Structure-based design of inhibitors specific for bacterial thymidylate synthase

Thymidylate synthase is an attractive target for antiproliferative drug design because of its key role in the synthesis of DNA. As such, the enzyme has been widely targeted for anticancer applications. In principle, TS should also be a good target for drugs used to fight infectious disease. In practice, TS is highly conserved across species, and it has proven to be difficult to develop inhibitors that are selective for microbial TS enzymes over the human enzyme. Using the structure of TS from Lactobacillus casei in complex with the nonsubstrate analogue phenolphthalein, inhibitors were designed to take advantage of features of the bacterial enzyme that differ from those of the human enzyme. Upon synthesis and testing, these inhibitors were found to be up to 40-fold selective for the bacterial enzyme over the human enzyme. The crystal structures of two of these inhibitors in complex with TS suggested the design of further compounds. Subsequent synthesis and testing showed that these second-round compounds inhibit the bacterial enzyme at sub-micromolar concentrations, while the human enzyme was not inhibited at detectable levels (selectivities of 100-1000-fold or greater). Although these inhibitors share chemical similarities, X-ray crystal structures reveal that the analogues bind to the enzyme in substantially different orientations. Site-directed mutagenesis experiments suggest that the individual inhibitors may adopt multiple configurations in their complexes with TS.     

Determinants of research follow-up participation in an alcohol treatment outcome trial

We have previously reported a common variation in the liver promoter of the human glucokinase, which is regulated by insulin, in the patients with non-insulin-dependent diabetes mellitus (NIDDM). The variation occurred within a 10-bp region completely conserved between human and rat. Its basic motif was almost identical to the insulin regulatory element of the phosphoenolpyruvate carboxykinase gene. In vitro transfection experiment showed that the G-to-A variation causes a 58% reduction in the promoter activity. After oral glucose challenge, the homozygous A/A subjects had the highest stimulated insulin levels at 60 and 90 minutes and the highest insulin area under the curve as compared to the subjects with other genotypes, which suggested the homozygous A/A subjects were more insulin resistant. As insulin resistance is a risk factor of NIDDM, we concluded that this promoter variation is a risk factor for NIDDM.     

Enhanced negative priming in Parkinson's disease.

In the ignored repetition paradigm, negative priming (NP) is defined as the increase in response time that occurs when the current target stimulus served as a distractor stimulus in the previous trial. In this study, 25 Parkinson's disease (PD) participants and 17 age-matched healthy controls (HCs) were tested using a touchscreen version of the ignored repetition task that allowed response time to be partitioned into response initiation and response execution segments. In both groups, NP effects were stronger in the response execution than in the response initiation segments. The most striking result was that the PD group showed larger NP effects overall than the HC group. In PD, clinical ratings of bradykinesia, but not tremor, were related to larger NP effects. Results indicate that in PD, disruption of dopamine neuromodulation diminishes response efficiency when action must be directed toward previously ignored information. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Status and cockpit dynamics: A review and empirical study.

One of the most troublesome dynamics evident in the airplane cockpit is related to patterns of authority relations between the captain and the first officer. Too often, captains fail to listen and first officers fail to speak. The authors propose that many instances of superordinate and subordinate behavior in the cockpit—the captain's tendency to reject input from other team members and the first officer's hesitancy to question the captain—represent cases of status generalization. First, the authors describe the theory of status generalization and show support for the operation of the theory by presenting examples of flightcrew behavior that the theory predicts. Second, an initial empirical test was conducted to instantiate the claim that captain–first officer differences can be seen as status differences. Finally, the significance and implications of this perspective are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Design data for externally pressurised spherical gas bearings

Radial and thrust loading characteristics are given for both single row entry and double row entry bearings. The design data presented gives a basis for comparison of the two bearing configurations. The paper shows that the double entry configuration reduces the negative stiffness regions which occur in single entry bearings. Finally a procedure is given to enable the designer to select a bearing to meet his particular requirements.     

Massive haemoptysis death and other morbidity associated with high dose rate intraluminal radiotherapy for carcinoma of the bronchus

Fluctuation analysis allows for the determination of mutation rates in cell cultures in vitro. As originally described by Luria and Delbruck and extended by Lea and Coulson and by Capizzi and Jameson, this analysis has been useful in estimating mutation rates in cultured cells where the frequency of mutational events is low. However. in cultures where high mutation rates and multiple independent mutation events occur, leading to the accumulation of many mutant cells, these standard methods may not apply. Here, we present a new method for the estimation of mutation rates based on the assumption that multiple events may contribute to the accumulation of mutant cells. We compared mutation rates determined by Lea and Coulson's and by Capizzi and Jameson's methods with those determined by our method using experimental and stimulated data from our studies of immunoglobulin gene mutation and isotype switching in B lymphocyte cultures. The three methods resulted in very different calculated rates when many mutants were present in the culture, such as when mutation rates were high, while only small differences in calculated rates were found when mutants were rare. Unlike previous fluctuation analysis calculations, our method is applicable for the estimation of both low and high rates.     

Controlling Legionella in hospital water systems: experience with the superheat-and-flush method and copper-silver ionization

OBJECTIVE: To evaluate the effect of copper-silver ionization on Legionella colonization and nosocomial legionnaires' disease and to compare the efficacy of metal ions versus the superheat-and-flush method of disinfection. DESIGN: Prospective determination over a 36-month period of copper and silver ion concentrations in the recirculating hot-water system, Legionella colonization of the hospital water distribution system, and cases of nosocomial legionnaires' disease. Retrospective comparison of results with the previous 13 years, during which the superheat-and-flush method was used. SETTING: The Pittsburgh Veterans' Affairs Health Care System (University Drive Division) acute-care hospital. INTERVENTION: Three copper-silver ionization systems were installed on the hot-water distribution system in November 1994. RESULTS: The average number of cases of legionnaires' disease per year and the percentage of distal sites positive for Legionella pneumophila for the superheat-and-flush method versus the copper-silver ionization method was six cases with 15% positivity versus two cases with 4% positivity, respectively. The reduction in Legionella colonization after copper-silver ionization was significant (P<.05) compared to the superheat and flush. Mean copper and silver ion concentrations (mg/L) were 0.29 and 0.054 from hot-water tanks, and 0.17 and 0.04 from distal outlets, respectively. CONCLUSIONS: We conclude that a properly maintained and monitored copper-silver ionization system was more effective than the superheat-and-flush method for reducing the recovery of Legionella from the hospital water distribution system.