Elevated levels of a high molecular weight antigen detected by antibody W1 in sera from breast cancer patients.

A novel screening assay was used to test 13 previously described antibreast cancer antibodies for those which recognize antigens elevated in serum of breast cancer patients. Binding of three of these antibodies to breast or lung carcinoma cells was inhibited to a significantly greater extent by tumor patient serum than by normal serum, suggesting that the antigens might be useful serum markers. Two of these antibodies, W1 and W9, were shown to recognize nonoverlapping epitopes on a high molecular weight molecule(s) purified from serum from breast cancer patients. A sensitive double determinant immunoassay was developed to measure W1 antigen levels in sera from a total of 389 cancer patients and controls. Forty seven % (37 of 79) of individuals having breast cancer showed elevated serum levels of the W1 antigen, whereas only 4% (1 of 25) of normal controls and 2% (1 of 47) of patients hospitalized for nonmalignant disorders showed elevated levels. These differences were statistically significant (P less than 0.001). The percentage of breast cancer patients showing elevated serum levels was greater for individuals with metastatic disease. Statistically significant numbers of lung, ovarian, and prostate, but not colon, cancer patients also had elevated serum levels of the W1 antigen. These data suggest that measurement of the W1 antigen in serum might provide clinically useful information on the course of metastatic breast and other cancers.

Two modes of sulfite oxidation in the extremely thermophilic and acidophilic archaeon acidianus ambivalens

Sulfite-oxidizing enzyme activities were analyzed in cell-free extracts of aerobically grown cells of Acidianus ambivalens, an extremely thermophilic and chemolithoautotrophic archaeon. In the membrane and cytoplasmic fractions, two distinct enzyme activities were found. In the membrane fraction, a sulfite:acceptor oxidoreductase activity was found [530 mU (mg protein)(-1); apparent K(m) for sulfite, 3.6 mM]. In the cytoplasmic fraction the following enzyme activities were found and are indicative of an oxidative adenylylsulfate pathway: adenylylsulfate reductase [138 mU (mg protein)(-1)], adenylylsulfate:phosphate adenyltransferase ["ADP sulfurylase"; 86 mU (mg protein)(-1)], adenylate kinase [650 mU (mg protein)(-1)], and rhodanese [thiosulfate sulfur transferase, 9.2 mU (mg protein)(-1)]. In addition, 5',5"'-P(1),P(4)-di(adenosine-5') tetraphosphate (Ap(4)A) synthase and Ap(4)A pyrophosphohydrolase activities were detected.

Reconstruction of New Orleans after Hurricane Katrina: a research perspective.

Four propositions drawn from 60 years of natural hazard and reconstruction research provide a comparative and historical perspective on the reconstruction of New Orleans after Hurricane Katrina. Decisions taken over its 288-year history that have made New Orleans so vulnerable to Katrina reflect a long-term pattern of societal response to hazard events--reducing consequences to relatively frequent events, and increasing vulnerability to very large and rare events. Thus Katrina's consequences for New Orleans were truly catastrophic--accounting for most of the estimated 1,570 deaths of Louisiana residents and $40-50 billion in monetary losses. A comparative sequence and timing of recovery provides a calendar of historical experience against which to gauge progress in reconstruction. Using this calendar, the emergency post-disaster period appears to be longer in duration than that of any other studied disaster. The restoration period, the time taken to restore urban services for the smaller population, is in keeping with or ahead of historical experience. The effort to reconstruct the physical environment and urban infrastructure is likely to take 8-11 years. Conflicting policy goals for reconstruction of rapid recovery, safety, betterment, and equity are already evident. Actions taken demonstrate the rush to rebuild the familiar in contrast to planning efforts that emphasize betterment. Because disasters tend to accelerate existing economic, social, and political trends, the large losses in housing, population, and employment after Katrina are likely to persist and, at best, only partly recover. However, the possibility of breaking free of this gloomy trajectory is feasible and has some historical precedent.

Improved purification of the membrane-bound hydrogenase-sulfur-reductase complex from thermophilic archaea using epsilon-aminocaproic acid-containing chromatography buffers.

A hydrogenase-sulfur reductase (SR) complex was purified from membrane preparations of the extremely thermophilic, acidophilic archaeon Acidianus ambivalens using a combination of sucrose density gradient centrifugation and column chromatography (FPLC). All chromatographic steps were performed in the presence of 0.5% epsilon-aminocaproic acid resulting in the elution of the SR complex as a sharp peak. In contrast, chromatography using buffers without epsilon-aminocaproic acid, or in the presence of detergents, were not successful. The purified A. ambivalens SR complex consisted of at least four subunits with relative molecular masses of 110000, 66000, 39000 and 29000, respectively. A similar procedure was applied to purify the membrane-bound hydrogenase from Thermoproteus neutrophilus, a non-related extremely thermophilic but neutrophilic archaeon, which consisted of only two subunits with relative molecular masses of 66000 and 39000, respectively.

Evaluation of a Treponema pallidum western immunoblot assay as a confirmatory test for syphilis.

Tests for the detection of antibodies to Treponema pallidum are recommended for the confirmation of reactive nontreponemal test results and the accurate diagnosis of syphilis. The present-day use of Western blot (immunoblot) technology for the diagnosis of retroviruses prompted the development and evaluation of a Western blot assay with whole-cell T. pallidum as the antigen. The assay detected antibodies in syphilitic serum or plasma from dilutions of specimens incubated overnight with test strips. A test was considered positive when at least three of four major antigens having molecular masses of 15.5, 17, 44.5, and 47 kDa were detected. The Western blot assay had 93.8% sensitivity and 100% specificity for clinically defined samples. The Western blot assay was compared with double-staining fluorescent treponemal antibody absorption [FTA-ABS (DS)], which had a sensitivity and a specificity of 91.7 and 92.0%, respectively. Dilution series studies of syphilis-positive specimens indicated that the Western blot assay has an endpoint of reactivity at least 3 to 4 serial dilutions greater than that for FTA-ABS (DS). Overall, the greater than 95% agreement between the Western blot assay and FTA-ABS (DS) for clinically defined specimens indicates that the sensitivity of the Western blot assay is equal to or greater than that of FTA-ABS (DS). The Western blot assay demonstrated no false-positive or equivocal reactivities for nonsyphilitic specimens, including normal specimens (both plasma and serum), biological false-positives, and specimens with elevated gamma globulin or antinuclear antibody. We conclude that the high sensitivity and specificity of the T. pallidum Western blot assay, together with its simplicity and objectivity, make it a good confirmatory test for syphilis.

Improved serologic detection of hepatitis C virus with a paramagnetic microparticle assay using multiple antigenic sequences.

A paramagnetic microparticle (MP) assay for antibody to hepatitis C virus (anti-HCV) was developed, in which the probe for antibody consisted of synthetic peptides corresponding to HCV capsid and nonstructural c-100 regions, as well as a recombinant protein corresponding to the nonstructural c33c region. Assay performance was evaluated by testing serum from 108 geographically diverse patients with non-A, non-B hepatitis (NANBH). The frequency of anti-HCV reactivity detected with the MP assay and with an enzyme-linked immunosorbent assay (ELISA) for c-100 was 91 and 70 percent, respectively. All c-100 HCV ELISA-reactive specimens also reacted on the MP assay. In addition, anti-HCV seroconversion in three plasma donors was detected one to two blood collection dates earlier by the MP assay than by the c-100 HCV ELISA and at similar blood collection dates by the MP assay and a second-generation anti-HCV ELISA. Serologic responses to the three distinct antigenic regions of HCV in NANBH patients varied: reactivity to all three antigens was most common (49%), reactivity to both capsid and c33c (40%) was next most common, and single-antigen reactivity was rare (4%). MP assay reactivity of 825 volunteer donors was 0.1 percent. These results demonstrate both the utility of additional HCV antigens for an effective anti-HCV screening assay and the application of paramagnetic MP technology to serologic testing for HCV infection.

A novel semi-automated paramagnetic microparticle based enzyme immunoassay for hepatitis C virus: its application to serologic testing.

A new rapid serologic enzyme immunoassay for antibodies to hepatitis C virus (HCV) is described. The assay combines synthetic peptide and recombinant antigens representing putative structural and non structural HCV gene products with paramagnetic microparticle assay (MP assay) technology. Assay readout is based upon an enzymatically generated fluorescent product which is quantified with a novel semi-automated washer/reader instrument system. Assay sensitivity and specificity was determined to be greater than the first generation HCV C-100 EIA using a non-A, non-B hepatitis disease panel, an HCV performance panel, an HCV seroconversion panel, dilutions of HCV reactive sera, and random volunteer blood donor specimens.