Lung injury induced by antibody fragments to angiotensin-converting enzyme.

Rabbits given goat anti-rabbit angiotensin-converting enzyme antibodies or derived antibody fragments develop rapidly fatal pulmonary edema. Endothelial cell injury is manifested by bleb formation and the disintegration of cell membranes. Platelets are found along the injured endothelium and leukocytes block capillary lumens. The pathologic features are similar when immune IgG, F(ab')2, or Fab are given. In vitro studies of complement activation show that solubilized, purified angiotensin-converting enzyme alone activates C1, with consumption of C4 and C3. Addition of immune IgG plus converting enzyme enhances this activation. F(ab')2 plus enzyme enhances only C3 consumption, while Fab with enzyme produces no additional complement utilization. Thus, while complement activation may be involved in the pathogenesis of injury induced by IgG or F(ab')2, the mechanism of Fab-induced endothelial injury remains unclear.     

DNA fingerprinting of Pasteurella multocida recovered from avian sources

Repetitive sequence-based PCR (rep-PCR) and amplified fragment length polymorphism (AFLP) were used to characterize a sample of 43 field isolates and 4 attenuated vaccine strains of Pasteurella multocida recovered from multiple avian species. Both rep-PCR and AFLP assays were rapid and reproducible, with high indices of discrimination. Concordance analyses of rep-PCR and AFLP with somatic serotyping indicate that, in general, somatic serotyping is a poor indicator of genetic relatedness among isolates of P. multocida. In addition, the data provide evidence of host specificity of P. multocida clones. Overall, the results of our study indicate that the rep-PCR and AFLP techniques enable rapid fingerprinting of P. multocida isolates from multiple avian species and enhance the investigation of fowl cholera outbreaks.     

Immunologic mechanisms in hypersensitivity pneumonitis: I. Evidence for cell-mediated immunity and complement fixation in pigeon breeders' disease

Immunologic studies were performed on 5 patients with pigeon breeders' disease. Intradermal injection of pigeon serum produced an immediate wheal-and-flare reaction within 15 minutes and a secondary Arthus-type reaction within 4 to 8 hours. Immunofluorescent studies of the secondary reaction site showed IgG, C3, and C4 in 2 patients. Patients' sera produced multiple precipitin bands with pigeon serum when reacted by double diffusion in gel. IgG antibody isolated from each of the patients' serum formed precipitating immune complexes that fixed large amounts of complement (C4) when added to fresh human serum. Peripheral blood lymphocytes from 4 of the 5 patients produced macrophage migration inhibitory factor (MIF) when challenged with dilute pigeon serum. These studies are the first to show complement fixing antibodies and macrophage MIF production by lymphocytes from patients with hypersensitivity lung disease and suggest that both humoral and cellular immunity may be important in the pathogenesis of these disorders.     

Human observer detection experiments with mammograms and power-law noise

We determined contrast thresholds for lesion detection as a function of lesion size in both mammograms and filtered noise backgrounds with the same average power spectrum, P(f)=B/f3. Experiments were done using hybrid images with digital images of tumors added to digitized normal backgrounds, displayed on a monochrome monitor. Four tumors were extracted from digitized specimen radiographs. The lesion sizes were varied by digital rescaling to cover the range from 0.5 to 16 mm. Amplitudes were varied to determine the value required for 92% correct detection in two-alternative forced-choice (2AFC) and 90% for search experiments. Three observers participated, two physicists and a radiologist. The 2AFC mammographic results demonstrated a novel contrast-detail (CD) diagram with threshold amplitudes that increased steadily (with slope of 0.3) with increasing size for lesions larger than 1 mm. The slopes for prewhitening model observers were about 0.4. Human efficiency relative to these models was as high as 90%. The CD diagram slopes for the 2AFC experiments with filtered noise were 0.44 for humans and 0.5 for models. Human efficiency relative to the ideal observer was about 40%. The difference in efficiencies for the two types of backgrounds indicates that breast structure cannot be considered to be pure random noise for 2AFC experiments. Instead, 2AFC human detection with mammographic backgrounds is limited by a combination of noise and deterministic masking effects. The search experiments also gave thresholds that increased with lesion size. However, there was no difference in human results for mammographic and filtered noise backgrounds, suggesting that breast structure can be considered to be pure random noise for this task. Our conclusion is that, in spite of the fact that mammographic backgrounds have nonstationary statistics, models based on statistical decision theory can still be applied successfully to estimate human performance.     

Reticulate bodies as single antigen in Chlamydia trachomatis serology with microimmunofluorescence.

Formalin-fixed, purified reticulate bodies (RB) of Chlamydia trachomatis immunotype C/TW-3/OT were used as a serological test antigen in the microimmunofluorescence test. The sensitivity and specificity of the RB antigen were compared to elementary bodies (EB) used as antigens in the detection of C. trachomatis antibodies in human sera by microimmunofluorescence. RB reacted with all known C. trachomatis immunotypes with the same sensitivity as the homotypic EB. In routine serology with sera and endocervical secretions, the RB antigen had a sensitivity similar to that of the EB in detecting serum antibodies, endocervical secretion antibodies, and antibodies of immunoglobulin M and G classes. No false-positive reactions were detected with control sera. All positive reactions showed type-specific antibodies against an EB immunotype. RB seemed to demonstrate chlamydial group reactivity; sera from 10 psittacosis patients diagnosed clinically and serologically by complement fixation showed five positive, three equivocal, and two negative reactions. By immunofluorescence, RB appeared as distinct rings demonstrating uniform peripheral surface fluorescence at their rims. The EB appeared as pinpoint-sized dots. C/TW-3/OT RB used as a single test anitgen should provide a simple and sensitive serological assay for the detection of C. trachomatis antibody.     

Comparison of rapid,automated ribotyping and DNA macrorestriction analysis of Burkholderia pseudomallei

An automated ribotyping device (RiboPrinter) was used to determine the ribotypes of a collection of Burkholderia pseudomallei isolates. In a preliminary evaluation with the restriction enzymes BamHI and EcoRI, the protocol with EcoRI was more discriminating. The reproducibilities of the ribotypes obtained with EcoRI (EcoRI ribotypes) were determined by testing three levels of bacterial loads. The performance of the manufacturer's software was assessed by comparing the machine-optimized ribotypes with the type determined from the original gel image analyzed with Bionumerics software. The library of B. pseudomallei EcoRI ribotypes was then compared with the ribotypes obtained by DNA macrorestriction analysis of XbaI digests by pulsed-field gel electrophoresis. The typeability of B. pseudomallei by EcoRI ribotyping was 100%, and the discrimination index was 0.94. The slightly greater discrimination provided by DNA macrorestriction analysis (0.96) was achieved at the expense of a significantly longer processing time of 6 days, although the method was only half the cost of automated ribotyping. Typeability by macrorestriction analysis was lower (97%) unless a thiourea step was added to neutralize the action of Tris-dependent endonucleases. The digital record of B. pseudomallei isolates analyzed thus far provides a useful resource for future epidemiological studies and will help shorten the response time in the event of a further melioidosis outbreak or the deliberate release of B. pseudomallei as a biohazard.     

Genetic diagnoses and associated anomalies in fetuses prenatally diagnosed with esophageal atresia

Esophageal atresia (EA) is a congenital anomaly occurring in 2.3 per 10,000 live births. Due to advances in prenatal imaging, EA is more readily diagnosed, but data on the associated genetic diagnoses, other anomalies, and postnatal outcome for fetuses diagnosed prenatally with EA are scarce. We collected data from two academic medical centers (n = 61). Our data included fetuses with suspected EA on prenatal imaging that was confirmed postnatally and had at least one genetic test. In our cohort of 61 cases, 29 (49%) were born prematurely and 19% of those born alive died in the first 9 years of life. The most commonly associated birth defects were cardiac anomalies (67%) and spine anomalies (50%). A diagnosis was made in 61% of the cases; the most common diagnoses were vertebral defects, anal atresia, cardiac anomalies, tracheoesophageal fistula with esophageal atresia, radial or renal dysplasia, and limb anomalies association (43%, although 12% met only 2 of the criteria), trisomy 21 (5%), and CHARGE syndrome (5%). Our findings suggest that most fetuses with prenatally diagnosed EA have one or more additional major anomaly that warrants a more comprehensive clinical genetics evaluation. Fetuses diagnosed prenatally appear to represent a cohort with a worse outcome.     

Developmental trajectories of adolescents' depressive symptoms: predictors of change

In a sample of 240 adolescents assessed annually in Grades 6 through 11, the developmental trajectories of their depressive symptoms were examined using latent factor growth modeling. Growth in mother-reported adolescent depressive symptoms was quadratic; growth in adolescent-reported symptoms was linear. In the model with gender and maternal depression, girls reported a greater increase in depressive symptoms over time than boys, and adolescents of mothers with histories of mood disorders had higher initial levels of depressive symptoms than offspring of never-depressed mothers. After gender and maternal depression were controlled, initial levels of negative attributions and stressors significantly predicted initial levels of adolescent- and mother-reported depressive symptoms. Attributional styles that were increasingly negative across time were associated with significantly higher initial levels (mother reported) and increasing growth (adolescent reported) of depressive symptoms. Reciprocal models in which development of depressive symptoms predicted the development of attributions and stress also were examined.     

Expression of CD45 isoforms in lymph node reactive hyperplasia

The CD45 antigen family consists of multiple molecular isoforms ranging from 180 to 220 kDa. The highest Mr isoforms are recognized by monoclonal antibodies (MoAbs) designated CD45RA, while those recognizing the low Mr isoforms are designated CD45RO. T cells expressing CD45RA are "naive" or unprimed, while those expressing CD45RO have "memory." Further, stimulation of CD45RA+ T cells induces an isoform switch to the CD45RA-/CD45RO+ phenotype. The present study examined this in vitro process by determining the in vivo CD45 isoform expression of T cells from human hyperplastic lymph nodes. Hyperplastic, as opposed to nonhyperplastic, lymph nodes exhibited the expected CD45 isoform switch from CD45RA+ to CD45RO+ T cells that has been described in vitro. The percentage of CD45RO+ T cells did not correlate with other parameters of lymphoid activation. Thus, CD45RO expression probably represents a marker of differentiation and acquisition of "memory" or late cellular activation.