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The tumor microenvironment presents several therapeutic targets, with inflammation being one of them. In search of new drugs, plants have shown to be an effective source of potent anti-inflammatory and anticancer agents. This study aimed to evaluate the antitumoral and inflammatory activities of Boehmeria caudata aerial parts extract. Bioguided in vitro antiproliferative screening showed that phenanthroquinolizidine obtained from the aerial B. caudata ethanolic extract had a straight relationship with activity. Moreover, the orally administered ethanolic extract reduced Ehrlich solid tumor growth and displayed an anti-inflammatory effect in both evaluated experimental models (carrageenan-induced paw edema and croton oil-induced ear edema). These results suggest that the antitumor activity of the ethanolic extract could be explained by antiproliferative effects associated with anti-inflammatory action.

Food safety and quality assessment mechanisms are unmet needs that industries and countries have been continuously facing in recent years. Our study aimed at developing a platform using Machine Learning algorithms to analyze Mass Spectrometry data for classification of tomatoes on organic and non-organic. Tomato samples were analyzed using silica gel plates and direct-infusion electrospray-ionization mass spectrometry technique. Decision Tree algorithm was tailored for data analysis. This model achieved 92% accuracy, 94% sensitivity and 90% precision in determining to which group each fruit belonged. Potential biomarkers evidenced differences in treatment and production for each group.

Made available in DSpace on 2020-12-12T00:57:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-12-01 The recent outbreak of Zika virus (ZIKV) infection associated with microcephaly cases has elicited much research on the mechanisms involved in ZIKV-host cell interactions. It has been described that Zika virus impairs cell growth, raising a hypothesis about its oncolytic potential against cancer cells. ZIKV tumor cell growth inhibition was later confirmed for glioblastoma. It was also demonstrated that an inactivated ZIKV prototype (ZVp) based on bacterial outer membrane vesicles has antiproliferative activity upon other cancer cell lines, such as PC-3 prostate cancer cell. This study aims at understanding the pathways that might be involved with the antiproliferative effect of Zika virus against prostate cancer cells. A metabolomic approach based on high-resolution mass spectrometry analysis led to the identification of 21 statistically relevant markers of PC-3 cells treated with ZVp. The markers were associated with metabolic alterations that trigger lipid remodeling, endoplasmic reticulum stress, inflammatory mediators, as well as disrupted porphyrin and folate metabolism. These findings highlight molecular signatures of ZVp-induced response that may be involved on cellular pathways triggered by its antiproliferative effect. To our knowledge, this is the first reported metabolomic assessment of ZIKV effect on prostate cancer cells, a promising topic for further research. Innovare Biomarkers Laboratory School of Pharmaceutical Sciences University of Campinas (UNICAMP) Medical School São Paulo State University (UNESP) School of Pharmaceutical Sciences University of Campinas (UNICAMP) Department of Physiological Sciences Piracicaba Dental School University of Campinas (UNICAMP) Laboratory of Biotechnology School of Pharmaceutical Sciences University of Campinas (UNICAMP) Medical School São Paulo State University (UNESP)

In this study, we explore a series of novel potential antiprotozoal S-(−)-limonene-based benzaldehyde thiosemicarbazones were synthesised and their activity effective against the extracellular promastigote form of Leishmania amazonensis examined. Likewise, in parallel, a series of R-(+)-limonene-based thiosemicarbazones previously synthesised by our research group and thiosemicarbazones lacking the monoterpenic moiety, were also biologically evaluated. Here, we report the combination of theoretical and experimental approaches, as well as statistical analysis, to investigate the effect of the monoterpenic group and its stereochemistry in the biological activity of benzaldehyde thiosemicarbazone derivatives, for the identification of their structure-activity relationship. The terpenic thiosemicarbazones displayed the highest activities, confirming that the monoterpenic moiety is essential for activity. Notably, among the compounds tested, the S-(−)-enantiomer of the 4-nitro-derivative (8d) presented considerably lower cytotoxicity than its R-(+)-analogue, emphasizing the importance of the stereochemistry. The most active derivative (8d) exhibited a potent antiprotozoal activity (IC50 2.4 μM) and high selectivity (SI > 1147). Also, theoretical calculations were carried out at the density functional theory (DFT) level to show that the Gibbs free energy and LUMO orbitals present an excellent correlation with the experimental IC50 values. Finally, the combination of all these results may in principle be extremely advantageous to a deeper chemical understanding, as well as, allows a rational alternative for the future development of new drugs that act against leishmaniasis.

Ilex paraguariensis is a perennial plant used in the production of mate tea, "chimarrao" and "terere," cosmetics, and other food products. Its leaves are harvested every 12 or 18 months. Approximately 2 to 5 tons of residue are generated per hectare during the harvest. The bioactive composition of this residue has not been characterized to date. Therefore, this paper presents for the first time, the simultaneous characterization of the bioactive compounds of the leaves, thin branches, and thick branches (residue) from I. paraguariensis grown under two cultivation systems: "full sun" and "shaded." The identification and quantification of the compounds was performed using high-performance liquid chromatography coupled to electrospray ionisation and quadrupole time-of-flight mass spectrometer (HPLC-ESI-QTOF-MS). Consequently, 35 compounds were identified. The average dry weight of phenolic compounds in the residue was 4.1 g/100 g, whereas that in the bark of the residue was 12.9 g/100 g, which was similar to the content found in leaves. The same compounds were identified in the two cultivation systems but with a difference in their contents. While the "full sun" cultivation had a higher content of phenolic acids, the "shaded" cultivation had a higher content of flavonoids and saponins. It was found that the I. paraguariensis residue, particularly the bark, is rich in bioactive compounds, such as quinic, 3-caffeoylquinic, 5-cafefoylquinic, 3,5-dicaffeoylquinic, and 4,5-dicaffeoylquinic acids as well as rutin, and their contents vary according to the cultivation system. Therefore, this residue is an underutilized natural resource with a potential for industrial applications. PRACTICAL APPLICATION: Yerba mate producers will be able to choose the best cultivation system ("full sun" or "shaded") to increase the content of bioactive compounds. New products may be developed with yerba mate residue due to its high concentration of compounds that are beneficial to the human health. New destinations may be applied to yerba mate residue from the harvest pruning, adding commercial value to this unexplored natural resource.

COVID-19 is still placing a heavy health and financial burden worldwide. Impairment in patient screening and risk management plays a fundamental role on how governments and authorities are directing resources, planning reopening, as well as sanitary countermeasures, especially in regions where poverty is a major component in the equation. An efficient diagnostic method must be highly accurate, while having a cost-effective profile. We combined a machine learning-based algorithm with mass spectrometry to create an expeditious platform that discriminate COVID-19 in plasma samples within minutes, while also providing tools for risk assessment, to assist healthcare professionals in patient management and decision-making. A cross-sectional study enrolled 815 patients (442 COVID-19, 350 controls and 23 COVID-19 suspicious) from three Brazilian epicenters from April to July 2020. We were able to elect and identify 19 molecules related to the disease's pathophysiology and several discriminating features to patient's health-related outcomes. The method applied for COVID-19 diagnosis showed specificity >96% and sensitivity >83%, and specificity >80% and sensitivity >85% during risk assessment, both from blinded data. Our method introduced a new approach for COVID-19 screening, providing the indirect detection of infection through metabolites and contextualizing the findings with the disease's pathophysiology. The pairwise analysis of biomarkers brought robustness to the model developed using machine learning algorithms, transforming this screening approach in a tool with great potential for real-world application.

COVID-19 is still placing a heavy health and financial burden worldwide. Impairments in patient screening and risk management play a fundamental role on how governments and authorities are directing resources, planning reopening, as well as sanitary countermeasures, especially in regions where poverty is a major component in the equation. An efficient diagnostic method must be highly accurate, while having a cost-effective profile.We combined a machine learning-based algorithm with instrumental analysis using mass spectrometry to create an expeditious platform that discriminate COVID-19 in plasma samples within minutes, while also providing tools for risk assessment, to assist healthcare professionals in patient management and decision-making. A cross-sectional study with 728 patients (369 confirmed COVID-19 and 359 controls) was enrolled from three Brazilian epicentres (São Paulo capital, São Paulo countryside and Manaus) in the months of April, May, June and July 2020.We were able to elect and identify 21 molecules that are related to the disease’s pathophysiology and 26 features to patient’s health-related outcomes. With specificity >97% and sensitivity >83% from blinded data, this screening approach is understood as a tool with great potential for real-world application.

Paracoccidioidomycosis (PCM) is a fungal infection typically found in Latin American countries, especially in Brazil. The identification of this disease is based on techniques that may fail sometimes. Intending to improve PCM detection in patient samples, this study used the combination of two of the newest technologies, artificial intelligence and metabolomics. This combination allowed PCM detection, independently of disease form, through identification of a set of molecules present in patients’ blood. The great difference in this research was the ability to detect disease with better confidence than the routine methods employed today. Another important point is that among the molecules, it was possible to identify some indicators of contamination and other infection that might worsen patients’ condition. Thus, the present work shows a great potential to improve PCM diagnosis and even disease management, considering the possibility to identify concomitant harmful factors., Brazil and many other Latin American countries are areas of endemicity for different neglected diseases, and the fungal infection paracoccidioidomycosis (PCM) is one of them. Among the clinical manifestations, pneumopathy associated with skin and mucosal lesions is the most frequent. PCM definitive diagnosis depends on yeast microscopic visualization and immunological tests, but both present ambiguous results and difficulty in differentiating PCM from other fungal infections. This research has employed metabolomics analysis through high-resolution mass spectrometry to identify PCM biomarkers in serum samples in order to improve diagnosis for this debilitating disease. To upgrade the biomarker selection, machine learning approaches, using Random Forest classifiers, were combined with metabolomics data analysis. The proposed combination of these two analytical methods resulted in the identification of a set of 19 PCM biomarkers that show accuracy of 97.1%, specificity of 100%, and sensitivity of 94.1%. The obtained results are promising and present great potential to improve PCM definitive diagnosis and adequate pharmacological treatment, reducing the incidence of PCM sequelae and resulting in a better quality of life. IMPORTANCE Paracoccidioidomycosis (PCM) is a fungal infection typically found in Latin American countries, especially in Brazil. The identification of this disease is based on techniques that may fail sometimes. Intending to improve PCM detection in patient samples, this study used the combination of two of the newest technologies, artificial intelligence and metabolomics. This combination allowed PCM detection, independently of disease form, through identification of a set of molecules present in patients’ blood. The great difference in this research was the ability to detect disease with better confidence than the routine methods employed today. Another important point is that among the molecules, it was possible to identify some indicators of contamination and other infection that might worsen patients’ condition. Thus, the present work shows a great potential to improve PCM diagnosis and even disease management, considering the possibility to identify concomitant harmful factors.

Migration is a known phenomenon defined as the partitioning of chemical compounds from the packaging into food, and depends on several factors. Migration assays are generally time-consuming and require specific conditions in order to investigate the behavior of the packaging in different situations. Furthermore, these tests are often performed with food simulants, since the determination of migration under real conditions is highly impaired. Several methodologies have been designed to carry out this study, but an ideal approach should be capable of assessing the migration of compounds in real samples, providing fast and reliable results. Within this context, mass spectrometry can be considered a suitable and versatile technique that shows great potential to accurately characterize several contaminants in food by migration. Thus, in this work we present a mass spectrometry-based application for the detection of several compounds from plastic, directly from vacuum-packed meat samples. This preliminary and simple workflow can be easily applied in routine analyses for either quality control purposes or in the prospection of other potential bioactive contaminants in food.

Milk is an extremely complex food, capable of providing essential nutrients as well as being an important source of energy, and high-quality proteins and fats. Due to advances in technology, and to meet the increasing demand, production costs have increased, turning milk into a target of adulterations. Routine methods usually applied to certify the quality of the milk are restricted to microbiological tests, and assays that attest the nutritional composition within the expected values. However, potentially harmful byproducts generated by adulterating substances in general are not detected through these methodologies. In this contribution, we simulated the adulteration of freshly produced milk samples with four adulterants whose use already had reported for extended shelf life: formaldehyde, hydrogen peroxide, sodium hydroxide, and sodium hypochlorite. These samples were submitted to direct-infusion high-resolution mass spectrometry analysis and multivariate statistical analysis. This approach allows the characterization of a series of molecules modified by the adulterants, what demonstrates how these species affect the nutritious characteristics of this product.

A 78-year old male patient under active immunotherapy for high-grade non-muscle invasive bladder cancer presented severe COVID-19 A shorter hospitalization per

O piriproxifeno e um inseticida utilizado como estrategia de combate ao mosquito do genero Aedes e contra pragas em diversas culturas agricolas. Existem alguns estudos que demonstram que o pesticida tem efeito antiproliferativo em cerebro de mamiferos, alem de ser um agonista de PPAR?, mecanismo recentemente explorado para combate ao câncer. Dessa forma buscou-se elucidar possiveis marcadores que indiquem o mecanismo de acao do piriproxifeno sobre o glioblastoma. Verificou-se que ha mudancas no metabolismo lipidico das celulas, o que tambem ocorre quando o pesticida e associado ao Zika virus, que, por sua vez, ja demonstrou ter efeito citopatico em celulas de glioblastoma.

Weight gain is a metabolic disorder that often culminates in the development of obesity and other comorbidities such as diabetes. Obesity is characterized by the development of a chronic, subclinical systemic inflammation, and is regarded as a remarkably important factor that contributes to the development of such comorbidities. Therefore, laboratory methods that allow the identification of subjects at higher risk for severe weight-associated morbidity are of utter importance, considering the health, and safety of populations. This contribution analyzed the plasma of 180 Brazilian individuals, equally divided into a eutrophic control group and case group, to assess the presence of biomarkers related to weight gain, aiming at characterizing the phenotype of this population. Samples were analyzed by mass spectrometry and most discriminant features were determined by a machine learning approach using Random Forest algorithm. Five biomarkers related to the pathogenesis and chronicity of inflammation in weight gain were identified. Two metabolites of arachidonic acid were upregulated in the case group, indicating the presence of inflammation, as well as two other molecules related to dysfunctions in the cycle of nitric oxide (NO) and increase in superoxide production. Finally, a fifth case group marker observed in this study may indicate the trigger for diabetes in overweight and obesity individuals. The use of mass spectrometry combined with machine learning analyses to prospect and characterize biomarkers associated with weight gain will pave the way for elucidating potential therapeutic and prognostic targets.

Yeasts are able to reduce the levels of ochratoxin A in fermentative processes; and, through their enzymatic complex, these micro-organisms are also capable of forming modified mycotoxins. These mycotoxins are often underreported, and may increase health risks after ingestion of contaminated food. In this sense, this study aims to evaluate whether the presence of ochratoxin A influences yeast growth kinetic parameters and to elucidate the formation of modified ochratoxin by Saccharomyces cerevisiae strains during fermentation. Three S. cerevisiae strains (12 M, 01 PP, 41 PP) were exposed to OTA at the concentrations of 10, 20 and 30 μg/L. The Baranyi model was fitted to the growth data (Log CFU/mL), and the identification of modified ochratoxins was performed through High Resolution Mass Spectrometry. The presence of ochratoxin A did not influence the growth of S. cerevisiae strains. Four pathways were proposed for the metabolization of OTA: dechlorination, hydrolysis, hydroxylation, and conjugation. Among the elected targets, the following were identified: ochratoxin α, ochratoxin β, ochratoxin α methyl ester, ochratoxin B methyl ester, ethyl amide ochratoxin A, ochratoxin C, hydroxy-ochratoxin A, hydroxy-ochratoxin A methyl ester, and ochratoxin A cellobiose ester. These derivatives formed from yeast metabolism may contribute to the occurrence of underreporting levels of total mycotoxin in fermented products.

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Aspergillus carbonarius and Aspergillus niger are the main responsible fungi for the accumulation of ochratoxin A (OTA) in wine grapes. Some strains are able to convert the parent mycotoxin into other compounds by means of hydrolysis and/or conjugation reactions through their defense mechanisms and enzymatic activity, leading to the formation of a modified mycotoxin. Thus, the variability of growth and metabolite production are inherent to the strain, occurring distinctively even when submitted to similar conditions. In this sense, this contribution aimed at determining the variability in multiplication and production of OTA by strains of A. carbonarius and A. niger isolated from grapes, as well as investigating the formation of modified mycotoxins. Strains were incubated in grape-based medium, and the diameter of the colonies measured daily. The determination of OTA was performed by high-performance liquid chromatography and the identification of modified mycotoxins was carried out using high-resolution mass spectrometry. Variabilities in terms of growth and OTA production were assessed across five different strains. Peak production of OTA was detected on day 15, and a decline on day 21 was observed, indicating that the observed reduction may be associated with the degradation or modification of the OTA over time by the fungus. Ethylamide ochratoxin A, a modified mycotoxin identified in this study, provides evidence that there may be underreporting of total mycotoxin levels in food, increasing uncertainty concerning health risks to the population.

Dengue fever is a viral condition that has become a recurrent issue for public health in tropical countries, common endemic areas. Although viral structure and composition have been widely studied, the infection phenotype in terms of small molecules remains poorly established. This contribution providing a comprehensive overview of the metabolic implications of the virus-host interaction using a lipidomic-based approach through direct-infusion high-resolution mass spectrometry. Our results provide further evidence that lipids are part of both the immune response upon Dengue virus infection and viral infection maintenance mechanism in the organism. Furthermore, the species described herein provide evidence that such lipids may be part of the mechanism that leads to blood-related complications such as hemorrhagic fever, the severe form of the disease.

Statins are the preferred therapy to treat hypercholesterolemia. Their main action consists of inhibiting the cholesterol biosynthesis pathway. Previous studies report mitochondrial oxidative stress and membrane permeability transition (MPT) of several experimental models submitted to diverse statins treatments. The aim of the present study was to investigate whether chronic treatment with the hydrophilic pravastatin induces hepatotoxicity in LDL receptor knockout mice (LDLr-/-), a model for human familial hypercholesterolemia. We evaluated respiration and reactive oxygen production rates, cyclosporine-A sensitive mitochondrial calcium release, antioxidant enzyme activities in liver mitochondria or homogenates obtained from LDLr-/- mice treated with pravastatin for 3 months. We observed that pravastatin induced higher H2O2 production rate (40%), decreased activity of aconitase (28%), a superoxide-sensitive Krebs cycle enzyme, and increased susceptibility to Ca2+-induced MPT (32%) in liver mitochondria. Among several antioxidant enzymes, only glucose-6-phosphate dehydrogenase (G6PD) activity was increased (44%) in the liver of treated mice. Reduced glutathione content and reduced to oxidized glutathione ratio were increased in livers of pravastatin treated mice (1.5- and 2-fold, respectively). The presence of oxidized lipid species were detected in pravastatin group but protein oxidation markers (carbonyl and SH- groups) were not altered. Diet supplementation with the antioxidants CoQ10 or creatine fully reversed all pravastatin effects (reduced H2O2 generation, susceptibility to MPT and normalized aconitase and G6PD activity). Taken together, these results suggest that 1- pravastatin induces liver mitochondrial redox imbalance that may explain the hepatic side effects reported in a small number of patients, and 2- the co-treatment with safe antioxidants neutralize these side effects.

Falsified, counterfeit and adulterated medicines are an endemic problem worldwide that results in both monetary and health-related losses. Developing fast and reliable methods that are able to present a timely result based on the drug's spectral profile is an effort that is sure to benefit those involved in the whole distribution chain. We propose herein a Laser Desorption/Ionization imaging-based method that provides simple and minimal sample preparation; this method is capable of providing specific markers that characterize adulterations, using as proof of concept one of the most adulterated drug products for oral use, sildenafil. Our approach is able to provide quality markers, which can be applied in the fast screening of any product within the same molecular class. This same strategy may be a useful alternative to provide accurate measurements with high specificity for unraveling contaminants and/or byproducts in virtually any given pharmaceutical product.

Cachaça is a popular spirit produced in Brazil, obtained by distillation of fermented sugar cane. Among the contaminants arising from production, ethyl carbamate is a carcinogenic compound that occurs naturally in fermented foods and beverages; in Brazil, the maximum limit established by current legislation is 150 µg L−1. Quality control is usually performed using gas chromatography; however, robustness and reproducibility of quantitative results may be severely impaired, as the addition of 6–30 g L−1 of sucrose is a common procedure for taste standardization, directly interfering in the results. This work describes the development of a novel method to improve ethyl carbamate quantification in cachaças using a new approach of QuEChERS extraction based on salting-out phenomenon, to effectively separate ethanol from sugar-containing water. Eighteen different brands of cachaça were analyzed. The proposed methodology was able to eliminate components that contaminate the sample flow path in the gas chromatography system, while improving precision and accuracy by using a triple-quadrupole approach, in comparison with the methodology usually employed: direct analysis of cachaça samples with no sample prep. Results indicate that this approach is more effective due to the removal of sugar content, with no impact in costs per analysis.

Zika virus (ZIKV) has emerged as one of the most medically relevant viral infections of the past decades; the devastating effects of this virus over the developing brain are a major matter of concern during pregnancy. Although the connection with congenital malformations are well documented, the mechanisms by which ZIKV reach the central nervous system (CNS) and the causes of impaired cortical growth in affected fetuses need to be better addressed. We performed a non-invasive, metabolomics-based screening of saliva from infants with congenital Zika syndrome (CZS), born from mothers that were infected with ZIKV during pregnancy. We were able to identify three biomarkers that suggest that this population suffered from an important inflammatory process; with the detection of mediators associated with glial activation, we propose that microcephaly is a product of immune response to the virus, as well as excitotoxicity mechanisms, which remain ongoing even after birth.

Finding specific molecular targets and the mechanism of action of praziquantel in the treatment of schistosomiasis remains a challenging task. Our efforts were focused on obtaining further information on worm composition before and after exposure to praziquantel in the treatment of schistosomiasis to elucidate the potential sites of action of this drug. Evidence indicates that the lipid bilayer is changed by treatment with praziquantel. Following this rationale, we employed a mass spectrometry imaging-based approach that helped to characterise lipids in specific locations, which are directly involved in the biochemical pathways of the BH strain of Schistosoma mansoni, as well as differentiating the molecular response that each worm sex presents in vivo. Our findings demonstrated significant differences between the chemical markers found in adult worms before and after praziquantel exposure, especially in phospholipids, which were predominantly identified as chemical markers in all samples. Results also indicate that distinct molecular pathways in both male and female worms could be differentially affected by praziquantel treatment. These data shine new light on the mechanism of action of praziquantel, taking a further step towards its full understanding.

Sunlight exposure causes several types of injury to humans, especially on the skin; among the most common harmful effects due to ultraviolet (UV) exposure are erythema, pigmentation and lesions in DNA, which may lead to cancer. These long-term effects are minimized with the use of sunscreens, a class of cosmetic products that contains UV filters as the main component in the formulation; such molecules can absorb, reflect or diffuse UV rays, and can be used alone or as a combination to broaden the protection on different wavelengths. Currently, worldwide regulatory agencies define which ingredients and what quantities must be used in each country, and enforce companies to conduct tests that confirm the Sun Protection Factor (SPF) and the UVA (Ultraviolet A) factor. Standard SPF determination tests are currently conducted in vivo, using human subjects. In an industrial mindset, apart from economic and ethical reasons, the introduction of an in vitro method emerges as an interesting alternative by reducing risks associated to UV exposure on tests, as well as providing assertive analytical results. The present work aims to describe a novel methodology for SPF determination directly from sunscreen formulations using the previously described cosmetomics platform and mass spectrometry as the analytical methods of choice.

Background: Cystic fibrosis is a disabling genetic disease with an increased prevalence in European heritage populations. Currently, the most used technique for collection of cystic fibrosis samples and diagnosis is provided through uncomfortable tests, with uncertain results, mostly based on chloride concentration in sweat. Since cystic fibrosis mutation induces many metabolic changes in patients, exploring these alterations might be an alternative to visualize potential biomarkers that could be used as interesting tools for further diagnostic upgrade, prioritizing simplicity, low cost and quickness. Methods: This contribution describes an accurate strategy to provide potential biomarkers related to cystic fibrosis, which may be understood as a potential tool for new diagnostic approaches and/or for monitoring disease evolution. Therefore, the present proposal consists of using skin imprints on silica plates as a way of sample collection, followed by direct-infusion high-resolution mass spectrometry and multivariate data analysis, intending to identify metabolic changes in skin composition of cystic fibrosis patients. Results: Metabolomics analysis allowed identifying chemical markers that can be traced back to cystic fibrosis in patients’ skin imprints, differently from control subjects. Seven chemical markers from several molecular classes were elected, represented by bile acids, a glutaric acid derivative, thyrotropin releasing hormone, an inflammatory mediator, a phosphatidic acid, and diacylglycerol isomers, all reflecting metabolic disturbances that occur due to of cystic fibrosis. Conclusion: The comfortable method of sample collection combined with the identified set of biomarkers represent potential tools that open the range of possibilities to manage cystic fibrosis and follow the disease evolution. This exploratory approach points to new perspectives about the development of diagnostic assay using biomarkers and the management cystic fibrosis.

Follicular fluid (FF) protects the oocyte against proteolysis and extrusion during ovulation, providing an appropriate microenvironment that favors proper embryonic development; thereby, FF plays a key role in embryo quality. Being directly related to cattle breeding, studying FF is extremely important in livestock science to measure cattle fertility. This may eventually help to assess the quality of both meat and milk, products widely consumed worldwide. There is an important commercial interest in the evaluation and characterization of compounds present in the FF of livestock that present greater likelihood of pregnancy. Mass spectrometry is a great ally for this type of analysis and can provide quick and efficient screening for molecular markers in biological samples. The present study demonstrated the potential of high-resolution mass spectrometry in analyzing FF samples from two distinct groups of Nellore cows (Bos indicus): high and low fertility, as determined by the number of oocytes produced. We were able to delineate markers of interest for each group, which may ultimately be related to biochemical pathways that lead to higher or lower reproductive performance.

Cheeses have always been targeted for frauds. Among the most common is the adulteration of highly-priced milk (goat and sheep) with less valuable cow milk, which is sold as a raw material for the cheese industry. The need for sensitive techniques to assess authenticity encourages the development and improvement of analytical methods. Herein we present an approach employing Direct Imprinting in Glass Surface Mass Spectrometry (DIGS-MS) for qualitative cheese analysis in a MALDI instrument, featuring easy sample preparation and fast data acquisition/interpretation. It is possible to readily identify complex lipids in different types of cheese, associating them as potential quality markers. Controlling productive stages may also be feasible using this technology by integrating analytical and statistical data, resulting in a powerful combination for sample discrimination based on lipid profiles. This approach presents great signal cleanliness in mass spectra, with easy data workup and interpretation.

Chocolate is a popular food bearing a number of different classifications that are differentiated by proportions of cocoa solids, milk and cocoa butter. Literature brings evidence that chocolates with a high percentage of cocoa solids contribute to good health maintenance due to the presence of phenolic compounds. On the other hand, it is known that the productive process, including pre-processing, may influence the level of these substances in the finished product. Thus, accurate strategies to measure the levels of this class of molecules that can be highly adaptable throughout the manufacturing process are important to ensure high-quality products. Mass spectrometry is an analytical tool of high sensitivity and specificity that is leading the research in food analysis towards new directions. By using mass spectrometry imaging in direct food analysis, this contribution developed an effective methodology for comparatively establishing the levels of catechin/epicatechin as phenolics content markers for cocoa content in a series of commercial chocolates from a single manufacturer, rendering a versatile tool that can be applied in fast screening of cocoa content in finished products and during manufacturing.

Statins are efficient cholesterol-lowering medicines utilized worldwide. However, 10% of patients suffer from adverse effects specially related to skeletal muscle function. Pro- or anti-oxidant effects of statins have been reported. Here we hypothesized that statins induce muscle mitochondrial oxidative stress leading to mitochondrial permeability transition (MPT) which may explain statin muscle toxicity. Thus, our aims were to investigate the effects of statin chronic treatment on muscle mitochondrial respiration rates, MPT and redox state indicators in the context of hypercholesterolemia. For this purpose, we studied muscle biopsies of the hypercholesterolemic LDL receptor knockout mice (LDLr-/-) treated with pravastatin during 3 months. Plantaris, but not soleus muscle of treated mice showed significant inhibition of respiration rates induced by ADP (–14%), oligomycin (–20%) or FCCP (–40%). Inhibitions of respiratory rates were sensitive to EGTA (Ca2+ chelator), cyclosporin A (MPT inhibitor), ruthenium red (inhibitor of mitochondria Ca2+ uptake) and coenzyme Q10 (antioxidant), indicating that pravastatin treatment favors Ca2+ induced MPT. Diet supplementation with creatine (antioxidant) also protected treated mice against pravastatin sensitization to Ca2+ induced MPT. Among several antioxidant enzymes analyzed, only catalase activity was increased by 30% in plantaris muscle of pravastatin treated mice. Oxidized lipids, but not proteins biomarkers were identified in treated LDLr-/- plantaris muscle. Taken together, the present results suggest that chronic pravastatin administration to a model of familial hypercholesterolemia promotes mitochondrial dysfunctions in plantaris muscle that can be counteracted by antioxidants administered either in vitro (CoQ10) or in vivo (creatine). Therefore, we propose that inhibition of muscle mitochondrial respiration by pravastatin leads to an oxidative stress that, in the presence of calcium, opens the permeability transition pore. This mitochondrial oxidative stress caused by statin treatment also signals for cellular antioxidant system responses such as catalase upregulation. These results suggest that the detrimental effects of statins on muscle mitochondria could be prevented by co-administration of a safe antioxidant such as creatine or CoQ10.

Schistosomiasis is a common tropical disease caused by Schistosoma species Schistosomiasis' pathogenesis is known to vary according to the worms' strain. Moreover, high parasitical virulence is directly related to eggs release and granulomatous inflammation in the host's organs. This virulence might be influenced by different classes of molecules, such as lipids. Therefore, better understanding of the metabolic profile of these organisms is necessary, especially for an increased potential of unraveling strain virulence mechanisms and resistance to existing treatments. In this report, direct-infusion electrospray high-resolution mass spectrometry (ESI(+)-HRMS) along with the lipidomic platform were employed to rapidly characterize and differentiate two Brazilian S. mansoni strains (BH and SE) in three stages of their life cycle: eggs, miracidia and cercariae, with samples from experimental animals (Swiss/SPF mice). Furthermore, urine samples of the infected and uninfected mice were analyzed to assess the possibility of direct diagnosis. All samples were differentiated using multivariate data analysis, PCA, which helped electing markers from distinct lipid classes; phospholipids, diacylglycerols and triacylglycerols, for example, clearly presented different intensities in some stages and strains, as well as in urine samples. This indicates that biochemical characterization of S. mansoni may help narrowing-down the investigation of new therapeutic targets according to strain composition and aggressiveness of disease. Interestingly, lipid profile of infected mice urine varies when compared to control samples, indicating that direct diagnosis of schistosomiasis from urine may be feasible.

In an attempt to develop potent and selective antitumor agents, a series of novel thiosemicarbazones derived from a natural monoterpene R-(+)-limonene was synthesized and their antitumor activity was evaluated. Overall, the majority of tested compounds exhibited considerable inhibitory effects on the growth of a wide range of cancer cell lines. Almost all of tested thiosemicarbazones were especially sensitive to prostate cells (PC-3). Derivatives 5, 6, 8, 9, 10, 11 and 13 presented the most potent antitumor activity against PC-3 cells. These compounds showed lower value of GI50 (0.04–0.05 μM) than the reference drug paclitaxel, besides a high selectivity for the same cell line. The 4-fluorobenzaldehyde derivative 10 was the most selective compound for prostate cells, while 2-hydroxybenzaldehyde derivative 8 was the most active compound, with potent antitumor activity against all tested cell lines.

The analysis of impurities and degradation products in pharmaceutical preparations are usually performed by chromatographic techniques such as high-performance liquid chromatography (HPLC). This approach demands extensive analysis time, mostly due to extraction and separation phases. These steps must be carried out in samples in order to adapt them to the requirements of the analytical method of choice. In the present contribution, matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was employed to quantify an important degradation product in atorvastatin calcium 80 mg tablets: the atorvastatin lactone. Through the standard of the impurity, it was possible to perform quantitative analysis directly on the drug tablet, using a quick and novel approach, suitable for quality control processes in the pharmaceutical industry.

Schistosomiasis is a neglected disease with large geographic distribution worldwide. Among the several different species of this parasite, S. mansoni is the most common and relevant one; its pathogenesis is also known to vary according to the worms' strain. High parasitical virulence is directly related to granulomatous reactions in the host's liver, and might be influenced by one or more molecules involved in a specific metabolic pathway. Therefore, better understanding the metabolic profile of these organisms is necessary, especially for an increased potential of unraveling strain virulence mechanisms and resistance to existing treatments. In this report, MALDI-MSI and the metabolomic platform were employed to characterize and differentiate two Brazilian S. mansoni strains: males and females from Belo Horizonte (BH) and from Sergipe (SE). By performing direct analysis, it is possible to distinguish the sex of adult worms, as well as identify the spatial distribution of chemical markers. Phospholipids, diacylglycerols and triacylglycerols were located in specific structures of the worms' bodies, such as tegument, suckers, reproductive and digestive systems. Lipid profiles were found to be different both between strains and males or females, giving specific metabolic fingerprints for each group. This indicates that biochemical characterization of adult S. mansoni may help narrowing-down the investigation of new therapeutic targets according to worm composition, molecule distribution and, therefore, aggressiveness of disease. Copyright © 2014 John Wiley & Sons, Ltd.

Grapes are known for their health benefits and high antioxidant activity due to phenolic content. Our work provides metabolic fingerprints of three grape products, namely grape juice (GJ), grape juice concentrate (GJC) and grape skin extract powder (GSE). Using direct infusion electrospray ionization mass spectrometry (ESI–MS) and evaluating the relationship between total lphenolic content and antioxidant activity, it is possible to compare which product has the best effectiveness. Fingerprints of GJC and GSE showed similar characteristic distributions of resveratrol and pterostilbene in relatively significant abundances. GJC provided higher antioxidant activity/phenolic ratio; therefore, the identified phenolic compounds from GJC may offer enhanced antioxidant potential when compared to the other two samples (GJ and GSE).

This work reports, for the first time, a validated methodology for qualitative and quantitative melatonin analyses directly from a complex matrix, namely rat milk, using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI), in a useful strategy for fast assessment of this molecular species. This novel approach uses the imaging feature of a MALDI instrument to create bidimensional images of a selected area. Results for sample concentration are expressed as a function of pixel-by-pixel intensity in grayscale, which are quantified according to pixel intensity through ImageJ open source software. For the extent of this work, two conditions were monitored: light- and dark-obtained milk. Quantification was performed using the selected reaction monitoring (SRM) transitions m/z 233→174 for melatonin. Limit of detection and limit of quantification have presented lower values (picomolar) when compared to other melatonin quantification strategies, indicating that this method presents high sensitivity. Furthermore, this method has a total analysis time of less than 1 min. Melatonin concentrations are significantly higher in samples obtained from dark conditions, consistent with previous results, and thus proving that MALDI-MSI is a suitable, simple, and highly sensitive methodology for direct melatonin analysis from complex biological samples.

Herein we present a novel approach for the quantification of active pharmaceutical ingredients (APIs) using mass spectrometry imaging. This strategy uses a filter paper previously “eluted” with a MALDI matrix solution as a support for analyte application. Samples are submitted to mass spectrometry imaging (MSI) and quantification through characteristic fingerprints is ultimately performed. Results for the content of rosuvastatin from a known formulation are comparable to those obtained with a validated HPLC method.

Made available in DSpace on 2018-12-11T16:44:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-10-01 Recent outbreaks of Zika virus in Oceania and Latin America, accompanied by unexpected clinical complications, made this infection a global public health concern. This virus has tropism to neural tissue, leading to microcephaly in newborns in a significant proportion of infected mothers. The clinical relevance of this infection, the difficulty to perform accurate diagnosis and the small amount of data in literature indicate the necessity of studies on Zika infection in order to characterize new biomarkers of this infection and to establish new targets for viral control in vertebrates and invertebrate vectors. Thus, this study aims at establishing a lipidomics profile of infected mosquito cells compared to a control group to define potential targets for viral control in mosquitoes. Thirteen lipids were elected as specific markers for Zika virus infection (Brazilian strain), which were identified as putatively linked to the intracellular mechanism of viral replication and/or cell recognition. Our findings bring biochemical information that may translate into useful targets for breaking the transmission cycle. Innovare Biomarkers Laboratory School of Pharmaceutical Sciences University of Campinas Animal Viruses Laboratory Department of Genetics Evolution and Bioagents Institute of Biology University of Campinas Emerging Viruses Study Laboratory Department of Genetics Evolution and Bioagents Institute of Biology University of Campinas Vector Functional Genomics and Microbiology Laboratory UNESP Institute of Biotechnology São Paulo State University, Alameda das Tecomarias s/n Department of Bioprocesses and Biotechnology Faculty of Agronomical Sciences São Paulo State University, Rua José Barbosa 1780 Vector Functional Genomics and Microbiology Laboratory UNESP Institute of Biotechnology São Paulo State University, Alameda das Tecomarias s/n Department of Bioprocesses and Biotechnology Faculty of Agronomical Sciences São Paulo State University, Rua José Barbosa 1780