Elucidation of a PTS–Carbohydrate Chemotactic Signal Pathway in Escherichia coli Using a Time-resolved Behavioral Assay

Chemotaxis of Escherichia coli toward phosphotransferase systems (PTSs)–carbohydrates requires phosphoenolpyruvate-dependent PTSs as well as the chemotaxis response regulator CheY and its kinase, CheA. Responses initiated by flash photorelease of a PTS substrates d-glucose and its nonmetabolizable analog methyl α-d-glucopyranoside were measured with 33-ms time resolution using computer-assisted motion analysis. This, together with chemotactic mutants, has allowed us to map out and characterize the PTS chemotactic signal pathway. The responses were absent in mutants lacking the general PTS enzymes EI or HPr, elevated in PTS transport mutants, retarded in mutants lacking CheZ, a catalyst of CheY autodephosphorylation, and severely reduced in mutants with impaired methyl-accepting chemotaxis protein (MCP) signaling activity. Response kinetics were comparable to those triggered by MCP attractant ligands over most of the response range, the most rapid being 11.7 ± 3.1 s−1. The response threshold was <10 nM for glucose. Responses to methyl α-d-glucopyranoside had a higher threshold, commensurate with a lower PTS affinity, but were otherwise kinetically indistinguishable. These facts provide evidence for a single pathway in which the PTS chemotactic signal is relayed rapidly to MCP–CheW–CheA signaling complexes that effect subsequent amplification and slower CheY dephosphorylation. The high sensitivity indicates that this signal is generated by transport-induced dephosphorylation of the PTS rather than phosphoenolpyruvate consumption.

Organization and expression of the double-stranded RNA genome of Helminthosporium victoriae 190S virus, a totivirus infecting a plant pathogenic filamentous fungus.

The complete nucleotide sequence, 5178 bp, of the totivirus Helminthosporium vicotoriae 190S virus (Hv190SV) double-stranded RNA, was determined. Computer-assisted sequence analysis revealed the presence of two large overlapping ORFs; the 5'-proximal large ORF (ORF1) codes for the coat protein (CP) with a predicted molecular mass of 81 kDa, and the 3'-proximal ORF (ORF2), which is in the -1 frame relative to ORF1, codes for an RNA-dependent RNA polymerase (RDRP). Unlike many other totiviruses, the overlap region between ORF1 and ORF2 lacks known structural information required for translational frameshifting. Using an antiserum to a C-terminal fragment of the RDRP, the product of ORF2 was identified as a minor virion-associated polypeptide of estimated molecular mass of 92 kDa. No CP-RDRP fusion protein with calculated molecular mass of 165 kDa was detected. The predicted start codon of the RDRP ORF (2605-AUG-2607) overlaps with the stop codon (2606-UGA-2608) of the CP ORF, suggesting RDRP is expressed by an internal initiation mechanism. Hv190SV is associated with a debilitating disease of its phytopathogenic fungal host. Knowledge of its genome organization and expression will be valuable for understanding its role in pathogenesis and for potential exploitation in the development of biocontrol measures.

Genomic DNA fingerprinting by restriction fragment end labeling.

A typing method for bacteria was developed and applied to several species, including Escherichia coli and Actinobacillus actinomycetemcomitans. Total genomic DNA was digested with a restriction endonuclease, and fragments were enabled with [alpha-32P]dATP by using the Klenow fragment of DNA polymerase and separated by electrophoresis in 6% polyacrylamide/8 M urea (sequencing gel). Depending on the restriction endonuclease and the bacterium, the method produced approximately 30-50 well-separated fragments in the size range of 100-400 nucleotides. For A. actinomycetemcomitans, all strains had bands in common. Nevertheless, many polymorphisms could be observed, and the 31 strains tested could be classified into 29 distinct types. Furthermore, serotype-specific fragments could be assigned for the three serotypes investigated. The method described is very sensitive, allowing more distinct types to be distinguished than other commonly used typing methods. When the method was applied to 10 other clinically relevant bacterial species, both species-specific bands and strain-specific bands were found. Isolates from different locations of one patient showed indistinguishable patterns. Computer-assisted analysis of the DNA fingerprints allowed the determination of similarity coefficients. It is concluded that genomic fingerprinting by restriction fragment end labeling (RFEL) is a powerful and generally applicable technique to type bacterial species.

Quantitative analysis of senile plaques in Alzheimer disease: observation of log-normal size distribution and molecular epidemiology of differences associated with apolipoprotein E genotype and trisomy 21 (Down syndrome).

The discovery that the epsilon 4 allele of the apolipoprotein E (apoE) gene is a putative risk factor for Alzheimer disease (AD) in the general population has highlighted the role of genetic influences in this extremely common and disabling illness. It has long been recognized that another genetic abnormality, trisomy 21 (Down syndrome), is associated with early and severe development of AD neuropathological lesions. It remains a challenge, however, to understand how these facts relate to the pathological changes in the brains of AD patients. We used computerized image analysis to examine the size distribution of one of the characteristic neuropathological lesions in AD, deposits of A beta peptide in senile plaques (SPs). Surprisingly, we find that a log-normal distribution fits the SP size distribution quite well, motivating a porous model of SP morphogenesis. We then analyzed SP size distribution curves in genotypically defined subgroups of AD patients. The data demonstrate that both apoE epsilon 4/AD and trisomy 21/AD lead to increased amyloid deposition, but by apparently different mechanisms. The size distribution curve is shifted toward larger plaques in trisomy 21/AD, probably reflecting increased A beta production. In apoE epsilon 4/AD, the size distribution is unchanged but the number of SP is increased compared to apoE epsilon 3, suggesting increased probability of SP initiation. These results demonstrate that subgroups of AD patients defined on the basis of molecular characteristics have quantitatively different neuropathological phenotypes.

Detección automática con snakes y representación 3D sobre imágenes cerebrales

Comunicación presentada en la VI Conferencia de la Asociación Española para la Inteligencia Artificial (CAEPIA'95), Alicante, 15-17 noviembre 1995.

Spanish Nursing Students' Practical Experience of Computer Applications in Nursing

This study aimed to determine the level of computer practical experience in a sample of Spanish nursing students. Each student was given a Spanish language questionnaire, modified from an original used previously with medical students at the Medical School of North Carolina University (USA) and also at the Education Unit of Hospital General Universitario del Mar (Spain). The 10-item self-report questionnaire probed for information about practical experience with computers. A total of 126 students made up the sample. The majority were female (80.2%; n=101). The results showed that just over half (57.1%, n=72) of the students had used a computer game (three or more times before), and that only one third (37.3%, n=47) had the experience of using a word processing package. Moreover, other applications and IT-based facilities (e.g. statistical packages, e-mail, databases, CD-ROM searches, programming languages and computer-assisted learning) had never been used by the majority of students. The student nurses' practical experience was less than that reported for medical students in previous studies.

Aplicación del dibujo arquitectónico en 3D para el análisis histórico de los sistemas constructivos industrializados

La condición tridimensional de la construcción edificatoria precisa del uso del dibujo en 3D como la mejor herramienta de proyecto y transmisión de conocimientos técnicos y formales. El objetivo de esta comunicación es mostrar la aplicación de la expresión gráfica en 3D en un análisis histórico sobre la evolución de la envolvente industrializada en arquitectura, identificando sus principales condicionantes técnicos y constructivos. El estudio compara la evolución del uso de sistemas constructivos industrializados mediante un análisis gráfico de las soluciones constructivas más destacables. La metodología empleada se basa en la identificación y estudio de determinados sistemas constructivos industrializados compuestos por materiales ligeros así como de obras de arquitectura representativas por su influencia en la evolución de la envolvente arquitectónica en la segunda mitad del siglo XX. La representación gráfica en 3D ayuda a comparar las obras analizadas desde aspectos tecnológicos y formales, constatándose la utilidad del dibujo asistido por ordenador en el análisis constructivo realizado. En conclusión, el uso del dibujo arquitectónico en 3D contribuye, por la mejor comprensión de las características espaciales de las soluciones constructivas, al análisis de las propiedades materiales y funcionales de los sistemas constructivos industrializados y su aplicación al diseño arquitectónico, ayudando a perfeccionar su conocimiento e incrementando la calidad constructiva y compromiso social de las propuestas arquitectónicas.

A user's manual for the CALINE-2 computer program. Interim report.

Mode of access: Internet.

RASCEL: a programmable analog computer based on a regular array of stochastic computing element logic

Thesis--University of Illinois.

The role of the university in the computer age,

Includes bibliographical references.

The design of an economically viable large-scale computer based education system /

"February, 1969"

Computer access in higher education for students with disabilities : a practical guide to the selection and use of adapted computer technology /

Mode of access: Internet.

New Techniques in Deep Representation Learning

Thesis (Ph.D.)--University of Washington, 2016-06

Erectile Dysfunction Drug Use, Risk Behavior, and Sexually Transmitted Diseases

Thesis (Master's)--University of Washington, 2016-06

Constructing and evaluating alternative prediction models for identifying clinically relevant cases in an STD clinic

Thesis (Master's)--University of Washington, 2016-06