Evaluation of DapE Inhibitors Utilizing the DapE Ninhydrin and Thermal Shift Assays Toward the Discovery of Novel Antibiotics

An ongoing effort continues against the rising tide of antibiotic resistant bacteria, underscoring the urgent need to discover antibiotics with a novel mechanism of action. To this end, we have focused on the inhibition of the bacterial enzyme N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE). Guided by docking with the computational suite Molecular Operating Environment (MOE) and lead molecules obtained through a HiTS (High Throughput Screening) assay, several lead molecules and analogs were identified, synthesized, and optimized by our research group. These potential new inhibitors are tested in our recently described and updated biochemical DapE ninhydrin assay, with IC50 data obtained for inhibitors including cyclobutanone, tetrazole, pyrazole, sultam, and indoline sulfonamide analogs. To gain a greater understanding of the thermodynamic effects of these inhibitors on the binding of these synthesized analogs with the DapE enzyme, we have employed a Thermal Shift Assay (TSA)

Evaluation of Various Analogs of High-Throughput Screening Target Molecules as DapE Inhibitors Using the DapE Ninhydrin Assay for Novel Antibiotic Discovery

In our medicinal organic chemistry research lab, I am performing a ninhydrin-based enzyme inhibition assay, previously developed in our lab, to discover novel antibiotics to combat antibiotic resistance that poses an imminent public health crisis. These drugs target the bacterial enzyme N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE), a crucial enzyme to bacterial survival due to its involvement in the biosynthetic pathway of lysine, an amino acid essential to protein synthesis and mDAP, which is essential for bacterial cell wall construction. This pathway is exclusive to bacteria and is not found in mammalian cells, thus inhibition of this enzyme should be selectively toxic to bacteria. The assay enables testing of potential new inhibitors of DapE, done by spectrophotometric analysis of the products of the in vitro enzyme assay, quantitatively analyzing the amount of enzyme substrate converted to product. The goal is to evaluate several newly discovered inhibitor series involving indoline sulfonamides, tetrazoles, and N-aryl sulfonamides. Studying analogs of these compounds will greatly aid our understanding of the enzyme dynamics involved in the inhibition process. This information will broaden our understanding of the enzyme’s catalytic machinery to significantly aid in our future endeavors of drug synthesis and drug discovery for the optimization of new inhibitors

A forward speed effects study on jet noise from several suppressor nozzles in the NASA/Ames 40- by 80-foot wind tunnel

A test program was conducted in a 40 by 80 foot wind tunnel to evaluate the effect of relative velocity on the jet noise signature of a conical ejector, auxiliary inlet ejector, 32 spokes and 104 tube nozzle with and without an acoustically treated shroud. The freestream velocities in the wind tunnel were varied from 0 to 103.6 m/sec (300 ft/sec) for exhaust jet velocities of 259.1 m/sec (850 ft/sec) to 609.6 m/sec (2000 ft/sec). Reverberation corrections for the wind tunnel were developed and the procedure is explained. In conjunction with wind tunnel testing the nozzles were also evaluated on an outdoor test stand. The wind tunnel microphone arrays were duplicated during the outdoor testing. The data were then extrapolated for comparisons with data measured using a microphone array placed on a 30.5 meter (100 ft) arc. Using these data as a basis, farfield to nearfield arguments are presented with regards to the data measured in the wind tunnel. Finally, comparisons are presented between predictions made using existing methods and the measured data

Diaconia e edificação de comunidade a partir de Balsas — Nordeste

This text retrieves part of what took place in the Lutheran Congregation in Balsas, state of Maranhão, between 1987 and 1995, which is the period when diaconal workers were active there. This is a case study that is used as a basis for reflection on issues related to diaconia, particularly in the realm of community/congregation building. Diaconia as a dimension of the church’s being inserts the diaconic ministry in the building and maintenance of Christian communities.Este trabajo rescata un poco de lo que sucedió en la Comunidad Evangélica de Confesión Luterana de Balsas, en Maranhão, entre los anos de 1987 y 1995, período cuando obreras diaconales actuaron allí. Se trata de un estudio de caso a partir del cual se quiere reflexionar algunas cuestiones vinculadas a la diaconía, específicamente en el ámbito de la edificación de comunidades. La diaconía, como una dimensión de ser iglesia, instala el ministerio diaconal en la construcción y manutención de comunidades cristianas.Este trabalho resgata um pouco do que aconteceu na Comunidade Evangélica de Confissão Luterana de Balsas, no Maranhão, entre os anos de 1987 e 1995, período em que obreiras diaconais atuaram ali. Trata-se de um estudo de caso a partir do qual se quer refletir sobre algumas questões ligadas à diaconia, especialmente no âmbito da edificação de comunidades. A diaconia como dimensão do ser Igreja insere o ministério diaconal na construção e manutenção de comunidades cristãs

The Effectiveness of Psychotherapy for Schizophrenia Spectrum Disorders in Community Residential Settings

The purpose of this research was to analyze the effectiveness of psychotherapy for individuals diagnosed with schizophrenia spectrum disorders who reside in community residential settings. The present body of literature did not address the utility of psychotherapy treatment for this population. A key area of focus for this research was whether psychotherapy has an impact on psychiatric hospitalization rates for the target population. An additional research question was whether significant differences exist in psychiatric hospitalization rates between males and females for the target population. Data analyses were conducted using archival data from the Blossom Hill Corporation and Sunrise Farm Corporation in the State of Minnesota. Research questions were analyzed with a 2x2 factorial analysis of variance (ANOVA). Results indicated no significant differences in hospitalization rates for individuals in the target population who received psychotherapy (n = 60) compared to those who did not (n = 76). Hospitalization rates also did not differ between gender in psychotherapy treatment response for individuals diagnosed with schizophrenia spectrum disorders in community residential settings. This study has implications for social change because it informs community residential providers in Minnesota serving individuals in the target population about the impact of psychotherapy on reducing psychiatric hospitalizations. Social change is further affected by providing data about how psychotherapy and theory can be used to better treat and understand the target population\u27s mental health stability

O preço e o valor percebido: uma abordagem mercadológica.

O valor percebido pelo consumidor é um importante fator a ser considerado na formação do preço de venda de produtos/mercadorias e serviços. Ele tende a estabelecer o teto do preço, a partir do qual não há mais demanda, uma vez que, acima desse, o valor monetário da transação passa a ser maior que o valor percebido pelo consumidor. O valor percebido abrange inúmeros elementos, objetivos e subjetivos, que lhe dão diversas dimensões. O artigo pretende também apresentar uma metodologia para a identificação do valor percebido, assim como uma descrição das aplicações mercadológicas do mesmo

Long-term experiments to investigate irreversibility in sorption of pesticides to soil

Experiments investigated irreversibility in pesticide sorption to soil. Sorption behaviour under abiotic conditions was quantified for chlorotoluron, prometryn and hexaconazole in three soils over periods of up to 274 days. An isotope-exchange procedure was used whereby sorption of 12C- and 14C-pesticide isotopes in shaken suspensions of three soils (7-168 days shaking) was followed by substitution of the isotopes in the liquid phase and a 14-day exchange phase. This was followed by forced isotope exchange where the sorbed 14C material was desorbed by adding an excess of non-radiolabelled compound. Experiments were concluded with solvent extraction and soil combustion to determine remaining radioactivity. The pesticide-soil systems took around four months to approach sorption equilibrium, resulting in strong asymmetry between the profiles of exchange for isotopes of all three compounds. Under the steep concentration gradient of forced isotope exchange, physically entrapped residues were released back into solution and small amounts of radioactivity were still being released at the termination of the experiment. Results were shown not to deviate markedly from ideal behaviour based on the assumption that sorption is fully reversible. Whilst the timescales for release of sorbed residues back into solution were very long, soil combustion at study termination only yielded <1-2% of applied radioactivity; this confirms that sorption processes under abiotic soil conditions were overwhelmingly reversible for these compounds and set of soils

Proporcionando atualizações em nutrição para a comunidade - 53955031

Trabalho apresentado no 31º SEURS - Seminário de Extensão Universitária da Região Sul, realizado em Florianópolis, SC, no período de 04 a 07 de agosto de 2013 - Universidade Federal de Santa Catarina.A nutrição desportiva visa orientar para que a alimentação e a hidratação realizadas antes, durante e após as modalidades desportivas sejam de boas escolhas, favorecendo o bem-estar do indivíduo. Os escolares que apresentam melhor rendimento desportivo são aqueles que têm garantidos, em seus organismos, adequados suprimentos de energia, nutrientes e, de líquidos corporais, contribuindo para com o desenvolvimento de jogos escolares e as modalidades em aulas de educação física, que requerem esforço físico. É usual que muitas crianças e adolescentes não realizem o desjejum antes de irem à escola e, com isso, seu rendimento na atividade desportiva ficará aquém de seu potencial, caso não ingiram alimentos e tomem líquidos antes das aulas de educação física. O mesmo se aplica em situações de preparo de alunos para campeonatos estudantis, nos turnos inversos ao horário de aulas. A Nutrição se dedica a orientar para que as referidas escolhas alimentícias por escolares sejam saudáveis, apropriadas ao processo digestório do momento, promovam o aporte energético e de hidratação requeridos ao desporto, antes, durante e após a atividade física