Psychometric Properties and Gender Invariance of the Portuguese (European) Version of the Aging Sexual Knowledge and Attitudes Scale.

The sexuality of older people was understood as non-existent or as something outside the prevailing norm. In this sense, analysing people's sexual knowledge and attitudes towards older people is a challenge for theory and practice. The aim of this study is to translate and validate the Aging Sexual Knowledge and Attitudes Scale (ASKAS) for the Portuguese population. A sample of 994 Portuguese adults (70.9% women, n = 705) completed the ASKAS-PT along with a series of self-report measures. Confirmatory factor analysis and the psychometric properties of the Portuguese (European) version of ASKAS-PT were investigated, particularly reliability, temporal stability, and convergent and critical validity. This study also examined the gender measurement invariance of the ASKAS-PT. After confirmatory factor analysis, a two-factor model fit the Portuguese version best. Reliability and validity results also showed good results, and the ASKAS-PT appears to be a gender-invariant measure. Overall, the Portuguese version of the ASKAS showed good psychometric properties and appears to be a valid and reliable measure for assessing knowledge and attitudes about aging.

Drug Repurposing for COVID-19: A Review and a Novel Strategy to Identify New Targets and Potential Drug Candidates.

In December 2019, the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19) was first identified in the province of Wuhan, China. Since then, there have been over 400 million confirmed cases and 5.8 million deaths by COVID-19 reported worldwide. The urgent need for therapies against SARS-CoV-2 led researchers to use drug repurposing approaches. This strategy allows the reduction in risks, time, and costs associated with drug development. In many cases, a repurposed drug can enter directly to preclinical testing and clinical trials, thus accelerating the whole drug discovery process. In this work, we will give a general overview of the main developments in COVID-19 treatment, focusing on the contribution of the drug repurposing paradigm to find effective drugs against this disease. Finally, we will present our findings using a new drug repurposing strategy that identified 11 compounds that may be potentially effective against COVID-19. To our knowledge, seven of these drugs have never been tested against SARS-CoV-2 and are potential candidates for in vitro and in vivo studies to evaluate their effectiveness in COVID-19 treatment.

Identification of aminopyrimidine-sulfonamides as potent modulators of Wag31-mediated cell elongation in mycobacteria.

There is an urgent need to discover new anti-tubercular agents with novel mechanisms of action in order to tackle the scourge of drug-resistant tuberculosis. Here, we report the identification of such a molecule - an AminoPYrimidine-Sulfonamide (APYS1) that has potent, bactericidal activity against M. tuberculosis. Mutations in APYS1-resistant M. tuberculosis mapped exclusively to wag31, a gene that encodes a scaffolding protein thought to orchestrate cell elongation. Recombineering confirmed that a Gln201Arg mutation in Wag31 was sufficient to cause resistance to APYS1, however, neither overexpression nor conditional depletion of wag31 impacted M. tuberculosis susceptibility to this compound. In contrast, expression of the wildtype allele of wag31 in APYS1-resistant M. tuberculosis was dominant and restored susceptibility to APYS1 to wildtype levels. Time-lapse imaging and scanning electron microscopy revealed that APYS1 caused gross malformation of the old pole of M. tuberculosis, with eventual lysis. These effects resembled the morphological changes observed following transcriptional silencing of wag31 in M. tuberculosis. These data show that Wag31 is likely not the direct target of APYS1, but the striking phenotypic similarity between APYS1 exposure and genetic depletion of Wag31 in M. tuberculosis suggests that APYS1 might indirectly affect Wag31 through an as yet unknown mechanism.

Synergy between Circular Bacteriocin AS-48 and Ethambutol against Mycobacterium tuberculosis.

The increasing incidence of multidrug-resistant Mycobacterium tuberculosis strains and the very few drugs available for treatment are promoting the discovery and development of new molecules that could help in the control of this disease. Bacteriocin AS-48 is an antibacterial peptide produced by Enterococcus faecalis and is active against several Gram-positive bacteria. We have found that AS-48 was active against Mycobacterium tuberculosis, including H37Rv and other reference and clinical strains, and also against some nontuberculous clinical mycobacterial species. The combination of AS-48 with either lysozyme or ethambutol (commonly used in the treatment of drug-susceptible tuberculosis) increased the antituberculosis action of AS-48, showing a synergic interaction. Under these conditions, AS-48 exhibits a MIC close to some MICs of the first-line antituberculosis agents. The inhibitory activity of AS-48 and its synergistic combination with ethambutol were also observed on M. tuberculosis-infected macrophages. Finally, AS-48 did not show any cytotoxicity against THP-1, MHS, and J774.2 macrophage cell lines at concentrations close to its MIC. In summary, bacteriocin AS-48 has interesting antimycobacterial activity in vitro and low cytotoxicity, so further studies in vivo will contribute to its development as a potential additional drug for antituberculosis therapy.

Total Synthesis of Ripostatin B and Structure-Activity Relationship Studies on Ripostatin Analogs.

Described is the total synthesis of the myxobacterial natural product ripostatin B and of a small number of analogs. Ripostatin B is a polyketide-derived 14-membered macrolide that acts as an inhibitor of bacterial RNA-polymerase, but is mechanistically distinct from rifamycin-derived RNA-polymerase inhibitors that are in use for tuberculosis treatment. The macrolactone ring of ripostatin B features two stereocenters and a synthetically challenging doubly skipped triene motif, with one of the double bonds being in conjugation with the ester carbonyl. Appended to the macrolactone core are an extended hydroxy-bearing phenylalkyl side chain at C13 and a carboxymethyl group at C3. The triene motif was established with high efficiency by ring-closing olefin metathesis, which proceeded in almost 80% yield. The side chain-bearing stereocenter α to the ester oxygen was formed in a Paterson aldol reaction between a methyl ketone and a ß-chiral ß-hydroxy aldehyde with excellent syn selectivity (dr >10:1). The total synthesis provided a blueprint for the synthesis of analogs with modifications in the C3 and C13 side chains. The C3-modified analogs showed good antibacterial activity against efflux-deficient Escherichia coli but, as ripostatin B, were inactive against Mycobacterium tuberculosis, in spite of significant in vitro inhibition of M. tuberculosis RNA-polymerase.

A Genetic Lab-on-Phone Test for Point-of-Care Diagnostic of Lactose Intolerance near Patient and in less than 90 Minutes.

BACKGROUND: The -13910 C/T single nucleotide polymorphism located within the MCM6 gene, an enhancer region located upstream of the lactase-phlorizin hydrolase gene, is associated with lactase persistence/non-persistence traits among the Caucasian population. The performance of a new point-of-care CE-IVD (In Vitro Diagnostic) marked isothermal lab-on-phone lactose intolerance assay, using crude samples, was assessed in comparison with Sanger sequencing using purified DNA, as reference method. METHODS: The study was conducted following a non-probability sampling using direct buccal swab (n = 63) and capillary blood (n = 43) clinical samples from a total of 63 volunteers. A 3 × 3 confusion matrix/contingency table was used to evaluate the performance of the isothermal lab-on-phone lactose intolerance assay. RESULTS: The isothermal lab-on-phone lactose intolerance assay successfully detected the -13910 C/T variant with a limit of detection of 5 cells/assay and demonstrated an overall accuracy of 98.41% (95% CI, 91.47%-99.96%) for buccal swab samples and 100% (95% CI, 91.19%-100%) for capillary blood, taking just 90 min from sample to result, with only 2 min hands-on. CONCLUSIONS: The lab-on-phone pocket-sized assay displayed good performance when using direct buccal swab and capillary blood samples, enabling a low-cost, real-time, and accurate genotyping of the -13910 C/T region for the rapid diagnosis of primary lactose intolerance at point-of-care, which enables a prompt implementation of appropriate diet habits and/or intolerance therapies. To our knowledge, this is the first point-of-care genetic test for lactose intolerance to be made available on the market.

Association between lifestyle-related risk behaviors, chronic diseases, and body image distortion: gender differences in follow-up 1 of the ELSA-Brasil cohort.

Aim: To determine the prevalence of body image accuracy/distortion in Brazilian men and women and to investigate sociodemographic and lifestyle-related factors, and the presence of chronic diseases associated with body image distortion. Methods: Data from 6,357 men and 7,657 women participating in the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil) were collected using a multidimensional questionnaire covering sociodemographic characteristics, health behaviors, heath conditions and body image perception. Results: Most participants (53.5% of the women and 54.7% of the men) were found to have an accurate self-perception of their body. When the factors associated with the perception of being heavier than reality were investigated, adopting weight loss measures and not being hypertensive proved protective against this distortion, both in women and men. Conversely, the perception of being lighter than reality was associated, in both women and men, with better education, being black or of mixed race, adopting weight loss measures and not being hypertensive or diabetic. Additional factors associated with underestimating weight were not consuming alcohol (only in women) and belonging to a lower social class (only in men). Conclusion: These findings may contribute to the implementation of public health policies and interventions to promote health and well-being in the Brazilian population.

Role of the Mmr efflux pump in drug resistance in Mycobacterium tuberculosis.

Efflux pumps are membrane proteins capable of actively transporting a broad range of substrates from the cytoplasm to the exterior of the cell. Increased efflux activity in response to drug treatment may be the first step in the development of bacterial drug resistance. Previous studies showed that the efflux pump Mmr was significantly overexpressed in strains exposed to isoniazid. In the work to be described, we constructed mutants lacking or overexpressing Mmr in order to clarify the role of this efflux pump in the development of resistance to isoniazid and other drugs in M. tuberculosis. The mmr knockout mutant showed an increased susceptibility to ethidium bromide, tetraphenylphosphonium, and cetyltrimethylammonium bromide (CTAB). Overexpression of mmr caused a decreased susceptibility to ethidium bromide, acriflavine, and safranin O that was obliterated in the presence of the efflux inhibitors verapamil and carbonyl cyanide m-chlorophenylhydrazone. Isoniazid susceptibility was not affected by the absence or overexpression of mmr. The fluorometric method allowed the detection of a decreased efflux of ethidium bromide in the knockout mutant, whereas the overexpressed strain showed increased efflux of this dye. This increased efflux activity was inhibited in the presence of efflux inhibitors. Under our experimental conditions, we have found that efflux pump Mmr is mainly involved in the susceptibility to quaternary compounds such as ethidium bromide and disinfectants such as CTAB. The contribution of this efflux pump to isoniazid resistance in Mycobacterium tuberculosis still needs to be further elucidated.

Trans(gender) journeys: rights and the (non-)recognition of "human".

A human right paradigm has been challenging the biomedical perspectives that tend to be normalized in the Western context concerning the lives of trans people. The aim of this study is to understand how trans people in Portugal and Brazil perceive the (non-)recognition of their socio-cultural, economic and political rights. Specifically, the study intends to know in what extent these perceptions influence the processes of identity (de)construction. For this purpose, 35 semi-structured interviews were conducted with people self-identified as trans, transsexuals and transvestites in Brazil and Portugal. The narratives of the participants were analyzed according to the thematic analysis method and the following six main themes emerged: (i) Who are the rights for; (ii) Types of rights; (iii) Paradigm of distribution of rights; (iv) Local or global rights; (v) Non-recognition of the "human"; and, (vi) Transphobias (and cissexism). The results allowed the knowledge of rights and the non-recognition of the "human" which is the central organizer of the analysis. Among the main conclusions of this study, we emphasize the circumscription of rights to certain international, regional and/or national contexts; the existence of local instead of global rights, since they are influenced by regional and international law, but they depend on the legislation in force in each country; and the way human rights can also be understood as a platform of invisibility and exclusion of other people. Based on a commitment to social transformation, this article also contributes to rethinking the violence that is exercised on trans people as a continuum, whether through 'normalizing devices' by medical contexts, family contexts, public space, or even through internalized transphobia. Social structures produce and sustain transphobias and, simultaneously, are responsible for fighting them by changing the paradigm about the conception of transsexualities.

Ethidium bromide transport across Mycobacterium smegmatis cell-wall: correlation with antibiotic resistance.

BACKGROUND: Active efflux systems and reduced cell-wall permeability are considered to be the main causes of mycobacterial intrinsic resistance to many antimicrobials. In this study, we have compared the Mycobacterium smegmatis wild-type strain mc2155 with knockout mutants for porins MspA (the main porin of M. smegmatis) and MspC, the efflux pump LfrA (the main efflux pump system of M. smegmatis) and its repressor LfrR for their ability to transport ethidium bromide (EtBr) on a real-time basis. This information was then correlated with minimum inhibitory concentrations (MICs) of several antibiotics in the presence or absence of the efflux inhibitors chlorpromazine, thioridazine and verapamil. RESULTS: In the absence of porins MspA and MspC, accumulation of ethidium bromide decreased and the cells became more resistant to several antibiotics, whereas the knockout mutant for the LfrA pump showed increased accumulation of EtBr and increased susceptibility to EtBr, rifampicin, ethambutol and ciprofloxacin. Moreover, the efflux inhibitors caused a reduction of the MICs of streptomycin, rifampicin, amikacin, ciprofloxacin, clarithromycin and erythromycin in most of the strains tested. CONCLUSIONS: The methodology used in this study demonstrated that porin MspA plays an important role in the influx of quaternary ammonium compounds and antibiotics and that efflux via the LfrA pump is involved in low-level resistance to several antimicrobial drugs in M. smegmatis. The results obtained with this non-pathogenic mycobacterium will be used in future studies as a model for the evaluation of the activity of the same efflux inhibitors on the susceptibility of multidrug resistant strains of Mycobacterium tuberculosis to isoniazid and rifampicin.

Measuring Efflux and Permeability in Mycobacteria.

Mycobacteria are intrinsically resistant to most antimicrobials, which is generally attributed to the impermeability of their cell wall that considerably limits drug uptake. Moreover, like in other pathogenic bacteria, active efflux systems have been widely characterized from diverse mycobacterial species in laboratory conditions, showing that they can promote resistance by extruding noxious compounds prior to their reaching their intended targets. Therefore, the intracellular concentration of a given compound is determined by the balance between permeability, influx, and efflux.Given the urgent need to discover and develop novel antimycobacterial compounds in order to design effective therapeutic strategies, the contributions to drug resistance made by the controlled permeability of the cell wall and the increased activity of efflux pumps must be determined. In this chapter, we will describe a method that allows (1) the measuring of permeability and the quantification of general efflux activity of mycobacteria, by the study of the transport (influx and efflux) of fluorescent compounds, such as ethidium bromide; and (2) the screening of compounds in search of agents that increase the permeability of the cell wall and efflux inhibitors that could restore the effectiveness of antimicrobials that are subject to efflux.

Psychomedical Interventions with Transgender People in Portugal and Brazil: A Critical Approach.

This study aims to analyze biopsychomedical interventions with transgender people. For this purpose, we carried out 35 semi-structured interviews with people who self-identify as transsexuals and transvestites in Brazil and Portugal. The responses of the study participants were systematized according to a thematic analysis, which led to the emergence of the following three main themes: "institutional power", "expectations of trans-bodies", and "experiences in health services". This study demonstrates how some trans people perform bodily modifications to fight the transphobia they experience throughout their lives. In addition, they believe that, by making their bodies conform to each other, they may become more attractive and desirable. The process of cisnormativity is, furthermore, conveyed by the idea present in the answers of some respondents: that having "integrated" bodies means facing less discrimination and that they will, therefore, obtain more satisfactory ways of personally and socially experiencing their identities. This study contributes to a deepening critical reflection on the experiences/exclusions of trans people, especially in the psychomedical context of "normalization" devices. Hence, just as social structures produce and sustain transphobia, the same structures are responsible for combating it.

Violence in the affective-sexual trajectories of young gay men: "new" settings and "old" challenges. / Violências nas trajetórias afetivo-sexuais de jovens gays: "novas" configurações e "velhos" desafios.

This paper discusses the phenomenon of violence in the affective-sexual trajectories of young, cisgender gay men, from popular strata, in the metropolitan region of Rio de Janeiro, Brazil. Brazilian literature on violence against gay men generally focuses on the discrimination suffered by this population. However, the violence they are submitted to among family relationships due to their sexual orientation, or even their relationships known as "dating" or "hookup", is hardly discussed. This qualitative study used in-depth interviews based on a semi-structured guide to discuss violence during the young gay men's affective-sexual trajectory. The results evidenced multiple faces of violence during childhood and adolescence in family relationships, spanning their affective-sexual relationships in adolescence and youth, including sexual, physical, psychological, and institutional violence. Support networks are limited, for example, to a few friends and access to blogs on the Internet. No health professional was cited as a helping source. There is a need to discuss the prevention of violence and discuss health promotion of this social group, expanding the perspective on the various contemporary ways of relating intimately.

Este artigo aborda o fenômeno da violência nas trajetórias afetivo-sexuais de jovens gays cisgênero, pertencentes a camadas populares da região metropolitana do Rio de Janeiro, Brasil. A literatura brasileira sobre violência contra homens gays, geralmente, tem como foco a discriminação sofrida por esta população, mas pouco se problematiza a violência que eles sofrem nas relações familiares devido à sua orientação sexual, ou ainda, em suas relações ditas como "namoro" ou "ficar". Com o objetivo de discutir a presença de violências durante a trajetória afetivo-sexual dos jovens, este estudo qualitativo realizou entrevistas em profundidade a partir de um roteiro semiestruturado. Os resultados mostraram que há múltiplas faces de violência que ocorrem durante a infância e adolescência nas relações familiares, perpassando nos seus relacionamentos afetivo-sexuais na adolescência e juventude, incluindo violências sexuais, físicas, psicológicas e institucionais. As redes de apoio dos jovens são limitadas, como exemplo, a poucos amigos e ao acesso de blogs na internet. Nenhum profissional de saúde foi citado pelos jovens como fonte de ajuda. Destaca-se também a necessidade de debater a prevenção da violência e promoção da saúde destes jovens, ampliando o olhar para as várias formas contemporâneas de se relacionar intimamente.

Priority effects of stream eutrophication and assembly history on beta diversity across aquatic consumers, decomposers and producers.

Priority effects are stochastic processes that consider the influence of the order of arrival of species on community dynamics and structure. We evaluated the short-term effects of stream eutrophication and colonization time in freshwater benthic communities (primary producers - periphytic algae, decomposers - fungi, and consumers - macroinvertebrates) to test whether (i) beta diversity is higher in eutrophic streams due to priority effects driven by stochastic community formation processes (ecological drift or random dispersal), and (ii) in the early stages of colonization, priority effects drive the history of the formation and the initial establishment of the community in the stream, resulting in higher beta diversity. The present study was conducted in situ over 28 days in temperate streams along a trophic gradient, with colonization being evaluated every seven days. The study identified 84 species of alga, 43 families of macroinvertebrates, and 44 species of aquatic fungi. Our results demonstrated that deterministic processes were responsible for the formation of aquatic producers, while priority effects (stochasticity) were more important for the aquatic decomposers and consumers. In the case of the producers, beta diversity was highest in the hypertrophic stream, but did not vary significantly over colonization time. The beta diversity of the decomposers was highest in the hypertrophic stream and in the later stages of succession, due primarily to mechanisms of facilitation. The beta diversity of the consumers was lowest in the hypertrophic stream due primarily to the priority and inhibitory effects of the predominant groups, and highest at seven and 21 days of colonization. As these three taxonomic groups differ in their intrinsic biological characteristics, and in their functional role in the ecosystem, our short-term field study demonstrated that both stochastic and deterministic processes combine to influence the configuration of the community, and that the relative importance of the two processes varies systematically along a trophic gradient.

Low-Phytotoxic Deep Eutectic Systems as Alternative Extraction Media for the Recovery of Chitin from Brown Crab Shells.

The versatility of chitin and its derivatives has allowed their utilization in a wide range of applications, from wastewater treatment to pharmaceutical or biomedical industries. However, even though the extraction method used industrially is extremely efficient, it involves the use of strong acids and bases and results in the disposal of large quantities of toxic effluents. Deep eutectic systems (DESs) have emerged as a promising new class of alternative solvents, including for chitin recovery. Yet, the assessment of their toxicity has often been neglected. Therefore, in this work, the phytotoxicity of choline chloride (ChCl)/organic acid-based DESs toward wheat seeds was evaluated by measuring different growth parameters and stress biomarkers. DESs were then explored for the efficient recovery of chitin contained in brown crab shell residues at varying conditions of temperature and processing time as well as with and without water addition. The obtained chitin was then characterized through different analytical techniques and compared to a standard as well as to chitin obtained by a conventional acid/alkaline hydrolysis. Results have shown that by applying a ChCl/lactic acid-based DES (which was the system that showed the least phytotoxic effects on wheat; EC50 ≥ 1.6 mg/mL) at 130 °C, it was possible to obtain pure chitin (up to 98%) with characteristics similar to those presented by commercial chitin or chitin recovered by conventional hydrolysis in a shorter time (more than 8-fold faster), thus suggesting that ChCl/organic acid-based DESs can truly represent a low-phytotoxic alternative extraction media for the recovery of chitin from the crab shell biomass.

Phenothiazines, bacterial efflux pumps and targeting the macrophage for enhanced killing of intracellular XDRTB.

Phenothiazines have their primary effects on the plasma membrane of prokaryotes and eukaryotes. Among the components of the prokaryotic plasma membrane affected are efflux pumps, their energy sources, energy providing enzymes such as ATPases, and genes that regulate and code for permeability aspects of the bacterium. The responses of multi-drug (MDR) and extensively drug resistant (XDR) Mycobacterium tuberculosis to the neuroleptic phenothiazine thioridazine are reviewed. The information collated suggests that this phenothiazine has the potential to cure XDR and MDR tuberculosis infections, a potential that has been recently demonstrated by its ability to cure 10 patients who presented with XDR TB infections. The mechanism by which this phenothiazine produces the desired effects within the infected macrophage is also discussed.

Efflux pump inhibitors as a promising adjunct therapy against drug resistant tuberculosis: a new strategy to revisit mycobacterial targets and repurpose old drugs.

INTRODUCTION: In 2018, an estimated 377,000 people developed multidrug-resistant tuberculosis (MDR-TB), urging for new effective treatments. In the last years, it has been accepted that efflux pumps play an important role in the evolution of drug resistance. Strategies are required to mitigate the consequences of the activity of efflux pumps. AREAS COVERED: Based upon the literature available in PubMed, up to February 2020, on the diversity of efflux pumps in Mycobacterium tuberculosis and their association with drug resistance, studies that identified efflux inhibitors and their effect on restoring the activity of antimicrobials subjected to efflux are reviewed. These support a new strategy for the development of anti-TB drugs, including efflux inhibitors, using in silico drug repurposing. EXPERT OPINION: The current literature highlights the contribution of efflux pumps in drug resistance in M. tuberculosis and that efflux inhibitors may help to ensure the effectiveness of anti-TB drugs. However, despite the usefulness of efflux inhibitors in in vitro studies, in most cases their application in vivo is restricted due to toxicity. In a time when new drugs are needed to fight MDR-TB and extensively drug-resistant TB, cost-effective strategies to identify safer efflux inhibitors should be implemented in drug discovery programs.

Demonstration of the activity of P-glycoprotein by a semi-automated fluorometric method.

BACKGROUND: We have developed a semi-automated fluorometric method that utilizes ethidium bromide (EB), a common substrate of bacterial efflux pumps. The method is sufficiently sensitive to characterize the efflux pump systems of bacteria. Because EB is also recognized and extruded by ATP-binding cassette (ABC) transporters and these have similarity to P-glycoprotein (P-gp), the method has been extended for the evaluation of agents that can inhibit the extrusion of EB on a real-time basis by mouse lymphoma cells containing the human ABCB1 (mdr1) gene. MATERIALS AND METHODS: Monitoring of uptake and extrusion of EB was assessed using the Rotor-Gene 3000 (Corbett Research) under different conditions. RESULTS: Whereas extrusion of EB took place readily, the addition of known inhibitors of efflux pumps (verapamil, reserpine) caused retention of EB. CONCLUSION: This method is inexpensive and allows the detection of neoplastic cells with increased efflux activity as well as the screening of large numbers of compounds for inhibition of the P-gp.

Characterization of intrinsic efflux activity of Enterococcus faecalis ATCC29212 by a semi-automated ethidium bromide method.

Enterococcus faecalis is recognized as a multidrug-resistant nosocomial pathogen. The phenotypic basis for this is largely uncharacterized. The intrinsic efflux system of the antibiotic-susceptible E. faecalis ATCC29212 strain was studied using a semi-automated method that assesses accumulation and efflux of the universal efflux pump substrate ethidium bromide (EB). The results show that the intrinsic efflux system of this Enterococcus strain is controlled by energy derived from the catabolism of glucose and the proton concentration of the medium. At pH 5, agents that inhibit efflux pumps in Gram-positive organisms and the proton gradient un-coupler CCCP do not increase accumulation nor inhibit efflux of EB. In contrast, at pH 8, where the proton concentration is 1,000-fold lower, these agents increase accumulation and efflux of EB. These results are relevant to infections produced by E. faecalis and subsequent antibiotic therapy with antibiotics to which the organism is known to be intrinsically resistant.

Mycobacterial Aminoglycoside Acetyltransferases: A Little of Drug Resistance, and a Lot of Other Roles.

Aminoglycoside acetyltransferases are important determinants of resistance to aminoglycoside antibiotics in most bacterial genera. In mycobacteria, however, aminoglycoside acetyltransferases contribute only partially to aminoglycoside susceptibility since they are related with low level resistance to these antibiotics (while high level aminoglycoside resistance is due to mutations in the ribosome). Instead, aminoglycoside acetyltransferases contribute to other bacterial functions, and this can explain its widespread presence along species of genus Mycobacterium. This review is focused on two mycobacterial aminoglycoside acetyltransferase enzymes. First, the aminoglycoside 2'-N-acetyltransferase [AAC(2')], which was identified as a determinant of weak aminoglycoside resistance in M. fortuitum, and later found to be widespread in most mycobacterial species; AAC(2') enzymes have been associated with resistance to cell wall degradative enzymes, and bactericidal mode of action of aminoglycosides. Second, the Eis aminoglycoside acetyltransferase, which was identified originally as a virulence determinant in M. tuberculosis (enhanced intracellular survival); Eis protein in fact controls production of pro-inflammatory cytokines and other pathways. The relation of Eis with aminoglycoside susceptibility was found after the years, and reaches clinical significance only in M. tuberculosis isolates resistant to the second-line drug kanamycin. Given the role of AAC(2') and Eis proteins in mycobacterial biology, inhibitory molecules have been identified, more abundantly in case of Eis. In conclusion, AAC(2') and Eis have evolved from a marginal role as potential drug resistance mechanisms into a promising future as drug targets.