Cloning and characterization of the genes of the CeqI restriction-modification system

Two genes from Corynebacterium equii, a Gram-positive bacterium producing the CeqI restriction-modification enzymes were cloned and sequenced. In vivo restriction experiments, DNA and amino acid sequence data suggest that the two genes code for the endonuclease and the methyltransferase enzymes. However, when the two genes are expressed in E. coli, practically no enzyme activity can be detected in the supernatants of sonicated cells. Based on the DNA sequence data CeqI restriction endonuclease (an EcoRV izoschizomer) consists of 270 amino acid residues with a predicted molecular mass of 31.6 kDa, in good agreement with the previously measured 32 +/- 2 kDa. The methyltransferase is 517 residues long (approx. 60 kDa). The two genes are in opposite orientation and overlap by 37 base pairs on the chromosome. The deduced amino acid sequence of the putative endonuclease gene revealed long stretches of hydrophobic amino acids, that may form the structural basis of the unusual aggregation properties of the restriction endonuclease. The amino acid sequence of the methylase shows homologies with other type II methyltransferases.     

Two-dimensional map of Haemophilus influenzae following protein enrichment by heparin chromatography

Two-dimensional gel electrophoresis separates several hundred protein molecules in one single experiment and is efficiently used to study the products expressed by different genomes. Low-copy-number gene products are invisible on a stained two-dimensional map and must be enriched such that sufficient amounts are present for visualization and identification. We investigated the enrichment of proteins of the bacterium Haemophilus influenzae by chromatography on immobilized heparin which has affinity for growth and protein biosynthesis factors. Total soluble proteins of the microorganism were fractionated on Heparin-Actigel which resulted in enrichment of approximately 160 proteins. The eluates, representing about 40% of the applied proteins, were analyzed by two-dimensional gel electrophoresis and the protein spots were characterized by amino acid composition analysis and matrix-assisted laser desorption ionization mass spectrometry. The proteins enriched by chromatography on the heparin gel were not exclusively low-copy-number gene products and they did not exclusively belong to one single class of proteins. The proteins that bound to the heparin gel are indicated in a two-dimensional protein map which includes more than 110 newly identified proteins.     

Career development in adolescence as a family project.

This study conceptualizes and investigates career-relevant parent–child conversations and other actions over time as a family project. Dyads composed of a parent and an adolescent from 20 families participated in a videotaped career-related conversation to determine a family career-development project that was subsequently monitored for a 6-month period and followed up with a 2nd videotaped conversation. On the basis of a systematic qualitative analysis, several dimensions were identified as facilitating the family career-development project, including joint goals, communication, goals-steps congruence, and individuation. These family career-development projects were organized as part of broader relationship, identity, parenting, and cultural projects that also played a decisive role in the success of the family career-development projects themselves. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Lipid peroxidation and antioxidant system changes in acute L-arginine pancreatitis in rats

The World Wide Web-based form is a promising method for the construction of an on-line data collection system for clinical and epidemiological research. It is, however, laborious to prepare a common gateway interface (CGI) program for each project, which the World Wide Web server needs to handle the submitted data. In medicine, it is even more laborious because the CGI program must check deficits, type, ranges, and logical errors (bad combination of data) of entered data for quality assurance as well as data length and meta-characters of the entered data to enhance the security of the server. We have extended the specification of the hypertext markup language (HTML) form to accommodate information necessary for such data checking and we have developed software named AUTOFORM for this purpose. The software automatically analyzes the extended HTML form and generates the corresponding ordinary HTML form, 'Makefile', and C source of CGI programs. The resultant CGI program checks the entered data through the HTML form, records them in a computer, and returns them to the end-user. AUTOFORM drastically reduces the burden of development of the World Wide Web-based data entry system and allows the CGI programs to be more securely and reliably prepared than had they been written from scratch.     

Orientation-Dependent Developments in Misorientation and Residual Stress in Rolled Aluminum: The Defining Role of Dislocation Interactions

Orientation-dependent developments in misorientation and residual stress, in rolled aluminum, were quantified experimentally and simulated numerically. The latter involved analysis using a crystal plasticity finite element model, accounting for anisotropies in slip system hardening but neglecting near-neighbor interactions, and discrete dislocation dynamics of the single crystals. Both were successful in capturing the experimental patterns of orientation dependence. Numerical simulations, without slip transfer across the neighboring grains, thus established the defining role of dislocation interactions in establishing orientation-sensitive microstructural evolution.     

Contingent Use of Negotiators’ Tactics in Construction Dispute Negotiation

In the course of negotiation, negotiators’ tactics should be responsive to the situational factors. This is commonly described as the contingent use of negotiators’ tactics. This study examines this concept in construction dispute negotiation and has three stages of work. Stage 1 develops taxonomies of the three construction dispute negotiation dimensions: dispute sources, negotiators’ tactics, and negotiation outcomes by exploratory factor analysis. A structural equation modeling is also used to confirm the taxonomies. Stage 2 examines the contingent use of negotiators’ tactics on outcomes respective to the dispute sources through the use of moderated multiple regression (MMR). Stage 3 discusses the findings. The dispute source, “Delay” is found to be a universal moderator in the MMR analysis of the tactic-outcome relationships. That means when the dispute source is delay, a wide range of negotiators’ tactics can be used, respective to outcome intended. It is also found that the most versatile tactics are those that seek progress. This group of tactics is effective in almost every group of dispute source and, in general, positive results can be expected. However, aggressive and assertive tactics should be used restrictively, as they will only be useful against a compromising negotiation counterpart.     

Using H3+ and H2D+ as probes of star-forming regions

The H3+ and H2D+ ions are important probes of the physical and chemical conditions in regions of the interstellar medium where new stars are forming. This paper reviews how observations of these species and of heavier ions such as HCO+ and H3O+ can be used to derive chemical and kinematic properties of nearby pre-stellar cores and the cosmic ray ionization rate towards more distant regions of high-mass star formation. Future prospects in the field are outlined at the end.     

Determination of small angular displacement by moiré fringes of matched radial-parallel gratings

We investigate the properties of the moiré fringes of matched radial-parallel (MRP) gratings that are useful for the fine visual indication of angular displacements. Moiré patterns formed by overlapping two different MRP gratings of angular pitches, P(theta) and P(theta)? (相似文献   

Microscopic measurement of cellular infiltration in the rheumatoid arthritis synovial membrane: a comparison of semiquantitative and quantitative analysis

BACKGROUND: A significant proportion of burn patients with inhalation injuries incur difficulties with airway protection, dysphagia, and aspiration. In assessing the need for intubation in burn patients, the efficacy of fiberoptic laryngoscopy was compared with clinical findings and the findings of diagnostic tests, such as arterial blood gas analysis, measurement of carboxyhemoglobin levels, pulmonary function tests, and radiography of the lateral aspect of the neck. OBJECTIVE: To determine if these patients were at risk for aspiration or dysphagia, barium-enhanced fluoroscopic swallowing studies were performed. DESIGN: Prospective study. SETTINGS: Burn intensive care unit in an academic tertiary referral center. MAIN OUTCOME MEASURES: Need for endotracheal intubation and potential for aspiration. RESULTS: Six (55%) of 11 patients had clinical findings and symptoms that indicated, under traditional criteria, endotracheal intubation for airway protection. Visualization of the upper airway with fiberoptic laryngoscopy obviated the need for endotracheal intubation in all 11 patients. These patients also failed to evidence an increased risk of aspiration or other swallowing dysfunction. CONCLUSIONS: In comparison with other diagnostic criteria, fiberoptic laryngoscopy allows differentiation of those patients with inhalation injuries who, while at risk for upper airway obstruction, do not require intubation. These patients may be safely observed in a monitored setting with serial fiberoptic examinations, thus avoiding the possible complications associated with intubation of an airway with a compromised mucosalized surface. In these patients, swallowing abnormalities do not manifest.