bluuebunny/crossref_metadata_2025_split huggingface dataset:
Assay for prostate specific antigen (PSA): Problems and possible solutions
Authors: Wu James T. 
Published: March 2007

AbstractThe absolute tissue specificity of prostate specific antigen (PSA) allows the use of PSA test not only for detecting recurrence or metastasis at an early stage after radical prostatectomy but also for screening prostate cancer if combined with digital rectal examination. There is also a need to improve the current PSA test to better differentiate between prostate cancer and benign prostate hyperplasia (BPH). Because of these clinical applications, a much greater demand was placed on PSA test for extra sensitivity, accuracy, and precision even within the normal PSA concentration range. However, the current commercial assay kits for PSA do not provide correct PSA values. Many factors contributing to the problem include the specificity of the anti‐PSA antibodies, the composition of the calibrator, the PSA values assigned to the calibrator, the PSA isoform used for anti‐PSA antibody preparation, the test design, and the composition of the diluent. Most problems were derived from the failure of realizing earlier that the majority of the PSA exists in serum not as free PSA but as complexes with protease inhibitors. Other problems, such as constantly changing composition of various forms of PSA in serum specimens, and different clearance rates for various forms of PSA make almost impossible to develop an ideal assay for PSA. Therefore, we suggest that test should be designed for measuring PSA‐ACT (PSA‐α1‐antichymotrypsin) complex only. Changing the focus from the measurement of total PSA of various forms to the PSA‐ACT complex alone may improve the differentiation between prostate cancer and BPH but may also simplify the selection of anti‐PSA antibodies and the preparation of calibrator for the assay. © 1994 Wiley‐Liss, Inc.
Use of micro samples of finger prick blood dried on filter paper for a quick and simple dipstick dot‐EIA for diagnosis of amebic liver abscess (ALA)
Authors: Sharma Manoj , Ghosh S. , Singal A. K. , Anand B. S. , Talwar G. P. 
Published: March 2007

AbstractFilter paper was used as a support to absorb micro samples of finger prick blood for detection of antibodies to Entamoeba histolytica (causative organism of amebiasis) by a rapid dipstick dot‐EIA technique; 8 μl of blood was sufficient to saturate discs (diameter 6 mm) of Whatman paper No. 3. Conditions of elution of blood from the discs were optimized and the best results were obtained when 0.4 ml of buffer was used for elution for 30 minutes at room temperature. The filter paper technique is extremely useful for field use and for diagnosis of amebic liver abscess on a large scale since it does not involve centrifugation of blood and use of sterile vials because blood dried on paper can be stored in polythene bags at room temperature for up to 3 months and for a week at 42°C without significant loss of antibody activity, thereby eliminating the need of refrigeration of samples for storage or transportation to a reference laboratory. Elution of blood from different discs of the same patient was found to be highly reproducible without appreciable loss of sensitivity or specificity. Twenty‐four confirmed ALA, 29 non‐ALA, and 25 apparently normal healthy controls, with no previous history of amebiasis, were tested. Sensitivity and specificity of the test was found to be 96% and 92%, respectively. © 1994 Wiley‐Liss, Inc.
Modulation of suppressor cell activities by cyclophosphamide in breast cancer patients
Authors: Kuroi Katsumasa , Sato Yukio , Yamaguchi Yoshiyuki , Toge Tetsuya 
Published: March 2007

AbstractWe investigated the immunomodulatory effect of cyclophosphamide on the induction of suppressor cell activities in peripheral blood lymphocytes. The sera from advanced breast cancer patients as well as concanavalin‐A (Con‐A) induced suppressor cell activities in lymphocytes from healthy volunteers. Pretreatment of these lymphocytes, including CD4+ T cells with 4‐hydroperoxycyclophosphamide (4‐HC), an active form of cyclophosphamide, abrogated the induction of suppressor cell activities by either cancer sera or Con‐A. A decrease in CD4+CD45RA+ suppressor/inducer T cells and reduction of Con‐A induced suppressor cell activities were observed in breast cancer patients with metastases that were treated with cyclophosphamide (CPM). These results suggest that cyclophosphamide may modulate the immune responses of breast cancer patients by interference with suppressor/inducer T cells. © 1994 Wiley‐Liss, Inc.
Anti‐HTLV‐I IgG in urine detected by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using a synthetic peptide, Cys‐env gp46(188‐224), as antigen
Authors: Hashida Seiichi , Hirota Kouichi , Kohno Takeyuki , Ishikawa Eiji 
Published: March 2007

AbstractAntibody IgG to human T‐cell leukemia virus type I (HTLV‐I) in urine was detected by a sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using a synthetic peptide, Cys‐env gp46(188–224), as antigen, the sensitivity and specificity of which were 100 and 98.5%, respectively, using serum samples. Anti‐HTLV‐I IgG in urine 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 (Escherichia coli) conjugate. The complex formed, consisting of the three components, was trapped onto polystyrene balls coated with affinity‐purified (anti‐2,4‐dinitrophenyl group) IgG, eluted with ϵN‐2,4‐dinitrophenyl‐L‐lysine and transferred to polystyrene balls coated with affinity‐purified (anti‐human IgG γ‐chain) IgG. Finally, bound β‐D‐galactosidase activity was assayed by fluorometry. Thirty‐one urine samples from seropositive subjects and 100 urine samples from seronegative subjects were tested. The sensitivity and specificity were 87 and 100%, respectively, with unconcentrated urine samples and 94 and 100%, respectively, with approximately 10‐fold concentrated urine samples. These results were superior to those by the conventional ELISA and gelatin particle agglutination test. © 1994 Wiley‐Liss, Inc.
Failure of in vitro T‐cell assays to predict clinical outcome after human kidney transplantation
Authors: Steinmann J. , Kaden J. , May G. , Schröder K. , Herwartz C. , Müller‐Ruchholtz W. 
Published: March 2007

AbstractAllotransplant rejection is a T‐cell‐dependent reaction. Functional in vitro T‐cell assays are being used widely for donorrecipient matching in bone marrow transplantation and have recently been used in some centres for transplant monitoring. In order to assess tolerance induction after clinical transplantation, we measured the T‐cell response of the host against donor spleen cells of 33 kidney transplant patients before and every 3 months after transplantation over a period of 18 months. The T‐cell reactivity before transplantation was not significantly different in any of the assays in rejecting and nonrejecting patients. In the classical mixed lymphocyte culture (MLC), a donor‐specific loss of reactivity was seen only in a patient with a CMV‐associated irreversible transplant rejection. One patient with chronic rejection acquired a very high MLC response against donor spleen cells and a high response against third‐party cells. Little or nonspecific changes were seen in the MLCs of all other patients. Using the method of limiting dilution analysis (LDA), we found a significant reduction of donor‐specific cytotoxic T‐cell precursors (CTL‐p) within the first 3 months after transplantation in most patients with high antidonor CTL‐p frequencies before transplantation. The reduction of donor‐specific CTL‐p was seen in patients with rejection episodes as well as in patients without. Thus we conclude, in contrast to others, that MLC and CTL‐p LDA have no predictive value on the outcome of clinical transplantation. © 1994 Wiley‐Liss, Inc.
Detection of glycosylated protein in glomeruli of STZ‐induced diabetic rats using the nitroblue tetrazolium (NBT) reaction
Authors: Saitoh Akemi , Tomino Yasuhiko , Kuramoto Takao , Fukui Mitsumine , Ohmuro Hiroyuki , Shirato Isao , Koide Hikaru , Itoh Kiichi 
Published: March 2007

AbstractDetection of glycosylated protein in renal tissues was determined in streptozotocin (STZ)‐induced diabetic rats using the nitroblue tetrazolium (NBT) reaction. The glycosylation of extra‐cellular matrix (ECM) components such as laminin and fibronectin was examined in vitro using the same method. Immunofluorescence staining of laminin or type IV collagen was also performed in renal tissues of STZ‐induced diabetic rats. There was no significant difference in the intensity of NBT in renal tissues between 4 week STZ‐induced diabetic rats and control rats of the same age. The intensity of NBT staining in the glomerular mesangial areas and capillary walls was marked in 12 week diabetic rats. The mean values of fructosamine measured by the NBT reaction in the glycosylated‐laminin and fibronectin were increased dose dependently. In immunofluorescence, laminin and type IV collagen were observed significantly in the glomerular mesangial areas and capillary walls of 12 week diabetic rats. However, there was no significant change in renal histopathology in 4 and 12 week diabetic rats. It appears that the non‐enzymatic glycosylation and expression of ECM components in glomeruli increased in the early stage of diabetic nephropathy prior to the appearance of marked histologic alterations. In conclusion, non‐enzymatic glycosylation of glomerular structural components may play an important role in the initiation of the early stage of renal injuries in diabetes. © 1994 Wiley‐Liss, Inc.
Measurement of free and total hydroxyproline by automated flow injection of serum or urine samples from maintenance hemodialysis patients with renal osteodystrophy
Authors: Uji Yoshinori , Karmen Arthur , Okabe Hiroaki , Hata Keishi , Miura Masakazu , Ozaki Kazuyuki , Minamizaki Mitsuo , Shibata Tetsushi , Inayama Seiichi 
Published: March 2007

AbstractAn automated measurement of total and free hydroxyproline in serum or urine is presented that uses flow injection analysis. After exclusion of nonspecific substances, hydroxyproline was oxidized by chloramine‐ T and L‐cysteine with Ehrlich's reagent.The linearity obtained was from 3.8μmole/ L to 1.22 mmole/L with good precision (CV <3%). Comparison of the proposed method with HPLC yielded r = 0.939 as the correlation coefficient.Reference intervals of free and total hydroxyproline are 1.4–9.7 μmole/L, 3.8–27.2 μmole/L for serum, and 10.0–72.5 μmole/L, 25.2–303.6 μmole/L for urine, respectively. Serum free and total hydroxyproline levels in renal osteodystrophy patients on maintenance hemodialysis (N = 71) were significantly higher than in controls (P<0.0001). This method is superior to the use of HPLC with regard to stability of the color reaction. The measurement of serum free and total hydroxyproline is a useful marker for therapeutic observation of renal osteodystrophy patients. © 1994 Wiley‐Liss, Inc.
Diagnosis and prediction of pediatric HIV‐1 infection and AIDS: Current status
Authors: Ugen Kenneth E. , Von Feldt Joan M. , Weiner David B. , Ziegner Ulrike H. M. 
Published: March 2007

AbstractThe increase in the incidence of HIV‐1 infection in women of child bearing age has resulted in a surge in the number of cases of pediatric AIDS. The World Health Organization (WHO) has estimated that the number of cases of pediatric AIDS worldwide will be at least 10 million by the year 2000. This alarming statistic underscores the need for accurate prediction and diagnosis of pediatric HIV‐1 infection which is of paramount importance for the initiation of effective therapeutic interventions. Since circulating maternal anti‐HV‐1 antibody persists in the baby for up to 21 months, early conventional serological diagnosis of infection is not possible. Other methods for diagnosis of HIV‐1 infection in a child less than 2 years of age have been utilized including the polymerase chain reaction (PCR), measurements of the HIV‐1 p24 core protein and anti‐HIV‐1 IgA, as well in vitro measurements of antibody producing cells. In addition, the ability to predict HIV‐1 infection in the child based upon maternal humoral immune responses to the envelope glycoprotein has also been suggested. This review summarizes the recent serological, biological and molecular methodologies used to predict and diagnose pediatric HIV‐1 infection and AIDS. © 1994 Wiley‐Liss, Inc.
Use of microplates and fluororeader for ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) of anti‐HTLV‐I IgG
Authors: Ishikawa Setsuko , Hashida Seiichi , Kohno Takeyuki , Ishikawa Eiji 
Published: March 2007

AbstractPreviously, an ultrasensitive solid phase enzyme immunoassay (immune complex transfer enzyme immunoassay) was described to detect low levels of anti‐HTLV‐I IgG in serum below those detectable by conventional methods. In this method, polystyrene balls as solid phase were transferred from test tube to test tube with tweezers. This was not only tedious but also causative of false‐positivity by carryover, unless tips of the tweezers were washed carefully after each transfer of polystyrene balls. Bound enzyme activities for many samples were measured one by one by fluorometry using a spectrofluorophotometer. As a result, the assay of many samples was difficult. In the present study, microplates and a fluororeaderwere used in place of test tubes and a spectrofluorophotometer. Polystyrene balls were transferred quickly and easily from well to well by placing a microplate upside down on that containing polystyrene balls, and turning the two plates together upside down. Tweezers were not used for transfer of polystyrene balls, minimizing the possibility of false‐positivity. Fluorescence intensities of bound enzyme activities for 96 samples were measured within a minute by using a fluororeader. Thus, it became easy to test many samples, although the sensitivity was lowered to some extent. © 1994 Wiley‐Liss, Inc.
Measurement of serum Lp(a) by COBAS MIRA using a latex immunoturbidimetric assay kit
Authors: Fujita Seiichi , Sano Takahiro , Katayama Yoshiaki 
Published: March 2007

AbstractWe had an opportunity to test a reagent kit for serum lipoprotein (a) [Lp(a)] assay, which was based on the latex immunoturbidimetric assay method, and applied it to COBAS MIRA (Roche). Reproducibility of the assay procedure was 0.92–4.0% CV of three samples which contained 12.58–60.4 mg/dl of Lp(a). Minimum detectable concentration was 1.5–2.0 mg/dl. It was confirmed that interference of co‐existing substances, i.e. hemoglobin, bilirubin, and intralipid, was negligible. No cross reactivity was seen with plasminogen. Correlation with a ELISA method was excellent. Frequency distribution of Lp(a) in healthy Japanese was similar to that found in white population. © 1994 Wiley‐Liss, Inc.
Magnetophoresis: I. detection of magnetically labeled cells
Authors: Winoto‐Morbach S. , Tchikov V. , Müller‐Ruchholtz W. 
Published: March 2007

AbstractA new method for the detection of magnetically labeled cells in suspension using an optical microscope is presented. The movement of individual cells in a nonuniform magnetic field (magnetophoresis) can be studied and the direction and velocity of their movement can be determined. Cell‐associated magnetic particles (e.g., attached magnetic immunomicrospheres) influence these parameters. We developed a magnetophoresis chamber for measuring the movement of single cells in a high‐gradient magnetic field. Paramagnetic erythrocytes were used to monitor the level of magnetic force in the chamber. Bw6 antigen‐positive REH cells were used as a biological model. Magnetic microspheres coupled with anti‐Bw6 antibody were used as magnetic labels. The movement of magnetically labeled REH cells was investigated versus the intensity of magnetic field. The magnetization of microspheres was experimentally determined by the Faraday balance method. Experimental and theoretical results substantiate the feasibility of detecting a single cell associated with a single magnetic microsphere. Thus, magnetophoresis provides a sensitive biometrical system for magnetically labeled entities. © 1994 Wiley‐Liss, Inc.
Soluble MHC class I molecules in human body fluids
Authors: Zavazava N. , Wobst B. , Ferstl R. , Müller‐Ruchholtz W. 
Published: March 2007

AbstractApart from their well‐known function of antigen presentation in the form of peptides, major histocompatibility antigens (MHC) have been found to be unique markers of individual body odors in murine experimental models. In the current study we examined the nature and expression of soluble human MHC class I molecules in body fluids. Biochemical analysis of affinity purified serum class I molecules revealed a variety of molecules within the molecular weight region of 45 to 21 kD. SDS‐Western blotting of HLA derived from hepatocytes and spleen cells suggested that much of the small molecular mass fraction of sHLA (<45 kD) found in serum is derived from the liver. sHLA were detected and quantitated in serum, plasma, urine, cerebrospinal fluid, and sweat. No sHLA were detectable in cerebrospinal fluid (n = 20). In addition, sHLA was measured in serum of women during the menstrual cycle. A significant increase in sHLA was observed during the first half of the cycle, suggesting that sexual hormones may increase sHLA concentration. The observed increase was most prominent in women that were HLA‐A24. © 1994 Wiley‐Liss, Inc.
Cytokine production in whole blood cell cultures of patients with crohn's disease and ulcerative colitis
Authors: Elsässer‐Beile U. , von Kleist S. , Gerlach S. , Gallati H. , Mönting J. Schulte 
Published: March 2007

AbstractLevels of the cytokines interleukin‐1‐α, ‐1‐β, and ‐2 (IL‐1‐α, IL‐1‐β, IL‐2), tumor necrosis factor‐α (TNF‐α), and interferon‐γ (IFN‐γ) were measured in the mitogen‐stimulated whole blood cell cultures from 96 patients with Crohn's disease (48 untreated, 12 treated with sulfasalazine, 36 treated with corticosteroids), 74 patients with ulcerative colitis (21 untreated, 25 treated with sulfasalazine, 28 steroid treated), and 360 healthy controls. The cytokines were measured 4 days after induction by a sensitive immunoenzyme assay.In the blood cell cultures of the untreated and sulfasalazine treated patients with Crohn's disease and ulcerative colitis higher levels of TNF‐α, IL‐1‐α and IL‐1‐β were found whereas IL‐2 production was decreased and IFN‐γ‐production was not significantly different as compared to the controls. Leukocytes of the corticosteroid‐treated patients with both diagnoses showed a lower production of all measured cytokines compared to the untreated patients.The same results were obtained, when the somewhat different counts of mononuclear cells in the peripheral blood of the patients and controls were taken into account. The elevated production of proinflammatory cytokines in the blood cell cultures suggests a systemic immune activation in patients with inflammatory bowel disease. © 1994 Wiley‐Liss, Inc.
Solid‐Phase polymerase chain reaction
Authors: Kohsaka Hitoshi , Carson Dennis A. 
Published: March 2007

AbstractThe polymerase chain reaction (PCR) has facilitated the diagnosis of infectious diseases and genetic disorders, because of its ability to amplify minute amounts of nucleic acids. To distinguish genes of interest from nonspecifically amplified DNA, PCR products commonly are fractionated by electrophoresis, transferred to membranes, and then probed with a labeled internal sequence‐specific oligonucleotide. Alternatively, the PCR products have been labeled directly, and hybridized to immobilized oligonucleotide probes. These methods require the tedious physical transfer of the PCR products. In order to amplify and immobilize genes simultaneously, we have developed a simple solid‐phase PCR method. The technique enabled us to detect the HIV envelope gene readily without any transfer of amplified DNA. © 1994 Wiley‐Liss, Inc.
Racial differences in serum immunoglobulin levels: Relationship to cigarette smoking, t‐cell subsets, and soluble interleukin‐2 receptors
Authors: Tollerud David J. , Weiss Scott T. , Brown Linda Morris , Blattner William A. , Maloney Elizabeth M. , Kurman Carol C. , Nelson David L. , Hoover Robert N. 
Published: March 2007

AbstractTo investigate the influence of race, cigarette smoking, and immunologic parameters on serum immunoglobulins, we analyzed serum IgG, IgA, and IgM levels in 455 healthy adults. The study population ranged in age from 20 to 69 years, including 282 whites and 173 blacks, 181 never‐smokers, 93 ex‐smokers, and 181 current smokers. Race and smoking were independently associated with alterations in serum IgG levels. Blacks had significantly higher IgG levels than whites (1,587 vs. 1,209 mg/dl; P<0.001), and never smokers had significantly higher levels than current smokers (1,426 vs. 1,287 vs. mg/dl; P<0.001). IgA and IgM levels were unrelated to race or smoking. Serum IgG was also found to be directly related to the proportion of HLA‐DR+ cells and the level of soluble interleukin‐2 receptors (sIL‐2R) and inversely related to the proportion of CD4+ cells. Investigation of this racial heterogeneity may provide insights into the pathogenesis of immunologic diseases that exhibit unexplained racial variation.©1995 wiley‐Liss, inc.
Magnetophoresis: II. quantification of iron and hemoglobin content at the single erythrocyte level
Authors: Winoto‐Morbach S. , Müller‐Ruchholtz W. , Tchikov V. 
Published: March 2007

AbstractThe iron and hemoglobin content of individual erythrocytes was determined using a method based on parallel velocity measurements during magnetophoresis and gravitational sedimentation of individual erythrocytes in suspension. In previous publications we have suggested employing cell magnetophoresis, a biophysical phenomenon characterized by cell movement in a fluid under magnetic field influence, for cytometry. The paramagnetic ferric iron in methemoglobin is used as a magnetic label. The iron content is estimated from the magnetophoresis velocity, and hemoglobin content from the gravitational sedimentation velocity of erythrocytes. Blood samples are also analyzed in a Coulter counter to determine their mean corpuscular hemoglobin. The time course of the reaction of methemoglobin reduction is quantified at the single erythrocyte level. The methemoglobin content in individual erythrocytes is determined following the oxidation reaction. Erythrocytes from patients with normo‐, hypo‐, or hyperchromic anemia exhibit magnetophoresis and gravitational sedimentation velocities that correlate closely with mean corpuscular hemoglobin. We propose the utilization of magnetophoretic cytometry for detailed diagnostic studies at the single erythrocyte level. Furthermore, the magnetophoresis velocity to gravitational sedimentation velocity ratio is proposed as a standard value for comparative study of magnetically labeled cells in future investigations, as it was found to be constant and independent of hemoglobin content.©1995 wiley‐Liss, inc.