摘要:

AbstractWe sought to determine whether measurement ofHistoplasma capsulatumvar.capsulatumantigen concentration in tissues and blood provided a marker for antifungal effect of itraconazole in a nonlethal murine model of histoplasmosis. Treatment with itraconazole (Sporanox), in cyclodextrin, was evaluated in a pulmonary model of histoplasmosis. Mice infected with 4.0 × 107yeast‐phase organisms by endotracheal inoculation were treated with itraconazole, 1.5 mg twice daily by gavage, for 10 consecutive days, beginning on day 4 of infection. All mice were sacrificed on day 15 of infection. Blood, spleen, and lung tissues were removed for culture and quantification of antigen. Numbers of organisms were significantly lower in spleens from the treated group: 20.8 ± 41.8 vs. 65.8 ± 39.1 in the control group,P= 0.017. Numbers of organisms in lung were 9.6 ± 27.3 colonyforming units in treated versus 24.2 ± 36.3 in control animals,P= 0.267. Antigen concentrations in spleen tissue and serum were lower in treated versus control mice: spleen, 1.8 ± .6 units in treated versus 11.0 ± 2.3 in controls,P<0.001; serum, 0.8 ± 0.2 units in treated versus 2.2 ± 1.0 units in controls,P<0.001. Lung antigen concentrations were similar in the two groups, 19.2 ± 1.4 units in treated compared to 17.9 ± 3.0 units in control mice,P= 0.142. The cyclodextrin formulation of itraconazole (Sporanox) demonstrated antifungal activity in experimental histoplasmosis. Antigen detection was a useful marker for antifungal effect. © 1994 Wi

ISSN:0887-8013    

DOI:10.1002/jcla.1860080102

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractThe distribution of the major VLsubgroups (VλI, VλII, VλIII, VλIV, VλVI, and VλVIII) among λ‐type immunoglobulins (Igs) in normal serum was determined by a sandwich enzymelinked immunosorbent assay (ELISA) using a panel of murine anti‐human Vλ‐subgroup‐specific monoclonal antibodies (MoAbs) and appropriate reference standard proteins. The mean concentration of λI, λII, λIII, λIV, λVI, and λVIII Igs in serum specimens obtained from 23 adults was 2158, 162, 1958, 264, 225, and 169 μg/mL and represented 44, 3, 40, 5, 5, and 3% of the total Igλ population, respectively. The low percentage of λII Igs in normal serum was in marked contrast to the ∼40% incidence of this Vλsubgroup found among λ‐type Bence Jones proteins and monoclonal serum Igs obtained from patients with multiple myeloma and AL amyloidosis, or ∼60% in Waldenström's macroglobulinemia. The non‐random expresson of the VλII subgroup in these diseases implies a relationship between Vλ‐gene usage and plasma cell, as well as certain types of lymphoc

ISSN:0887-8013    

DOI:10.1002/jcla.1860080103

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractA simple, rapid assay for the pancreatic isoenzyme of human serum amylase was developed. The assay utilized an immunoabsorbent prepared by coating latex beads with a monoclonal antibody specific for pancreatic amylase. Treatment of patient serum with immunoabsorbent removed pancreatic amylase, and measurement of residual amylase activity with standard total amylase methodology allowed estimation of the pancreatic amylase content. Extraction efficiency of pancreaticamylase was consistent at amylase concentrations up to 1,000 U/L (y = 0.97 × + 16.7 U/L; r = 0.9995). The assay was standardized with purified pancreatic amylase added to neonatal serum (low endogenous activity). A comparison of patient specimen results with the results of a standard technique (cellulose acetate electrophoresis) yielded an excellent correlation (immunoabsorption result = 0.96 electrophoresis result + 1.2 U/L; r = 0.987). Salivary amylase did not interfere with the assay until levels exceeded 1,000 U/L. Daily analysis of a frozen serum pool yielded a coefficient of variation of 9.2% at mean pancreatic amylase value of 54 U/L (±5 U/L). A normal range study found a strong influence of age, with pancreatic amylase levels increasing dramatically in the first 3 years of life, to stabilize at a range of 0–66 U/L thereafter. © 1994 Wiley‐Lis

ISSN:0887-8013    

DOI:10.1002/jcla.1860080104

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractImmunoassays have been recently developed that measure the avidity of IgG antibodies to complex microbial antigens and are suitable for serologic diagnosis of infectious diseases. In these avidity ELISAs, proteindenaturing agents are applied either as diluents of patient sera to prevent the immune complexing of IgG (diluting principle), or the preformed complexes are treated with the protein denaturants (eluting principle). We compared four protein denaturants previously used in such assays, in a diagnostic avidity ELISA for rubella IgG. Diethylamine, guanidine, thiocyanate, or urea were applied, by either principle at various concentrations, and thiocyanate, or urea were applied, by either principle at various concentrations, and thiocyanate at an optimum pH. Patient sera obtained recently after primary infection were distinguished from sera representing past rubella immunity by any protein denaturant tested by the eluting principle, which was superior to the diluting principle. © 1994 Wiley‐Liss, I

ISSN:0887-8013    

DOI:10.1002/jcla.1860080105

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractWe studied 134 “healthy persons” with no past history of any serious or latent diseases, and observed differences in their 25 lymphocyte subsets by conducting two‐color analyses using 12 useful combinations of dye‐labeled monoclonal antibodies.CD4+Leu8+cells and CD25+CD3+cells were significantly higher in males than in females, whereas CD4+Leu8−cells and CD23+CD19+cells were significantly higher in females. In comparison of the 25 lymphocyte subsets among four age groups, CD45RA−CD4+cells and CDw29+CD4+cells significantly increased with age, and CD8+CD11b−cells and CD57−CD8+cells significantly decreased with age.In females, helper T cells significantly increased and helper inducer T cells increased, while cytotoxic T cells decreased with age. These are important findings that should be considered in studies of immune function and autoimmune disorders. © 1994

ISSN:0887-8013    

DOI:10.1002/jcla.1860080106

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractThe present study was undertaken to determine whether monoclonal antibodies (moABs TKH‐2, MA54, MA61, B72.3, and CC49) directed toward O‐linked mucin‐type glycoprotein detect the NeuAcα2‐6GalNAc (sialyl Tn) epitope in meconium‐ and amniotic fluidderived mucin. Fetal colonic mucosal cells express the sialyl Tn antigen, particularly in goblet cell mucin. The reactivities of these moABs with a perchloric acid extract of meconium (meconium extract) and different native and neuraminidase treated glycoproteins were examined by solid‐phase enzyme‐linked immunosorbent assay (ELISA). All the moABs react with the meconium supernatant and meconium extract. The reactivities of TKH‐2, MA54, and MA61 are neuraminidase sensitive, and the reactivity of TKH‐2 with meconium extract was specifically inhibited by ovine submaxillary mucin (OSM). A NeuAcα 2‐6GalNAc epitope is the characteristic component in meconium. Mucin released from the fetal respiratory tract could, in part, provide an alternative source in the amniotic fluid. TKH‐2 is the most sensitive antibody directed to sialyl Tn antigen in meconium supernatant. The likelihood of TKH‐2 serving as the basis for a sensitive assay to detect sialyl Tn in meconium‐ and amniotic fluid‐derived mucin is pro

ISSN:0887-8013    

DOI:10.1002/jcla.1860080107

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

ISSN:0887-8013    

DOI:10.1002/jcla.1860080108

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractWe developed a method to quantitate hepatitis B virus (HBV) DNA in serum by ammonium sulfate fractionation and DNA hybridization. Serum samples were precipitated with 45% saturated ammonium sulfate, resuspended in buffer, and spotted on a nylon membrane. Following denaturation in alkali, HBV DNA sequences on the membrane were detected by hybridization with a32P‐labeled DNA probe of the entire HBV genome. Bound radioactivity was measured with liquid scintillation counting. Ammonium sulfate fractionation of positive samples increased assay sensitivity by 10–30‐fold compared to no treatment. Sensitivity for detection of cloned HBV DNA added to negative serum was 0.2 pg. Recovery of cloned HBV DNA added to negative serum before fractionation was equivalent to direct spotting of DNA onto the membrane in the absence of serum. This method enhanced HBV DNA recovery from serum into small volumes, thereby expanding the potential analytic range of spot hybridization assays. © 1994 Wiley‐L

ISSN:0887-8013    

DOI:10.1002/jcla.1860080109

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

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

ISSN:0887-8013    

DOI:10.1002/jcla.1860080110

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

ISSN:0887-8013    

DOI:10.1002/jcla.1860080101

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY