matrilysin through an AP‐1‐dependent pathway in colon cancer cells.
Comparison of cathepsin d determinations in human carcinomas by enzyme immunoassay and immunoradiometric assay
Authors: Shaheen Raymond M. , Miseljic Slobodan , Doering David L. , Wittliff James L. 
Published: March 2007

AbstractOverexpression of cathepsin D in several types of carcinoma in women appears to be associated with a poor clinical course. In this prospective investigation, cathepsin D levels in 170 specimens of normal and neoplastic human tissues were determined simultaneously by enzyme immunoassay (EIA) and immunoradiometric assay (IRMA) to allow comparisons in multicentric studies, such as cooperative clinical trials. Nonmalignant uteri and specially prepared reference powders were also evaluated. Linear regression analysis between the two assays for all specimens [EIA = 0.87(IRMA) – 3.18] demonstrated a correlation coefficient (r) of 0.99 (P <0.001). When malignancies were categorized by the tissue origin (i.e., breast, uterus, ovary, lymph node, and colon), highly significant correlations were also observed (regressions slopes ranged from 0.58 to 1.02). Intra‐ and interassay controls conducted for the new EIA procedure gave CV% ranging from 4.4 to 10.2, which was similar to the IRMA test for cathepsin D. The results of both assays correlated well and were highly reproducible. Either assay may be used with confidence that comparable cathepsin D values will be obtained in a wide range of tissue biopsies.
Problems in determining levels of free digoxin in patients treated with digoxin immune fab
Authors: Honda Stacey A. A. , Rios Carlos N. , Murakami Lourna , Morita Thelma , Scottolini Alfred G. , Bhagavan Nadhipuram V. 
Published: March 2007

AbstractDetermination of free digoxin levels in patients treated with digoxin immune FAb has long been a problematic area in clinical laboratory testing. The older radioimmunoassays resulted in inaccurate and variable results due to the competition of the administered drug with the radioactively labelled forms. The 1995 Physicians' Desk Reference continues to state that digoxin immune FAb will interfere with digitalis immunoassay measurements. This statement, however, is based primarily on the RIA methods. We evaluated the Stratus digoxin assay and the TDX digoxin II assay. Increasing amounts of immune FAb were added in a stepwise fashion to 12 patient samples containing high normal to elevated digoxin levels. Results showed a progressive decrease in digoxin levels when assayed with the Stratus kit. However, five patient samples tested with the TDX kit resulted in constant digoxin values despite the presence of increasing digibind levels. These results suggest that the Stratus method measures free digoxin, whereas the TDX method measures the total digoxin. Measurement of digoxin by the Stratus method is simple and quick. The Stratus digoxin assay may be an accurate and objective way of measuring free digoxin levels in patients on digoxin immune FAb.