Role of GSH in estrone sulfate binding and translocation by the multidrug resistance protein 1 (MRP1/ABCC1)

Multidrug resistance protein 1 (MRP1/ABCC1) is an ATP-dependent efflux pump that can confer resistance to multiple anticancer drugs and transport conjugated organic anions. Unusually, transport of several MRP1 substrates requires glutathione (GSH). For example, estrone sulfate transport by MRP1 is stimulated by GSH, vincristine is co-transported with GSH, or GSH can be transported alone. In the present study, radioligand binding assays were developed to investigate the mechanistic details of GSH-stimulated transport of estrone sulfate by MRP1. We have established that estrone sulfate binding to MRP1 requires GSH, or its non-reducing analogue S-methyl GSH (S-mGSH), and further that the affinity (Kd) of MRP1 for estrone sulfate is 2.5-fold higher in the presence of S-mGSH than GSH itself. Association kinetics show that GSH binds to MRP1 first, and we propose that GSH binding induces a conformational change, which makes the estrone sulfate binding site accessible. Binding of non-hydrolyzable ATP analogues to MRP1 decreases the affinity for estrone sulfate. However, GSH (or S-mGSH) is still required for estrone sulfate binding, and the affinity for GSH is unchanged. Estrone sulfate affinity remains low following hydrolysis of ATP. The affinity for GSH also appears to decrease in the post-hydrolytic state. Our results indicate ATP binding is sufficient for reconfiguration of the estrone sulfate binding site to lower affinity and argue for the presence of a modulatory GSH binding site not associated with transport of this tripeptide. A model for the mechanism of GSH-stimulated estrone sulfate transport is proposed.

Schools and the formation of Black identity during the Civil Rights Movement: Change and resistance in Holly Springs, Mississippi, 1964--1974

Desegregation of social and public spaces was the most visible result of the Civil Rights Movement. After 1960, the integration of schools in Mississippi became a source of conflict. The social change of Civil Rights attacked the social order of White Resistance that supported the state superstructure. The public schools were a place for the discovery of identity for Blacks. The integrated on of the schools caused many Whites to leave rather than be integrated with Blacks. Desegregation of schools was also a slow process because the local and state government could not enforce the decisions of the US Courts, leading Blacks to realize their place in American society could only be secured through individual action. ^ This work explains the role of schooling during the integration of the Holly Springs Separate School System. The process of forging a new identity by local Blacks is examined against the forces of social change and resistance. I addition, this work examines the perils for the Blacks as they faced the uncertainty of change in the crucial Civil Rights years between 1964 and 1974. ^ This work analyzes how the Black community dealt with the problems triggered by the desegregation of the school system in Holly Springs, of a constructed social condition, a psychological state of being, the realities of racism and segregation, and the change and resistance between the individual and the collective. It is based on six months of field work investigation. Although the schools were a crucial aspect of community life for Blacks and Whites, Blacks did form their identity in them. Other institutions, such churches were more crucial. Second, the aspect of politeness and belief in law made the experience in Holly Springs unique to that place, and thus, warrants further study to determine its place within the Civil Rights Movement. Finally, while the political and economic control of Holly Springs remained with Whites, desegregation led to the resegregation of the public schools: as Whites left to private schools. ^

Multiplex PCR for Assessment of Tetracycline Resistance (tet) Genes in Antibiotic-Resistant Soil Bacteria and the Ecological Implications

Antibiotics are becoming increasingly prevalent in bacterial communities due to clinical and agricultural misuse and overuse in their environment. As exposure increases, so does the incidence of microbial resistance. Such is the case with bacterial resistance to tetracyclines, a phenotype often acquired through the horizontal gene transfer of tet genes between bacteria. The objective of this project was to analyze the bacterial diversity of tet resistance genes in soil from Miami-Dade County. Bacterial isolates were Gram-stained and the Kirby-Bauer antibiotic disk diffusion test was performed to determine each bacterium’s degree of resistance. The 16S rRNA gene from antibiotic-resistant isolates was amplified by PCR and sequenced to identify the isolates. All isolates’ tet genes were amplified by multiplex PCR, sequenced, and compared. Among eight isolates, three distinct species were positively identified based on their 16S rRNA sequences and four distinct tet genes were identified, though all tested susceptible to tetracycline via the Kirby-Bauer test. This project clarifies some aspects of the ecology of antibiotic resistance genes, their natural ecological function and the potential for the expansion of intrinsic multi-antibiotic resistance into new ecosystems and/or hosts.

The Spiritual Resistance of Edith Stein and Regina Jonas

Emily Leah Silverman of San Jose State University gives a lecture on the spiritual resistance and religious vision of Edith Stein and Regina Jonas, two German Jewish women who engaged in "deviant" religious desires while working for their communities during the Holocaust. Event held on November 14, 2014 at the Jewish Museum of Florida-Florida International University.

The Antibiotic Resistance and Prescribing in European Children Project: A Neonatal and Pediatric Antimicrobial Web-based Point Prevalence Survey in 73 Hospitals Worldwide

BACKGROUND: The neonatal and pediatric antimicrobial point prevalence survey (PPS) of the Antibiotic Resistance and Prescribing in European Children project (http://www.arpecproject.eu/) aims to standardize a method for surveillance of antimicrobial use in children and neonates admitted to the hospital within Europe. This article describes the audit criteria used and reports overall country-specific proportions of antimicrobial use. An analytical review presents methodologies on antimicrobial use.

METHODS: A 1-day PPS on antimicrobial use in hospitalized children was organized in September 2011, using a previously validated and standardized method. The survey included all inpatient pediatric and neonatal beds and identified all children receiving an antimicrobial treatment on the day of survey. Mandatory data were age, gender, (birth) weight, underlying diagnosis, antimicrobial agent, dose and indication for treatment. Data were entered through a web-based system for data-entry and reporting, based on the WebPPS program developed for the European Surveillance of Antimicrobial Consumption project.

RESULTS: There were 2760 and 1565 pediatric versus 1154 and 589 neonatal inpatients reported among 50 European (n = 14 countries) and 23 non-European hospitals (n = 9 countries), respectively. Overall, antibiotic pediatric and neonatal use was significantly higher in non-European (43.8%; 95% confidence interval [CI]: 41.3-46.3% and 39.4%; 95% CI: 35.5-43.4%) compared with that in European hospitals (35.4; 95% CI: 33.6-37.2% and 21.8%; 95% CI: 19.4-24.2%). Proportions of antibiotic use were highest in hematology/oncology wards (61.3%; 95% CI: 56.2-66.4%) and pediatric intensive care units (55.8%; 95% CI: 50.3-61.3%).

CONCLUSIONS: An Antibiotic Resistance and Prescribing in European Children standardized web-based method for a 1-day PPS was successfully developed and conducted in 73 hospitals worldwide. It offers a simple, feasible and sustainable way of data collection that can be used globally.

Neoliberal Capitalism, Social Reproduction Crisis, Gender Implications and Forms of Resistance: The case of Greece

Neoliberal capitalism and its accompanied austerity measures has had a profound effect on social reproduction throughout Europe. Social reproduction, ie, the process that makes it possible for individuals, families, and society to reproduce itself – is in crisis. The concept of social reproduction utilized in this presentation is framed within the feminist and Marxist tradition. Historically, social reproduction was met through both waged and unwaged work with women playing a predominant role either as waged, predominantly public sector, workers (social workers, teachers, carers, nurses, etc) or as unpaid workers (unpaid carers: mothers, wives, etc.) but now both forms of work are destabilized and undermined. Thus, the crisis in social reproduction. Within this crisis women’s unequal position is further undermined but at the same time progressive women and men are organizing to present alternatives forms of social reproduction. This presentation by utilizing the case of Greece will first outline the neoliberal processes that have caused the crisis in social reproduction. Second, it will present the consequences of social reproduction by highlighting the gender implications. Third, it will discuss the reorganization of social reproduction which is taking place outside the logic of capitalist society and is manifested through the creation of solidarity health clinics and pharmacies, communal kitchens, creation of various forms of bartering etc. This presentation will argue that these forms present glimpses of another society , a society based on principles of solidarity and cooperation. A society worth fighting for. And finally the presentation will conclude by discussing at which level can this crisis be resolved and the role of the social work profession.

A comparative study of the corrosion resistance of incoloy MA 956 and PM 2000 superalloys

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Deciphering the role of plant microbiome on strawberry and raspberry growth, resistance, fruit quality and aroma

Besides their own adaptation strategies, plants might exploit microbial symbionts for overcoming both biotic and abiotic stresses and increase fitness. The current scenario of rapid climate change is demanding more sustainable agricultural management practices. The application of microbe-based products as a valid alternative to synthetic pesticides and fertilizers and their use to overcome stresses exacerbated by climate change, have been reviewed in the first part of this thesis. Berry fruits are widely cultivated and appreciated for their aromatic and nutraceutical properties. This thesis is focused on the role of plant and fruit microbiome on strawberry and raspberry growth, resistance, fruit quality and aroma. A taxonomical and functional description of the microbiome of different organs of three strawberry genotypes was performed both by traditional cultural dependent method and Next Generation Sequencing technique, highlighting a significant role of plant organs and genotype in determining the composition of microbial communities. Additionally, a selection of bacteria native of strawberry plants were isolated and screened for their plant growth promoting abilities and tested under the biotic stress of Xanthomonas fragariae infection and the abiotic stress of induced salinity. The monitoring of biometric parameters allowed the selection of a more restricted panel of bacterial strains, whose beneficial potential was tested in coordinated inoculations, or singularly. Raspberry plant was used for investigating the effect of cultivation method in determining fruit microbiome, and its consequent influence of berry quality and aroma. Interestingly, the cultivation method strongly influenced fruit nutraceutical traits, aroma and epiphytic bacterial biocoenosis. The involvement of the bacterial microbiota in fruit aroma determination was evaluated by performing GC–MS analysis of VOCs occurring in control, sterile and artificially reinoculated berries and by characterizing control and reinoculated berry microbiome. Differently treated berries showed significantly different aromatic profile, confirming the role of bacteria in fruit aroma development.

Steel hopper silos filled with granular material: experimental tests on the tensile resistance of corrugated sheets and on the filling and discharging pressures

This thesis addresses various aspects related to silos, from the strength of some structural parts to internal actions due to grain. Two hopper silo models were mainly studied, so the thesis is divided into two parts. The first part focuses only on the silo cylinder and deals with the collapse of a silo due to failure of the vertical walls. We had the opportunity to access data from a real silo and perform tensile tests on corrugated sheets. The theoretical and experimental resistance of the corrugated sheet forming the silo cylinder was studied. The resistance was then compared with the internal actions due to grain prescribed by various standards. The second part, however, focused on the hopper of a silo in which a load test (loading and unloading of the silo) was performed. Through the test data, an attempt was made to reproduce the pressures normal to the hopper through analytical reasoning. The experimental pressures were then compared with the theoretical pressures predicted by the standards. In addition, with mathematical reasoning, an attempt was made to reproduce the horizontal pressure on the vertical walls of the silo from the experimental normal pressure in the hopper. In fact, the test was related only to the hopper part and not to the silo cylinder.

Isolation of extraintestinal pathogenic Escherichia coli from diarrheic dogs and their antimicrobial resistance profile

From January to December 2006, 92 Escherichia coli isolates from 25 diarrheic dogs were analyzed by screening for the presence of adhesin-encoding genes (pap, sfa, afa), hemolysin and aerobactin genes. Virulence gene frequencies detected in those isolates were: 12% pap, 1% sfa, 10% hemolysin and 6.5% aerobactin. Ten isolates were characterized as extraintestinal pathogenic E. coli (ExPEC) strains; all showed a multidrug resistance phenotype that may represent a reason for concern due the risk of dissemination of antimicrobial resistant genes to the microbiota of human beings.

Emergence of Klebsiella pneumoniae carrying the novel extended-spectrum 'beta'-lactamase gene variants 'bla IND. SHV-40', 'bla IND. TEM-116' and the class I integron-associated 'bla IND. GES-7' in Brazil

A clinical Klebsiella pneumoniae isolate carrying the extended-spectrum beta-lactamase gene variants bla(SHV-40), bla(TEM-116) and bla(GES-7) was recovered. Cefoxitin and ceftazidime activity was most affected by the presence of these genes and an additional resistance to trimethoprim-sulphamethoxazole was observed. The bla(GES-7) gene was found to be inserted into a class 1 integron. These results show the emergence of novel bla(TEM) and bla(SHV) genes in Brazil. Moreover, the presence of class 1 integrons suggests a great potential for dissemination of bla(GES) genes into diverse nosocomial pathogens. Indeed, the bla(GES-7) gene was originally discovered in Enterobacter cloacae in Greece and, to our knowledge, has not been reported elsewhere

Stepped and smooth spillways: resistance effects on stilling basin lengths

The Darcy-Weisbach equation was used in the analysis of flow over spillways, furnishing theoretical tools to design stilling basins. Predictions for the length of hydraulic jump stilling basins downstream of stepped and smooth spillways are presented, together with ranges of values for the Darcy-Weisbach friction factor of both spillways. The experimental data were compared with results of the theoretical solution of the gradually varied flow equation. All comparisons were made in non-dimensional form. The values of the Darcy-Weisbach friction factor were roughly five times smaller for smooth spillways than for stepped spillways. The theoretical predictions and the experimental data allow to present approximate equations for a preliminary evaluation of the length and the bed level of hydraulic jump stilling basins. In the same way, approximate equations were presented for the evaluation of the friction factor in smooth and stepped spillways, as a function of the Froude number at the downstream cross-section.

Effect of the processing conditions and the addition of trans-polyoctenylene rubber on the properties of natural rubber/styrene-butadiene rubber blends

In this study, the influence of the processing conditions and the addition of trans-polyoctenylene rubber (TOR) on Mooney viscosity, tensile properties, hardness, tearing resistance, and resilience of natural rubber/styrene-butadiene rubber blends was investigated. The results obtained are explained in light of dynamic mechanical and morphological analyses. Increasing processing time produced a finer blend morphology, which resulted in an improvement in the mechanical properties. The addition of TOR involved an increase in hardness, a decrease in tear resistance, and no effect on the resilience. It resulted in a large decrease in the Mooney viscosity and a slight decrease in the tensile properties if the components of the compounds were not properly mixed. The results indicate that TOR acted more as a plasticizer than a compatibilizer. (c) 2008 Wiley Periodicals, Inc.