bluuebunny/crossref_metadata_2025 huggingface dataset:
Peripheral blood mononuclear cells from aids patients release but do not respond to antigen‐stimulated NK‐activating lymphokines
Authors: Lapham Cheryl K. , Sacks Leonard V. , Tomar Russell H.
Published: March 2007

We have previously shown that streptococcal product (SP) extracted from the culture supernatants of Streptococcus pyogenes ATCC 19615 augments NK activity, partly by inducing the release of soluble factors from mononuclear cells. In this study we use SP as an NK stimulant to investigate the mechanism of NK depression in AIDS.

We demonstrate that the NK activity of AIDS patients is lower than normal controls and can be significantly enhanced with SP, although not to control levels. The decreased cytotoxicity in AIDS is not due to a depletion of NK cells, nor to the depletion of lymphokines from CD4 cells that are directly involved in NK cell activation. PBMCs from patients with AIDS respond to SP, producing normal levels of NK‐en‐hancing substances in their supernatants. However, upon examining the ability of SP‐induced supernatants from control cells to augment the NK activity of cells from AIDS patients, we find significantly less activity in AIDS cells than in control cells, suggesting an intrinsic effector dysfunction of NK cells in AIDS.
Evaluation of enhanced luminescence immunoenzymometric assays (lia) for ferritin and free T4
Authors: Armbruster David A. , Jirinzu David C. , Williams John V.
Published: March 2007

The Amerlite enhanced luminescence immunoenzymometric assays (LIA) for ferritin and free T4 (FT4) have been evaluated. The LIA assay system employs a two‐site binding approach in which one antibody is coated on the interior of plastic microtiter wells and another antibody, or antigen, is tagged with horseradish peroxidase (HRPO). Signal, proportional to the amount of patient analyte, is produced when the HRPO catalyzes the oxidation of luminol resulting in light production. The light signal is increased in magnitude by the incorporation of an enhancing agent which also greatly prolongs the lifetime of the signal. Light production is monitored by an automated luminometer which uses a stored calibration curve to calculate patient results.

FT4 within‐run coefficients of variation (CVs) ranged from 2.7% to‐ 6.6%; day‐to day CVs from 6% to 15.5% for three levels of control. Sensitivity is 0.03 ng/dl. T3 concentrations up to 8 ng/ml produced no cross‐reactivity. Dilutional linearity was exhibited. Lipemia and icterus produced positive interferences. Linear regression analysis of Amerlite and radioimmunoassay (RIA) FT4 values yielded the following values: Amerlite = 0.63 RIA ‐ 0.2, r = .87, n = 79.

Ferritin within‐run CVs ranged from 4.9% to 10.5%; day‐to‐day CVs from 5.8‐12.5% for three levels of control. Sensitivity is less than 1 ng/ml. Recoveries ranged from 93% to 106% (av. = 99.4%). Dilutional linearity was exhibited. Lipemia caused a positive interference; hemoglobin caused a negative interference. Linear regression analysis of Amerlite and RIA ferritin values yielded the following values: Amerlite = 1.0 RIA + 11.1, r = .96, n = 55.
Functional and metabolic heterogeneity among normal neutrophil subpopulations
Authors: Smith Duane M. , Hanes Beverly , Johnson June A. , Turner Robert A.
Published: March 2007

In this study, normal human peripheral blood neutrophils were separated into sub‐populations by centrifugation on discontinuous Percoll density gradients and further characterized according to various functional and metabolic capabilities. Chemotaxis was measured in response to optimal concentrations (20 nM) of the chemotactic peptide N‐formyl‐methionyl‐leucyl‐phenylalanine (fMLP). According to a population ratio analysis, the majority of cells that contain the highest proportion of fast migrating cells are isolated between the densities of 1.093 and 1.096 gm/ml. Arachidonic acid (AA) metabolism was also determined in neutrophil subpopulations. On initial density gradients, cells isolated from 1.084–1.087 gm/ml and incubated in the presence of exogenous [3H]AA produce 50% of the total [3H]LTB4 produced in response to ionophore A23187 stimulation; whereas after prelabeling with [3H]AA for 2 hr at 37°C the cells between 1.087 and 1.093 gm/ml are the most active at both releasing [3H]AA from the cellular phospholipids and synthesizing [3H]LTB4 (dpm/cell). These studies demonstrate the significant functional and metabolic heterogeneity that exists among normal neutrophil subpopulations.
Suggested guidelines for the use of emergency tests in clinical biochemistry
Authors: Bhatnagar D. , Weinkove C. , McMurray J. R.
Published: March 2007

We have discussed the need for guidelines as a means of reducing out‐of‐hours investigations in clinical biochemistry, and suggest protocols that can be modified to suit local needs.
Serum level of cryptic tumor antigens in breast cancer patients as determined by two monoclonal antibodies (m85/f36) and its comparison with ca 15‐3
Authors: Ming T. , Constantine Rodric , Nemoto Takuma
Published: March 2007

An enzyme immunoassay system that measures cryptic epitopes on breast cancer mucinlike antigens (BCM‐EIA) was evaluated in a double‐blind manner in sera from 58 normal blood donors, 36 sex‐ and age‐matched controls, 36 patients with benign breast diseases, and 47 patients with breast cancer. Two murine monoclonal antibodies, M85 (IgM) as the solid‐phase and F36122 (lgG3) as the probe, were used in the configuration of the assay kit. The assay additionally utilized neuraminidase to remove terminal sialic acid from carbohydrate side‐chains to expose cryptic epitopes that were masked in serum specimens. BCM‐EIA monoclonal assay from the normal healthy blood donors resulted in 17.34 ± 7.04 units/ml (mean ± 1 S.D.) with an upper normal cutoff of 31.4 units/ml. The distributions of serum BCM in the sex‐ and age‐matched controls (17.77 ± 11.17) and benign breast diseases (14.34 2 11.46) were similar to that of normal blood donors. A mean value of 66.04 units/ml and 27.74 units/ml was obtained from breast cancer patients with active disease and without evidence of disease, respectively, a level much greater than those of normals, controls, and benign breast diseases. Simultaneous analysis of CA 15‐3, a putative breast tumor marker, in the normal donors and breast cancer patients revealed correlation regression of (CA 15‐3) = 0.876 (BCM) + 1.972, r = 0.856; and (CA 15‐3) = 0.66 (BCM) + 16.094, r = 0.730, respectively. These data showed that there is a statistically significant correlation of CA 15‐3 and BCM in normal blood donors' specimens. However, such a strong and positive correlation canot be unequivocally established in breast cancer patients' specimens. In conclusion, the double‐blind evaluation of breast cancer patients revealed the feasibility and clinical potential of the assay. The favorable comparison of BCM with CA 15‐3 was encouraging. In addition, an excellent repeatability was obtained by 24 different paired and blind specimens supporting the technical reliability of the assay system. These initial results demonstrated that BCM is a potential serum marker parameter for breast cancer.
Quantitation of hiv‐1 rna in blood cells of arc and aids patients
Authors: Solomon Robert , Thompson James , Gillespie David
Published: March 2007

Peripheral blood mononuclear cells from HIV‐1‐infected persons were mixed with 5 M guanidine thiocyanate, and HIV‐1‐specific probes were hybridized with target RNA directly in the lysate. No RNA purification was needed. Hybrids were purified by repeated capture on superparamagnetic beads coated with oligo(dT) in a device‐assisted format, a procedure termed “reversible target capture.” Blood mononuclear cells from 70 ARC and AIDS patients were examined and found to have an average of 1.8 × 105 molecules of HIV‐1 RNA per 2 × 106 cells.
Prolactin as a protective factor in stress‐induced biological changes
Authors: Drago Filippo , D'Agata Velia , Iacona Teresa , Spadaro Francesco , Grassi Michele , Valerio Carmela , Astuto Carmelo , Lauria Nunzio , Raffaele Rocco , Vitetta Matteo
Published: March 2007

The adenohypophyseal hormone prolactin (PRL) is released during stress of physical and psychological nature. In animals, this hormone facilitates adaptive behavior, induces analgesia, and enhances grooming behavior. It also reduces corticosterone secretion and the incidence of gastric ulcers induced by physical stress. It is possible that PRL plays a protective role against stress‐induced biological modifications in animals.
Evaluation of the ELISA test for detection of entamoeba histolytica in feces
Authors: Merino Elizabeth , Glender Walter , del Muro Ruben , Ortiz‐Ortiz Librado
Published: March 2007

The clinical utility of an ELISA test with monoclonal antibodies to detect antigen of Entamoeba histolytica in feces was evaluated in 150 patients with gastrointestinal symptoms. Each subject was examined by rectosigmoidoscopy with rectal smear and/or a triple stool search for ova‐bacteria‐parasite (OBP); in addition, one stool sample was collected for the ELISA test. All the tests were independent and double blind. E. histolytica was detected by OBP and/or rectosigmoidoscopy in 66 patients; 61 patients had other parasites; and in 23, no parasites were identified. Of all patients, 116 were positive for the ELISA test. Of these, E. histolytica was identified in 52. In 47, other parasites were identified and in 17, no parasites were found. The ELISA test with a monoclonal antibody against E. histolytica antigen showed higher sensitivity than the standard diagnostic methods: the ability to detect the presence of E. hisrolytica antigen regardless of the destruction of the parasite or of the error due to misidentification of the parasite resulting from faulty preparation of the samples.
An enzyme‐linked immunosorbent assay for the detection of antitetanus toxoid antibody using aluminum‐absorbed coating antigen
Authors: Thiele G. M. , Rogers J. , Collins M. , Smith D. , McDonald T. L. , Yasuda N.
Published: March 2007

An enzyme‐linked immunosorbent assay (ELISA) for the detection of antitetanus antibodies was developed using aluminum‐absorbed tetanus toxoid as the coating antigen. The assay was tested by measuring antitetanus antibody levels in serum obtained from subjects before and after immunization with the aluminum‐absorbed tetanus suspension. The specificity of the antibodies for tetanus antigen was tested both before and after antitetanus activity was removed with tetanus antigen–coated beads. Also, the activities of the sera were compared with those of a commercially available test. Our results indicated that the aluminum‐absorbed tetanus suspension can be coated onto standard polystyrene ELISA plates and used to measure antibody titers to tetanus toxoid.
Prostate‐specific antigen and prostate gland volume: Correlation and clinical application
Authors: Babaian R. Joseph , Evans Robert B. , Fritsche Herbert A.
Published: March 2007

We studied 103 patients seen in our Prostate Cancer Detection Clinic to determine whether a correlation exists between serum prostate‐specific antigen (PSA) values and ultrasound‐calculated prostate gland volume. Seventy men (68%) had a PSA value ≤ ng/ml (our upper limit of normal). The men were subclassified by prostate gland volume at arbitrary break points. Twenty‐five men (24%) had a prostate gland volume ≤ 25 cm3; in 96%, the PSA value was ≤ 4 mg/ml. Further analysis revealed that the percentage of men with a normal serum PSA value decreased as the prostate gland volume increased; 65.6% of the group with a gland volume between 25 and 50 cm3 (40 of 61) and 35.5% of the group whose prostate volume exceeded 50 cm3 (6 of 17) had PSA values ≤4 ng/ml. Four men had PSA values ≥20 ng/ml; all had prostate cancer. Cancer was diagnosed in four additional patients, three with PSA values between 5 and 10 ng/ml and one with a PSA value <4 ng/ml. There appears to be a direct relationship between prostate gland volume and PSA value, as well as a cancer value threshold. The clinical implications of these findings are discussed.
Mitogenic effect of interleukin‐2 on unstimulated human T cells: An editorial review
Authors: Mookerjee Basab K. , Pauly John L.
Published: March 2007

The most salient feature of the lymphokine interleukin‐2 (IL‐2), a hormone‐like protein, is its ability to sustain the proliferation of immunocompetent T cells. Results of initial studies characterizing IL‐2 in vitro led investigators to conclude that IL‐2 had no known effects on lymphocytes that had not been previously activated by exposure to a mitogen or antigen. Several groups postulated that T cell growth required two signals. The first signal, delivered by a mitogen or antigen, induced T cell activation. Resting T cells, which were thought to lack the membrane receptor for interleukin‐2 (IL‐2R), progressed from the G0‐G1 phase to the S phase, during which time they converted from IL‐2R− to IL‐2R+ cells. Thereafter, the second signal, served by IL‐2, induced T cell proliferation of the tL‐2R+ cells. Recently, a number of investigators have demonstrated that highly purified preparations of both natural and recombinant IL‐2 induced high levels of T cell proliferation in the absence of any known mitogens or antigens. Presented herein is a review of these studies and an overview of the hypotheses of the mechanisms whereby IL‐2 alone induces T cell activation and proliferation.
Clinical assessment of the hitachi 736‐30 chemistry analyzer
Authors: Schotters Susan B. , McBride James H. , Rodgerson Denis O. , McGinley Margaret H. , Pisa Marilyn
Published: March 2007

We assessed the Hitachi 736‐30 as a possible replacement for the SMAC I and as a laboratory cost‐saving measure. For 24 analytes, both intra‐ and interassay precisions were acceptable; they also had good measuring ranges. Essentially no interference from lipemia was observed, while minimal interference from bilirubin was demonstrated. Hemoglobin interfered in the measurement of 12 of the analytes. Correlation with the SMAC I, Demand, Astra‐8, ACA, and Varian Atomic Absorption Spectrophotometer was found to be acceptable, except for chloride which showed poor correlation with SMAC I and Astra‐8 (Hitachi = 0.888 [SMAC] + 11.102, r = 0.9652; Hitachi = 0.885 [Astra] + 10.264, r = 0.9136). The Hitachi 736‐30 offers reagent and method flexibility, high volume capability, and “walk‐away” operation.
Assay of follitropin and lutropin by fluorescence enzyme immunoassay
Authors: Armbruster David A. , Armbruster F. , Hawes Leah C.
Published: March 2007

Recently developed fluorescent enzyme immunoassays (FEls) for follitropin (FSH) and lutropin (LH) designed for the Stratus analyzer were compared with a typical dual FSH‐LH radioimmunoassay (RIA) procedure. The FEls use monoclonal antibodies in a sandwich reaction scheme similar to that of immunoradiometric procedures but with filter paper tabs rather than polystyrene beads and alkaline phosphatase as a tag instead of a radioisotope. Precision of the FEI was comparable with that of the RIA tests with CVs typically under 10%. The detection limit of the FSH test is 0.4 IU/L and that of the LH test is 0.6 IUIL. Measurable carryover exists but is clinically inconsequential. Neither assay exhibits total specificity, but the degree of cross‐reactivity is unlikely to be problematic in most situations. Recovery with the FSH assay was 103%, with the LH assay, it averaged 108%. Moderate amounts of bilirubin and triglycerides and gross hemolysis caused no significant interferences. Method comparison studies yielded the following: FEI for FSH, 1.16 RIA + 3.1, r = 0.97, n = 114; FEI for LH, 0.77 RIA ‐ 0.23, r = 0.96, n = 112. Difference analysis of the method comparison data revealed relatively poor agreement between the FEI tests and the dual RIA procedure. With the adoption of method‐specific normal reference ranges, the FEI tests offer attractive alternatives to radioisotopic FSH and LH assays.
Simplified solid‐phase immunobead assay for detection of ciguatoxin and related polyethers
Authors: Hokama Yoshitsugi
Published: March 2007

This study examined the development of a highly simplified solid‐phase colored latex immunobead assay for the detection of ciguatoxin and related polyethers. This procedure was compared with the stick enzyme immunoassay previously reported. Chi‐square analysis of two separate experiments on 153 and 283 fish of various species gave χ2 values of p < 0.001 and p < 0.005, respectively Agreement between the two procedures with 26 fish implicated in ciguatera poisoning was 100%. A preliminary assessment in the field showed encouraging results. The procedure appears to be simple and applicable to field use. Furthermore, this procedure should be applicable to other antibody‐antigen detections, especially low dalton determinations.
Detection of a case of pseudolymphocytosis due to cryoglobulins
Authors: Banfi G. , Grazioli V. , Dolci A. , Murone M. , Bonini P. A.
Published: March 2007

A new type of interference of cryoglobulins on hemocytometric tests is described. The precipitation of temperature‐dependent proteins produced a pseudolymphocytosis on a three‐part differential leukocyte count of Coulter S‐Plus VI, whereas unaffected results, identical to the microscopical count, were obtained using the cytometer Coulter VCS. The laboratory detection of cryoglobulin interference on hematological data is very important in patients with underlying diseases, where the accuracy of absolute and differential leukocyte counts is critical for follow‐up. Histograms from the Coulter S‐Plus VI can help detect these cases.
Myeloid leukemia differentiation by phorbol ester and retinoic acid: A practical approach
Authors: Marques‐Silva V. M. , De Souza M. H. O. , Teieira M. C. L. , Arcuri R. A. , Rumjanek V. M.
Published: March 2007

The effects of TPA (12‐0‐tetradecanoyl‐phorbol‐13‐acetate) and RA (retinoic acid) were investigated on the cell lines HL60 (acute promyelocytic leukemia) and K562 (erythro‐leukemia) and on cells from patients with several kinds of leukemia. There were 14 cases of acute lymphocytic leukemia (ALL), 2 cases of chronic lymphocytic leukemia (CLL), 23 cases of acute myeloid leukemia (M1‐M7), 5 cases of chronic myelocytic leukemia in blast crisis (CML‐BC) and 2 mixed leukemias. In almost all of the cases examined, after TPA exposure cells from patients with proven myeloid leukemia became adherent to the substrate, while lymphoid leukemia cells remained in suspension, allowing the differentiation of lymphoid from myeloid blasts. The only exception was in one case of CLL, which had cells that became adherent with long filamental projections. In addition, increased phagocytosis following TPA exposure permitted characterization of M7 as this was the only myeloid leukemia negative for phagocytosis. Further discrimination between the subtypes of myeloid leukemia could be based on the increased lysozyme production seen after TPA in M4 and M5. Es‐terase positivity allowed the discrimination of MI cells, which were negative before and after TPA treatment. In agreement with the results of other authors, TPA and RA led to independent ways of differentiation, granulocytic‐like lineage and monocytic‐like cells being favored by RA and TPA, respectively. The capacity of the same cell to differentiate into more than one lineage, depending on whether RA or TPA was used, was only seen in the present study with M3 cells. Our results show the usefulness of using differentiation inducers and functional tests together with morphology to elucidate the nature of leukemic cells.
Antibodies to viral antigens, xenoantigens, and autoantigens in alzheimer's disease
Authors: Ounanian A. , Seigneurin J.‐M. , Guilbert B. , Avrameas S. , Renverez J.‐C.
Published: March 2007

Sera from 19 patients with Alzheimer's disease (AD) and 21 control subjects were studied by immunofluorescence and enzyme immunnoassay for antibody activity against various viruses and 12 self‐ and non‐self‐antigens. Total IgG mean level was significantly higher in the AD group; the IgG level was above 15 g/L in 52.8% of AD patients versus 14.3% of control subjects. Antiviral antibody titers showed no significant differences except for antibodies to herpes simplex virus‐1, which were increased in control group. In contrast, autoantibodies were more frequently found in AD patients, and the prevalence of antibodies to spectrin, peroxidase, and thyroglobulin was significantly increased. Thus, in our series, autoimmune but not antiviral responses were heightened in at least 42% of AD patients (versus 9% of the control group) suggesting the existence of two sub‐populations in the AD group.
A high‐affinity human monoclonal igm antibody reacting with multiple strains of mycoplasma hominis
Authors: Möller Susanna A. , Borrebaeck Carl A. K. , Birkelund Svend
Published: March 2007

Human mknoclonal antibodies were produced against Mycoplasma hominis by in vitro immunization of peripheral blood lymphocytes from a healthy seropositive donor using low amounts of antigen (5 ng/ml). The immune B lymphocytes were subsequently immortalized by Epstein‐Barr virus transformation followed by somatic cell fusion. Using this procedure, several specific anti‐M. hominis monoclonal antibodies of μ isotype were obtained exhibiting apparent affinity constants as high as 3 × 109 M−1. The productivity of a number of different hybridomas was determined to be in the range 7‐22 μg × (24 hr × 106 cells)−1. The specificity of one antibody, 129.1, was further characterized by enzyme‐linked immunosorbent assay against several different species of Mycoplasma and by Western blot analysis. The antibody specifically bound to a protein of M, 100,000 present in 10 of 12 different strains of M. hominis.
Measurement of serum unconjugated estriol and estradiol by high‐performance liquid chromatography
Authors: Kondo Zenjiro , Makino Tsunehisa , Lizuka Rihachi
Published: March 2007

Measurement of Estriol (E3) and estradiol (E2) within 22 min by high‐performance liquid chromatography (HPLC) was achieved in this study, and the values were compared with those of a radioimmunoassay (RIA). A totally computerized HPLC method was developed for measuring unconjugated E3(u‐E3) and E2 (u‐E2) in the sera of pregnant women. The serum samples were injected directly into the apparatus and transferred to a pre‐treatment column where the estrogens were absorbed while hydrophilic components such as proteins and carbohydrates were excluded. The estrogens were then passed through another separation column containing a new type of polymer gel. The mobile phase consisted of an acetonitrile‐water mixture, and separation was achieved by means of a reversed‐phase procedure. The eluate was monitored for fluorescence. All procedures were monitored and controlled with a built‐in microcomputer. Serum samples from 97 normal pregnant women at 20–41 wk gestation were simultaneously assayed by HPLC and RIA. The correlations obtained by HPLC and RIA were as follows: u‐E3 y = 0.905x–0.385, with a coefficient of correlation of r = 0.912; for u‐E2, y = 0.964x + 5.024, with a coefficient of correlation of r = 0.841 (y, RIA value; x, HPLC value). The quick measurement of u‐E3 and u‐E2 by HPLC can be a useful method for evaluating fetoplacental function.
Interleukin‐4 induces both lgg4 and ige secretion by peripheral blood b cells
Authors: Nüsslein Hubert G. , Spiegelberg Hans L.
Published: March 2007

Peripheral blood mononuclear cells (PBMCs) from healthy nonallergic donors were cultured with recombinant interleukin‐4 (rIL‐4), and the Ig of different isotypes was quantitated in the culture supernatants by radioimmunoassays. Recombinant IL‐4 induced IgG4 and IgE secretion in a dose‐dependent manner, whereas it had no consistent effect on the secretion of the other isotypes. In the absence of rIL‐4, B cells in the PBMC preparations secreted <1 ng IgE/ml and a mean of 5 ng IgG4/ml. In the presence of the optimal dose of 100 U rIL‐4/ml, PBMCs from five donors secreted a mean ± SEM of 37 2 8ng IgE/ml and 66 ± 25 ng IgG4/ml. In kinetic studies, no IgG4 or IgE secretion was detected during the first 5 days of culture, and approximately 50% of the IgG4 and IgE secreted by day 15 was detected in supernatants on day 7. Cycloheximide, actinomycin‐D, and mytomycin‐C completely inhibited the rIL‐4‐induced IgG4 and IgE secretion, indicating that de novo protein, RNA, and DNA synthesis was required. As shown by Percoll buoyant density centrifugation, rIL‐4 induced B cells in the high‐density fraction to secrete IgG4 and IgE, whereas it inhibited spontaneous IgG4 secretion by low‐density B cells. Interferon‐r inhibited IL‐4‐induced lgG4 and IgE secretion. The data demonstrate that 11‐4 induces small, dense, peripheral blood B cells to secrete not only IgE but also lgG4, which paralells the IL‐4‐induced IgE and IgG1 secretion by murine B cells. Our in vitro data, the elevation of IgE and lgG4 in the serum of patients with helminth parasitic infections, and the prevalence of IgE and IgG4 antibodies to allergens suggest that the formation of these two isotypes is likely to share a common IL‐4‐dependent regulatory pathway.
Immune complex transfer enzyme immunoassay for (anti‐human t‐cell leukemia virus type i) igg in serum using a synthetic peptide, env gp46(188–209), as antigen
Authors: Kohno Takeyuki , Ishikawa Eiji , Sakoda Iwane
Published: March 2007

An immune complex transfer enzyme immunoassay for (anti‐human T‐cell leukemia virus type I) IgG (anti‐HTLV‐I IgG) in serum using a chemically and safely synthesized peptide, env gp46(188–209), 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 dinitrophenyl bovine serum albumin‐env gp46(188–209)‐β‐D‐galactosidase conjugate. The complex formed of the three components was trapped onto polystyrene balls coated with affinity‐purified (anti‐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 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 sensitive and detected anti‐HTLV‐I IgG in serum samples which were negative by the conventional enzyme immunoassay and Western blotting. And the specificity of this assay was confirmed by preincubation of test serum with excess of env gp46(188–209). However, some disadvantages were also noted.
Quantitation of anti‐tetanus and anti‐diphtheria antibodies by enzymoimmunoassay: Methodology and applications
Authors: Virella G. , Hyman Barbara
Published: March 2007

We have developed enzymoimmunoassays (EIA) for the quantitation of antibodies (Ab) to tetanus and diphtheria toxoids (TT, DT) using lmmulon I plates coated with the appropriate toxoid. A preparation of human tetanus immunoglobulin with a known concentration of anti‐TT Ab was used as calibrator of the anti‐TT antibody assay. The assay of anti‐DT Ab is calibrated with a pool of human sera whose anti‐DT Ab concentration was determined by quantitative immunoelectrophoresis, using a horse anti‐DT with known Ab concentration as calibrator. A peroxidase‐conjugated anti‐human IgG was used in both assays. ABTS was used as substrate, and the reaction was stopped after 1 min incubation with citric acid and the OD measured at 414 nm on a Vmax reader. The assays have been applied to a variety of clinical situations. In patients suspected of having tetanus, the quantitation of antibodies has been helpful in establishing a diagnosis. In patients with a history of hypersensitivity to tetanus toxoid, verification of the levels of anti‐TT antibody may prevent unnecessary and potentially harmful immunizations. The assays have also been used for the diagnostic evaluation of the humoral immune response to TT and DT, both in pediatric patients and in immunosuppressed patients. Several non‐responders have been detected, and we have recently used the assay to monitor the effects of fish oil administration on the humoral immune response. Finally, the sensitivity of the assay for anti‐TT antibody has been proven to be adequate for the assay of anti‐TT Ab produced in vitro by stimulated human B cells. In conclusion, the assay of anti‐TT and anti‐DT antibodies by EIA is relatively simple and has a variety of clinical and research applications.
Detection of anti‐nuclear antibodies from patients with systemic rheumatic diseases by elisa using hep‐2 cell nuclei
Authors: Jitsukawa Tomofumi , Nakajima Satoko , Usui Junko , Watanabe Hiroshi
Published: March 2007

A microtiter plate was coated with cell nuclei from HEp‐2 cells, and the stabilized nuclei were fixed with acetone for an enzyme‐linked immunosorbent assay (ELISA). Auto‐antibodies against nuclear antigens were detected from the sera of patients with various systemic rheumatic disease but not from healthy individuals by means of the nucleus ELISA procedure. Ninety‐one percent of anti‐nuclear antibody (ANA)—positive sera as demonstrated by immunofluorescence (IF) test were also judged as positive for ANAs by the nucleus ELISA and 5% of them as psudo‐positive. Patient's sera with homogeneous and fine‐speckled IF patterns displayed the highest ELISA titers. A large portion of ss‐A/Ro antigen is localized in the cytoplasm; therefore, anti‐ss‐A/Ro antibody was hardly detected by nucleus ELISA. The population of ANAs detectable by nucleus ELISA included anti‐ss‐B/La, anti‐DNA, anti‐histone, anti‐Sm, anti‐RNP, and anti‐scl‐70.
Evaluation of the kodak ektachem slide assays for iron and lactate
Authors: Marx Alfred M. , Gressner Axel M.
Published: March 2007

We evaluated the analytical performance of two clinical chemistry slides developed recently by Eastman Kodak Company for the analysis of lactate and iron with the Kodak Ektachem 700 XR analyzer. Tested with supplemented serum pool materials we found imprecisions of the lactate analysis between 0.7% and 1.3% (within‐run), resp., 1.0% and 1.5% (between‐run). The imprecision of the iron analysis was between 1.0% and 2.2%, resp., 1.2% and 2.7%. Linearity of the methods was proven between 0.7 mmol/L and 13.0 mmol/L (lactate analysis) and between 5.4 μmol/L and 82.4 μmmol/L (iron analysis), respectively. We found no significant interferences by hemolysis, lipemia, or bilirubin. The Kodak lactate results (x) correlated well with those of enzymatic methods (y) of Boehringer Mannheim (r = 0.995; y = 1.095x − 0.128) and of DuPont (r = 0.990; y = 1.015x − 0.242). The Kodak iron analysis (x) was correlated with the FerroZine® methods (y) of Boehringer Mannheim (r = 0.988; y = 1.031x + 0.091), Hoffman LaRoche (r = 0.989; y = 0.868x + 1.111), and with atomic absorption (y) (r = 0.960; y = 0.826x + 3.416). We conclude that the newly developed methods for the analysis of lactate and iron with the Kodak Ektachem analyzer are acceptable alternatives to other common methods.
Quantitative changes in t helper inducer (cd4+ cd45ra−), t suppressor inducer (cd4+ cd45ra+), t suppressor (cd8+ cd11b+), and t cytotoxic (cd8+ cd11b−) subsets in human immunodeficiency virus infection
Authors: Reddy Mohan M. , Grieco Michael H.
Published: March 2007

To determine changes in subsets of CD4 and CD8 in relation to HIV infection and progression to AIDS, we quantitated peripheral blood lymphocytes obtained from 17 heterosexual controls, 22 asymptomatic HIV‐seronegative and 18 HIV‐seropositive intravenous drug users, 50 patients with AIDS‐related complex (ARC), and 9 patients with AIDS using an EPICS “C” flow cytometer by twocolor analysis. Both T helper inducer (CD4+ CD45RA−) and suppressor inducer (CD4+ CD45RA+) lymphocytes were decreased significantly in patients with ARC or AIDS. In contrast, T cytotoxic (CD8+ CD11b−) cells were significantly increased and accompanied by a significant decrease in the T suppressor (CD8+ CD11b+) subset in patients with ARC or AIDS. These results suggest that both T helper inducer and T suppressor inducer subsets of the CD4+ population and the T suppressor subset of CD8+ are depleted after HIV infection, while the T cytotoxic subset of CD8+ was increased after HIV infection.
In vitro sensitivity assays in cancer: A review, analysis, and prognosis
Authors: Hoffman Robert M.
Published: March 2007

Tumors are complex systems consisting of heterogeneous cancer cells as well as normal cells with each exhibiting unique drug sensitivity spectra. There have been many attempts to design in vitro systems to determine drug response of tumors. The most widely used system is the clonogenic assay, which has demonstrated some clinical predictivity. However, the clonogenic assay has been shown to have negative aspects, including low frequency of evaluation, clump artifacts, lack of cytotoxic end‐points and disruption of normal cell‐cell interactions existing in a true tissue environment. Newer models are described utilizing cytotoxic as well as cell‐proliferation end‐points, and maintenance of three‐dimensional tissue architecture in vitro. It is concluded that less artifactual, more realistic models can be used to select more tumor‐specific drugs which themselves in turn will make in vitro chemosensitivity assays more useful for cancer patients.
Purification of soluble human t lymphocyte receptors for sheep erythrocytes
Authors: Itano Eiko N. , Ono Mario A. , Sumigawa Mari , Longo Leda M. , Moura Nayla C. , Mendes Nelson F.
Published: March 2007

Using a polyclonal heterologous anti‐soluble E‐receptor serum, we identified molecules of molecular weight circa 58,000 and 150,000

The soluble receptor molecule with molecular weight of approximately 58,000 (Rs1) was initially purified from supernatant of heated lymphocytes through chromatography on Sephadex G‐200 and/or DEAE‐cellulose. The soluble receptor molecule with molecular weight of approximately 150,000 (Rs2) is detected at high levels in the serum of patients with cancer and uremia. Rs1 and Rs2 present in serum from cancer patients were purified by chromatography on Sephadex G‐200 and by affinity chromatography using anti‐Rs1 IgG.

131I‐labelled supernatant of heated lymphocytes binds to sheep erythrocytes and the elution and analysis of the molecules adsorbed showed bands of molecular weights approximately 58,000 and 150,000, confirming the receptor activity of these molecules.
Diagnostic value of lactate dehydrogenase isoenzymes in cerebrospinal fluid
Authors: Chatterley Scott , Sun Tsieh , Lien York
Published: March 2007

To evaluate the diagnostic value of lactate dehydrogenase (LD) isoenzymes in cerebrospinal fluid (CSF), 93 consecutive CSF specimens were analyzed. These specimens were from patients of four categories tumors, infections, hemorrhages, and others. It was found that the isoenzyme patterns overlapped among different categories, but they differed within each category and were thus helpful in differential diagnosis. For instance, metastatic tumors showed prominent LD‐5, whereas a primary brain tumor demonstrated an increase in all fractions. Viral encephalitis revealed an increase in the first three isoenzymes and bacterial meningitis, the last two. In acquired immune deficiency syndrome (AIDS) cases, however, LD isoenzyme changes were demonstrated in CSF when only cryptococcal meningitis and not when encephalitis was present. Both subdural and subarachnoid hemorrhages showed elevation of all fractions in our study. Elevation of the first three fractions was usually due to brain tissue damage or hemorrhage, as proven by our isoenzyme study of hemolysate mixed with CSF. The prominence of the last two fractions was related to anaerobic metabolism in the central nervous system or to granulocytic infiltration. In conclusion, LD isoenzyme analysis in CSF is helpful in differential diagnosis of various CNS disorders, although its sensitivity awaits further improvement.
Monoclonal antibody to human cartilage cells and its reactivities to chondrocytic tumors
Authors: Eisenbrey A. Bradley , Chen Jenn C. , Dyer Charissa A. , Bernstein Jay , Poulik Miroslav D.
Published: March 2007

A murine monoclonal antibody (E10) was made against cultured cartilage cells. The E10 antibody binding is localized to the surface of cultured cartilage cells in suspension and is present in the cytoplasm in paraffin embedded sections. There is no reactivity with cartilage matrix, or with the matrix of cartilaginous tumors. Reactivity is removed by treatment with trypsin and hyaluronidase, but not by treatment with heparinase, neuraminidase, and chondroitinase.

Regeneration of E10 antigen after trypsinization takes 48 hours in chondrocytes in tissue culture. SDS‐polyacrylamide gel electrophoresis of an E10 immune precipitate of cultured chondrocytes results in two peaks: one at a very high molecular weight and a small fragment at approximately 250 kd.

Specificity has been demonstrated by cytofluorometry, immunofluorescence, and immunohistochemistry, in both frozen and paraffin‐embedded tissues. Positive reactivity was seen in cultured cartilage cells, chondrocytes in fetal and adult cartilage, chondrosarcomas, and chordomas. Minimal reactivity was found in a chondromyxoid liposarcoma. Acinar cells of salivary and sweat glands and mast cells in various tissues and tumors were also positive. There was no reactivity with other tissues and tumors, including myxoid and mucinous tumors and epithelial tissues.
Sensitive competitive‐binding elisas for quantifying free kappa and lambda light chains in cerebrospinal fluid
Authors: Stanescu Gabriel L. , Swick Alan R. , Tuohy Vincent K. , Rudick Richard A.
Published: March 2007

Simple, sensitive, and fully standardized solid‐phase enzyme‐linked competitive binding immunoassays to quantify free kappa and lambda immunoglobulin light chains are described. The assays were developed to measure the concentration of free light chains in cerebrospinal fluid (CSF), in part because elevated levels of free kappa light chains have utility as a diagnostic marker for multiple sclerosis (MS). Polyclonal rabbit antibodies raised against pooled Bence Jones proteins are bound to solid‐phase Staphylococcal protein A and used as the primary antisera in this assay. A pool of Bence Jones proteins isolated from the urine of 10 individuals with multiple myeloma are used as a biotin‐labeled ligand and to develop a standard curve. The assays as described are sensitive to the low nanogram range and are specific for free kappa or lambda light chains. The assays were found to have acceptable precision, and results correlated highly with concentrations determined using competitive‐binding radioimmunoassays previously described.
Effect of different test designs of immunoassays on “hook effect” of ca 19‐9 measurement
Authors: Wu James T. , Christensen Sonia E.
Published: March 2007

Determining serum CA 19‐9 using regular Centocor kit frequently requires numerous repeats when specimens contain highly elevated levels of CA 19‐9. We found that the number of repeats could be reduced if the Centocor kit with an extended concentration range was used. Using an extended Centocor kit could also eliminate falsely low CA 19‐9 results. However, when a Biomira RIA kit, based on a competitive binding principle, was used, falsely low CA 19‐9 values could be completely avoided and, in most cases, only a single repeat was usually required to arrive at the true result. This is because the level of CA 19‐9 in the specimen can be approximated from the radioactivity count. Therefore, when designing an immunoassay for tumor markers, one needs to take into consideration the concentration range of the tumor marker in the specimens if one would like to avoid repeats and falsely low values.
Enzyme‐Linked immunosorbent assay to detect mycobacterium tuberculosis antigen 5 and antimycobacterial antibody in cerebrospinal fluid of patients with tuberculous meningitis
Authors: Krishnan V. V. Radha , Mathai A.
Published: March 2007

Inhibition ELISA and indirect ELISA was standardised to detect Mycobacterium tuberculosis antigen5 and antimycobacterial antibody in cerebrospinal fluid (CSF) specimens of 75 patients with tuberculous meningitis, (TBM) and 75 patients with non‐tuberculous neurological diseases (control group). In both ELISAs, no false‐negative results were observed in 15 culture proven patients with TBM. Detection of M. tuberculosis antigen 5 is more sensitive than detection of antimycobacterial antibody. However, both ELISAs showed 100% specificity for tuberculous aetiology in culture‐negative patients with TBM. ELISA should be considered as one of the approaches in the laboratory diagnosis of TBM, particularly when standard bacteriological methods are unable to demonstrate M. tuberculosis in CSF specimens of patients with TBM.
Cryopreservation and long‐term liquid nitrogen storage of peripheral blood mononuclear cells for flow cytometry analysis effects on cell subset proportions and fluorescence intensity
Authors: Brown L. Morris , Clark J. W. , Neuland C. Y. , Mann D. L. , Pankiw‐Trost L. K. , Blattner W. A. , Tollerud D. J.
Published: March 2007

The effect of cryopreservation and longterm liquid nitrogen storage on peripheral blood mononuclear cell (PBMC) subsets was prospectively analyzed using monoclonal antibodies and flow cytometry. Brief cryopreservation did not significantly alter the proportion of positively stained cells for CD3+, CD4+, CD8+, CD14+, CD16+, and CD19+, cells. A small but statistically significant increase in the proportion of positive cells was observed for HLA‐DR+ and HLe‐1 + cells. Brief cryopreservation was associated with a decrease in the mean fluorescence intensity (MFl) values for CD3+, CD4+, and CD8+ cells; an increase in MFl values for CD14+ and HLA‐DR+ cells; and no change for CD16+, CD19+, and HLe‐1+ cells. There was no significant change in the proportion of CD3+, CD4+, or CD16+ cells during 20 months of storage in liquid nitrogen. Small but statistically significant decreases in the proportion of CD8+ and CD19+ cells were observed over the same interval, and the proportion of CD14+ cells (monocytes) was highly variable. Chronologic changes in fluorescence intensity during longterm storage were observed for all cell subsets except CD16+ and CD19+ cells. Cryopreservation is a valuable technique for longterm storage of viable cells. For many laboratory applications, the small changes noted in the present study will have no practical importance. However, for clinical and epidemiological investigations encompassing large numbers of samples, statistical techniques to adjust for small changes during storage should be considered.
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

A 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

The 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

The 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

Increased 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.