Uncovering the Cryptic Gene Cluster ahb for 3-amino-4-hydroxybenzoate Derived Ahbamycins, by Searching SARP Regulator Encoding Genes in the Streptomyces argillaceus Genome.

Genome mining using standard bioinformatics tools has allowed for the uncovering of hidden biosynthesis gene clusters for specialized metabolites in Streptomyces genomes. In this work, we have used an alternative approach consisting in seeking "Streptomyces Antibiotic Regulatory Proteins" (SARP) encoding genes and analyzing their surrounding DNA region to unearth cryptic gene clusters that cannot be identified using standard bioinformatics tools. This strategy has allowed the unveiling of the new ahb cluster in Streptomyces argillaceus, which had not been retrieved before using antiSMASH. The ahb cluster is highly preserved in other Streptomyces strains, which suggests a role for their encoding compounds in specific environmental conditions. By combining overexpression of three regulatory genes and generation of different mutants, we were able to activate the ahb cluster, and to identify and chemically characterize the encoded compounds that we have named ahbamycins (AHBs). These constitute a new family of metabolites derived from 3-amino-4-hydroxybenzoate (3,4-AHBA) known for having antibiotic and antitumor activity. Additionally, by overexpressing three genes of the cluster (ahbH, ahbI, and ahbL2) for the synthesis and activation of 3,4-AHBA, a new hybrid compound, AHB18, was identified which had been produced from a metabolic crosstalk between the AHB and the argimycin P pathways. The identification of this new BGC opens the possibility to generate new compounds by combinatorial biosynthesis.

Expanding the utility of Marfey's analysis by using HPLC-SPE-NMR to determine the Cß configuration of threonine and isoleucine residues in natural peptides.

The determination of amino acid chirality in natural peptides is typically addressed by Marfey's analysis. This approach relies on the complete hydrolysis of the peptide followed by the reaction of the resulting amino acid pool with Marfey's reagent, a chiral derivatizing agent which turns amino acid enantiomers into diastereomeric pairs which can be resolved by conventional reversed-phase HPLC. However, for certain amino acids possessing a second chiral centre at Cß, the discrimination between the two possible epimers may still be challenging due to the lack of chromatographic resolution. Such is the case of isoleucine and threonine which can also be found in natural nonribosomal peptides as their allo-diastereomers. We describe a new approach based on the extension of Marfey's analysis using HPLC-SPE-NMR to sort out this challenge. Marfey's derivatives of these epimeric amino acids at Cß can be differentiated by their distinct NMR spectra. Thus, simple comparison of the NMR spectra of trapped HPLC peaks with the corresponding spectra of standards enables the unambiguous assignment of the absolute configuration at the second chiral centre in such cases. The general applicability of this approach is showcased for two model cyclic peptides bearing L-Ile and L-Thr.

Biosynthesis of Largimycins in Streptomyces argillaceus Involves Transient ß-Alkylation and Cryptic Halogenation Steps Unprecedented in the Leinamycin Family.

Largimycins A1 and A2 are key members of a recently identified family of hybrid nonribosomal peptide polyketides belonging to the scarcely represented group of antitumor leinamycins. They are encoded by the gene cluster lrg of Streptomyces argillaceus. This cluster contains a halogenase gene and two sets of genes for the biosynthesis and incorporation of ß branches at C3 and C9. Noticeably, largimycins A1 and A2 are nonhalogenated compounds and only contain a ß branch at C3. By generating mutants in those genes and characterizing chemically their accumulated compounds, we could confirm the existence of a chlorination step at C19, the introduction of an acetyl-derived olefinic exomethylene group at C9, and a propionyl-derived ß branch at C3 in the biosynthesis pathway. Since the olefinic exomethylene group and the chlorine atom are absent in the final products, those biosynthetic steps can be considered cryptic in the overall pathway but essential to generating keto and epoxide functionalities at C9 and C18/C19, respectively. We propose that chlorination at C19 is utilized as an activation strategy that creates the precursor halohydrin to finally yield the epoxy functionality at C18/C19. This represents a novel strategy to create such functionalities and extends the small number of natural product biosynthetic pathways that include a cryptic chlorination step.

Grapevine Xylem Sap Is a Potent Elicitor of Antibiotic Production in Streptomyces spp.

Streptomyces bacteria produce a wide number of antibiotics and antitumor compounds that have attracted the attention of pharmaceutical and biotech companies. In this study, we provide evidence showing that the xylem sap from grapevines has a positive effect on the production of different antibiotics by several Streptomyces species, including S. ambofaciens ATCC 23877 and S. argillaceus ATCC 12596 among others. The production of several already known compounds was induced: actinomycin D, chromomycin A3, fungichromin B, mithramycin A, etc., and four compounds with molecular formulas not included in the Dictionary of Natural Products (DNP v28.2) were also produced. The molecules present in the xylem sap that acts as elicitors were smaller than 3 kDa and soluble in water and insoluble in ether, ethyl acetate, or methanol. A combination of potassium citrate and di-D-fructose dianhydrides (related to levanbiose or inulobiose) seemed to be the main effectors identified from the active fraction. However, the level of induction obtained in the presence of these compounds mix was weaker and delayed with respect to the one got when using the whole xylem sap or the 3 kDa sap fraction, suggesting that another, not identified, elicitor must be also implied in this induction.

Euglenatides, Potent Antiproliferative Cyclic Peptides Isolated from the Freshwater Photosynthetic Microalga Euglena gracilis.

By limiting the nitrogen source to glutamic acid, we isolated cyclic peptides from Euglena gracilis containing asparagine and non-proteinogenic amino acids. Structure elucidation was accomplished through spectroscopic methods, mass spectrometry and chemical degradation. The euglenatides potently inhibit pathogenic fungi and cancer cell lines e.g., euglenatide B exhibiting IC50 values of 4.3 µM in Aspergillus fumigatus and 0.29 µM in MCF-7 breast cancer cells. In an unprecedented convergence of non-ribosomal peptide synthetase and polyketide synthase assembly-line biosynthesis between unicellular species and the metazoan kingdom, euglenatides bear resemblance to nemamides from Caenorhabditis elegans and inhibited both producing organisms E. gracilis and C. elegans. By molecular network analysis, we detected over forty euglenatide-like metabolites in E. gracilis, E. sanguinea and E. mutabilis, suggesting an important biological role for these natural products.

Cross-cultural adaptation and validation of a questionnaire on eating habits and physical activity of university students in confinement due to coronavirus disease.

OBJECTIVE: The purpose of the current study was to cross-culturally adapt and validate an online questionnaire to assess eating habits and physical activity of university students under confinement due to coronavirus disease (COVID-19). DESIGN: Generation of a cross-sectional online survey to university students conducted during confinement due to COVID-19. The study was divided into two phases. SETTINGS: Students, Chile. PARTICIPANTS: Phase 1 considered the process of translation and back translation, expert panel, cultural adaptation and the generation of a pilot to validate a preliminary format of the questionnaire. In Phase 2, information from the instrument was collected from two hundred and sixty-eight university students, ages 16 to 30 years old, with a mean age of 21·6 (3·3) The major proportion of participants were female (82 %). RESULTS: The adapted questionnaire was statistically validated in three dimensions: (A) eating habits and behaviours during quarantine, (B) perception of risk and (C) physical activity changes during the quarantine. The reliability of Cronbach's α for dimensions A, B and C was 0·59, 0·85 and 0·97, respectively. The complete questionnaire obtained 0·61 in internal consistency and 0·61 (0·58-0·67) ICC reliability. A statistically significant positive correlation matrix was observed. CONCLUSIONS: This questionnaire is a practical tool to obtain accurate information about the relation of COVID-19 confinement on people's eating habits and physical activity. Therefore, it could contribute to establishing appropriate strategies to prevent negative effects on people's health.

Review of transition of care literature: Epidermolysis bullosa-A paradigm for patients with complex dermatologic conditions.

BACKGROUND: Transition from pediatric to adult care is a critical component of health care for children with long-term needs. The characteristics of epidermolysis bullosa (EB) demand higher than average levels of provider support. There is consensus among health care professionals regarding the importance of transition; however, there is a scarcity of practical information regarding models for patients with EB. OBJECTIVE: To review transition of care programs in varying specialties. Highlight practical considerations to facilitate the development of programs for patients with EB and other complex dermatologic conditions. METHODS: Articles were identified via MEDLINE and EMBASE health literature databases and screened for relevance to transition of care. RESULTS: Various models for transition exist. A well-executed formal transition program, early introduction, interdisciplinary collaboration, and psychosocial support were themes associated with successful outcomes. LIMITATIONS: Transition of care programs that have not been described in the literature are not reflected in this review. CONCLUSIONS: Patients with EB have unique needs that affect transition and span expertise across traditional boundaries, such as dependency on others for daily skin care, failure to thrive, and risk of squamous cell carcinoma. Given the rarity of the disease, patients with EB will benefit from collaborative efforts to develop programs to optimize successful transition.

Anti-CD44-Conjugated Olive Oil Liquid Nanocapsules for Targeting Pancreatic Cancer Stem Cells.

The latest trends in cancer research and nanomedicine focus on using nanocarriers to target cancer stem cells (CSCs). Specifically, lipid liquid nanocapsules are usually developed as nanocarriers for lipophilic drug delivery. Here, we developed olive oil liquid NCs (O2LNCs) functionalized by covalent coupling of an anti-CD44-fluorescein isothiocyanate antibody (αCD44). First, O2LNCs are formed by a core of olive oil surrounded by a shell containing phospholipids, a nonionic surfactant, and deoxycholic acid molecules. Then, O2LNCs were coated with an αCD44 antibody (αCD44-O2LNC). The optimization of an αCD44 coating procedure, a complete physicochemical characterization, as well as clear evidence of their efficacy in vitro and in vivo were demonstrated. Our results indicate the high targeted uptake of these αCD44-O2LNCs, and the increased antitumor efficacy (up to four times) of paclitaxel-loaded-αCD44-O2LNC compared to free paclitaxel in pancreatic CSCs (PCSCs). Also, αCD44-O2LNCs were able to selectively target PCSCs in an orthotopic xenotransplant in vivo model.

Long-Term Outcomes of 496 Anatomical Cementless Modular Femoral Stems: Eleven to Twenty Years of Follow-Up.

BACKGROUND: The aim of the study is to assess the long-term outcomes of this specific stem (anatomical cementless modular stem ESOP), to review the survivorship, complication rate, and radiographic and clinical outcomes. METHODS: Descriptive and analytical retrospective longitudinal observational study of patients was operated on total hip arthroplasty between 1998 and 2007. Four hundred ninety-six prostheses corresponding to 447 patients were reviewed, mean age was 65.8 years (standard deviation [SD] ±11.6 years), and median follow-up time was 13.4 years (range 1-20). The most used cups were cementless (75.8%). The most frequent friction pairs were metal-polyethylene (53.1%) and ceramic-polyethylene (24.2%). Main variables analyzed were stem survival, subsidence, coronal orientation, osteolysis, reintervention, and Oxford Hip Score. RESULTS: From 496 implants, there were 22 lost to follow-up (4.4%). Stem revision was performed in 51 patients: 26 periprosthetic joint infections (2-stage revision), 16 periprosthetic fractures, and 8 one-stage revisions (6 real aseptic loosening with negative culture after revision). The stem survivorship at more than 15 years for any reason was 89.2% and for aseptic loosening 97.97%. No specific complications were found due to modularity. The mean subsidence and orientation was 2.06 mm (SD ±5.11 mm) and 0.41° varus (SD ±2.20°) respectively. Subsidence >5 mm or varus >5° was associated with a higher revision rate. Osteolysis was found in 110 patients (zone I = 79, VII = 57), associated with zirconium-polyethylene, without relationship to the stem revision rate or Oxford Hip Score. CONCLUSION: To our knowledge, this study represents the largest series of this stem, revealing an excellent survival rate and long-term clinical outcomes similar to the best results of classical cementless stems published in the literature.

Co-occurring Congeners Reveal the Position of Enantiomeric Amino Acids in Nonribosomal Peptides.

The absolute configuration of the constituent amino acids in microbial nonribosomal peptides is typically determined by Marfey's method after total hydrolysis of the peptide. A challenge to structure elucidation arises when both d and l enantiomeric configurations of an amino acid are present. Determining the actual position of each amino acid enantiomer within the peptide sequence typically requires laborious approaches based on peptide partial hydrolysis or even total synthesis of the possible diastereomers. Herein, an alternative solution is discussed based on the homogeneous backbone chirality that governs all peptides biosynthesized by a common nonribosomal peptide synthetase. The information on configuration provided by Marfey's analysis of co-occurring minor congeners can reveal unequivocally the stereochemical sequence of the whole peptide family.

A newly described cutaneous reaction at sites of insulin pump use in a child with Type 1 diabetes.

Continuous subcutaneous insulin infusion (CSII), or insulin pumps, with or without continuous glucose monitoring (CGM) devices have become the standard of care for patients with type 1 diabetes. While increasingly popular, a wide range of reported skin reactions to CSII and CGM devices was found. We present this case of a pyogenic granuloma-like neutrophilic and granulomatous response to an insulin pump to increase awareness of a previously uncharacterized cutaneous adverse reaction at insulin pump infusion sites.

Prácticas de salud de dominación y exclusión: visiones de activistas, profesionales e investigadoras sobre situación de esterilizaciones forzadas en mujeres y niñas con discapacidad en España / Health practices of domination and exclusion: views of activists, professionals and researchers on the situation of forced sterilization of women and girls with disabilities in Spain

Resumen En 2017, las Naciones Unidas declaran que 38 países, entre ellos España, continuaban realizando esterilizaciones forzadas en mujeres y niñas con discapacidad, a pesar de tratarse de una práctica ampliamente condenada por diversos organismos internacionales de derechos humanos. Este estudio analizó la situación de esta práctica en España, desde la perspectiva de activistas, profesionales e investigadoras con experiencias vinculadas a este colectivo. La metodología empleada fue de tipo cualitativa basada en la Teoría Fundamentada Constructivista. Se aplicaron entrevistas semiestructuradas en profundidad a 22 informantes, que representaron a 6 comunidades autónomas del país. Las participantes identificaron un modelo de prácticas de salud de dominación y exclusión, donde la sexualidad y reproducción de mujeres con discapacidad ha sido objeto de expropiación, alienación y desprecio por parte de sistemas, estructuras y políticas diferenciadas, con escasa transferencia del marco global de derechos humanos a su realidad. La violencia sexual ha pasado inadvertida, naturalizándose por el entorno, han promovido mecanismos de exclusión social e inequidades en salud, al privarles de sus derechos humanos fundamentales. Al tratarse de una situación que se replica en diversos lugares del mundo, debiese considerarse un tema de relevancia para la salud pública internacional.

Abstract In 2017, the United Nations declared that 38 countries, including Spain, continued to practice forced sterilizations on women and girls with disabilities, despite it being a practice widely condemned by various international human rights organizations. This study analyzed the situation of this practice in Spain, from the perspective of activists, professionals and researchers with experiences related to this group. The methodology used was qualitative, based on Constructivist Grounded Theory. Semi-structured in-depth interviews were applied to 22 informants, representing 6 autonomous communities of the country. The participants identified a model of health practices of domination and exclusion, where the sexuality and reproduction of women with disabilities has been the object of expropriation, alienation and contempt by systems, structures and differentiated policies, with little transfer of the global framework of human rights to their reality. Sexual violence has gone unnoticed, becoming naturalized by the environment, and has promoted mechanisms of social exclusion and health inequities, depriving them of their fundamental human rights. As it is a situation that is replicated in various parts of the world, it should be considered an issue of relevance for international public health.

Prácticas de salud de dominación y exclusión: visiones de activistas, profesionales e investigadoras sobre situación de esterilizaciones forzadas en mujeres y niñas con discapacidad en España / Health practices of domination and exclusion: views of activists, professionals and researchers on the situation of forced sterilization of women and girls with disabilities in Spain

Resumen En 2017, las Naciones Unidas declaran que 38 países, entre ellos España, continuaban realizando esterilizaciones forzadas en mujeres y niñas con discapacidad, a pesar de tratarse de una práctica ampliamente condenada por diversos organismos internacionales de derechos humanos. Este estudio analizó la situación de esta práctica en España, desde la perspectiva de activistas, profesionales e investigadoras con experiencias vinculadas a este colectivo. La metodología empleada fue de tipo cualitativa basada en la Teoría Fundamentada Constructivista. Se aplicaron entrevistas semiestructuradas en profundidad a 22 informantes, que representaron a 6 comunidades autónomas del país. Las participantes identificaron un modelo de prácticas de salud de dominación y exclusión, donde la sexualidad y reproducción de mujeres con discapacidad ha sido objeto de expropiación, alienación y desprecio por parte de sistemas, estructuras y políticas diferenciadas, con escasa transferencia del marco global de derechos humanos a su realidad. La violencia sexual ha pasado inadvertida, naturalizándose por el entorno, han promovido mecanismos de exclusión social e inequidades en salud, al privarles de sus derechos humanos fundamentales. Al tratarse de una situación que se replica en diversos lugares del mundo, debiese considerarse un tema de relevancia para la salud pública internacional.

Abstract In 2017, the United Nations declared that 38 countries, including Spain, continued to practice forced sterilizations on women and girls with disabilities, despite it being a practice widely condemned by various international human rights organizations. This study analyzed the situation of this practice in Spain, from the perspective of activists, professionals and researchers with experiences related to this group. The methodology used was qualitative, based on Constructivist Grounded Theory. Semi-structured in-depth interviews were applied to 22 informants, representing 6 autonomous communities of the country. The participants identified a model of health practices of domination and exclusion, where the sexuality and reproduction of women with disabilities has been the object of expropriation, alienation and contempt by systems, structures and differentiated policies, with little transfer of the global framework of human rights to their reality. Sexual violence has gone unnoticed, becoming naturalized by the environment, and has promoted mechanisms of social exclusion and health inequities, depriving them of their fundamental human rights. As it is a situation that is replicated in various parts of the world, it should be considered an issue of relevance for international public health.

Viennamycins: Lipopeptides Produced by a Streptomyces sp.

Extracts from Streptomyces sp. S4.7 isolated from the rhizosphere of edelweiss, an alpine medicinal plant, exhibited activity against Gram-positive bacteria. LC-HRMS analyses of the extracts resulted in the detection of two unknown, structurally related lipopeptides that were assumed to be responsible for the antibiotic activity. LC-MS guided isolation and structure elucidation of viennamycins A and B (1 and 2) by HR-MS/MS, 1D and 2D NMR, and Marfey's analyses revealed them to be novel compounds, with viennamycin A containing cysteic acid, a unique feature for lipopeptides. Tests for antibacterial, antifungal, and cytotoxic activities of purified viennamycins, both with and without divalent cations, did not reveal any bioactivity, suggesting that their biological function, which could not be determined in the tests used, is atypical for lipopeptides. The genome of Streptomyces sp. S4.7 was sequenced and analyzed, revealing the viennamycin biosynthetic gene cluster. Detailed bioinformatics-based analysis of the viennamycin gene cluster allowed elucidation of the biosynthetic pathway for these lipopeptides.

Iron and Heme Metabolism at the Leishmania-Host Interface.

Species of the protozoan Leishmania are causative agents of human leishmaniasis, a disease that results in significant death, disability, and disfigurement around the world. The parasite is transmitted to a mammalian host by a sand fly vector where it develops as an intracellular parasite within macrophages. This process requires the acquisition of nutritional iron and heme from the host as Leishmania lacks the capacity for de novo heme synthesis and does not contain cytosolic iron-storage proteins. Proteins involved in Leishmania iron and heme transport and metabolism have been identified and shown to be crucial for the parasite's growth and replication within the host. Consequently, a detailed understanding of how these parasites harness host pathways for survival may lay the foundation for promising new therapeutic intervention against leishmaniasis.

Leishmania heme uptake involves LmFLVCRb, a novel porphyrin transporter essential for the parasite.

Leishmaniasis comprises a group of neglected diseases caused by the protozoan parasite Leishmania spp. As is the case for other trypanosomatid parasites, Leishmania is auxotrophic for heme and must scavenge this essential compound from its human host. In mammals, the SLC transporter FLVCR2 mediates heme import across the plasma membrane. Herein we identify and characterize Leishmania major FLVCRb (LmFLVCRb), the first member of the FLVCR family studied in a non-metazoan organism. This protein localizes to the plasma membrane of the parasite and is able to bind heme. LmFLVCRb levels in Leishmania, which are modulated by overexpression thereof or the abrogation of an LmFLVCRb allele, correlate with the ability of the parasite to take up porphyrins. Moreover, injection of LmFLVCRb cRNA to Xenopus laevis oocytes provides these cells with the ability to take up heme. This process is temperature dependent, requires monovalent ions and is inhibited at basic pH, characteristics shared by the uptake of heme by Leishmania parasites. Interestingly, LmFLVCRb is essential as CRISPR/Cas9-mediated knockout parasites were only obtained in the presence of an episomal copy of the gene. In addition, deletion of just one of the alleles of the LmFLVCRb gene markedly impairs parasite replication as intracellular amastigotes as well as its virulence in an in vivo model of cutaneous leishmaniasis. Collectively, these results show that Leishmania parasites can rescue heme through plasma membrane transporter LFLVCRb, which could constitute a novel target for therapeutic intervention against Leishmania and probably other trypanosomatid parasites in which FLVCR genes are also present.

Executive Summary of the Consensus Statement of the Spanish Society of Infectious Diseases and Clinical Microbiology (SEIMC), the Spanish Society of Tropical Medicine and International Health (SEMTSI), the Spanish Association of Surgeons (AEC), the Spanish Society of Pneumology and Thoracic Surgery (SEPAR), the Spanish Society of Thoracic Surgery (SECT), the Spanish Society of Vascular and Interventional Radiology (SERVEI), and the Spanish Society of Paediatric Infectious Diseases (SEIP), on the Management of Cystic Echinococcosis.

The Spanish Society of Infectious Diseases and Clinical Microbiology (SEIMC), the Spanish Society of Tropical Medicine and International Health (SEMTSI), the Spanish Association of Surgeons (AEC), the Spanish Society of Pneumology and Thoracic Surgery (SEPAR), the Spanish Society of Thoracic Surgery (SECT), the Spanish Society of Vascular and Interventional Radiology (SERVEI), and the Spanish Society of Paediatric Infectious Diseases (SEIP) considered it pertinent to issue a consensus statement on the management of cystic echinococcosis (CE) to guide healthcare professionals in the care of patients with CE. Specialists from several fields (clinicians, surgeons, radiologists, microbiologists, and parasitologists) identified the most clinically relevant questions and developed this Consensus Statement, evaluating the available evidence-based data to propose a series of recommendations on the management of this disease. This Consensus Statement is accompanied by the corresponding references on which these recommendations are based. Prior to publication, the manuscript was open for comments and suggestions from the members of the SEIMC and the scientific committees and boards of the various societies involved.

Silica nano supra-assembly for the targeted delivery of therapeutic cargo to overcome chemoresistance in cancer.

Cancer cells become resistant over the period to chemotherapeutic drugs and pose a challenging impediment for oncologists in providing effective treatment. Nanomedicine allows to overcome chemoresistance and is the focus of our investigation. Silica nanostructures have been highlighted as an interesting drug delivery platform in vitro and in vivo applications. Here we show the validity of nanomedicine approach for targeted chemotherapeutic cargo delivery to overcome chemoresistance in cancer cells both in vitro and in vivo. For demonstrating the concept, we functionalised ∼100 nm long porous silica nanoparticles (∼20 nm diameter ordered pore structure) by conjugating anticancer drug, cytochrome c enzyme and dual-function anticancer aptamer AS1411 in single supra-assembled nanocargos. The supra-assembly on the porous silica nanostructure allows for a high loading of catalytic enzyme cytochrome c, anticancer drug and aptamer. The silica supra-assembly is characterized by transmission electron microscopy (TEM) and Brunauer-Emmett-Teller (BET) surface area analysis. Conjugation of cargoes has been monitored at each step by UV-vis and Fluorescence spectroscopy. Finally, the constructed supra-assembled nanocarrier tested on chemoresistance colon cancer (HCT116) cells. A pH-responsive, intracellular theranostic cargo delivery has been achieved and the triple action of the nanocargo made an efficient killing of drug resistance colon cancer cells in vitro (∼ 92% cell death) through triplex therapy effects by supressing the P-glycoprotein (P-gp) level. Furthermore, in vivo animal toxicity studies demonstrated, the supra-assembled nanocargos have encouraging safety index to be used in cancer therapy and drug delivery applications.

The Gender Violence - Implicit Association Test to measure attitudes toward intimate partner violence against women

Attitudes play a central role in intimate partner violence against women and are related to its origin, to the responses of women who suffer violence, and to the settings where it occurs. In fact, these attitudes are recognized as one of the risk factors linked to violent perpetration and to public, professional, and victim responses to this type of violence. However, even though available research generally shows a broad rejection of this violence, it remains a serious social and health problem that has reached epidemic proportions. This suggests that the information available about these attitudes (obtained through explicit and direct measures, i.e., self-reports) may be distorted or influenced by factors such as social desirability. In this context, the overall objective of our research project is to provide multi-method measures (explicit and implicit) of attitudes toward intimate partner violence against women, and the main goal of this paper is to propose an instrument for the implicit measurement of these attitudes. In this regard, the Implicit Association Test (IAT) is the most common procedure used, providing a superior predictive validity compared to explicit measures for socially sensitive topics. We will present an exploratory study that describes its adaptation for our purposes, and the development of the Gender Violence - Implicit Association Test (GV-IAT) to use among Spanish-speaking populations, and discuss the strengths and limitations of this proposal.

Heme synthesis through the life cycle of the heme auxotrophic parasite Leishmania major.

Heme is an essential molecule synthetized through a broadly conserved 8-step route that has been lost in trypanosomatid parasites. Interestingly, Leishmania reacquired by horizontal gene transfer from γ-proteobacteria the genes coding for the last 3 enzymes of the pathway. Here we show that intracellular amastigotes of Leishmania major can scavenge heme precursors from the host cell to fulfill their heme requirements, demonstrating the functionality of this partial pathway. To dissect its role throughout the L. major life cycle, the significance of L. major ferrochelatase (LmFeCH), the terminal enzyme of the route, was evaluated. LmFeCH expression in a heterologous system demonstrated its activity. Knockout promastigotes lacking lmfech were not able to use the ferrochelatase substrate protoporphyrin IX as a source of heme. In vivo infection of Phlebotomus perniciosus with knockout promastigotes shows that LmFeCH is not required for their development in the sandfly. In contrast, the replication of intracellular amastigotes was hampered in vitro by the deletion of lmfech. However, LmFeCH-/- parasites produced disease in a cutaneous leishmaniasis murine model in a similar way as control parasites. Therefore, although L. major can synthesize de novo heme from macrophage precursors, this activity is dispensable being an unsuited target for leishmaniasis treatment.-Orrego, L. M., Cabello-Donayre, M., Vargas, P., Martínez-García, M., Sánchez, C., Pineda-Molina, E., Jiménez, M., Molina, R., Pérez-Victoria, J. M. Heme synthesis through the life cycle of the heme auxotrophic parasite Leishmania major.