Recent developments in genomics opened unprecedented opportunities to study the interface among ecology, evolution, and genetics, and made possible to understand adaptations of organisms in detail. We use natural populations of the model organism Drosophila melanogaster and its relatives and combine modern next-generation sequencing technologies integrated with laboratory and field studies to understand natural selection and adaptations. The main goal of this project is to consolidate my research group on ecological genomics of insects. We will expand our studies on climate adaptations and adaptations to ecological interactions. First, we will use historical samples to study genome wide long-term stability of clines, response to global warming, and the relative importance of natural selection versus admixture. Second, we will use a highly integrative approach to study clinal variation in South American D. melanogaster populations. Third, we will develop a mapping panel with high statistical power and fine resolution to dissect the genetic architecture of climate adaptation. Forth, we will use an integrative approach to study climate adaptation in Neotropical drosophilid species collected in the Atlantic Forest and the Cerrado. Finally, we will test other factors, beyond climate adaptation, that affect the interaction between Neotropical drosophilid species and the endosymbiont Wolbachia, including virus protection, cytoplasmatic incompatibility, and phylogenetic origins. We will train postdocs, graduate, and undergraduate students in a modern theme, consolidate a network of nine associated researchers from eight universities in four different countries, and address broad interest questions that will result in publications with high potential impact. (AU)

The alarming rate of unintended pregnancies (almost 50%) contributes to the unsustainable world population growth and can have serious health and socioeconomic consequences for women and their families. This scenario underscores the need for novel contraceptives, especially male options, since they are limited to condoms and vasectomy. Druggable proteins with crucial roles in regulating sperm function are compelling pharmacological targets for non-hormonal male contraceptives. The sperm-binding protein EPPIN fits into this male contraception profile. Under normal physiological conditions, EPPIN interaction with the seminal plasma protein SEMG1 on the human sperm surface leads to transient inhibition of sperm function after ejaculation. The ability of EPPIN ligands targeting the SEMG1-binding site to inhibit sperm motility triggered the development of fully innovative spermostatic drugs. We recently showed that EPPIN binds SVS2 (mouse SEMG1 ortholog) in mouse spermatozoa. These facts indicate that EPPIN is evolutionarily conserved in terms of protein-binding partners and roles in sperm function between humans and mice. Indeed, both SEMG1 and SVS2 are seminal plasma inhibitory factors of sperm motility and capacitation-associated events (e.g., hyperactivation and acrosome reaction) and essential for sperm survival in the uterus. These findings pave the way for using mouse models to unravel EPPIN roles on male fertility in vivo. Here, we aim to unfold the mechanisms underlying EPPIN modulation of sperm function, the relevance of its protein-protein interactions in such events using mice as translational models, and to search for novel peptide-based EPPIN ligands displaying spermostatic activity. We will perform protein-protein interaction assays to determine EPPIN/SVS2 binding sequences crucial for their interaction. To test the hypothesis that EPPIN/SVS2 binding is a key event for the regulation of sperm function, we will employ state-of-the-art functional, molecular, and live single-cell imaging methodologies to evaluate whether recombinant SVS2 truncations containing EPPIN-binding sequences inhibit functional parameters (motility, acrosome reaction, and fertilization rates) and signaling pathways (cAMP production, intracellular Ca2+ levels, intracellular pH, and protein phosphorylation profile) governing sperm fertility potential. We will also determine the functional requirement for Eppin in male fertility in vivo generating transgenic Eppin knock-out mice using the CRISPR/Cas9 technology. We will build on this knowledge to rationally design novel peptide-based EPPIN ligands using state-of-the-art microwave-assisted solid-phase peptide synthesis and test their effects on human and mouse sperm motility. This proposal creates a network of scientists from Brazil, Argentina, Portugal, and the United Kingdom in a multidisciplinary team to generate novel insights on EPPIN as a sperm drug target for male contraception, thus providing solid grounds for innovation in Reproductive Pharmacology. (AU)

Colorants have been used for centuries to enhance or restore the original appearance of different products and ensure their uniformity. The global market of colorants includes artificial and natural molecules; however, due to the consumer concerns regarding their health, the market for natural products have been raising in several industrial segments, especially, increasing the studies for the microbial production of natural colorants. These bioproducts exhibit further advantages because of their biological activities such as antioxidant and antimicrobial activity. Therefore, this project aims to develop a biorefinery of microbial colorants towards a sustainable circular bioeconomy, following the development of 2 sub-projects: 1) production of azaphilone colorants by Talaromyces amestolkiae and; 2) production of astaxanthin by Phaffia rhodozyma. For both processes, agroindustrial residues, namely, citrus waste, sugarcane bagasse and straw, will be valorized through their polysaccharide's hydrolysis, which will be then employed as carbon source in the stage of submerged cultivation in stirred tank bioreactor. After, the extraction of colorants using biosolvents-based liquid-liquid extraction platforms will be studied, to ensure an ecologically and environmentally sustainable process. A scale-up of the best process for extracting each colorant will be performed using centrifugal partition chromatography. The toxicity of azaphilone colorants and their application in hydrogels and textile matrices will be also evaluated. As the final stage of the project, the integration of the entire process will combine main upstream and downstream processing aiming to maximize the yield of target colorants. The long-term sustainability of technologies will be assessed through both techno-economic and Life Cycle Analysis (LCA). The present proposal intends to apply a holistic and integrative approach of chemical and biological engineering for the microbial production of natural colorants, aiming to develop an environmentally and economically sustainable process, with a positive impact on the environment and helping to democratize the access to natural and healthier additives. (AU)

Microbial biofilms are ubiquitous and fundamental for several natural and industrial processes and disease development. Dental caries results from dysbiosis in biofilm caused by microbial metabolism of dietary carbohydrates. These carbohydrates are substrates to produce extracellular matrix components and organic acids. These acids are trapped by the matrix inside the biofilm and at the interface of biofilm/teeth. Over time, these acids demineralize the surfaces and lead to teeth structure loss, affecting oral and systemic health. The main matrix components are exopolysaccharides (soluble and insoluble), extracellular DNA, and lipoteichoic acids. The association of these components restricts the diffusion of substances in and out of biofilms and renders them resistant. The main goal is to unravel the biology of the extracellular matrix for the coordinated construction of cariogenic biofilm and develop strategies for controlling them. Thus, we propose to: 1) determine how Streptococcus mutans (main known matrix producer species) cell wall and membrane turnover affects the bridging of extracellular matrix components and consequent configuration (creation of microniches); 2) evaluate how non-matrix producers' species benefit from the matrix assembled by S. mutans and how localized alkali production by early colonizers interferes with the matrix; 3) determine the contribution of species (other than S. mutans) to the matrix build-up during the development of in situ biofilms; these species (their pathways) could be targeted to control dysbiosis; 4) assess the efficacy of drugs associated or not with nanotechnology for the prevention of dental caries in vivo, including the evaluation of the microbiota (diversity), matrix (matrixome), aspects associated with antimicrobial tolerance and resistance (resistome), and demineralization. (AU)

Brazil is the third country in the world in total tons of pesticides use per year. Pesticides are toxic substances or a mixture of substances that are naturally or chemically synthesized. They are widely used in agriculture to control plagues; however, since they are toxic to life, they are inevitably toxic to humans. Heavy use of pesticides has contaminated ecosystems. In Brazil, at least 27 pesticides are found in potable water, with a range of concentrations. Ministry of Health set concentration threshold levels for each pesticide, in theory not toxic, allowed to be found in potable water. However, in the literature, there is no systematic investigation of heart and nervous system toxicity of human tissue by these compounds at concentrations set by Ministry of Health. Also, there is no information if a mixture of these pesticides potentiates toxicity. Thus, to address these questions, we had the following aims: 1) evaluate in vitro single or mixture of pesticides in acute and chronic toxicity for cardiomyocytes and neurons derived from human-induced pluripotent stem cells, using the concentration approved by the Ministry of Health; 2) using mice determine in vivo if long-term low dose of single or a mixture of pesticides impact the functioning of the heart and central nervous system; 3) investigate in vivo how long-term low-dose pesticides impact the functioning of the heart and central nervous system in offspring of pregnant mice exposed to pesticides during pregnancy; 4) if SCN5A polymorphisms enhance the potency and toxicity of pesticides in vitro. At the end we will be able to determine the safety of potable water of Brazil in terms of pesticide contamination or if changes are needed in the guidelines of Ministry of Health. (AU)

Extracellular Vesicles (EVs) play an important role in export systems. They act as vehicles for the transference of complex cargoes with broad biological functions. These vesicles cross the fungal cell wall to reach the extracellular space and transport its major virulence factors contributing to fungal infection outcomes. Since we had the FAPESP young investigator grant phase 1, our group has demonstrated that fungal EVs exerts immunomodulatory functions during fungus-host interactions and mediate intra-species cellular communication. Furthermore, we have verified that fungal EVs may also plays crucial roles regarding to inter-species cellular communication. These observations motivate the proposal to study the fungal inter-species cellular communication, and to better understand the role of fungal EVs in immunomodulation and intra-species cellular communication. More specifically, we propose to do the following aims: (1) to clarify the immunomodulatory properties of EVs from C. neoformans, P. brasiliensis, A. fumigatus, and C. albicans; (2) to dissect the responsible interactions of EVs from C. neoformans, P. brasiliensis, A. fumigatus, and C. albicans in intra- and inter-species communication by several approaches. Our expectation is that fungal EVs are responsible for important functions on fungal infection process. Thus, the development of this young investigators grants phase 2 will consolidate the EVs research line initiated by FAPESP and will provide opportunities to develop new strategies against fungal infections. (AU)

Heart failure is a public health problem and is one of the leading causes of death worldwide. Despite current advances, the prognosis is poor, and there remains no cure for heart failure. Heart failure is characterised by breathlessness (respiratory irregularities), bronchoconstriction, leg oedema, a progressive loss in cardiac output, left ventricular hypertrophy, sleep apnoea, and raised sympathetic activity. This proposal aims to determine a treatment strategy to reduce sympathetic hyperactivity, respiratory irregularities, and bronchoconstriction in heart failure. A clue to a driver of heart failure arises from the strong correlation between peripheral chemoreceptor reflex sensitivity and survival post-myocardial infarction. Of particular interest is the purinergic signalling (ATP and P2X3 receptors), which plays an essential role in both the sensitisation (increase in tonicity) and hyperreflexia of peripheral sensory afferent neurones in several pathological conditions. Activation of carotid body produces release of ATP from type I (glomus) and type II (glial) cells and stimulates purinergic receptors on the terminals of petrosal ganglion afferents, which form a basket like endings on these cells. In the present research project, we will evaluate whether purinergic signalling and glial cells mediate the afferent tonicity and sensitisation within the carotid body in heart failure and explore the potential therapeutic benefit of abolishing aberrant purinergic signalling in a small animal model of heart failure. The following hypotheses will be tested: I) the intrinsic excitability is enhanced and the expression of P2X3 receptors are upregulated in the chemoreceptive petrosal ganglion neurones in rats after myocardial infarction; II) upregulation of P2X3 receptors induces sensitisation of the peripheral chemoreflex responses (respiratory, sympathetic and chemoreceptive neurones) after myocardial infarction; III) ATP bioavailability increased in the carotid body after myocardial infarction, and; IV) following myocardial infarction, chronic genetic manipulation of glomus and glial cells purinergic signalling in the carotid body will lower sympathetic activity, improve cardiac function, reduce respiratory irregularities, bronchoconstriction, left ventricular mass and myocardial fibrosis. We propose that aberrant ATP signalling triggers hypertonicity in the carotid body that via P2X3 receptors plays a crucial role in heart failure progression and offers a therapeutic angle to reverse multiple components (sympathetic hyperactivity, respiratory irregularities, and bronchoconstriction) of its pathogenesis. (AU)

Covering less than 0.1% of the ocean flor, warm, shallow-water coral reefs are recognized for supporting over a quarter of the worlds' marine biodiversity and also for the critical ecosystem services they provide to human kind. Nonetheless, due to the ongoing climate change and other anthropogenic challenges, about 75% of the coral reefs are considered threatened, with several already severely degraded or under phase-shift to algal dominated habitats. Even though some hope has been put forward - the extinction risk of most coral species is lower than previously thought and, as a lineage, scleractinian corals have persisted to at least four of the five great mass extinction events -, it is a common sense that coral reefs are not going to be the same for the future generations. In the Southwestern Atlantic (SWA) the panorama is not different, with stronger and more frequent ocean thermal anomalies taking its toll on coral cover. Nevertheless, although displaying low diversity (but high endemicity), corals from this region have been purported to be more resistant and even more resilient to the warming ocean effects. Such "shielding" being the result of the more nutrient rich, turbid waters that our corals are adapted. However, we know virtually nothing on the genetic structure and genetic diversity of the SWA coral populations. Overall, genetic diversity is an essential source for biodiversity, providing the raw material for evolution. When interconnected, populations tend to preserve greater genetic diversity compared to more isolated populations, which in turn are more susceptible to genetic drift effects. Thus, endeavoring to fill such gaps and, consequently, improve our understanding in how climate change has and is shaping SWA coral evolution as a lineage and as a system, the present proposal consists of consolidating an interdisciplinary research line cored on RNA/DNA deep sequencing and its usefulness at the Department of Marine Science of the Federal University of São Paulo (DCMar-UNIFESP). Overall, advances in high-throughput sequencing and SNP genotyping technologies have greatly expanded our ability to identify signatures of positive selection. Such signatures help us to delimit regions of the genome that are, or have been, functionally important, poiting towards the identification of genetic variation that contributes to phenotypic diversity and genotype selection. Examining such diversity in the light of phylogeny / phylogeography / holobiont will open horizons to explore where, when and how traits - of any nature - have changed over different time scales. Retrieved biological patterns at population and specific levels will also enable the proposition of evolutionary processes and subsequent transformations of corals in light of environment variables. Therefore, at the onset of the Decade of the Ocean, and understanding that such knowledge is essential to define efficient management and conservation strategies, the present proposal will persue long lasting questions regarding the major architects of shallow-water coral reefs in times not just of major scientific interest, but also intense public and media concern because of the uncertain fate of these ecosystems in the face of ever increasing anthropogenic challenges. (AU)

Despite recent scientific advances and the growth of the Gluten-Free (GF) market, important issues such as the improvement of the sensory and nutritional quality of GF bakery foods, shelf life, convenience, and variety all still need to be addressed. All these factors are essential for people with gluten-related disorders to follow a more satisfying diet, in addition to improving their quality of life and health through nutrition. To overcome these challenges, the main goal of this proposal is applying innovative technological approaches to improve the nutritional quality of GF bakery products and to assess their potential health benefits from in vitro, in vivo, and clinical studies. The proposal has a multifaceted approach with objectives focused on the interface between food science, nutrition, and health to develop new approaches and overcome the tremendous scientific, technological and market challenges regarding GF foods. GF bread is the most challenging product, and therefore the focus of the proposal. Provide realistic and useful technological solutions to meet the consumers and producers demands, demonstrating the social and economic impact of the project. Additionally, the results will allow the understanding of the health benefit effects of these specially designed GF products, which can support a GF diet with products with low glycemic response, antioxidant, and anti-inflammatory potential and help the intestinal microbiota to reach homeostasis, all that provide better nutrition and health for consumers. In addition, the knowledge created will narrow the gap regarding the composition of Brazilian GF products, as well as producers' knowledge and consumer´s opinions, perceptions, and behavior about GF bakery foods. (AU)

The applicant is a young outstanding scientist who has published 45 articles (1569 citations, H-index = 15), supervised 5 PhD thesis, coordinated successful project grants (over 500,000 EURO), and became quite early an Associate Professor. His studies were pioneer in developing spermatogonial stem cell (SSC) transplantation in zebrafish, and showing the role of crucial growth factors, Igf3, Amh and Gnih, on fish SSC regulation. The overarching goal of this research is to investigate new paracrine factors and indentify long non-coding RNAs (lncRNAs) that regulate SSC activity, and develop biotechnological strategies to delay puberty in fish. Early sexual maturation of males is still one the major problems limiting the further development of aquaculture in the world. Sexual precocity is an undesirable trait in fish which is correlated with negative aspects such as reduced growth and lower feed conversion. In males, the onset of puberty is generally associated with a switch in SSC activity, from self-renewal to differentiation towards meiosis. Therefore, understanding the mechanisms controlling the switch in SSC activity is the basis for developing approaches to control precocious puberty in males. Focusing on new growth factors and lncRNAs, functional studies that combine in vivo approaches and gain-of-function ex vivo models will be employed. Functional studies will be evaluated with a combination of histomorphometric, biochemical and molecular approaches. In this project, lambari (Astyanax altiparanae), a Neotropical model and promising species for Brazilian aquaculture, and zebrafish will be used as experimental models. Based on the knowledge acquired on the new gonadal factors, we propose to employ somatic gene transfer to delay the start of puberty in fish. The applicant is considered fully capable to coordinate the current project, and certainly will generate important knowledge to improve aquaculture. (AU)