Pulmonary lesions in experimental ophidian paramyxovirus pneumonia of Aruba Island rattlesnakes, Crotalus unicolor

Histologic and ultrastructural changes were observed in the respiratory portions of lung in five 29-40-month-old Aruba Island rattlesnakes, Crotalus unicolor, that were inoculated with an Aruba Island Rattlesnake virus (AIV) strain of ophidian paramyxovirus (OPMV) isolated from an Aruba Island rattlesnake. Lungs from one non-infected and three mock-infected Aruba Island rattlesnakes were examined also. From 4 to 22 days following intratracheal inoculation, progressive microscopic changes were seen in the lung. Initially, increased numbers of heterophils were observed in the interstitium followed by proliferation and vacuolation of epithelial cells lining faveoli. The changes appeared to progress from cranial to caudal portions of the respiratory lung following inoculation. Beginning at 4 days postinoculation, viral antigen was demonstrated in epithelial cells lining faveoli with an immunofluorescent technique using a rabbit anti-AIV polyclonal antibody. Electron microscopy revealed loss of type I cells, hyperplasia of type II cells, and interstitial infiltrates of heterophils and mononuclear cells. Viral nucleocapsid material was seen within the cytoplasm and mature virus was seen budding from cytoplasmic membranes of infected type I and type II cells from 8 to 19 days after infection. A virus consistent with AIV was isolated from lung tissues of infected rattlesnakes, thus fulfilling Koch's postulates.     

New insights into the co-evolution of cytochrome c reductase and the mitochondrial processing peptidase

The mitochondrial processing peptidase (MPP) is a heterodimeric enzyme that forms part of the cytochrome c reductase complex from higher plants. Mitochondria from mammals and yeast contain two homologous enzymes: (i) an active MPP within the mitochondrial matrix and (ii) an inactive MPP within the cytochrome c reductase complex. To elucidate the evolution of MPP, the cytochrome c reductase complexes from lower plants were isolated and tested for processing activity. Mitochondria were prepared from the staghorn fern Platycerium bifurcatum, from the horsetail Equisetum arvense, and from the colorless algae Polytomella, and cytochrome c reductase complexes were purified by a micro-isolation procedure based on Blue-native polyacrylamide gel electrophoresis and electroelution. This is the first report on the subunit composition of a respiratory enzyme complex from a fern or a horsetail. The cytochrome c reductase complexes from P. bifurcatum and E. arvense are shown to efficiently process mitochondrial precursor proteins, whereas the enzyme complex from Polytomella lacks proteolytic activity. An evolutionary model is suggested that assumes a correlation between the presence of an active MPP within the cytochrome c reductase complex and the occurrence of chloroplasts.     

Translocation of human calcitonin in respiratory nasal epithelium is associated with self-assembly in lipid membrane

We studied the mechanisms involved in the translocation of human calcitonin (hCT) through excised bovine nasal mucosa (net mucosal-to-serosal permeability approximately 10(-)5 cm s-1). To determine structural requirements for the suggested vesicular internalization two carboxyfluorescein-labeled (fl) hCT fragments, the C-terminal fragment [Nalpha-fl]hCT(9-32) and the N-terminal fragment [Lys(fl)18]hCT(1-24) were synthesized. In presence of the endocytosis inhibitor cytochalasin D mucosal-to-serosal and serosal-to-mucosal hCT permeabilities were equal. Pathway visualization by confocal laser scanning microscopy showed punctated fluorescence indicating vesicular internalization of both hCT and [Nalpha-fl]hCT(9-32). In contrast, the N-terminal fragment lacking the beta-sheet forming C-terminus (25-32) was not internalized. Circular dichroism showed that, when interacting with neutral and negatively charged liposomes, hCT adopts beta-sheet conformation. In a concentrated aqueous solution, beta-sheet formation induces hCT self-assembly and fibrillation. High partitioning of hCT into lipid bilayer membranes was reflected by an apparent partition coefficient log D(pH 7.4) = 2.5 (liposome-buffer equilibrium dialysis). We propose that the high lipid partitioning and beta-sheet formation result in C-terminus-restricted supramolecular self-assembly of hCT and [Nalpha-fl]hCT(9-32) in lipid membranes. Vesicular internalization is suggested to be associated with self-assembly induced perturbation of the lipid bilayer. Condensed hCT self-assemblies may explain the high capacity of net mucosal-to-serosal hCT permeation, which compares favorably with the low transport capacity of receptor-mediated endocytosis.     

Undetermined family diagnosis--uncertain diagnosis

The results of testing convergence- and discriminance-validity of the central dimensions "cohesion" and "control" for family diagnostics and therapy are presented in this paper by means of a multitrait-multimethod matrix which includes five known family diagnostic instruments from seven studies published up until the end of 1995. The main result demonstrates that both discriminance- and convergence-validity cannot be considered safe from "cohesion" or "control".     

Mathematics learning disabilities: a view from developmental psychology

U.S. education suffers from shortcomings that put even children possessing adequate intellectual abilities at risk for low mathematics achievements. Consequently, identifying and understanding children whose academic failure is influenced by a genuine learning disability requires a complex "developmental" research agenda. This perspective suggests the use of sensitive research methods--clinical interviews, ethnographies--to examine the development of children's construction of knowledge in the context of schooling. Researchers should consider such factors as the adequacy of classroom instruction, the availability in children of informal knowledge, the role of motivation, the effects of specific interventions, the role and operation of different cognitive processes in constructing mathematical understanding, children's difficulties across different areas of mathematics, and the development of children's thinking throughout the school years.     

Classification of compressed breast shapes for the design of equalization filters in x-ray mammography

We are developing an external filter method for equalizing the x-ray exposure in mammography. Each filter is specially designed to match the shape of the compressed breast border and to preferentially attenuate the x-ray beam in the peripheral region of the breast. To be practical, this method should require the use of only a limited number of custom built filters. It is hypothesized that this would be possible if compressed breasts can be classified into a finite number of shapes. A study was performed to determine the number of shapes. Based on the parabolic appearance of the outer borders of compressed breasts in mammograms, the borders were fit with the polynomial equations y = ax2 + bx3 and y = ax2 + bx3 + cx4. The goodness-of-fit of these equations was compared. The a,b and a,b,c coefficients were employed in a K-Means clustering procedure to classify 470 CC-view and 484 MLO-view borders into 2-10 clusters. The mean coefficients of the borders within a given cluster defined the "filter" shape, and the individual borders were translated and rotated to best match that filter shape. The average rms differences between the individual borders and the "filter" were computed as were the standard deviations of those differences. The optimally shifted and rotated borders were refit with the above polynomial equations, and plotted for visual evaluation of clustering success. Both polynomial fits were adequate with rms errors of about 2 mm for the 2-coefficient equation, and about 1 mm for the 3-coefficient equation. Although the fits to the original borders were superior for the 3-coefficient equation, the matches to the "filter" borders determined by clustering were not significantly improved. A variety of modified clustering methods were developed and utilized, but none produced major improvements in clustering. Results indicate that 3 or 4 filter shapes may be adequate for each mammographic projection (CC- and MLO-view). To account for the wide variations in exposures observed at the peripheral regions of breasts classified to be of a particular shape, it may be necessary to employ different filters for thin, medium and thick breasts. Even with this added requirement, it should be possible to use a small number of filters as desired.