Multivariate analysis of chemical parameters of cocoa: a contribution to origin designation / Análise multivariada de parâmetros químicos do cacau: uma contribuição para a denominação de origem

Brazil has been trying to become a key player in the specialized market through the production of fine chocolate. However, there is still no rules about the chemical quality of cocoa beans that allows it to be grouped by micro-regions. In this context, the objective of this work was to determine some chemical parameters of cocoa samples, aiming to contribute to the creation of the designation of origin for the cocoa produced in the south of Bahia. For this propose, proteins, lipids, total minerals, and fatty acid profile analysis were performed. The results obtained were correlated using multivariate statistical tests. The chemical composition of the cocoa beans, allowed to differentiate samples of cocoa, revealing the formation of three groups of samples. The two main components (lipids and proteins) were analysed together, characterizing the producing region. Regarding the analysis of the fatty acid profile, they showed that the cocoa harvested in the main season presents a higher influence of saturated fatty acids, while in the early season the higher influence is of unsaturated fatty acids. Multivariate techniques were able to group the different types of cocoa according to their chemical profile, helping to create an origin denomination for those produced in Southern Bahia

AGRONOMIC EFFICIENCY OF FERTILIZERS BASED ON HUMUS, ROCK POWDER, AND MINERALS ON SOYBEAN YIELD IN PARAGUAY-PY

Soybean is the most commercially cultivated crop in Paraguay, and obtaining high yields requires the application of large amounts of fertilizers, raising the cost of production. Developing strategies for the efficient use of applied nutrients is necessary. Therefore, the study aimed to evaluate the agronomic efficiency of combinations of mineral fertilizers with organic matter and rock powder in the development, nutrition, and yield of soybean. The experiment was carried out in Hernandarias, Paraguay, in the 2016-2017 harvest. The different fertilizers influenced the absorption of Zn, Mg and K, and grain yield. Among the treatments, the highlight was the agronomic efficiency index obtained by the replacement of 30% of mineral fertilizer by humus, presenting grain yield of 3219, 67 kg ha-1. However, it was equal to the mixing 30% of humus + rock powder with 70% of NPK formulation 04:40:10 with grain yield of 3206.50 kg ha-1, and the mixing 20% of humus + rock powder with 80% of NPK formulation 04:40:10 with grain yield of 3165.17 kg ha-1. Thus, it is recommended to use rock powder and humus in soybean production in Paraguay, especially in Latossolos (Oxisols) that have little organic matter and low CTC

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear understanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5,6,7 vast areas of the tropics remain understudied.8,9,10,11 In the American tropics, Amazonia stands out as the world's most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepresented in biodiversity databases.13,14,15 To worsen this situation, human-induced modifications16,17 may eliminate pieces of the Amazon's biodiversity puzzle before we can use them to understand how ecological communities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple organism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region's vulnerability to environmental change. 15%–18% of the most neglected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lost

Lipid nanoparticles as triterpene carriers for the treatment of experimental visceral leishmaniasis

A leishmaniose é uma doença causada por protozoários flagelados, e que afeta milhões de pessoas em todo o mundo. Os tratamentos disponíveis possuem eficácia limitada e induzem efeitos colaterais variando de leve a grave, dificultando a aderência de pacientes. Assim, torna-se essencial o desenvolvimento de novos medicamentos e estratégias relacionadas ao tratamento desta doença tropical negligenciada. O presente estudo avaliou as atividades leishmanicidas in vitro e/ou in vivo dos triterpenos ácido maslínico (AM), ácido ursólico (AU), betulina (Be) e lupeol (Lu) carreados ou não em nanosistemas. Os triterpenos AM, AU, Be e Lu foram testados em formas promastigotas e amastigotas intracelulares de Leishmania (Leishmania) infantum. Todos os triterpenos, exceto Be, foram ativos contra as formas promastigotas e amastigotas de L. (L.) infantum. As principais alterações ultraestruturais dos parasitos tratados com AM, AU e Lu estiveram relacionadas à formação de compartimentos vesiculares com ou sem figuras de mielina ou restos de membranas, além de desestruturação da mitocôndria do parasito, que se mostrou com volume aumentado e fragmentada. Adicionalmente, formas promastigotas de L. (L.) infantum apresentaram alteração no potencial de membrana após 15 minutos de incubação com AU e Lu. A cromatina dos parasitos também se mostrou fragmentada, sugerindo que os triterpenos induzem morte celular programada. Para maximizar o potential leishmanicida dos triterpenos Au e Lu, nanopartículas lipídicas sólidas (SLN) e carreadores lipídicos nanoestruturados (NLC) contendo estes triterpenos foram produzidos pelas técnicas homogeneização à alta pressão (HP) a quente e por ultrasonicação (Son), com diferentes concentrações de lipídeo (5 e 10%), tensoativo (1 e 2%) e fármaco (0,10%). Análises físico-quimicas mostraram que SLN e NLC exibiram forma esférica com superfícies lisas e com tamanho médio abaixo dos 272 nm; o potencial zeta das nanopartículas apresentaram se negativos e elevados, variando entre -26,11 a -37,22 mV, sugerindo que as nanoformulações são estáveis. Além disso, se apresentaram com índice de polidispersão menor que 0,25. Estudos de Calorimetria exploratória diferencial (DSC) e espectroscopia no infravermelho com transformada de Fourier (FTIR) mostraram que o AU e Lu foi solubilizado na matriz lipídica, tanto nas SLN como nos NLC, uma vez que não houve quaisquer eventos de fusão de droga nessas técnicas. Estudos in vitro mostraram que NLC carreando AU ou Lu apresentaram elevada atividade leishmanicida quando comparado às SLNc carreando os triterpenos. De acordo com os dados físicos e de potencial leishmanicida, NLC carreando AU (NLC-AU) ou Lu (NLC-Lu) foram selecionadas para realização de estudos in vivo. Em hamsters saudáveis, foi visto que NLC-AU ou NLC-Lu não alteraram a estrutura histológica de baço, fígado, rim, pulmão e coração, entretanto foi observado leve infiltrado inflamatório no espaço porta nos grupos tratados com o Lu livre. Animais infectados e tratados com NLC-AU ou NLC-Lu tiveram diminuição significativa do parasitismo hepático e esplênico e a eficácia das nanoformulação foi superior àquela do AU, Lu e anfotericina B administrados livremente. A atividade terapêutica se correlacionou com elevação da expressão de IFN-e/ou iNOS nos animais infectados e tratados com NLC-AU ou NLC-Lu. Os dados sugerem que os NLCs são potenciais sistemas para a entrega de triterpenos com atividade anti-Leishmania aprimoradaLeishmaniasis is a disease caused by flagellated protozoa and affects millions of people around the world. Available treatments have limited efficacy and induce side effects ranging from mild to severe, affecting the compliance of patients. Thus, it is essential to develop new drugs and strategies related to the treatment of this neglected tropical disease. The present study evaluated the in vitro and/or in vivo leishmanicidal activities of the triterpenes maslinic acid (MA), ursolic acid (UA), betulin (Be) and lupeol (Lu) carried or not in nanosystems. The triterpenes MA, UA, Be and Lu were tested in promastigote and intracellular amastigote forms of Leishmania (Leishmania) infantum. All of them, except Be, were active on the promastigote and amastigote forms of L. (L.) infantum. The main ultrastructural alterations of the parasites treated with MA, UA, and Lu were related to the formation of vesicular compartments with or without myelin figures or membrane debris; in the treated parasites, the mitochondria were enlarged and fragmented. Additionally, parasites treated with UA and Lu exhibited an altered mitochondria membrane potential after 15 minutes of incubation. Fragmentation of chromatin was observed in treated parasites, suggesting that triterpenes induce programmed cell death. To maximize the leishmanicidal potential of UA and Lu, solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) containing triterpenes were produced by hot homogenization (HP) and ultrasonication (Son) techniques with different lipid concentrations (5 and 10%), surfactant (1 and 2%) and drug (0.10%). Physicochemical analysis showed that SLN and NLC exhibited a spherical shape with smooth surfaces and an average size below 272 nm; the zeta potential of the nanoparticles was negative and elevated, ranging from -26.11 to -37.22 mV, suggesting that the nanoformulations are stable. Furthermore, they had a polydispersion index lower than 0.25. Differential Scanning Calorimetry (DSC) and Fourier Transform Infrared Spectroscopy (FTIR) studies showed that UA and Lu were solubilized in the lipid matrix, both in SLN and NLC, since there were no drug fusion events in these techniques. In vitro studies showed that NLC carrying UA or Lu showed elevated leishmanicidal activity compared to SLN carrying the same triterpenes. Thus, according to physical and leishmanicidal data, NLC carrying UA (UA-NLC) or Lu (Lu-NLC) were selected for in vivo studies. In healthy golden hamsters, it was observed that UA-NLC and Lu-NLC did not alter the morphology of the spleen, liver, kidney, lung, and heart, however, mild inflammatory infiltrate was observed in the portal space in the groups treated with Lu. Animals infected and treated with UA-NLC or Lu-NLC exhibited a significant decrease in the number of parasites in the liver and spleen and and the effectiveness of the nanoformulations was superior to that of freely administered AU, Lu and amphotericin B. The therapeutic activity correlated with increased expression of IFN- and/or iNOS in animals infected and treated with UA-NLC or Lu-NLC. The data suggest that NLCs are potential systems to deliver triterpenes with enhanced anti-Leishmania activit

AGRONOMIC EFFICIENCY OF FERTILIZERS BASED ON HUMUS, ROCK POWDER, AND MINERALS ON SOYBEAN YIELD IN PARAGUAY-PY

Soybean is the most commercially cultivated crop in Paraguay, and obtaining high yields requires the application of large amounts of fertilizers, raising the cost of production. Developing strategies for the efficient use of applied nutrients is necessary. Therefore, the study aimed to evaluate the agronomic efficiency of combinations of mineral fertilizers with organic matter and rock powder in the development, nutrition, and yield of soybean. The experiment was carried out in Hernandarias, Paraguay, in the 2016-2017 harvest. The different fertilizers influenced the absorption of Zn, Mg and K, and grain yield. Among the treatments, the highlight was the agronomic efficiency index obtained by the replacement of 30% of mineral fertilizer by humus, presenting grain yield of 3219, 67 kg ha-1. However, it was equal to the mixing 30% of humus + rock powder with 70% of NPK formulation 04:40:10 with grain yield of 3206.50 kg ha-1, and the mixing 20% of humus + rock powder with 80% of NPK formulation 04:40:10 with grain yield of 3165.17 kg ha-1. Thus, it is recommended to use rock powder and humus in soybean production in Paraguay, especially in Latossolos (Oxisols) that have little organic matter and low CTC

Leishmanicidal Activity and Ultrastructural Changes of Maslinic Acid Isolated from Hyptidendron canum

The therapeutic arsenal for the treatment of leishmaniasis is limited and has serious obstacles, such as variable activity, high toxicity, and costs. To overcome such limitations, it becomes urgent to characterize new bioactive molecules. Plants produce and accumulate different classes of bioactive compounds, and these molecules can be studied as a strategy to combat leishmaniasis. The study presented herein evaluated the leishmanicidal effect of maslinic acid isolated from the leaves of Hyptidendron canum (Lamiaceae) and investigated the morphological that occurred on Leishmania (Leishmania) infantum upon treatment. Maslinic acid was active and selective against promastigote and amastigote forms in a dose-dependent manner. Additionally, it was not toxic to peritoneal macrophages isolated from golden hamsters, while miltefosine and amphotericin B showed mild toxicity for macrophages. Morphological changes in promastigotes of L. (L.) infantum treated with maslinic acid were related to cytoplasmic degeneration, intense exocytic activity, and blebbing in the kDNA; disruption of mitochondrial cristae was observed in some parasites. The nucleus of promastigote forms seems to be degraded and the chromatin fragmented, suggesting that maslinic acid triggers programmed cell death. These results indicate that maslinic acid may be an interesting molecule to develop new classes of drugs against leishmaniasis

Nanoemulsified Butenafine for Enhanced Performance against Experimental Cutaneous Leishmaniasis

The production of ergosterol lipid involves the activity of different enzymes and is a crucial event for the Leishmania membrane homeostasis. Such enzymes can be blocked by azoles and allylamines drugs, such as the antifungal butenafine chloride. This drug was active on parasites that cause cutaneous and visceral leishmaniasis. Based on the leishmanicidal activity of butenafine chloride and considering the absence of reports about the therapeutic potential of this drug in cutaneous leishmaniasis, the present work is aimed at analyzing the efficacy of butenafine formulated in two different topical delivery systems, the self-nanoemulsifying drug delivery systems (BUT-SNEDDS) and in a SNEDDS-based nanogel (BUT-SNEDDS gel) as well as in the free form in experimental cutaneous leishmaniasis. Physical studies showed that both formulations were below 300 nm with low polydispersity (<0.5) and good colloidal stability (around -25 mV). Increased steady-state flux was reported for nanoenabled butenafine formulations with reduced lag time in Franz cell diffusion assays across Strat-M membranes. No toxic or inflammatory reactions were detected in animals treated with BUT-SNEDDS, BUT-SNEDDS gel, or butenafine. Animals topically treated with butenafine (free or nanoformulated) showed small dermal lesions and low tissue parasitism. Furthermore, BUT-SNEDD gel and butenafine presented similar efficacy than the standard drug Glucantime given by the intralesional route. Increased levels of IFN-γ were observed in animals treated with BUT-SNEDDS gel or butenafine. Based on these data, the antifungal drug butenafine chloride can be considered an interesting repurposed drug for the treatment of cutaneous leishmaniasis

Related Pentacyclic Triterpenes Have Immunomodulatory Activity in Chronic Experimental Visceral Leishmaniasis

Leishmaniasis is a neglected tropical disease caused by the flagellated protozoa of the genus Leishmania that affects millions of people around the world. Drugs employed in the treatment of leishmaniasis have limited efficacy and induce local and systemic side effects to the patients. Natural products are an interesting alternative to treat leishmaniasis, because some purified molecules are selective toward parasites and not to the host cells. Thus, the aim of the present study was to compare the in vitro antileishmanial activity of the triterpenes betulin (Be), lupeol (Lu), and ursolic acid (UA); analyze the physiology and morphology of affected organelles; analyze the toxicity of selected triterpenes in golden hamsters; and study the therapeutic activity of triterpenes in hamsters infected with L. (L.) infantum as well as the cellular immunity induced by studied molecules. The triterpenes Lu and UA were active on promastigote (IC50=4.0±0.3 and 8.0±0.2 μM, respectively) and amastigote forms (IC50=17.5±0.4 and 3.0±0.2 μM, respectively) of L. (L.) infantum, and their selectivity indexes (SI) toward amastigote forms were higher (≥13.4 and 14, respectively) than SI of miltefosine (2.7). L. (L.) infantum promastigotes treated with Lu and UA showed cytoplasmic degradation, and in some of these areas, cell debris were identified, resembling autophagic vacuoles, and parasite mitochondria were swelled, fragmented, and displayed membrane potential altered over time. Parasite cell membrane was not affected by studied triterpenes. Studies of toxicity in golden hamster showed that Lu did not alter blood biochemical parameters associated with liver and kidney functions; however, a slight increase of aspartate aminotransferase level in animals treated with 2.5 mg/kg of UA was detected. Lu and UA triterpenes eliminated amastigote forms in the spleen (87.5 and 95.9% of reduction, respectively) and liver of infected hamster (95.9 and 99.7% of reduction, respectively); and UA showed similar activity at eliminating amastigote forms in the spleen and liver than amphotericin B (99.2 and 99.8% of reduction). The therapeutic activity of both triterpenes was associated with the elevation of IFN-γ and/or iNOS expression in infected treated animals. This is the first comparative work showing the in vitro activity, toxicity, and therapeutic activity of Lu and UA in the chronic model of visceral leishmaniasis caused by L. (L.) infantum; additionally, both triterpenes activated cellular immune response in the hamster model of visceral leishmaniasis