Non-coding nucleotides and amino acids near the active site regulate peptide deformylase expression and inhibitor susceptibility in Chlamydia trachomatis

Chlamydia trachomatis, an obligate intracellular bacterium, is a highly prevalent human pathogen. Hydroxamic-acid-based matrix metalloprotease inhibitors can effectively inhibit the pathogen both in vitro and in vivo, and have exhibited therapeutic potential. Here, we provide genome sequencing data indicating that peptide deformylase (PDF) is the sole target of the inhibitors in this organism. We further report molecular mechanisms that control chlamydial PDF (cPDF) expression and inhibition efficiency. In particular, we identify the σ66-dependent promoter that controls cPDF gene expression and demonstrate that point mutations in this promoter lead to resistance by increasing cPDF transcription. Furthermore, we show that substitution of two amino acids near the active site of the enzyme alters enzyme kinetics and protein stability

Design Optimization of Structures with Shape Memory Alloy Member Actuators

Thesis (Master's)--University of Washington, 2013A capability was developed to build a perform mass-minimization of a truss with SMA actuation to determine optimal element diameters and actuator applied temperature changes. The Nonlinear Programming/Approximation Concepts (NLP/AC) approach was used. This method is based on using a sequence of approximate optimization problems rather than optimizing with the full nonlinear analyses. Some special approximations based on new intermediate variables were used. Results are presented and analyzed

A New GUI for Global Orbit Correction at the Als Using Matlab

Orbit correction is a vital procedure at particle accelerators around the world. The orbit correction routine currently used at the Advanced Light Source (ALS) is a bit cumbersome and a new Graphical User Interface (GUI) has been developed using MATLAB. The correction algorithm uses a singular value decomposition method for calculating the required corrector magnet changes for correcting the orbit. The application has been successfully tested at the ALS. The GUI display provided important information regarding the orbit including the orbit errors before and after correction, the amount of corrector magnet strength change, and the standard deviation of the orbit error with respect to the number of singular values used. The use of more singular values resulted in better correction of the orbit error but at the expense of enormous corrector magnet strength changes. The results showed an inverse relationship between the peak-to-peak values of the orbit error and the number of singular values used. The GUI interface helps the ALS physicists and operators understand the specifi c behavior of the orbit. The application is convenient to use and is a substantial improvement over the previous orbit correction routine in terms of user friendliness and compactness

U1A Inhibits Cleavage at the Immunoglobulin M Heavy-Chain Secretory Poly(A) Site by Binding between the Two Downstream GU-Rich Regions

The immunoglobulin M heavy-chain locus contains two poly(A) sites which are alternatively expressed during B-cell differentiation. Despite its promoter proximal location, the secretory poly(A) site is not expressed in undifferentiated cells. Crucial to the activation of the secretory poly(A) site during B-cell differentiation are changes in the binding of cleavage stimulatory factor 64K to GU-rich elements downstream of the poly(A) site. What regulates this change is not understood. The secretory poly(A) site contains two downstream GU-rich regions separated by a 29-nucleotide sequence. Both GU-rich regions are necessary for binding of the specific cleavage-polyadenylation complex. We demonstrate here that U1A binds two (AUGCN(1-3)C) motifs within the 29-nucleotide sequence and inhibits the binding of cleavage stimulatory factor 64K and cleavage at the secretory poly(A) site

RE-ENGINEERING THE WELFARE SYSTEM - A STUDY OF ADMINISTRATIVE CHANGES TO THE FOOD STAMP PROGRAM: FINAL REPORT

All States in a recent study undertook at least one "re-engineering" activity in their Food Stamp Programs (FSPs) as a result of the Personal Responsibility and Work Opportunity Reconciliation Act of 1996 (PRWORA). In addition, 35 States implemented changes in 3 or more re-engineering categories, while 24 States planned changes for fiscal year (FY) 2000 in 2 or more categories. PRWORA dramatically changed the systems that provide cash assistance and food stamps to low-income Americans. Along with mandatory changes in food stamp eligibility, States were given greater flexibility to administer their programs to meet their unique needs. While States had begun changing the way program services were delivered before passage of welfare reform legislation, PRWORA provided additional opportunities for them to "re-engineer" FSPs. The purpose of the study was to examine State-level administrative changes to FSPs as a result of PRWORA, both those made before FY 2000 and those planned for FY 2000. This report provides the study results. For the State data collection instrument, see Re-engineering the Welfare System-A Study of Administrative Changes to the Food Stamp Program: State Data Collection Instrument at http: //www.ers.usda.gov/publications/efan01009

STATE USE OF FUNDS TO INCREASE WORK SLOTS FOR FOOD STAMP RECIPIENTS: REPORT TO CONGRESS

Since passage of the Balanced Budget Act of 1997, most States have retargeted services in the Food Stamp Employment and Training (E&T) Program to able-bodied adults without dependents (ABAWDs). The Balanced Budget Act more than doubled funding to States for USDA's Food Stamp E&T Program. The law requires States to send at least 80 percent of the funding on services that can fulfill the work requirement imposed by welfare reform legislation in 1996 on ABAWDs. ABAWDs must either work at least 20 hours per week or participate in a qualifying employment and trainig activity. This report fulfills the Act's mandate to USDA to examine and report on how States use the new funds to create work opportunities for ABAWDs and if this is done in an efficient and effective manner. The results show that total E&T program spending increased, though States used less of their Federal grant allocations and more of State matching funds. Nationwide participation in the E&T program dropped sharply after the Balanced Budget Act, with variations among States and among E&T component types. States have specific recommendations for improving the program. This publication provides the findings of the study. For more information on the survey instruments used for the study, see State Use of Funds To Increase Work Slots for Food Stamp Recipients: State Data Collection Instruments at htt://www.ers.usda.gov/publications/efan01007

RE-ENGINEERING THE WELFARE SYSTEM - A STUDY OF ADMINISTRATIVE CHANGES TO THE FOOD STAMP PROGRAM: FINAL REPORT

All States in a recent study undertook at least one "re-engineering" activity in their Food Stamp Programs (FSPs) as a result of the Personal Responsibility and Work Opportunity Reconciliation Act of 1996 (PRWORA). In addition, 35 States implemented changes in 3 or more re-engineering categories, while 24 States planned changes for fiscal year (FY) 2000 in 2 or more categories. PRWORA dramatically changed the systems that provide cash assistance and food stamps to low-income Americans. Along with mandatory changes in food stamp eligibility, States were given greater flexibility to administer their programs to meet their unique needs. While States had begun changing the way program services were delivered before passage of welfare reform legislation, PRWORA provided additional opportunities for them to "re-engineer" FSPs. The purpose of the study was to examine State-level administrative changes to FSPs as a result of PRWORA, both those made before FY 2000 and those planned for FY 2000. This report provides the study results. For the State data collection instrument, see Re-engineering the Welfare System-A Study of Administrative Changes to the Food Stamp Program: State Data Collection Instrument at http://www.ers.usda.gov/publications/efan01009.PRWORA, service delivery, structure, casework, accessibility, client tracking, monitoring, conforming, Food Consumption/Nutrition/Food Safety, Food Security and Poverty,

The Central Role of cAMP in Regulating Plasmodium falciparum Merozoite Invasion of Human Erythrocytes

International audienceAll pathogenesis and death associated with Plasmodium falciparum malaria is due to parasite-infected erythrocytes. Invasion of erythrocytes by P. falciparum merozoites requires specific interactions between host receptors and parasite ligands that are localized in apical organelles called micronemes. Here, we identify cAMP as a key regulator that triggers the timely secretion of microneme proteins enabling receptor-engagement and invasion. We demonstrate that exposure of merozoites to a low K + environment, typical of blood plasma, activates a bicarbonate-sensitive cytoplasmic adenylyl cyclase to raise cytosolic cAMP levels and activate protein kinase A, which regulates microneme secretion. We also show that cAMP regulates merozoite cytosolic Ca 2+ levels via induction of an Epac pathway and demonstrate that increases in both cAMP and Ca 2+ are essential to trigger microneme secretion. Our identification of the different elements in cAMP-dependent signaling pathways that regulate microneme secretion during invasion provides novel targets to inhibit blood stage parasite growth and prevent malaria