Mechanism of irreversible inactivation of phosphomannose isomerases by silver ions and flamazine

Silver ions and silver-containing compounds have been used as topical antimicrobial agents in a variety of clinical situations. We have previously shown that the enzyme phosphomannose isomerase (PMI) is essential for the biosynthesis of Candida albicans cell walls. In this study, we find that PMI can be inhibited by silver ions. This process is shown to be irreversible, and is a two-step process, involving an intermediate complex with a dissociation constant, Ki, of 59 +/- 8 microM, and a maximum rate of inactivation of 0.25 +/- 0.04 min-1 in 50 mM Hepes buffer, pH 8.0 at 37 degrees C. The enzyme can be protected against this inactivation by the substrate mannose 6-phosphate, with a dissociation constant of 0.31 +/- 0.04 mM, close to its Km value. Flamazine (silver sulfadiazine) is a silver-containing antibiotic which is used clinically as a topical antimicrobial and antifungal agent. We compared the ability of silver sulfadiazine and two other silver-containing compounds to irreversibly inactivate C. albicans PMI. The addition of the organic moiety increased the affinity of the compounds, with silver sulfadiazine showing a Ki of 190 +/- 30 nM. In all cases, the maximum inhibition rate was similar, implying a similar rate-determining step. Silver sulfadiazine does not inhibit Escherichia coli PMI, and this suggests a role of the only free cysteine, Cys-150, in the inactivation process. To confirm this, we mutated this residue to alanine in C. albicans PMI. The resultant Cys150 --> Ala mutant protein showed similar Vm and Km values to the wild-type enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)     

Peak expiratory flow rate control chart in asthma care: chart construction and use in asthma care

We have analyzed X-chromosome inactivation patterns in lymphocytes of 264 females from 38 families not known to have any genetic disease. Quantitative measures of X-inactivation showed strong sister-sister correlation in the degree of departure from equal numbers of cells having each X chromosome active, suggesting heritability of this phenotype. Strong sister-sister correlation was also observed for the fraction of cells having the same parent's X chromosome active, consistent with the possibility that this trait might be controlled by a cis-acting, X-linked gene. We used a sib-pair approach to determine whether X-inactivation phenotype was linked to loci in any region of the X chromosome. Both quantitative and discrete measures of X-inactivation phenotype showed evidence of linkage to markers in the region of the X inactivation center (XIC). The quantitative measure of X-inactivation phenotype used in our study also showed linkage to loci at Xq25-q26. This study provides the first evidence for X-linked inheritance of X chromosome inactivation phenotype derived from linkage analysis in phenotypically normal human families.     

Children's beliefs about drinking

This paper reports the results of a study in which age (grade level), racial/ethnic, and gender differences in beliefs and perceived norms about drinking were examined in a multi-ethnic urban sample of 4th through 7th grade children. Results showed that older children held beliefs and perceived norms that were more favorable toward drinking than younger children. The major difference between older and younger children lay in their differential estimates of the likelihood of certain consequences occurring and not in their evaluation of these consequences of drinking. Further, older children not only displayed less motivation to comply with their parents and greater motivation to comply with their peers, but they also perceived their parents, as well as their peers, as less disapproving of drinking than did younger children. There were few gender or race/ethnicity differences at these ages in children's beliefs and perceived norms about drinking.     

Efficient synthesis of gaussian filters by cascaded uniform filters

Gaussian filtering is an important tool in image processing and computer vision. In this paper we discuss the background of Gaussian filtering and look at some methods for implementing it. Consideration of the central limit theorem suggests using a cascade of ``simple' filters as a means of computing Gaussian filters. Among ``simple' filters, uniform-coefficient finite-impulse-response digital filters are especially economical to implement. The idea of cascaded uniform filters has been around for a while [13], [16]. We show that this method is economical to implement, has good filtering characteristics, and is appropriate for hardware implementation. We point out an equivalence to one of Burt's methods [1], [3] under certain circumstances. As an extension, we describe an approach to implementing a Gaussian Pyramid which requires approximately two addition operations per pixel, per level, per dimension. We examine tradeoffs in choosing an algorithm for Gaussian filtering, and finally discuss an implementation.     

Investigating T Cell Immunity in Cancer: Achievements and Prospects

T cells play a key role in tumour surveillance, both identifying and eliminating transformed cells. However, as tumours become established they form their own suppressive microenvironments capable of shutting down T cell function, and allowing tumours to persist and grow. To further understand the tumour microenvironment, including the interplay between different immune cells and their role in anti-tumour immune responses, a number of studies from mouse models to clinical trials have been performed. In this review, we examine mechanisms utilized by tumour cells to reduce their visibility to CD8⁺ Cytotoxic T lymphocytes (CTL), as well as therapeutic strategies trialled to overcome these tumour-evasion mechanisms. Next, we summarize recent advances in approaches to enhance CAR T cell activity and persistence over the past 10 years, including bispecific CAR T cell design and early evidence of efficacy. Lastly, we examine mechanisms of T cell infiltration and tumour regression, and discuss the strengths and weaknesses of different strategies to investigate T cell function in murine tumour models.     

Premodification of Pt/γAl2O3 with Cinchonidine for the Enantioselective Hydrogenation of Ethyl Pyruvate: Effect of Premodification Conditions on Reaction Rate and Enantioselection

The premodification of a 5 wt% Pt/-Al₂O₃ catalyst with cinchonidine (0.01 and 0.2 g g^-1 _catalyst) is described and discussed. Premodification is carried out by treating the catalyst with a solution of cinchonidine followed by solvent removal. Catalysts premodified in this way give the same ee and initial rate of reaction for the enantioselective hydrogenation of ethyl pyruvate as those using the standard in situ modification procedure. Investigations of different solvents for premodification and reaction (dichloromethane, ethanol) show that it is the solvent used for the reaction that controls the observed enantioselection. Premodified catalysts also display the initial transient behavior typically observed with in situ modified catalysts in which the ee increases with conversion in the early part of the reaction. Premodified catalysts show an enhanced rate of reaction when ethanol is used as the reaction solvent compared with in situ modified catalysts under the same conditions. Premodification using aerobic conditions gives the best results and premodified catalysts can be stored prior to use for up to a week without loss of catalytic performance.     

Evidence of an oviposition stimulant for peachtree borer

Peachtree borer (PTB) cocoons, peach tree bark, and a mixture of frass and gum obtained arount PTB wounds were extracted with organic solvents of increasing polarity. Female PTB moths,Synanthedon exitiosa (Say), laid the greatest number of eggs on filter paper substrates treated with acetone extracts of PTB cocoons, ethyl acetate extracts of peach tree bark and acetone extracts of frass and gum mixtures. The extracts of cocoons or frass and gum mixture were separated by liquid chromatography and thin layer chromatography.Synanthedon exitiosa (Say).Mention of a commercial or proprietory product does not constitute an endorsement of this product by the USDA.