Publications by authors named "Alexandra Christine Helena Frankland Sawaya"

26 Publications

  • Page 1 of 1

Systemic antioxidant and anti-inflammatory effects of yellow passion fruit bagasse extract during prostate cancer progression.

J Food Biochem 2021 Aug 2:e13885. Epub 2021 Aug 2.

Faculty of Food Engineering, Department of Food and Nutrition, University of Campinas (UNICAMP), Campinas, Brazil.

We evaluated the impact of yellow passion fruit (Passiflora edulis sp.) bagasse extract (PFBE) administration in systemic oxidative and inflammatory parameters in vivo, considering prostate cancer progression in transgenic mice (TRAMP). Piceatannol, scirpusin-B, dicaffeoylquinic acid, citric acid, and (+)-catechin were identified in PFBE, and the extract showed high in vitro antioxidant capacity. Some alterations in systemic parameters were verified during prostate cancer progression, as the increase in ALT and MDA levels, and SOD and GPx activities in the plasma. In the liver, higher MDA, TNF-α, and NF-κB levels, and GR and GPx activities were verified. Compared to their respective controls, the short- and long-term PFBE administration reduced MDA levels in the liver and plasma. The long-term treatment increased the catalase activity in the plasma, while the short-term treatment increased the hepatic SOD and catalase activities. Still, a reduction in hepatic TNF-α and NF-κB levels was verified after long-term treatment. PRACTICAL APPLICATIONS: Prostate cancer progression is associated with changes in systemic redox status and inflammation markers. Moreover, the intake of polyphenols with antioxidant properties, besides delaying prostate carcinogenesis, may improve the systemic antioxidant defenses and inflammatory response. In vitro studies pointed to a promising antioxidant and anti-inflammatory potential of yellow passion fruit bagasse. However, in vivo studies are scarce. Our results provided information about in vivo impacts of PFBE oral consumption on antioxidant defense and inflammation, indicating its potential as an adjuvant during the initial steps of prostate cancer.
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http://dx.doi.org/10.1111/jfbc.13885DOI Listing
August 2021

Lipid profile and high contents of cholesterol oxidation products (COPs) in different commercial brands of canned tuna.

Food Chem 2021 Aug 20;352:129334. Epub 2021 Feb 20.

Department of Food Technology, Institute of Technology, University Federal Rural of Rio de Janeiro, Rodovia Br 465, Seropédica, RJ, 23890-000, Brazil.

Canned fish is submitted to processes that may degrade its lipids and form harmful compounds called cholesterol oxidation products (COPs). Samples of Brazilian commercial canned tuna were analyzed to evaluate the influence of different liquid mediums (oil and brine) on the fatty acid composition and formation of COPs. The exchange between fish lipids and the constituents of the covering liquid was highlighted by the high levels of linoleic acid found in tuna conserved in oil. High amounts of COPs were found. However, higher contents of COPs were found in tuna in brine (933.14 to 1914.23 µg/g) than in oil (698.24 to 1167.88 µg/g). This result was mainly promoted by the presence of pro-oxidant elements such as salt, as well as greater heat transfer in brine than in oil. This study showed that canned tuna is a potential source of exogenous COPs, indicating the role of liquid mediums in oxidative processes.
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http://dx.doi.org/10.1016/j.foodchem.2021.129334DOI Listing
August 2021

Exudation of organic acid anions by tropical grasses in response to low phosphorus availability.

Sci Rep 2020 10 12;10(1):16955. Epub 2020 Oct 12.

Department of Crop Science, College of Agricultural Sciences, São Paulo State University, Botucatu, 18610-307, Brazil.

It has been suggested that some tropical grasses can acquire phosphorus (P) from hematite and gypsite by exuding organic acid anions (OAs). However, it remains to be determined exactly which OAs could be involved in each case. The objective of this study was to verify the exudation OAs by ruzigrass (Urochloa ruziziensis), palisade grass (U. brizantha), and Guinea grass (Megathyrsus maximus) as a response to P deficiency. The grasses were grown in leachate columns with adequate and deficient P nutrient solutions. The concentration of OAs in the leacheate and root surface, as well as shoot and root dry matter, and P uptake were determined. Citrate, isocitrate, and malate concentration in leachates and root surfaces increased with P starvation, mainly for the Urochloa grasses. Oxalate exudation was similar for the grasses under adequate P supply, but was lower in Guinea grass under P starvation. Palisade grass showed a higher concentration of total OAs in the root surface than the other species due to a great production of oxalate and isocitrate. Palisade grass showed greater dry matter yields regardless of P deficiency, and Guinea grass always had the higher shoot:root ratio. Urochloa grasses have a higher capacity to cope with low P availability by exuding OAs along with a lower shoot:root ratio than Guinea grass.
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http://dx.doi.org/10.1038/s41598-020-73398-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7552410PMC
October 2020

The anticholesterol oxidation effects of garlic (Allium sativum L.) and leek (Allium ampeloprasum L.) in frozen fish burgers submitted to grilling.

J Food Sci 2020 Aug 17;85(8):2416-2426. Epub 2020 Jul 17.

Department of Nutrition, School of Public Health, University of São Paulo (USP), São Paulo, SP, Brazil.

This study determined the total phenolic content and antioxidant capacity of garlic (Allium sativum L.) and leek (Allium ampeloprasum L.), as well as evaluated their anticholesterol oxidation potential in fish burgers. The total phenolic contents were 1.1 ± 0.1 mg GAE/g FW to garlic and 1.3 ± 0.4 mg GAE/g FW for leek. Leek extract showed antioxidant activity index (1.3 ± 0.01) in DPPH and β-carotene/linoleic acid assay (66.5 ± 1.6%); however, in ORAC assay, no statistic differences were observed (P > 0.05). Besides that, bioactive compounds of garlic and leek extracts were identified by ultra-high performance liquid chromatography-electrospray by ionization-mass spectrometry (UHPLC-ESI-MS). Fish burgers were prepared using different concentrations of leek and garlic and stored at -18 °C for 90 days. Thus, at days 0, 30, 60, and 90, the samples were grilled and analyzed as to their cholesterol and cholesterol oxidation products contents. Storage and grilling led to an increase in cholesterol oxidation products; however, addition of garlic and leek minimized cholesterol oxidation products formation. After 90 days, samples containing 3% leek + 0.5% garlic ware the most effective in inhibiting the cholesterol oxides formation during storage and showed the lowest increase in cholesterol oxidation products content (21.16%). Thus, the findings of this research indicate the potential application of garlic and leek as natural inhibitors of cholesterol oxidation in food. PRACTICAL APPLICATION: Garlic and leek have a set of bioactive compounds with a wide antioxidant capacity when used in meat foods such as fish burgers. Garlic and leek used as natural antioxidants perform well in the shelf life of fish burgers and can be substitutes for synthetic antioxidants in this type of product. The presence of both vegetables reduced the formation of prejudicial products to human health generated during the shelf life of the food.
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http://dx.doi.org/10.1111/1750-3841.15344DOI Listing
August 2020

Effect of aroeira (Schinus terebinthifolius Raddi) fruit against polyunsaturated fatty acids and cholesterol thermo-oxidation in model systems containing sardine oil (Sardinella brasiliensis).

Food Res Int 2020 06 14;132:109091. Epub 2020 Feb 14.

Department of Food Technology, Institute of Technology, University Federal Rural of Rio de Janeiro, Rodovia Br 465, Seropédica, RJ 23890-000, Brazil.

This study evaluated the protective effect of ground aroeira (Schinus terebinthifolius Raddi) fruit addition against fatty acids and cholesterol oxidation in model systems containing sardine oil (Sardinella brasiliensis) during heating (150 and 180 °C). High temperatures reduced the amount of essential polyunsaturated fatty acids and caused the formation of oxidized products. Total cholesterol oxides content increased from 58.9 ± 0.26 to 577.5 ± 2.14 μg/g oil, after heating at 180 °C. However, aroeira significantly protected lipids from oxidation. Although the synthetic antioxidant applied as standard (butylated hydroxytoluene) showed greater results, it was used in the maximum concentration permitted by Brazilian legislation (0.01%), suggesting that aroeira fruit could be used as a natural antioxidant for the food industry. The protective effect of aroeira may be correlated to its antioxidant capacity and the presence of bioactive compounds which were identified by UHPLC-ESI-MS in the aroeira extract.
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http://dx.doi.org/10.1016/j.foodres.2020.109091DOI Listing
June 2020

Aroeira fruit (Schinus terebinthifolius Raddi) as a natural antioxidant: Chemical constituents, bioactive compounds and in vitro and in vivo antioxidant capacity.

Food Chem 2020 Jun 22;315:126274. Epub 2020 Jan 22.

Department of Food Technology, Institute of Technology, University Federal Rural of Rio de Janeiro, Rodovia Br 465, Seropédica, RJ 23890-000, Brazil.

Schinus terebinthifolius Raddi fruit, known as Brazilian pepper or aroeira, is a natural source of bioactive compounds. However, studies about the antioxidant and nutritional contribution of this fruit in food systems are limited. Regarding the presence of bioactive compounds, flavonoids showed the highest level (10.33 ± 0.34 mg QE/g), and potential antioxidant components such biflavonoids were determined by ultra-high performance liquid chromatography/electrospray ionization mass spectrometry. The aroeira fruit extract showed antioxidant potential in DPPH (42.68 ± 0.05%), ORAC (43.40 ± 6.22 µM TE/g) and β- carotene/linoleic acid (61.41 ± 5.30%) assays. Besides that, in vivo analyses demonstrated the ability of aroeira extracts to decrease the damage caused by oxidative stress promoted by HO in Saccharomyces cerevisiae cells. Thus, the presence of phytochemicals with functional properties and the antioxidant capacity of aroeira fruit indicate its use as a potential natural antioxidant for the food industry.
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http://dx.doi.org/10.1016/j.foodchem.2020.126274DOI Listing
June 2020

Influence of environmental factors on the volatile composition of two Brazilian medicinal plants: Mikania laevigata and Mikania glomerata.

Metabolomics 2019 06 12;15(6):91. Epub 2019 Jun 12.

Faculty of Pharmaceutical Science, State University of Campinas, Campinas, SP, Brazil.

Introduction: Mikania laevigata Sch.Bip. ex Baker and Mikania glomerata Spreng. are medicinal plants popularly known as guaco, used for inflammatory diseases of the respiratory system, included in pharmaceutical formularies and often used without distinction. However, several studies show that the chemical composition varies between these species, as well as in plants are subjected to different environmental stresses. Few studies have been carried out with the volatile compounds of guaco, even less about the changes in volatile composition due to abiotic variation.

Objective: The aim of this work was to evaluate how volatile compounds vary according to the seasons and at different times of the day and if these compounds are influenced by the variations in the growth conditions such as: temperature, luminosity and water.

Methods: The headspace volatiles of the leaves were sampled by solid phase micro extraction and analyzed by gas chromatography-mass spectrometry. Untargeted metabolomic analysis of the resulting chromatograms and chemometrics was applied. The chemical profile of the volatiles of M. laevigata and M. glomerata were different; being clearly separated in the exploratory grouping analyzes (PCA), followed by analysis of variance of the marker compounds of both species.

Results: Only M. laevigata contained coumarin, considered to be the chemical marker of both species and to be responsible for the therapeutic activities. There was no significant difference between the morning and afternoon collections of either species. Coumarin, α-pinene and bicyclogermacrene were more intense in the volatiles of M. laevigata throughout the year and responsible for grouping the samples of this species. For M. glomerata, hexanal and 2-hexenal were responsible for grouping the samples and were more intense in all months. The growth conditions tested affected the intensity of specific compounds in the chromatograms. Some compounds were less intense with the increase of the temperature and in the plants subjected to full sunlight. However, certain volatile compounds-such as pinenes-were more intense in plants suffering drought.

Conclusion: The variation in composition between species of guaco was greater than those observed in the seasonal and cultivation studies, indicating that these species cannot be used indistinctly.
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http://dx.doi.org/10.1007/s11306-019-1546-xDOI Listing
June 2019

Propolis from Different Geographic Origins Decreases Intestinal Inflammation and Bacteroides spp. Populations in a Model of DSS-Induced Colitis.

Mol Nutr Food Res 2018 09 5;62(17):e1800080. Epub 2018 Aug 5.

College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.

Scope: Dietary supplementation with polyphenol-rich propolis can protect against experimentally induced colitis. We examined whether different polyphenol compositions of Chinese propolis (CP) and Brazilian propolis (BP) influence their ability to protect against dextran sulfate sodium (DSS)-induced colitis in rats.

Methods And Results: HPLC-DAD/Q-TOF-MS analysis confirmed that polyphenol compositions of CP and BP were dissimilar. Rats were given CP or BP by gavage (300 mg kg body weight) throughout the study, starting 1 week prior to DSS treatment for 1 week followed by 3 d without DSS. CP and BP significantly reduced the colitis disease activity index relative to controls not receiving propolis, prevented significant DSS-induced colonic tissue damage, and increased resistance to DSS-induced colonic oxidative stress as shown by reduced malonaldehyde levels and increased T-AOC levels. CP and BP significantly reduced DSS-induced colonic apoptosis. Colonic inflammatory markers IL-1β, IL-6, and MCP-1 were suppressed by CP and BP, whereas only BP-induced expression of TGF-β. CP, not BP, increased the diversity and richness of gut microbiota populations. Both forms of propolis significantly reduced populations of Bacteroides spp.

Conclusions: Despite the dissimilar polyphenol compositions of CP and BP, their ability to protect against DSS-induced colitis is similar. Nevertheless, some different physiological impacts were observed.
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http://dx.doi.org/10.1002/mnfr.201800080DOI Listing
September 2018

Metabolic responses of Eucalyptus species to different temperature regimes.

J Integr Plant Biol 2018 May 9;60(5):397-411. Epub 2018 Feb 9.

Bioscience, Wageningen Plant Research, Wageningen University and Research, Wageningen, The Netherlands.

Species and hybrids of Eucalyptus are the world's most widely planted hardwood trees. They are cultivated across a wide range of latitudes and therefore environmental conditions. In this context, comprehensive metabolomics approaches have been used to assess how different temperature regimes may affect the metabolism of three species of Eucalyptus, E. dunnii, E. grandis and E. pellita. Young plants were grown for 53 d in the greenhouse and then transferred to growth chambers at 10°C, 20°C or 30°C for another 7 d. In all three species the leaf chlorophyll content was positively correlated to temperature, and in E. pellita the highest temperature also resulted in a significant increase in stem biomass. Comprehensive metabolomics was performed using untargeted gas chromatography mass spectrometry (GC-MS) and liquid chromatography (LC)-MS. This approach enabled the comparison of the relative abundance of 88 polar primary metabolites from GC-MS and 625 semi-polar secondary metabolites from LC-MS. Using principal components analysis, a major effect of temperature was observed in each species which was larger than that resulting from the genetic background. Compounds mostly affected by temperature treatment were subsequently selected using partial least squares discriminant analysis and were further identified. These putative annotations indicated that soluble sugars and several polyphenols, including tannins, triterpenes and alkaloids were mostly influenced.
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http://dx.doi.org/10.1111/jipb.12626DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6220972PMC
May 2018

Comparison of the Morphology, Anatomy, and Chemical Profile of Mikania glomerata and Mikania laevigata.

Planta Med 2018 Feb 19;84(3):191-200. Epub 2017 Sep 19.

Faculty of Pharmaceutical Science, State University of Campinas, Campinas, Brazil.

and are medicinal plants popularly used in Brazil for colds and asthma. Although several studies report differences in their composition, they are frequently used indistinctly, which may be due to misidentification of the species or the plant part used. Herein, the chemical, anatomical, and morphological profiles of both species were evaluated, seeking to differentiate them. Due to the pattern of secondary metabolites found in and , the chemical profile of the leaves was the characteristic that best differentiated the two species. Coumarin was present in all the leaf samples of but absent or in low concentration in leaf samples of The stems presented coumarin in both species, albeit in different concentrations. The quantification of chlorogenic and dicaffeoylquinic acids was crucial for the separation of the two species, as higher concentrations are found in . The anatomic structure of the leaves, stem, and petioles of both species was conserved, even when exposed to different environments, but presented only subtle differences between species. The morphology of the leaves presented different characteristics for each species but altered when exposed to shade. Therefore, the chemical analysis together with the morphological characteristics of plants grown in full sunlight may be used for the correct identification of these species.
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http://dx.doi.org/10.1055/s-0043-119226DOI Listing
February 2018

Kinetic study on the inhibition of xanthine oxidase by acylated derivatives of flavonoids synthesised enzymatically.

J Enzyme Inhib Med Chem 2017 Dec;32(1):978-985

a Laboratory of Multidisciplinary Research , São Francisco University , Bragança Paulista , Brazil.

Studies have reported that flavonoids inhibit xanthine oxidase (XO) activity; however, poor solubility and stability in lipophilic media limit their bioavailability and applications. This study evaluated the kinetic parameters of XO inhibition and partition coefficients of flavonoid esters biosynthesised from hesperidin, naringin, and rutin via enzymatic acylation with hexanoic, octanoic, decanoic, lauric, and oleic acids catalysed by Candida antarctica lipase B (CALB). Quantitative determination by ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) showed higher conversion yields (%) for naringin and rutin esters using acyl donors with 8C and 10C. Rutin decanoate had higher partition coefficients (0.95), and naringin octanoate and naringin decanoate showed greater inhibitory effects on XO (IC of 110.35 and 117.51 μM, respectively). Kinetic analysis showed significant differences (p < .05) between the flavonoids before and after acylation regarding K values, whereas the values for V were the same, implying the competitive nature of XO inhibition.
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http://dx.doi.org/10.1080/14756366.2017.1347165DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6445226PMC
December 2017

Comparative Study of Chemical Composition and Biological Activity of Yellow, Green, Brown, and Red Brazilian Propolis.

Evid Based Complement Alternat Med 2016 21;2016:6057650. Epub 2016 Jul 21.

Departamento de Química, Universidade Estadual do Centro-Oeste (UNICENTRO), 85040-080 Guarapuava, PR, Brazil.

The chemical composition and biological activity of a sample of yellow propolis from Mato Grosso do Sul, Brazil (EEP-Y MS), were investigated for the first time and compared with green, brown, and red types of Brazilian propolis and with a sample of yellow propolis from Cuba. Overall, EEP-Y MS had different qualitative chemical profiles, as well as different cytotoxic and antimicrobial activities when compared to the other types of propolis assessed in this study and it is a different chemotype of Brazilian propolis. Absence of phenolic compounds and the presence of mixtures of aliphatic compounds in yellow propolis were determined by analysing (1)H-NMR spectra and fifteen terpenes were identified by GC-MS. EEP-Y MS showed cytotoxic activity against human tumour strain OVCAR-8 but was not active against Gram-negative or Gram-positive bacteria. Our results confirm the difficulty of establishing a uniform quality standard for propolis from diverse geographical origins. The most appropriate pharmacological applications of yellow types of propolis must be further investigated.
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http://dx.doi.org/10.1155/2016/6057650DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4972909PMC
August 2016

Characterization of anti-theft devices directly from the surface of banknotes via easy ambient sonic spray ionization mass spectrometry.

Sci Justice 2015 Sep 28;55(5):285-90. Epub 2015 Apr 28.

ThoMSon Mass Spectrometry Laboratory, Department of Analytical Chemistry, Institute of Chemistry, University of Campinas-UNICAMP, 13083-970 Campinas, SP, Brazil; Technical-Scientific Police Superintendency from São Paulo State, Criminalistic Institute, 05507-060, São Paulo, SP, Brazil. Electronic address:

Using Brazilian banknotes as a test case, forensic examination and identification of Rhodamine B dye anti-theft device (ATD) staining on banknotes were performed. Easy ambient sonic spray ionization mass spectrometry (EASI-MS) was used since it allows fast and simple analysis with no sample preparation providing molecular screening of the surface with direct desorption and ionization of the security dye. For a more accurate molecular characterization of the ATD dye, Q Exactive Orbitrap™ Fourier transform (tandem) mass spectrometry using eletrospray ionization (ESI-HRMS/MS) was also applied.
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http://dx.doi.org/10.1016/j.scijus.2015.04.004DOI Listing
September 2015

A simple protocol to determine lignin S/G ratio in plants by UHPLC-MS.

Anal Bioanal Chem 2015 Sep 14;407(23):7221-7. Epub 2015 Jul 14.

Department of Plant Biology, Institute of Biology, State University of Campinas, UNICAMP, 13083-970, Campinas, São Paulo, Brazil.

A simple extraction protocol and an ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) method for the determination of the syringyl/guaiacyl (S/G) ratio in lignin is reported herein. The method was entirely developed using stems of three Eucalyptus species, which were hydrolyzed with NaOH and partitioned with ethyl ether; vanillin (from the G monomer) and syringaldehyde (from S monomer) were quantified. The S/G ratios obtained were comparable to those usually reported for eucalyptus. The data for one of the eucalyptus species were compared with those obtained with a widely accepted method using thioacidolysis and gas chromatography-mass spectrometry (GC-MS). The method was also applied to sugarcane and showed to be reliable. The yield of the NaOH hydrolysis of the monolignols ranged from 89.94 to 95.69%, with more than 77.12% of recuperation in the liquid-liquid extraction. The whole analytical procedure was validated, achieving results with less than 4.38% of variation. The lowest LOD and LOQ were 0.01 and 0.05 μg/mL, respectively. In addition, the method combines reliability and a fast and direct quantification.
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http://dx.doi.org/10.1007/s00216-015-8886-9DOI Listing
September 2015

Unraveling the biosynthesis of pilocarpine in Pilocarpus microphyllus.

Nat Prod Commun 2015 May;10(5):721-4

Pilocarpine is found exclusively in species of Pilocarpus and the presence of other imidazole alkaloids has been reported in several species of the genus. Pilocarpine has several important pharmaceutical applications. Although several imidazole alkaloids related to pilocarpine have been reported in the previous years, little is still known about its biosynthetic route. At most, histidine has been reported as the precursor of pilocarpine. Based on our own previous reports and in an experiment where pilocarpine and related alkaloids (pilosine, trachyllophiline and anhydropilosine) were supplied to P. microphyllus leaves and the alkaloid profile analyzed by UPLC-MS, we suggest a biosynthesis pathway for pilocarpine. Further experiments using labeled precursors associated with transcriptome data may allow us to understand the whole biosynthesis pathway and its genetic control.
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May 2015

Chemoprotection of MNNG-initiated gastric cancer in rats using Iranian propolis.

Arch Iran Med 2015 Jan;18(1):18-23

Cancer Research Center, Tehran University of Medical Sciences, Tehran, Iran.

Background: Iranian propolis is a natural product of honeybees that has significant and varied anti-cancer benefits. The present study was designed to investigate the protective effects of Iranian propolis on gastric tissue carcinogenesis in an animal model. 

Methods: Propolis samples were collected from Hamadan and Taleghan districts of Iran, followed by ultra performance liquid chromatography mass spectrometry analysis. Fifty-five rats were divided into three groups; control, Taleghan propolis and Hamadan propolis. All the animals received N-methyl-N-nitro-N-nitrosoguanidine (MNNG, 100 μg/ml) in drinking water ad libitum for 34 weeks. In the treated groups, nutrition with propolis was started two weeks before MNNG administration. At the end of the study, the entire gastrointestinal tract was scrutinized for tumors, and the rest of the body was assessed for metastatic deposits. 

Results: Results indicated that the incidence and number of tumors were significantly decreased by propolis in comparison with the control group (P < 0.05). The nuclear/cytoplasmic ratio, epithelial stratification, nuclear dispolarity, structural abnormality, and Beta-catenin and Bcl-2 proteins expression were significantly reduced in the propolis group compared to the control group (P < 0.05). In addition, Bax protein expression was significantly increased in the propolis group in comparison with the control group (P < 0.05). 

Conclusion: The present study demonstrated the potential chemoprotective effects of the Iranian propolis against gastric cancer in a typical animal model. The results provide evidence for the hypothesis that Iranian propolis may exert a chemoprotective effect on MNNG-initiated gastric cancer through inhibition of cell proliferation and apoptosis induction.
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http://dx.doi.org/0151801/AIM.006DOI Listing
January 2015

A comparison between characterization and biological properties of Brazilian fresh and aged propolis.

Biomed Res Int 2014 3;2014:257617. Epub 2014 Nov 3.

Universidade Estadual do Centro Oeste (UNICENTRO), CEDETEG, Rua Simeão Camargo Varela de Sá, 03 Vila Carli, 85040-080 Guarapuava, PR, Brazil.

Objective: As propolis is a highly valued bee product, we aimed to verify the quality of aged propolis, investigating their phenolic and flavonoid composition, levels of toxic metals, radical scavenging and antimicrobial activities.

Material And Methods: Samples of fresh and aged propolis of six different beekeepers, from the same geographical location, were investigated in terms of their phenolic and flavonoid composition and levels of Pb, Cd, and Cr, as well as radical scavenging and antimicrobial activities.

Results: The two groups of propolis had similar qualitative composition by HPLC-PDA and ESI(-)-MS. Fresh propolis and aged propolis show no differences when average values of extraction yield, flavonoids, EC50, or MIC were compared and both types of propolis showed good antimicrobial activity at low concentrations. Only levels of phenolic compounds were higher in fresh propolis.

Conclusion: The propolis samples considered in this study, aged or fresh, had similar qualitative composition, although they were collected in different periods. Samples only differed in their levels of total phenolic content. Moreover, aged propolis conserves significant radical scavenging and antimicrobial properties. We suggest that aged propolis should not be discarded but explored for alternative applications.
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http://dx.doi.org/10.1155/2014/257617DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4235190PMC
August 2015

Characterisation of the membrane transport of pilocarpine in cell suspension cultures of Pilocarpus microphyllus.

J Plant Physiol 2015 Mar 20;175:37-47. Epub 2014 Nov 20.

Departamento de Biologia Vegetal, Instituto de Biologia, CP 6109, Universidade Estadual de Campinas, 13083-970 Campinas, SP, Brazil. Electronic address:

Pilocarpine is an alkaloid obtained from the leaves of Pilocarpus genus, with important pharmaceutical applications. Previous reports have investigated the production of pilocarpine by Pilocarpus microphyllus cell cultures and tried to establish the alkaloid biosynthetic route. However, the site of pilocarpine accumulation inside of the cell and its exchange to the medium culture is still unknown. Therefore, the aim of this study was to determine the intracellular accumulation of pilocarpine and characterise its transport across membranes in cell suspension cultures of P. microphyllus. Histochemical analysis and toxicity assays indicated that pilocarpine is most likely stored in the vacuoles probably to avoid cell toxicity. Assays with exogenous pilocarpine supplementation to the culture medium showed that the alkaloid is promptly uptaken but it is rapidly metabolised. Treatment with specific ABC protein transporter inhibitors and substances that disturb the activity of secondary active transporters suppressed pilocarpine uptake and release suggesting that both proteins may participate in the traffic of pilocarpine to inside and outside of the cells. As bafilomicin A1, a specific V-type ATPase inhibitor, had little effect and NH4Cl (induces membrane proton gradient dissipation) had moderate effect, while cyclosporin A and nifedipine (ABC proteins inhibitors) strongly inhibited the transport of pilocarpine, it is believed that ABC proteins play a major role in the alkaloid transport across membranes but it is not the exclusive one. Kinetic studies supported these results.
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http://dx.doi.org/10.1016/j.jplph.2014.11.002DOI Listing
March 2015

Pueraria tuberosa DC extract improves androgenesis and sexual behavior via FSH LH cascade.

ScientificWorldJournal 2013 30;2013:780659. Epub 2013 Dec 30.

Department of Pharmaceutical Sciences, Dr. H. S. Gour Vishwavidyalaya, Sagar, Madhya Pradesh 470003, India.

The aim of this study was to investigate the effects of ethanolic extract of Pueraria tuberosa (PT) on sexual behaviour and androgenic activity. Male albino rats were divided into four groups of six animals each: control group 1 (2% acacia solution), PT-treated group 2 (50 mg/Kg), PT-treated group 3 (100 mg/Kg), and PT-treated group 4 (150 mg/Kg). Sexual behavior of male rats in the presence of a female rat was recorded. The treated groups were evaluated for sexual parameters. The extract was characterized using LC-MS. The effect of treatment on anabolic and weight of secondary sexual organs was determined. The histological changes in section of testis and epididymis after treatment were observed. Sperm count in epididymis and fructose content in seminal vesicles were also measured. Levels of hormones like FSH, LH, and T were determined. A dose-dependent increase in sexual behaviors was evidenced in the animals of extract treated groups. Increase in testis weight was recorded in PT. At the highest dose PT also affects the hormones level. The four compounds namely puerarin, daidzein, biochanin-A and formononetin were identified in ethanolic extract using LC-MS. It concluded that PT extract possesses androgenic effect and it significantly increased the sexual behaviour and hormones level.
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http://dx.doi.org/10.1155/2013/780659DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3893017PMC
September 2014

Mass spectrometry imaging: an expeditious and powerful technique for fast in situ lignin assessment in Eucalyptus.

Anal Chem 2014 Apr 12;86(7):3415-9. Epub 2014 Mar 12.

Departamento de Biologia Vegetal, Instituto de Biologia, University of Campinas , CP 6109, 13083-970, Campinas, São Paulo, Brazil.

Plant biomass has been suggested as an alternative to produce bioethanol. The recalcitrance of plant biomass to convert cellulose into simpler carbohydrates used in the fermentation process is partially due to lignin, but the standard methods used to analyze lignin composition frequently use toxic solvents and are laborious and time-consuming. MS imaging was used to study lignin in Eucalyptus, since this genus is the main source of cellulose in the world. Hand-cut sections of stems of two Eucalyptus species were covered with silica and directly analyzed by matrix-assisted laser sesorption ionization (MALDI)-imaging mass spectrometry (MS). Information available in the literature about soluble lignin subunits and structures were used to trace their distribution in the sections and using a software image a relative quantification could be made. Matrixes routinely used in MALDI-imaging analysis are not satisfactory to analyze plant material and were efficiently substituted by thin layer chromatography (TLC) grade silica. A total of 22 compounds were detected and relatively quantified. It was also possible to establish a proportion between syringyl and guaiacyl monolignols, characteristic for each species. Because of the simple way that samples are prepared, the MALDI-imaging approach presented here can replace, in routine analysis, complex and laborious MS methods in the study of lignin composition.
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http://dx.doi.org/10.1021/ac500220rDOI Listing
April 2014

A model system to study the lignification process in Eucalyptus globulus.

Physiol Plant 2014 Sep 11;152(1):17-31. Epub 2014 Mar 11.

Departamento de Biologia Vegetal, Instituto de Biologia, CP 6109, Universidade Estadual de Campinas, 13083-970 Campinas, SP, Brazil.

Recalcitrance of plant biomass is closely related to the presence of the phenolic heteropolymer lignin in secondary cell walls, which has a negative effect on forage digestibility, biomass-to-biofuels conversion and chemical pulping. The genus Eucalyptus is the main source of wood for pulp and paper industry. However, when compared to model plants such as Arabidopsis thaliana and poplar, relatively little is known about lignin biosynthesis in Eucalyptus and only a few genes were functionally characterized. An efficient, fast and inexpensive in vitro system was developed to study lignification in Eucalyptus globulus and to evaluate the potential role of candidate genes in this biological process. Seedlings were grown in four different conditions, in the presence or absence of light and with or without sucrose in the growth medium, and several aspects of lignin metabolism were evaluated. Our results showed that light and, to a lesser extent, sucrose induced lignin biosynthesis, which was followed by changes in S/G ratio, lignin oligomers accumulation and gene expression. In addition, higher total peroxidase activity and differential isoperoxidase profile were observed when seedlings were grown in the presence of light and sucrose. Peptide sequencing allowed the identification of differentially expressed peroxidases, which can be considered potential candidate class III peroxidases involved in lignin polymerization in E. globulus.
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http://dx.doi.org/10.1111/ppl.12152DOI Listing
September 2014

Green tea extract supplementation induces the lipolytic pathway, attenuates obesity, and reduces low-grade inflammation in mice fed a high-fat diet.

Mediators Inflamm 2013 29;2013:635470. Epub 2013 Jan 29.

Departamento de Fisiologia, Disciplina de Fisiologia da Nutrição, Universidade Federal de São Paulo (UNIFESP), 04023-060 São Paulo, SP, Brazil.

The aim of this study was to evaluate the effects of green tea Camellia sinensis extract on proinflammatory molecules and lipolytic protein levels in adipose tissue of diet-induced obese mice. Animals were randomized into four groups: CW (chow diet and water); CG (chow diet and water + green tea extract); HW (high-fat diet and water); HG (high-fat diet and water + green tea extract). The mice were fed ad libitum with chow or high-fat diet and concomitantly supplemented (oral gavage) with 400 mg/kg body weight/day of green tea extract (CG and HG, resp.). The treatments were performed for eight weeks. UPLC showed that in 10 mg/mL green tea extract, there were 15 μg/mg epigallocatechin, 95 μg/mg epigallocatechin gallate, 20.8 μg/mg epicatechin gallate, and 4.9 μg/mg gallocatechin gallate. Green tea administered concomitantly with a high-fat diet increased HSL, ABHD5, and perilipin in mesenteric adipose tissue, and this was associated with reduced body weight and adipose tissue gain. Further, we observed that green tea supplementation reduced inflammatory cytokine TNFα levels, as well as TLR4, MYD88, and TRAF6 proinflammatory signalling. Our results show that green tea increases the lipolytic pathway and reduces adipose tissue, and this may explain the attenuation of low-grade inflammation in obese mice.
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http://dx.doi.org/10.1155/2013/635470DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3569937PMC
August 2013

Analytical methods applied to diverse types of Brazilian propolis.

Chem Cent J 2011 Jun 1;5(1):27. Epub 2011 Jun 1.

Plant Biology Department, Institute of Biology, UNICAMP, Campinas, SP, Brazil.

Propolis is a bee product, composed mainly of plant resins and beeswax, therefore its chemical composition varies due to the geographic and plant origins of these resins, as well as the species of bee. Brazil is an important supplier of propolis on the world market and, although green colored propolis from the southeast is the most known and studied, several other types of propolis from Apis mellifera and native stingless bees (also called cerumen) can be found. Propolis is usually consumed as an extract, so the type of solvent and extractive procedures employed further affect its composition. Methods used for the extraction; analysis the percentage of resins, wax and insoluble material in crude propolis; determination of phenolic, flavonoid, amino acid and heavy metal contents are reviewed herein. Different chromatographic methods applied to the separation, identification and quantification of Brazilian propolis components and their relative strengths are discussed; as well as direct insertion mass spectrometry fingerprinting.Propolis has been used as a popular remedy for several centuries for a wide array of ailments. Its antimicrobial properties, present in propolis from different origins, have been extensively studied. But, more recently, anti-parasitic, anti-viral/immune stimulating, healing, anti-tumor, anti-inflammatory, antioxidant and analgesic activities of diverse types of Brazilian propolis have been evaluated. The most common methods employed and overviews of their relative results are presented.
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http://dx.doi.org/10.1186/1752-153X-5-27DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3123264PMC
June 2011

Stimulation of acidic reduction of nitrite to nitric oxide by soybean phenolics: possible relevance to gastrointestinal host defense.

J Agric Food Chem 2011 May 29;59(10):5609-16. Epub 2011 Apr 29.

Departamento de Biologia Vegetal, Instituto de Biologia, Universidade Estadual de Campinas , P.O. Box 6109, Campinas, SP, Brazil, CEP 13083-970.

This study aimed to evaluate the potential of soybean-promoted acidic nitrite reduction and to correlate this activity with the content of phenolics and with the bactericidal activity against Escherichia coli O157:H7. Extracts of embrionary axes and cotyledons enriched in phenolics increased •NO formation at acidic pH at values that were 7.1 and 4.5 times higher, respectively, when compared to the reduction of the nonenriched extracts. Among the various phenolics accumulated in the soybean extracts, five stimulated nitrite reduction in the following decreasing order of potency: epicatechin gallate, chlorogenic acid, caffeic acid, galic acid and p-coumaric acid. Extracts of embrionary axes presented higher contents of epicatechin gallate and caffeic acid, compared to that of cotyledons, indicating a positive correlation between activity of the extracts and content of phenolics with regard to nitrite reducing activity. Soybean extracts enriched in phenolics interacted synergistically with acidified nitrite to prevent E. coli O157:H7 growth. The results suggest that soybean phenolics may interfere with the metabolism of •NO in an acidic environment by accelerating the reduction of nitrite, with a potential antimicrobial effect in the stomach.
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http://dx.doi.org/10.1021/jf201229xDOI Listing
May 2011

Synthesis and biological evaluation of cytotoxic properties of stilbene-based resveratrol analogs.

Eur J Med Chem 2009 Feb 15;44(2):701-7. Epub 2008 May 15.

Departamento de Química, Universidade Federal de Mato Grosso do Sul, CP 549, 79070-900 Campo Grande, MS, Brazil.

This work deals with the preparation of stilbene-based resveratrol analogs by employing the Perkin reaction, aiming at synthesizing potential antitumor lead compounds and evaluating their pharmacological activities. The proliferation inhibitor test against tumor cell lines identified analogs 9 and 11 as the most active among all synthesized derivatives, presenting IC(50) in micromolar range for certain cell lines. For study on the embryonic development, compounds 8 and 9 at the lowest tested concentration (41.7 microM) that inhibited sea urchin egg development, but only after third cleavage were used. Both the compounds inhibited 100% of normal development since first cleavage. These data partially corroborated the results obtained with MTT assay using tumor cell lines. None of the tested compounds revealed hemolytic action in assay with mouse erythrocytes.
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http://dx.doi.org/10.1016/j.ejmech.2008.05.003DOI Listing
February 2009

HPLC separation and determination of 12 cholesterol oxidation products in fish: comparative study of RI, UV, and APCI-MS detectors.

J Agric Food Chem 2006 Jun;54(12):4107-13

Department of Food Science, Faculty of Food Engineering, State University of Campinas, 13083-862 Campinas, Sao Paulo, Brazil.

A simple, fast, and sensitive method for the extraction through direct saponification, separation, quantification, and identification of 12 cholesterol oxidation products (COPs) and cholesterol in a single isocratic, normal-phase, high-performance liquid chromatography (HPLC) was developed. Three detectors were compared for determination of COPs and cholesterol in fish samples: refractive index (RI), ultraviolet (UV), and atmospheric pressure chemical ionization mass spectrometry (APCI-MS). The results did not show significant differences (p > 0.05) between the concentration of the cholesterol oxides and cholesterol obtained with these detectors. The present study demonstrated the presence of 19-hydroxycholesterol, 22"R"-hydroxycholesterol, 22"S"-hydroxycholesterol, 24"S"-hydroxycholesterol, and 25"R"-hydroxycholesterol for the first time in fish samples.
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http://dx.doi.org/10.1021/jf0532009DOI Listing
June 2006
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