Efflux pump of the proton antiporter family confers low-level fluoroquinolone resistance in Mycobacterium smegmatis.

Due to the resurgence of tuberculosis and the emergence of multidrug-resistant strains, fluoroquinolones (FQ) are being used in selected tuberculosis patients, but FQ-resistant strains of Mycobacterium tuberculosis have rapidly begun to appear. The mechanisms involved in FQ resistance need to be elucidated if the effectiveness of this class of antibiotics is to be improved and prolonged. By using the rapid-growing Mycobacterium smegmatis as a model genetic system, a gene was selected that confers low-level FQ resistance when present on a multicopy plasmid. This gene, lfrA, encodes a putative membrane efflux pump of the major facilitator family, which appears to recognize the hydrophilic FQ, ethidium bromide, acridine, and some quaternary ammonium compounds. It is homologous to qacA from Staphylococcus aureus, tcmA, of Streptomyces glaucescens, and actII and mmr, both from Streptomyces coelicoler. Increased expression of lfrA augments the appearance of subsequent mutations to higher-level FQ resistance.

Crystal structure of Pseudomonas aeruginosa catabolic ornithine transcarbamoylase at 3.0-A resolution: a different oligomeric organization in the transcarbamoylase family.

The crystal structure of the Glu-105-->Gly mutant of catabolic ornithine transcarbamoylase (OTCase; carbamoyl phosphate + L-ornithine = orthophosphate + L-citrulline, EC 2.1.3.3) from Pseudomonas aeruginosa has been determined at 3.0-A resolution. This mutant is blocked in the active R (relaxed) state. The structure was solved by the molecular replacement method, starting from a crude molecular model built from a trimer of the catalytic subunit of another transcarbamoylase, the extensively studied aspartate transcarbamoylase (ATCase) from Escherichia coli. This model was used to generate initial low-resolution phases at 8-A resolution, which were extended to 3-A by noncrystallographic symmetry averaging. Four phase extensions were required to obtain an electron density map of very high quality from which the final model was built. The structure, including 4020 residues, has been refined to 3-A, and the current crystallographic R value is 0.216. No solvent molecules have been added to the model. The catabolic OTCase is a dodecamer composed of four trimers organized in a tetrahedral manner. Each monomer is composed of two domains. The carbamoyl phosphate binding domain shows a strong structural homology with the equivalent ATCase part. In contrast, the other domain, mainly implicated in the binding of the second substrate (ornithine for OTCase and aspartate for ATCase) is poorly conserved. The quaternary structures of these two allosteric transcarbamoylases are quite divergent: the E. coli ATCase has pseudo-32 point-group symmetry, with six catalytic and six regulatory chains; the catabolic OTCase has 23 point-group symmetry and only catalytic chains. However, both enzymes display homotropic and heterotropic cooperativity.

Comparison of the defensive elicitation activity of cerato-populin and cerato-platanin: a structural model

Plants can defend themselves from potential pathogenic microorganisms relying on a complex interplay of signaling pathways: activation of the MAPK cascade, transcription of defense related genes, production of reactive oxygen species, nitric oxide and synthesis of other defensive compounds such as phytoalexins. These events are triggered by the recognition of pathogen’s effectors (effector-triggered immunity) or PAMPs (PAMP-triggered immunity). The Cerato Platanin Family (CPF) members are Cys-rich proteins secreted and localized on fungal cell walls, involved in several aspects of fungal development and pathogen-host interactions. Although more than hundred genes of the CPF have been identified and analyzed, the structural and functional characterization of the expressed proteins has been restricted only to few members of the family. Interestingly, those proteins have been shown to bind chitin with diverse affinity and after foliar treatment they elicit defensive mechanisms in host and non-host plants. This property turns cerato platanins into interesting candidates, worth to be studied to develop new fungal elicitors with applications in sustainable agriculture. This study focus on cerato-platanin (CP), core member of the family and on the orthologous cerato-populin (Pop1). The latter shows an identity of 62% and an overall homology of 73% with respect to CP. Both proteins are able to induce MAPKs phosphorylation, production of reactive oxygen species and nitric oxide, overexpression of defense’s related genes, programmed cell death and synthesis of phytoalexins. CP, however, when compared to Pop1, induces a faster response and, in some cases, a stronger activity on plane leaves. Aim of the present research is to verify if the dissimilarities observed in the defense elicitation activity of these proteins can be associated to their structural and dynamic features. Taking advantage of the available CP NMR structure, Pop1’s 3D one was obtained by homology modeling. Experimental residual dipolar couplings and 1H, 15N, 13C resonance assignments were used to validate the model. Previous works on CPF members, addressed the highly conserved random coil regions (loops b1-b2 and b2-b3) as sufficient and necessary to induce necrosis in plants’ leaves: that region was investigated in both Pop1 and CP. In the two proteins the loops differ, in their primary sequence, for few mutations and an insertion with a consequent diversification of the proteins’ electrostatic surface. A set of 2D and 3D NMR experiments was performed to characterize both the spatial arrangement and the dynamic features of the loops. NOE data revealed a more extended network of interactions between the loops in Pop1 than in CP. In addition, in Pop1 we identified a salt bridge Lys25/Asp52 and a strong hydrophobic interaction between Phe26/Trp53. These structural features were expected not only to affect the loops’ spatial arrangement, but also to reduce the degree of their conformational freedom. Relaxation data and the order parameter S2 indeed highlighted reduced flexibility, in particular for loop b1-b2 of Pop1. In vitro NMR experiments, where Pop1 and CP were titrated with oligosaccharides, supported the hypothesis that the loops structural and dynamic differences may be responsible for the different chitin-binding properties of the two proteins: CP selectively binds tetramers of chitin in a shallow groove on one side of the barrel defined by loops b1-b2, b2-b3 and b4-b5, Pop1, instead, interacts in a non-specific fashion with oligosaccharides. Because the region involved in chitin-binding is also responsible for the defense elicitation activity, possibly being recognized by plant's receptors, it is reasonable to expect that those structural and dynamic modifications may also justify the different extent of defense elicitation. To test that hypothesis, the initial steps of a protocol aimed to the identify a receptor for CP, in silico, are presented.

The Perpetration of Spousal Abuse by Male Combat Veterans: Recommendations for Individualized Group Domestic Violence Treatment for Veterans

Recent estimates suggest that spousal abuse is, in fact, on the rise in the U.S. military (The Miles Foundation, 2005). As research specific to the impact of posttraumatic stress disorder (PTSD) on U.S. soldiers has grown since the Vietnam War, clinicians and researchers have begun to investigate how combat-related trauma affects veterans in terms of aggression, hostility and social/emotional functioning. The training and stressors experienced by soldiers in the military are unique and affect all aspects of the veteran's functioning. This paper discusses questions related to why combat veterans may be at increased risk to commit spousal abuse (verbal, psychological, and physical), the relationship between PTSD, substance use, and violence, and the advantages to individualizing group domestic violence (DV) treatment programs for combat veterans. Recommendations will be made for a DV treatment program specifically for combat veterans who also suffer from PTSD.

Using Emotion to Foster Acceptance: A Treatment Approach for Family Therapists Working with Transgender Adolescents and their Families

Currently, there is limited research and clinical focus on family therapy with transgender adolescents. When an adolescent discloses his/her transgender identity to his/her family, the family can experience an array of emotions, such as fear, distrust, anger, and sadness, along with confusion and invalidating behavior that can threaten secure attachment among family members. The purpose of this paper is to present a family therapy treatment approach for therapists working with transgender adolescents that is both culturally sensitive to the needs of these families as well as based on a systemic family therapy model. Emotionally Focused Family Therapy (EFFT) is a systemic model that is grounded in attachment theory and focuses on using emotion as a key tool in restructuring problematic relational patterns and fostering more secure family bonds. Through the use of a hypothetical case study, this paper aims at illustrating how EFFT can help family members process feelings related to the transgender identity of an adolescent family member and restore their attachment in a manner that strengthens family relationships and bonds.

Exploring How AFFECT (A Family Focused Emotion Communication Training) and Self Psychology Are Mutually Enhancing

This paper implicitly advocates for a rapprochement between psychodynamic and behavioral approaches to psychotherapy, by exploring the similarities and differences between self psychology and A Family Focused Emotion Communication Training (AFFECT), a behavioral parent training model. Self psychology, a theory with broad applicability, has been applied to several modalities besides behavioral ones. Generally speaking, self psychology and AFFECT are both relational approaches to psychotherapy that emphasize the impact of parent responsiveness, more specifically empathic attunement, on a child's emotional development and emotion regulation. Differentiating aspects of each model are identified to enhance the other model. AFFECT has relevance for pushing self psychology theory more in the direction of operations, which has implications for enhancing the research potential of self psychology, as well as for the training of the self-psychologist. Conversely, self psychology has relevance for coaching the parent with low self-esteem and decreased self-efficacy in AFFECT, which has potential implications for AFFECT treatment outcomes.

La conciliación del empleo y del hogar: respuesta y reflejo de una organización del trabajo construida desde la institución del género

A la luz del debate en torno a la relación entre la vida laboral y familiar, este artículo propone un modelo compuesto por las variables: sexo, edad, estudios, parentesco y dos índices nombrados por vsp y vsf. El primero (vsp) trata de situar a la población ocupada en una escala que refleje la situación en términos de modalidad de contrato, jornadas, ingresos y supervisión. El segundo refleja las cargas familiares. Se plantean dos versiones distintas del modelo: vsp como dependiente o vsf. El objetivo de este ejercicio es doble: 1) observar hasta qué punto las cargas domésticas influyen en la posición dentro del puesto de trabajo remunerado, y 2) averiguar qué variable sociodemográfica tiene mayor influencia sobre ellas. Los datos han sido extraídos de la Encuesta de Calidad de Vida en el Trabajo (ECVT 2001, 2002 y 2003). A la luz de los resultados, la distribución por sexo en «vsp» y «vsf» es significativamente distinta: los hombres asalariados ocupan las mejores posiciones. Además, el modelo sólo actúa para las mujeres asalariadas principales de familia (cabezas de familia y cónyuges). Esta última observación es completamente coherente con una organización de trabajo construida desde la institución del género.

El Programa de Educación en Alimentación y Nutrición (1961-1982) y la capacitación de las amas de casa como responsables del bienestar familiar

El objetivo de este trabajo es analizar desde una perspectiva de género, a través del ejemplo del Programa de Educación en Alimentación y Nutrición (EDALNU) que se desarrolló en España en las décadas finales del siglo XX, los mensajes y estrategias de educación alimentaria nutricional que iban dirigidos a mejorar la capacitación de las amas de casa como garantes del bienestar familiar y como expertas en la gestión de los diversos aspectos del hogar. Su acción específica sobre las amas de casa y los argumentos sobre los que se sostenía dicha intervención afianzaron el modelo social de género en el que los hombres eran los únicos responsables de los recursos económicos del hogar y las mujeres las proveedoras de cuidados familiares y en concreto los relacionados con la alimentación y la nutrición.

Correlation between SD-OCT, immunocytochemistry and functional findings in an animal model of retinal degeneration

Purpose: The P23H rhodopsin mutation is an autosomal dominant cause of retinitis pigmentosa (RP). The degeneration can be tracked using different anatomical and functional methods. In our case, we evaluated the anatomical changes using Spectral-Domain Optical Coherence Tomography (SD-OCT) and correlated the findings with retinal thickness values determined by immunocytochemistry.Methods: Pigmented rats heterozygous for the P23H mutation, with ages between P18 and P180 were studied. Function was assessed by means of optomotor testing and ERGs. Retinal thicknesses measurements, autofluorescence and fluorescein angiography were performed using Spectralis OCT. Retinas were studied by means of immunohistochemistry. Results: Between P30 and P180, visual acuity decreased from 0.500 to 0.182 cycles per degree (cyc/deg) and contrast sensitivity decreased from 54.56 to 2.98 for a spatial frequency of 0.089 cyc/deg. Only cone-driven b-wave responses reached developmental maturity. Flicker fusions were also comparable at P29 (42 Hz). Double flash-isolated rod-driven responses were already affected at P29. Photopic responses revealed deterioration after P29.A reduction in retinal thicknesses and morphological modifications were seen in OCT sections. Statistically significant differences were found in all evaluated thicknesses. Autofluorescence was seen in P23H rats as sparse dots. Immunocytochemistry showed a progressive decrease in the outer nuclear layer (ONL), and morphological changes. Although anatomical thickness measures were significantly lower than OCT values, there was a very strong correlation between the values measured by both techniques.Conclusions: In pigmented P23H rats, a progressive deterioration occurs in both retinal function and anatomy. Anatomical changes can be effectively evaluated using SD-OCT and immunocytochemistry, with a good correlation between their values, thus making SD-OCT an important tool for research in retinal degeneration.

The Modelling of Photovoltaic, Solar Thermal, and Photovoltaic/Thermal Domestic Hot Water Systems

Solar heating of potable water has traditionally been accomplished through the use of solar thermal (ST) collectors. With the recent increases in availability and lower cost of photovoltaic (PV) panels, the potential of coupling PV solar arrays to electrically heated domestic hot water (DHW) tanks has been considered. Additionally, innovations in the SDHW industry have led to the creation of photovoltaic/thermal (PV/T) collectors, which heat water using both electrical and thermal energy. The current work compared the performance and cost-effectiveness of a traditional solar thermal (ST) DHW system to PV-solar-electric DHW systems and a PV/T DHW system. To accomplish this, a detailed TRNSYS model of the solar hot water systems was created and annual simulations were performed for 250 L/day and 325 L/day loads in Toronto, Vancouver, Montreal, Halifax, and Calgary. It was shown that when considering thermal performance, PV-DHW systems were not competitive when compared to ST-DHW and PVT-DHW systems. As an example, for Toronto the simulated annual solar fractions of PV-DHW systems were approximately 30%, while the ST-DHW and PVT-DHW systems achieved 65% and 71% respectively. With current manufacturing and system costs, the PV-DHW system was the most cost-effective system for domestic purposes. The capital cost of the PV-DHW systems were approximately $1,923-$2,178 depending on the system configuration, and the ST-DHW and PVT system were estimated to have a capital cost of $2,288 and $2,373 respectively. Although the capital cost of the PVT-DHW system was higher than the other systems, a Present Worth analysis for a 20-year period showed that for a 250 L/day load in Toronto the Present Worth of the PV/T system was approximately $4,597, with PV-DHW systems costing approximately $7,683-$7,816 and the ST-DHW system costing $5,238.

Exports and Capacity Constraints: A smooth transition regression model for six euro-area countries. CEPS Working Document No. 395/May 2014

The significant gains in export market shares made in a number of vulnerable euro-area crisis countries have not been accompanied by an appropriate improvement in price competitiveness. This paper argues that, under certain conditions, firms consider export activity as a substitute for serving domestic demand. The strength of the link between domestic demand and exports is dependent on capacity constraints. Our econometric model for six euro-area countries suggests domestic demand pressure and capacity-constraint restrictions as additional variables of a properly specified export equation. As an innovation to the literature, we assess the empirical significance through the logistic and the exponential variant of the non-linear smooth transition regression model. We find that domestic demand developments are relevant for the short-run dynamics of exports in particular during more extreme stages of the business cycle. A strong substitutive relationship between domestic and foreign sales can most clearly be found for Spain, Portugal and Italy, providing evidence of the importance of sunk costs and hysteresis in international trade.

An aided economy: the characteristics of the Transnistrian economic model. OSW Commentary No. 108, 2013-05-14

The economy of breakaway Transnistria is a peculiar combination of the command-and-distribution model inherited from the USSR with elements of a free-market economy which is heavily dependent on Russian energy and financial subsidies. The main pillars of the region’s economy are several large industrial plants, built in the Soviet era, which generate more than half of its GDP (in 2012, Transnistria’s GDP reached around US$1 billion). As a consequence, the condition of each of these companies, whose production is almost exclusively export- -oriented, has a huge impact on the economic situation in Transnistria. This makes the region extremely sensitive to any changes in the economic situation of its key trade partners. This problem is additionally aggravated by the extremely low diversification of Transnistrian exports. The only major economic entity in Transnistria which regularly yields profits and is not so heavily dependent on the external situation is Sheriff. This corporation controls the greater part of the local wholesale and retail trade, as well as a major part of the services sector on the domestic market.

Portfolio Entrepreneurship in Family Firms: A Resource-based Perspective

The phenomenon of portfolio entrepreneurship has attracted considerable scholarly attention and is particularly relevant in the family fi rm context. However, there is a lack of knowledge of the process through which portfolio entrepreneurship develops in family firms. We address this gap by analyzing four in-depth, longitudinal family firm case studies from Europe and Latin America. Using a resource-based perspective, we identify six distinct resource categories that are relevant to the portfolio entrepreneurship process. Furthermore, we reveal that their importance varies across time. Our resulting resource-based process model of portfolio entrepreneurship in family firms makes valuable contributions to both theory and practice.

Family Firm Incumbent’s Attitude toward Intra-family Succession: Antecedents and Effects on Intentions

Research shows that intention for intra-family succession is an important determinant of family firm behavior. To provide a systematic analysis of the antecedents of such intention, we use the theory of planned behavior to model the incumbent leader’s attitude toward intra-family succession because that particular attitude is idiosyncratic to family firms. Empirical tests using a sample of 271 Italian incumbent leaders of family firms show that, as predicted by planned behavior theory, attitude and self-efficacy are significant predictors of intention. They show further that attitude is affected by the number of children, emotional attachment, and need for control.

"Aunt Babette's" cook book : foreign and domestic receipts for the household : a valuable collection of receipts and hints for the housewife, many of which are not to be found elsewhere /

Includes sections on servants, grocery marketing, canning; household hints; samples of menus; recipes for German, French, Hungarian, Italian, Polish, Russian, Jewish dishes; some recipes include wine or liquor as an ingredient. Sample recipes: Filet de bœuf au champignons, Velvet potato puffs, Rhubarb custard pie, Bergamot ice-cream.