Identification and characterization of the PutP proline permease that contributes to in vivo survival of Staphylococcus aureus in animal models

Staphylococcus aureus is an important pathogen of humans and other animals, causing bacteremia, abscesses, endocarditis, and other infectious syndromes. A signature-tagged mutagenesis (STM) system was adapted for use in studying the genes required for in vivo survival of S. aureus. An STM library was ultimately created in S. aureus RN6390, with Tn917 being used to create the transposon mutations. Pools of S. aureus RN6390 mutants were screened in mouse abscess, bacteremia, and wound infection models for growth attenuation after in vivo passage. One of the mutants that was identified displayed marked attenuation following large-pool screening in all three animal models, which was confirmed in bacteremia and endocarditis models of infection with a smaller pool of mutants. Sequence analysis of the entire open reading frame showed a 99% identity to the high-affinity proline permease (putP) gene characterized in another strain of S. aureus. In wound and murine abscess infection models, the putP mutant was approximately 10-fold more attenuated than was wild-type strain RN6390. Another S. aureus strain transduced with the putP mutation also displayed an attenuated phenotype after passage in the wound model. A [3H]proline uptake assay showed that less proline was specifically transported into the putP mutant than into strain RN6390. The reduced viability of the bacteria possessing the mutation in the S. aureus high-affinity proline permease suggests that proline scavenging by the bacteria is important for in vivo growth and proliferation and that analogs of proline may serve as potential antistaphylococcal therapeutic agents.     

SEMI-ELLIPTICAL FATIGUE CRACK GROWTH UNDER ROTATING OR REVERSED BENDING COMBINED WITH STEADY TORSION

Abstract— An analysis of the influence of steady torsion loading on fatigue crack growth rates under rotating or reversed bending is presented. Mixed-mode (I + III) tests were carried out on cylindrical specimens in DIN Ck45k steel and results are compared for two different testing machines: rotary bending and reversed bending obtained by cyclic Mode I (Δ K ₁) with or without superimposed static Mode III ( K _III) loading, simulating the real conditions on power rotor shafts where many failures occur. The growth and shape evolution of semi-elliptical surface cracks, starting from a chordal notch on the cylindrical specimen surface, was measured for several Mode III/ Mode I ratios. Results have shown that the steady Mode III loading superimposed on the cyclic mode I leads to a significant reduction in the crack growth rates. It is suggested that this retardation is related to an increase of plastic zone size near the cylindrical surface in association with the interlocking of rough fracture surfaces, friction and fretting debris, leading to a decrease of the ΔK effective at the crack tip profile due to the "crack closure effect". This work provides a contribution to a better understanding of crack growth rates under mixed-mode load conditions thereby allowing one to predict remaining lifetimes and to estimate the risks of pre-cracked rotor shafts.     

Evaluation of the role of the Yersinia pestis plasminogen activator and other plasmid-encoded factors in temperature-dependent blockage of the flea

Yersinia pestis, the plague bacillus, has a plasminogen activator (pla) gene on the 9.5-kb plasmid pPla that is hypothesized to play a role in producing the foregut blockage in the flea vector that precedes transmission. In this study, however, Y. pestis that lacked pPla, the 70-kb virulence plasmid, or both plasmids, proved able to block Xenopsylla cheopis fleas normally. Blockage rates decreased with increasing environmental temperature for fleas infected with either wild type or pPla- Y. pestis. Thus, procoagulant ability of the Y. pestis pla gene product does not mediate blockage, nor does its ability to induce fibrinolysis at>28 degreesC account for failure to block at elevated temperatures. A Y. pestis strain that lacked all or part of the third plasmid of 110 kb, however, failed to colonize the flea midgut normally, indicating that one or more genes on the large plasmid may be required for vectorborne transmission.     

Natural attenuation of chlorinated organics in a shallow sand aquifer

This work presents the second phase of a groundwater remediation program for the migration control of a 1,2-dichloroethane (1,2-DCA) contaminated plume which includes natural attenuation at a distance downgradient from the source area. The conceived system for the plume migration control, implemented just after a major accidental release of 1,2-DCA in the soil, included a 300 m long physical barrier (cement-bentonite diaphragm wall) and 12 extraction wells. Results of field investigations have provided evidence that 1,2-DCA was naturally biodegrading into vinyl chloride as well as ethene under the natural anaerobic-reducing conditions at the site. In that case, source control measures were implemented to accelerate the overall remediation process. Although the results are favorable, the natural degradation of the 1,2-DCA does not guarantee acceptable levels of concentrations. Therefore, a pilot test to evaluate the enhancement of these processes is being carried out through the use of a biosparging system. This test is being implemented near the source to achieve sequential aerobic–anaerobic treatment zones.     

Improving quality and quantity of contributions: Two models for promoting knowledge exchange with shared databases

Shared databases are used for knowledge exchange in groups. Whether a person is willing to contribute knowledge to a shared database presents a social dilemma: Each group member saves time and energy by not contributing any information to the database and by using the database only to retrieve information which was contributed by others. But if all people use this strategy, then the database will be empty and, hence, useless for every group member. Based on theoretical approaches, two models for fostering the information-sharing behavior of database users are presented. One for enhancing the quality of database contents, and one for enhancing the quantity of those contents. The models take into account the following factors: the kinds of rewards the participants obtain for contributing information, the individual costs associated with this contribution, the prospective metaknowledge about the importance of one’s own information to the others, and the retrospective metaknowledge about how much others contributed to and retrieved from the database. These factors enhance the quantity of database contents as well as their quality.     

Performance Evaluation of a Novel Hybrid Multipulse Rectifier for Utility Interface of Power Electronic Converters

This paper presents an improved analysis of a novel programmable power-factor-corrected-based hybrid multipulse power rectifier (PFC-HMPR) for utility interface of power electronic converters. The proposed hybrid multipulse rectifier is composed of an ordinary three-phase six-pulse diode-bridge rectifier (Graetz bridge) with a parallel connection of single-phase switched converters in each three-phase rectifier leg. In this paper, the authors present a complete discussion about the controlled rectifiers' power contribution and also a complete analysis concerning the total harmonic distortion of current that can be achieved when the proposed converter operates as a conventional 12-pulse rectifier. The mathematical analysis presented in this paper corroborate, with detailed equations, the experimental results of two 6-kW prototypes implemented in a laboratory.     

Analysis of Scalar Maps for the Segmentation of the Corpus Callosum in Diffusion Tensor Fields

Diffusion tensor imaging (DTI) is a powerful technique for imaging axonal anatomy in vivo and its automatic segmentation is important for quantitative analysis and visualization. Application of the watershed transform is a recent approach for robustly segmenting diffusion tensor images. Since an important step of the watershed-based segmentation is the gradient computation, this paper investigates scalar maps from DTI and their ability to enhance borders and, therefore, their usefulness in gradient calculation. A comparison between existing scalar maps is conducted in the context of segmentation. New diffusion scalar maps, inspired by mathematical morphology concepts are proposed and included in the comparison. The watershed transform is then applied to segment the corpus callosum, based on the computed scalar maps.     

A genetic programming method for protein motif discovery and protein classification

Proteins can be grouped into families according to some features such as hydrophobicity, composition or structure, aiming to establish common biological functions. This paper presents MAHATMA—memetic algorithm-based highly adapted tool for motif ascertainment—a system that was conceived to discover features (particular sequences of amino acids, or motifs) that occur very often in proteins of a given family but rarely occur in proteins of other families. These features can be used for the classification of unknown proteins, that is, to predict their function by analyzing their primary structure. Experiments were done with a set of enzymes extracted from the Protein Data Bank. The heuristic method used was based on genetic programming using operators specially tailored for the target problem. The final performance was measured using sensitivity, specificity and hit rate. The best results obtained for the enzyme dataset suggest that the proposed evolutionary computation method is effective in finding predictive features (motifs) for protein classification.     

Global ESRD Costs Associated with a Short Daily Hemodialysis Program in the United States

In spite of the growing evidence that daily hemodialysis (DHD) improves clinical outcomes and quality of life, the additional dialysis costs are not currently reimbursed in the United States. Nor have there been reports of the effects of DHD on end-stage renal disease (ESRD) global costs, which would help predict the financial impact of DHD on the ESRD program. Since 1996, 22 patients (20 in-center, 2 home) have switched from conventional thrice-weekly dialysis to short, daily dialysis with six treatments per week. Eighteen patients started for medical indications, and four started for nonmedical reasons. Causes of ESRD were the following: diabetes mellitus (6), hypertension (4), glomerulonephritis (6), hereditary (2), and other (4). Mean age was 56 ± 16 years. Patients had an average of 3.3 major comorbidities. Weekly conventional HD dialysis times were divided into six DHD treatments, each 2.0 ± 0.3 hours. Weekly Kt/V remained unchanged. Twenty-two patients were followed on DHD for 220 patient-months: 7 patients died after 1.8 ± 1.3 months, 2 were transplanted at 4.3 ± 3.2 months, and 2 discontinued DHD at 3.6 ± 4.8 months. Eleven patients remain on DHD at 17.4 ± 8.3 months. Actual costs per extra dialysis session are as follows: $14.30 for supplies and $3.20 for labor for setup/cleanup time (15 minutes at $12.80/hour). Annualized DHD savings are based on comparison of doses of epoetin alpha (Epogen) and blood pressure medication at the start and after 12 months of DHD. Hospitalization rates include all enrolled patients, comparing rates for the 12 months prior to DHD with the first year on DHD, or annualized rates for those on DHD less than one year. Cost assumptions are $9/ 1000 U Epogen, $1/blood pressure pill, and $1200/per day of hospitalization. Extra transportation costs were covered by the patients. No increased access problems were observed. For patients on short DHD longer than 12 months, supply and labor costs increased to $2733/patient/year; however, Epogen use was reduced 55%, and blood pressure medications were reduced 40%. For all patients who switched to DHD, hospitalization rates were reduced 24%. This resulted in a net savings of about $4241/patient/ year after 12 months on DHD. Overall ESRD costs were substantially decreased on DHD. These cost savings must be passed on to providers before DHD becomes more widely available.