bluuebunny/crossref_metadata_2025_split huggingface dataset:
Relative quantification of collagen mrna in fibroblasts by a radioactive polymerase chain reaction technique
Authors: Goetzl Edward J. , Turck Christoph W. , Golden Jeffrey A. , Migchielsen Alexandra A. J. , Yu Yong Li 
Published: March 2007

AbstractA radioactive polymerase chain reaction (PCR) method has been developed for the relative quantification of the human α‐2 chain of type I collagen [hu α‐2 (I)] in cells. cDNAs generated by reverse transcription from the total pool of cytoplasmic RNA serve as a template for polymerase chain reaction amplification of a hu α‐2(I) cDNA primed by two sequence‐specific synthetic oligonucleotides. The distinctive 390 bp hu α‐2(I) cDNA and two Aval fragments of 220 and 170 bp are identified by agarose gel electrophoresis. α‐32P‐dCTP of defined specific activity is included in the PCR reaction and the 390 bp cDNA is excised from the electrophoresis gel to permit direct radioactive quantification of hu α‐2(I) mRNA. The amount of hu α‐2(I) mRNA expressed in as few as 111 fibroblasts was determined reliably. In contrast, the hu α‐2(I) mRNA from at least 5 × 105fibroblasts was required for detection by Northern blot analysis developed with the same cDNA probe radiolabelled with α‐32P‐dCTP by random priming. Human bronchoalveolar lavage (BAL) fluids of six patients with fibrosing lung diseases stimulated the level of expression of hu α‐2(I) mRNA in cultured human fibroblasts as determined by this technique. The radioactive PCR method thus quantifies hu α‐2(I) mRNA in fibroblasts with sufficient sensitivity to study fibroblast activation in vitro and detect fibroblast stimuli in human clinical samples.
Evaluation of direct agglutination test (dat) and elisa for serodiagnosis of visceral leishmaniasis in india
Authors: Pal Abhijit , Mukerji Krishna , Naskar Kshudiram , Ghosh Dilip K. , Basu Debashish , Mallick K. K. 
Published: March 2007

AbstractThe direct agglutination test (DAT) and the enzyme‐linked immunosorbent assay (ELISA) were used for serodiagnosis of parasitologically confirmed Indian visceral leishmaniasis (VL) cases. All the sera of VL cases were positive by both the methods. DAT titres of VL cases were ≥1:3,200, and ELISA values were ≥0.55 at 1:400 dilution. In the control group, sera of widely prevalent diseases of India, such as leprosy, tuberculosis, malaria, and liver cirrhosis, were included. Both tests could discriminate between VL and other patients of the control group. The sera of post‐kala‐azar dermal leishmaniasis (PKDL) patients gave OD values of >0:55 and had DAT titres of 1:1,600. Both tests are sensitive and specific for the diagnosis of VL cases. DAT, being simpler and more economical, will be suitable for diagnosis and epidemiological studies for VL under rural conditions of India.
Sensitized immunochemical method for the specific detection of human haemoglobin in dried biological samples
Authors: Ottó Szabolcs , Németh Mária Sz. , Kocsis György , Tulok István , Kelemen Katalin 
Published: March 2007

AbstractThe authors describe a modified version of counterimmunoelectrophoresis of easy performance and high sensitivity. With this method human haemoglobin can easily and specifically be determined either in fresh or dried biological samples. Glutaraldehyde pretreatment of the samples results in the development of complexes with highly favourable electrophoretic mobility and precipitating capacity. By this chemical modification and use of a double set of samples with varying antigen‐antibody, proportion, the sensitivity of haemoglobin detection in erythrocyte‐containing haemolysate and native blood was 300 ng/ml. The examination is easily performed, and without considerable outlay, even with conventional laboratory facilities. It is equally suitable for the detection of occult colorectal bleeding and for species‐specific study of blood stains of unknown origin.The authors succeeded in identifying human haemoglobin even in a 42‐month dried blood stain.
Increased serum alkaline phosphatase activity: A possible indicator of renal damage
Authors: Leibovitch Ilan , Ben‐Chaim Jacob , Ramon Jacob , Goldwasser Benad 
Published: March 2007

AbstractIncreased levels of serum alkaline phosphatase (ALP) (E.C.3.1.3.1.) were observed in 25 patients with various urological conditions involving the kidneys: malignancy, complicated nephrolithiasis, and surgical and percutaneous manipulations. Other possible sources for increased ALP level, mainly hepatic and osseous, were excluded by history, laboratory tests, and liver and bone imaging. Studies of isoenzymes of ALP did not show a distinctive pattern. ALP levels returned to the normal range by treating the underlying lesions involving the kidney: nephrectomy, complete removal of stones, or removal of nephrostomy. The increase in serum ALP activity may be derived from the injury to the brush border membrane of the renal tubular cells. Renal function impairment and contrast media induced nephrotoxicity may also be responsible for the increased serum ALP. Serum ALP may be a marker for involvement of the kidneys in pathological processes and an indicator of complete treatment. This clinical observation is worthy of further study.
Quick measurement of serum unconjugated estrogens, and its clinical application to abnormal pregnancies and ovarian follicle maturation
Authors: Kondo Zenjiro , Makino Tsunehisa , Lzumi Shun‐Ichiro , Lizuka Rihachi 
Published: March 2007

AbstractUsing an automated HPLC system reported recently by us (J Clin Lab Anal 4:410–413, 1990), serum unconjugated estriol and estradiol (u‐E3 and u‐E2) can be measured within 22 min. The clinical value of this method was evaluated in this study with abnormal pregnancies, twin pregnancies, and pregnancies associated with fetal anencephaly or complicated by gestational toxicosis or diabetes mellitus. A significant correlation was demonstrated between an abrupt decrease in serum u‐E3 level and an abnormal pattern on fetal heart rate monitor. Since urinary E3 was not sensitive enough to detect any depressed fetal condition, serum u‐E3 is better biochemical parameter for assessment of fetal well‐being.In addition, u‐E2 was measured in the sera of nonpregnant women with separation chromatography as a pretreatment in order to obtain higher sensitivity of the system. This modified HPLC system enabled us to determine the serum u‐E2 in about 60 min with the minimal detectable value of 80 pg/ml; the coefficient of correlation with RIA was 0.822. As a result, this quick measurement of u‐E3 and u‐E2 by our HPLC system revealed to be useful not only for evaluating fetoplacental function but also for monitoring ovarian follicle maturation.
Methodological pitfalls in serum IgG2 level measurements by immunoenzymatic assays with monoclonal antibodies
Authors: Aucouturier Pierre , Lacombe Corinne , Preud'homme Jean‐Louis 
Published: March 2007

AbstractFour anti‐IgG2 monoclonal antibodies (Mabs) were evaluated for their reactivity with purified myeloma IgG2 of different light chain types and Gm allotypes in three distinct immunoenzymatic assays (ELISA). The reactivity of three Mabs with solid‐phase antigens was similar whereas an anti‐Fab antibody (clone HP 6114) predominantly bound IgG2k. In competitive and immunometric (sandwich type) assays, the binding of the two anti‐IgG2 Mabs (HP 6014 and HP 6114) reacting with epitopes located on the Fab fragment was strongly influenced by the light chain type of IgG2 and by other factors (probably including differences in the variable regions): the Mab HP 6114 reacted virtually only with IgG2k whereas the Mab HP 6014 displayed a much stronger affinity for IgG2λ than for IgG2k; for both anti‐Fab Mabs, important differences were found in their binding to individual IgG2 proteins. In addition, the Mab HP 6014 seemed to show a slightly better affinity for IgG2 bearing the Gm(23) allotype. These results urge much caution in IgG2 level measurement, especially with commerical kits, most of which use the Mab 6014 as the single anti‐IgG2 reagent.
Microparticle‐enhanced nephelometric immunoassay for human C‐reactive protein
Authors: Montagne P. , Laroche P. , Cuillière M. L. , Varcin P. , Pau B. , Duheille J. 
Published: March 2007

AbstractPolyfunctional hydrophilic microspheres of 125‐nm diameter can be produced by copolymerization of acrylic monomers. Purified c‐reactive protein (CRP) was covalently bound to these new micropheres, and the conjugate obtained was used as reagent in a microparticle‐enhanced nephelometric immunoassay for human CRP. This assay was based on the measure, with a specially designed nephelometer, of the light scattered by aggregates formed during the immunoagglutination of the conjugate with anti‐CRP antiserum. Sensitive inhibition of this agglutination by free CRP (6 ng/ml) allowed CRP quantitation in highly diluted serum samples (1/500‐1/2,000), excluding any interference or sample pretreatment. The CRP assay was easy to perform (no washing or phase separation), reliable (coefficients of variation ranged from 1.3% to 9.3% for within‐run and between‐run determinations), and accurate (mean percentage of recovery: 104%; correlation coefficients with accepted analytical methods >0.97) over a large range of concentrations. The inhibition mode excluded errors in the antigen excess zone and provided total security at high concentrations.
Establishment of a sensitive radioimmunoassay for serum thymic factor with a special reference to extraction procedures
Authors: Mukaida Naofumi , Morimoto Taeko , Takanashi Naoki , Kawai Tadashi 
Published: March 2007

AbstractA radioimmunoassay (RIA) for serum thymic factor (facteur thymique serique) was established. This assay could detect <0.5pg of serum thymic factor without the interference by other thymic hormones, including thymopoietin ii and thymosin α1. Both zinc‐free and zinc‐bound serum thymic factor showed a similar standard curve, indicating that this RIA could detect both forms of serum thymic factor in the same way. We observed that two procedures were required prior to the measurement in order to measure serum thymic factor content in plasmas; the first, the extraction procedure to remove substances interfering with the assay nonspecifically; the second, the storage of the sample with ethylenediamine tetraacetic acid dissodium salt to inhibit the proteases in the plasma to degrade serum thymic factor. By employing this assay system, we observed that cord blood plasmas contained significantly higher level of serum thymic factor than did adult blood plasmas.
Oxidative metabolism of polymorphonuclear leukocytes (PMN) in patients with IgA nephropathy
Authors: Chen Hung‐Chun , Tomino Yasuhiko , Yaguchi Yutaka , Fukui Mitsumine , Yokoyama Ken‐ichi , Watanabe Asako , Koide Hikaru 
Published: March 2007

AbstractThe production of hydrogen peroxide (H2O2) by neutrophilic polymorphonuclear leukocytes (PMN) after stimulation and the infiltration of PMN in glomeruli were determined in 20 patients with primary IgA nephropathy. The H2O2 production of PMN after the stimulation was measured with a spectrophotometer using horseradish peroxidase as substrate. The results were as follows: (1) when PMN were pretreated with cytochalasine B, H2O2 production after stimulation with heataggregated IgG (IgG) or serum‐treated zymosan (STZ) was significantly higher in patients with IgA nephropathy than in controls, and (2) there was an increased amount of PMN localized in glomeruli in patients with IgA nephropathy using immunofluorescence of monoclonal anti‐PMN antibody.It appeared that the increased renal infiltration of PMN which have a high potential for production of reactive oxygen species might induce the glomerular injuries in patients with IgA nephropathy.
A monoclonal antibody‐based immunoassay for detecting tetrodotoxin in biological samples
Authors: Raybould T. J. G. , Bignami G. S. , Inouye L. K. , Simpson Samantha B. , Byrnes Jilanne B , Grothaus P. G. , Vann D. C. 
Published: March 2007

AbstractSpleen cells from mice hyperimmunized with a keyhole limpet hemocyanin‐tetrodotoxin‐formaldehyde conjugate were fused with murine P3X63Ag8.653 myeloma cells. A single hybridoma clone was identified that secretes an IgG1,k monoclonal antibody (MAb), designated T20G10, against tetrodotoxin (TTX), with an estimated affinity of 1.2 × 108 L/M. Competitive inhibition enzyme immunoassays (CIEIAs) for detecting TTX were developed using this MAb. A direct CIEIA using alkaline phosphatase‐labeled MAb detected TTX with sensitivities at IC50 and IC20 of 6‐7 ng/ml and 2‐3 ng/ml, respectively. The accuracy of the direct CIEIA was comparable with the high‐performance liquid chromatography (HPLC) and the mouse bioassay systems, but the direct CIEIA exhibited greater sensitivity. The direct CIEIA was also more cost effective, as it required less sample preparation, a shorter assay time, and reduced investment in equipment than either of the other assay systems.
Comparison of the use of bulk to micro culture of cell preparations for lymphokine‐activated cytotoxicity assays
Authors: Anderson Loraine H. , McDonald Thomas L. , Thiele Geoffrey M. , Klassen Lynell W. 
Published: March 2007

AbstractDifferent assay systems have been used to quantitate lymphokine‐induced natural cytotoxic activity as a measure of immune status. This study compares the effects of inducing cytotoxicity in a bulk culture system, where effector cells are transferred to a micro culture well for assay, to a micro culture system where the effector cells are not transferred. The effector/target ratio for both the bulk and micro culture systems was calculated using the number of viable effector cells present at the time of target cell addition. After over‐night incubation with interleukin‐2 (IL‐2), the lytic activity of murine spleen cells to targets using a micro culture system was increased two‐fold over the bulk culture method. This increase was amplified further after 5 days of activation with IL‐2, in that the micro culture system resulted in a four‐fold increase in cytotoxic activity. The loss of some adherent cells in the bulk culture system did not explain the overall decrease in recovered cytotoxicity. The difference appeared to be related to cell loss during centrifugation. Therefore, the E/T ratios are different in the two systems if not corrected for the number of viable cells. © 1992 Wiley‐Liss, Inc.
Delayed hypersensitivity skin tests in prognosis of human immunodeficiency virus infection
Authors: Zeballos R. S. , Cavalcante N. , Freire C. A. R. , Hernandez H. J. , Longo I. M. , Peixinho Z. F. , Moura N. C. , Mendes N. F. 
Published: March 2007

AbstractDelayed hypersensitivity skin tests (DHST) with recall antigens were investigated as prognostic markers in five different approaches. In the first study, 42 acquired immunodeficiency syndrome (AIDS) patients (IVb, IVcl, IVd, and IVe; MMWR 35:334–339, 1986) 26 AIDS‐related complex (ARC) patients (IVa and IVc2), and 98 asymptomatic patients (II and III) were evaluated with candidin, tricophytin, PPD and streptokinase‐streptodornase. In the second study, 10 patients (II and III) were evaluated sequentially with the same antigens. In the third, 45 patients with at least two positive skin tests (“reactors”) were followed for one year and evaluated every 6 months with the same antigens. In the fourth, 16 “reactors” were followed and evaluated every 3 months with the same antigens. We measured the interval from the time at which patients first presented with only one or no positive DHST until the development of ARC or AIDS. In the last study, the correlation between absolute number of CD4+ lymphocytes and the number of DHST was studied in 151 patients. We found that the decrease in reactiveness to DHST correlated directly with the progression to AIDS, demonstrating the usefulness of this simple procedure as a valid prognostic marker. © 1992 Wiley‐Liss, Inc.
Effect of heat inactivation and sheep erythrocyte adsorption on the titer of anticardiolipin antibodies in primary antiphospholipid syndrome and healthy blood donor' sera
Authors: Nava Arnulfo , Bañales José L. , Reyes Pedro A. 
Published: March 2007

AbstractThe standard enzyme linked immunosorbent assay (ELISA) currently in use for detection of anticardiolipin antibodies (ACA) was used to evaluate the influence of heat inactivation and sheep erythrocyte adsorption on individual optical density (OD) of sera from healthy blood donors or patients with primary antiphospholipid syndrome. Each sample was tested after single or combined maneuvers as follows: adsorbed, adsorbed and inactivated, only inactivated, and compared to basal readings.A significant increase of ACA titers did occur after inactivation of normal sera, but adsorption had no effect. In contrast, neither inactivation nor adsorption changed ACA titer in primary antiphospholipid syndrome sera as a group, although in certain sera there were changes.This observation may suggest the presence in normal serum of a thermolabile factor which modulates ACA binding to its antigen and the reactivity of the anticardiolipin antibodies of the primary antiphospholipid syndrome with sheep erythrocyte membrane phospholipids. © 1992 Wiley‐Liss, Inc.
Acid pH‐induced changes in the immunoreactivity of specific antigen and antibody in circulating immune complexes from tuberculosis sera
Authors: Kumar Uday , Saxena R. K. 
Published: March 2007

AbstractThe effect of acid pH treatment of circulating immune complexes (CIC) derived from tuberculosis sera was studied on the relative titers of specific antibody (CIC Ab) and mycobacterial antigen (CIC Ag) in the complexes. While the specific antibody titers increased, the titers of CIC Ag declined as a result of acid pH treatment of CIC, both changes being highly significant statistically. Direct exposure of TB sera to pH 2.8 also resulted in significant enhancements in the titers of antibodies directed against Mycobacterium tuberculosis (M.tb.). Competition experiments indicated that the acid pH‐treated antibodies retained their antigen specificity. TB sera treated with pH 2.8 for 30 min and the neutralized back to pH 7.4 retained the enhanced antibody reactivity even after 7 days of storage at 4°C. Our results indicate that acid pH treatment induces higher antigen binding ability in anti‐M.tb. antibodies, present in free or complexed form in TB sera. These changes appear to be irreversible. Dissociation of CIC and analyzing the titers of released antibody and antigen components offered no advantage with respect to the immunodiagnosis of tuberculosis, as compared to the levels of undissociated CIC components or the serum antibody. © 1992 Wiley‐Liss, Inc.
Specificity of a Tumor Marker (CA54/61) and Its Individual Epitopes Recognized by Monoclonal Antibodies, MA54 and MA61, in Human Tumor Patients
Authors: Mochizuki Hiroshi , Nagoya Kimiko , Yamauchi Tadakazu , Takao Yoshimi , Masuko Hideyuki , Sato Hiroshi , Sasaki Hirosuke , Itoh Kotaro , Tsukazaki Katsumi , Udagawa Yasuhiro , Kobayashi Toshifumi , Nozawa Shiro 
Published: March 2007

AbstractThe specificity of a tumor marker (CA54/61) and its individual epitopes (CA54 and CA61) recognized by monoclonal antibodies (MA54 and MA61) and expressed by the same tumor marker were studied. Serum levels of CA54 and CA61 were compared with that of CA54/61. In lung adenocarcinoma and ovarian carcinoma, the positive rates of CA61 (42% and 68%) were higher than those of CA54 (32% and 32%) and similar to those of CA54/61 (45% and 74%). The serum levels of CA54 and CA61 showed a significant correlation (r=0.78), but 22% of tested sera were positive for CA54 and negative for CA61 or negative for CA54 and positive for CA61. It was demonstrated that the tumor specificity between CA54 and CA61 was not same and that the tumor specificity of CA54/61 was similar to that of CA61 rather than CA54. Moreover, the difference in the tumor specificity between CA54 and CA61 was considered to be reflected in the difference in their epitope structure. © 1992 Wiley‐Liss, Inc.
Sera from simian immunodeficiency virus‐infected rhesus macaques inhibit lymphocyte proliferation
Authors: Tomar Russell H. , Hinds Paul , Katz David , Schultz Kevin 
Published: March 2007

AbstractRhesus macaque monkeys infected with the simian immunodeficiency virus (SIV) develop a syndrome mimicking acquired immunodeficiency syndrome (AIDS) in humans. We had demonstrated previously that sera from individuals infected with human immunodeficiency virus (HIV) inhibit the proliferation of lymphocytes from healthy non‐infected subjects and that this phenomenon was associated with the development of clinical AIDS. Thus, we sought to determine whether sera from SIV‐infected monkeys would also inhibit lymphocytes from healthy humans and SIV‐negative rhesus monkeys.Sera from SIV‐infected monkeys were compared with sera from uninfected animals and cultured with cells from healthy human volunteers or SIV‐negative monkeys in the presence or absence of phytohemagglutinin (PHA). Cell proliferation was determined by measuring the incorporation of radiolabeled thymidine into cellular DNA.Sera from SIV‐infected monkeys suppressed the proliferation of human and non‐human primate lymphocytes. This activity appears to be similar to that described for sera from HIV‐1‐infected humans. Therefore, rhesus macaques infected with SIV provide a model for the study of serum inhibitory factors previously reported in AIDS patients. © 1992 Wiley‐Liss, Inc.
Sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for (anti‐human T‐cell leukemia virus type I) IgG in serum using recombinant gag p24(14–214) as antigen
Authors: Kohno Takeyuki , Hirota Kouichi , Sakoda Iwane , Yamasaki Motoo , Yokoo Yoshiharu , Ishikawa Eiji 
Published: March 2007

AbstractA sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for (anti‐human T‐cell leukemia virus type 1) IgG (anti‐HTLV‐l IgG) in serum using recombinant gag p24 (14–214) of HTLV‐l is described. The recombinant gag p24(14–214) is soluble in the absence of detergents and allows the use of enzymes other than horseradish peroxidase as a lbel in teh assays.The usefulness of recombinant gag p24(14–214) was examined with 305 sera characterized by other methods including gelatin particle agglutination, enzyme‐linked immunosorbent assay (ELISA) using HTLV‐l, and Western blotting.This Assay was more sensitive than other methods using HTLV‐l as antigen. The specificity could be tested by preincubation of test serum with excess of the recombinant protein. Most of negative and positive sera were discriminated. However, some results appeared to be false‐positive or false‐negative, and recombinant gag p24(14–214) was suggested to be useful, when used with other recombinant proteins and/or peptides, for improving the reliability of serodiagnosis by separately demonstrating antibodies against as many different epitopes of HTLV‐l as possible.Anti‐HTLV‐l 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‐recombinant gag p24(14–214) conjugate and recombinant gag p24(14–214)‐β‐D‐galactosidase conjugate. The complex formed consisting of the three components was trapped onto polystyrene balls coated with affinity‐purified (anti‐2, 4‐dinintrophenyl 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‐purifiedc (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. © 1992 Wiley‐Liss, Inc.