Publications by authors named "Massuo J Kato"

58 Publications

Biosynthesis of Pellucidin A in (L.) HBK.

Front Plant Sci 2021 22;12:641717. Epub 2021 Mar 22.

Institute of Chemistry, University of São Paulo, São Paulo, Brazil.

(L.) HBK (Piperaceae) ("jabuti herb") is an herbaceous plant that is widespread in the tropics and has several ethnomedicinal uses. The phytochemical study of leaf extracts resulted in the isolation of 2,4,5-trimethoxycinnamic acid, 5,6,7-trimethoxyflavone, 2,4,5-trimethoxystyrene, 2,4,5-trimethoxybenzaldehyde, dillapiol, and sesamin in addition to pellucidin A. The co-occurrence of styrene and cyclobutane dimers suggested the formation of pellucidin A by a photochemical [2+2] cycloaddition of two molecules of 2,4,5-trimethoxystyrene. To investigate this biogenesis, analysis of plant leaves throughout ontogeny and treatments such as drought, herbivory and, exposure to jasmonic acid and UV light were carried out. Significant increases in the content of dillapiol (up to 86.0%) were found when plants were treated with jasmonic acid, whereas treatment under UV light increase the pellucidin A content (193.2%). The biosynthetic hypothesis was examined by feeding various C-labeled precursors, followed by analysis with GC-MS, which showed incorporation of L-(2-C)-phenylalanine (0.72%), (8-C)-cinnamic acid (1.32%), (8-C)-ferulic acid (0.51%), (8-C)-2,4,5-trimethoxycinnamic acid (7.5%), and (8-C)-2,4,5-trimethoxystyrene (12.8%) into pellucidin A. The enzymatic conversion assays indicated decarboxylation of 2,4,5-trimethoxycinnamic acid into 2,4,5-trimethoxystyrene, which was subsequently dimerized into pellucidin A under UV light. Taken together, the biosynthesis of pellucidin A in involves a sequence of reactions starting with L-phenylalanine, cinnamic acid, ferulic acid, 2,4,5-trimethoxycinnamic acid, which then decarboxylates to form 2,4,5-trimethoxystyrene and then is photochemically dimerized to produce pellucidin A.
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http://dx.doi.org/10.3389/fpls.2021.641717DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8020151PMC
March 2021

Chemical and Genotypic Variations in from the Brazilian Amazon Forest.

Molecules 2020 Dec 25;26(1). Epub 2020 Dec 25.

Institute of Chemistry, University of São Paulo, São Paulo 05508-000, SP, Brazil.

has been exploited since the end of the nineteenth century for its essential oil, a valuable ingredient in the perfumery industry. This species occurs mainly in Northern South America, and the morphological similarity among different species often leads to misidentification, which impacts the consistency of products obtained from these plants. Hence, we compared the profiles of volatile organic compounds (essential oils) and non-volatile organic compounds (methanolic extracts) of two populations of from the RESEX and FLONA conservation units, which are separated by the Tapajós River in Western Pará State. The phytochemical profile indicated a substantial difference between the two populations: samples from RESEX present α-phellandrene (22.8%) and linalool (39.6%) in their essential oil composition, while samples from FLONA contain mainly linalool (83.7%). The comparison between phytochemical profiles and phylogenetic data indicates a clear difference, implying genetic distinction between these populations.
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http://dx.doi.org/10.3390/molecules26010069DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7794742PMC
December 2020

Evaluation of DNA markers for molecular identification of three Piper species from Brazilian Atlantic Rainforest.

PLoS One 2020 19;15(10):e0239056. Epub 2020 Oct 19.

Institute of Chemistry, University of São Paulo, São Paulo, São Paulo, Brazil.

Piper is one of two large genera in the Piperaceae, and with ca. 2600 species, is one of the largest plant genera in the world. Species delimitation and evaluation of genetic diversity among populations are important requisites for conservation and adequate exploitation of economically important species. DNA barcoding has been used as a powerful tool and a practical method for species characterization and delimitation. The present work aims to evaluate molecular markers for barcoding three Piper species native to Brazil: P. gaudichaudianum ("jaborandi" or "pariparoba"), P. malacophyllum ("pariparoba-murta") and P. regnellii ("caapeba" or "pariparoba"). A reference DNA barcode library was developed using sequences of three candidate regions: ITS2, trnH-psbA and rbcL. Transferability of the microsatellite (SSR) primers Psol 3, Psol 6 and Psol 10, designed originally for Piper solmsianum, to the three Piper species was also evaluated. The discriminatory power of the markers was based on the determination of inter- and intraspecific distances, phylogenetic reconstruction, and clustering analysis, as well as BLASTn comparison. Sequences of ITS2 enabled efficient species identification by means of the BLASTn procedure. Based on these sequences, intraspecific divergence was lower than interspecific variation. Maximum Parsimony analyses based on ITS2 sequences provided three resolved clades, each corresponding to one of the three analysed species. Sequences of trnH-psbA and rbcL had lower discriminatory value. Analyses combining sequences of these regions were less effective toward the attainment of resolved and strongly supported clades of all species. In summary, robustly supported clades of P. regnellii were obtained in most of the analyses, based either on isolated or combined sequences. The SSRs primers Psol 3, Psol 6 and Psol 10 were shown to be transferable to P. gaudichaudianum and P. regnellii, but not to P. malacophyllum. Preliminary cluster analyses based on the polymorphism of the amplified products suggested that Psol 3 has lower potential than Psol 6 and Psol 10 for discrimination of Piper species.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0239056PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7571689PMC
November 2020

Effect of isoxazole derivatives of tetrahydrofuran neolignans on intracellular amastigotes of Leishmania (Leishmania) amazonensis: A structure-activity relationship comparative study with triazole-neolignan-based compounds.

Chem Biol Drug Des 2019 12 12;94(6):2004-2012. Epub 2019 Sep 12.

LASQUIM - Laboratório de Síntese e Química Medicinal, Faculdade de Ciências Farmacêuticas, Alimentos e Nutrição, Universidade Federal de Mato Grosso do Sul, Campo Grande, Brazil.

Isoxazole analogues derived from the neolignans veraguensin, grandisin, and machilin G were previously synthesized with different substitution patterns through the bioisosterism strategy. These compounds were tested on intracellular amastigotes of Leishmania (Leishmania) amazonensis; the derivatives proved to be active against intracellular amastigotes, with IC values ranging from 0.4 to 25 μM. The most active analogues were 4', 14', 15', and 18', with IC values of 0.9, 0.4, 0.7, and 1.4 μM, respectively, showing high selectivity indexes (SI = 277.0; 625.0; 178.5 and 357.1). Overall, the isoxazole analogues did not induce nitric oxide (NO) production by infected cells; there was no evidence that NO influences the antileishmanial mechanism of action, except for compound 4'. Trimethoxy groups as substituents seemed to be critical for antileishmanial activity. The SAR study demonstrated that the isoxazole compounds were more active than 1,2,3-triazole compounds with the same substitution pattterns, demonstrating the importance of the bioisosterism strategy in drug design.
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http://dx.doi.org/10.1111/cbdd.13609DOI Listing
December 2019

Editorial: The Ecology of Plant Chemistry and How it Drives Multi-Species Interactions.

Front Plant Sci 2019 30;10:967. Epub 2019 Jul 30.

Department of Fundamental Chemistry, Institute of Chemistry, University of São Paulo, São Paulo, Brazil.

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http://dx.doi.org/10.3389/fpls.2019.00967DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6682697PMC
July 2019

, an Endophyte That Establishes a Nutrient-Transfer Symbiosis With Banana Plants and Protects Against the Black Sigatoka Pathogen.

Front Microbiol 2019 7;10:804. Epub 2019 May 7.

Department of Chemistry, Universidad Autónoma de Guadalajara, Zapopan, Mexico.

Banana ( spp.) is an important crop worldwide, but black Sigatoka disease caused by the fungus threatens fruit production. In this work, we examined the potential of the endophytes of banana plants and , as antagonists of and support plant growth in nutrient limited soils by N-transfer. The two bacterial isolates were identified by MALDI-TOF mass spectrometry and corroborated by 16S rRNA sequence analysis. Both bacteria were positive for beneficial traits such as N-fixation, indole acetic acid production, phosphate solubilization, negative for 1-aminocyclopropane 1-carboxylic acid deaminase and were antagonistic to . To measure the effects on plant growth, the two plant bacteria and an strain (as non-endophyte), were inoculated weekly for 60 days as active cells (AC) and heat-killed cells (HKC) into plant microcosms without nutrients and compared to a water only treatment, and a mineral nutrients solution (MMN) treatment. Bacterial treatments increased growth parameters and prevented accelerated senescence, which was observed for water and mineral nutrients solution (MMN) treatments used as controls. Plants died after the first 20 days of being irrigated with water; irrigation with MMN enabled plants to develop some new leaves, but plants lost weight (-30%) during the same period. Plants treated with bacteria showed good growth, but AC treated plants had significantly greater biomass than the HKC. After 60 days, plants inoculated with AC showed intracellular bacteria within root cells, suggesting that a stable symbiosis was established. To evaluate the transference of organic N from bacteria into the plants, the 3 bacteria were grown with NHCl or NaNO as the nitrogen source. The N transferred from bacteria to plant tissues was measured by pheophytin isotopomer abundance. The relative abundance of the isotopomers 872.57, 873.57, 874.57, 875.57, 876.57 unequivocally demonstrated that plants acquired N atoms directly from bacterial cells, using them as a source of N, to support plant growth in restricted nutrient soils. might be a new alternative to promote growth and health of banana crops.
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http://dx.doi.org/10.3389/fmicb.2019.00804DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6513882PMC
May 2019

Design, synthesis and antitrypanosomatid activities of 3,5-diaryl-isoxazole analogues based on neolignans veraguensin, grandisin and machilin G.

Chem Biol Drug Des 2019 03 25;93(3):313-324. Epub 2018 Nov 25.

LASQUIM - Laboratório de Síntese e Química Medicinal, FACFAN - Faculdade de Ciências Farmacêuticas, Alimentos e Nutrição, Universidade Federal do Mato Grosso do Sul, UFMS, Campo Grande, MS, Brazil.

Using bioisosterism as a medicinal chemistry tool, 16 3,5-diaryl-isoxazole analogues of the tetrahydrofuran neolignans veraguensin, grandisin and machilin G were synthesized via 1,3-dipolar cycloaddition reactions, with yields from 43% to 90%. Antitrypanosomatid activities were evaluated against Trypanosoma cruzi, Leishmania (L.) amazonensis and Leishmania (V.) braziliensis. All compounds were selective for the Leishmania genus and inactive against T. cruzi. Isoxazole analogues showed a standard activity on both promastigotes of L. amazonensis and L. braziliensis. The most active compounds were 15, 16 and 19 with IC values of 2.0, 3.3 and 9.5 μM against L. amazonensis and IC values of 1.2, 2.1 and 6.4 μM on L. braziliensis, respectively. All compounds were noncytotoxic, showing lower cytotoxicity (>250 μM) than pentamidine (78.9 μM). Regarding the structure-activity relationship (SAR), the methylenedioxy group was essential to antileishmanial activity against promastigotes. Replacement of the tetrahydrofuran nucleus by an isoxazole core improved the antileishmanial activity.
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http://dx.doi.org/10.1111/cbdd.13417DOI Listing
March 2019

Natural History of a Sit-and-Wait Dipteran Predator That Uses Extrafloral Nectar as Prey Attractant.

Environ Entomol 2018 10;47(5):1165-1172

Department of Animal Biology, Universidade Estadual de Campinas, Campinas SP, Brazil.

Sit-and-wait predators use different strategies to encounter potential prey. Rhinoleucophenga myrmecophaga Vidal et (Vidal et Vilela; Diptera: Drosophilidae) larvae build sticky shelters on top of extrafloral nectaries (EFNs) of Qualea grandiflora Mart (Vochysiaceae), a common plant in the Brazilian cerrado savanna. Although larval shelters block the EFNs, nectar production is not obstructed and is used by the larvae to attract and trap nectar-gathering ants that are eventually eaten by the dipteran. Here we describe the natural history of R. myrmecophaga, its infestation pattern in Q. grandiflora, the ant assemblage at EFNs, and the insects used as prey. We use stable isotope composition (δ13C and δ15N) of R. myrmecophaga and potential food sources to infer its diet, and perform chemical analyses of the droplets found at shelter openings to determine whether nectar is used as a prey attractant. We found that Rhinoleucophenga larvae occur on the majority of Qualea plants and occupy active EFNs mainly in the rainy season. The two most frequent visiting species were also the most common insects found trapped at larval shelters. The stable isotope analyses confirmed that ants are the main food sources of R. myrmecophaga. Chemical analyses and field observations revealed that Rhinoleucophenga larvae use extrafloral nectar to attract prey to their shelters by pushing this liquid to the shelter opening where it forms a droplet. This is a rare case of sit-and-wait predator exploiting an ant-plant mutualism through the use of the very food reward produced by the plant to attract and capture potential ant mutualists.
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http://dx.doi.org/10.1093/ee/nvy097DOI Listing
October 2018

Structure⁻Activity Relationship of Piplartine and Synthetic Analogues against and Cytotoxicity to Mammalian Cells.

Int J Mol Sci 2018 06 19;19(6). Epub 2018 Jun 19.

Núcleo de Pesquisa em Biodiversidade e Biotecnologia, Biotec, Universidade Federal do Piauí, UFPI, Parnaíba-PI, 64202-020 Brazil.

Schistosomiasis, caused by helminth flatworms of the genus , is an infectious disease mainly associated with poverty that affects millions of people worldwide. Since treatment for this disease relies only on the use of praziquantel, there is an urgent need to identify new antischistosomal drugs. Piplartine is an amide alkaloid found in several species (Piperaceae) that exhibits antischistosomal properties. The aim of this study was to evaluate the structure–function relationship between piplartine and its five synthetic analogues (19A, 1G, 1M, 14B and 6B) against adult worms, as well as its cytotoxicity to mammalian cells using murine fibroblast (NIH-3T3) and BALB/cN macrophage (J774A.1) cell lines. In addition, density functional theory calculations and in silico analysis were used to predict physicochemical and toxicity parameters. Bioassays revealed that piplartine is active against at low concentrations (5⁻10 µM), but its analogues did not. In contrast, based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry assays, piplartine exhibited toxicity in mammalian cells at 785 µM, while its analogues 19A and 6B did not reduce cell viability at the same concentrations. This study demonstrated that piplartine analogues showed less activity against but presented lower toxicity than piplartine.
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http://dx.doi.org/10.3390/ijms19061802DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6032158PMC
June 2018

Secondary Metabolic Profiles of Two Cultivars of Piper nigrum (Black Pepper) Resulting from Infection by Fusarium solani f. sp. piperis.

Int J Mol Sci 2017 Dec 7;18(12). Epub 2017 Dec 7.

Programa de Pós-Graduação em Biotecnologia, Universidade Federal do Pará, Belém 66075-900, Brazil.

Bragantina and Cingapura are the main black pepper ( L.) cultivars and the Pará state is the largest producer in Brazil with about 90% of national production, representing the third largest production in the world. The infection of f. sp. , the causal agent of disease in black pepper, was monitored on the cultivars Bragantina (susceptible) and Cingapura (tolerant), during 45 days' post infection (dpi). Gas Chromatography-Mass spectrometry (GC-MS) analysis of the volatile concentrates of both cultivars showed that the Bragantina responded with the production of higher contents of α-bisabolol at 21 dpi and a decrease of elemol, mostly at 30 dpi; while Cingapura displayed an decrease of δ-elemene production, except at 15 dpi. The phenolic content determined by the Folin Ciocalteu method showed an increase in the leaves of plants inoculated at 7 dpi (Bragantina) and 7-15 dpi (Cingapura); in the roots, the infection caused a phenolic content decrease in Bragantina cultivar at 45 dpi and an increase in the Cingapura cultivar at 15, 30 and 45 dpi. High Performance Liquid Chromatography-Mass spectrometry (HPLC-MS) analysis of the root extracts showed a qualitative variation of alkamides during infection. The results indicated that there is a possible relationship between secondary metabolites and tolerance against phytopathogens.
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http://dx.doi.org/10.3390/ijms18122434DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5751101PMC
December 2017

Similarity in volatile communities leads to increased herbivory and greater tropical forest diversity.

Ecology 2017 Jul;98(7):1750-1756

Instituto de Química, Universidade de São Paulo, Av. Prof. Lineu Prestes, 748, Bloco 11 Térreo, São Paulo, São Paulo, 05508-000, Brasil.

A longstanding paradigm in ecology is that there are positive associations between herbivore diversity, specialization, and plant species diversity, with a focus on taxonomic diversity. However, phytochemical diversity is also an informative metric, as insect herbivores interact with host plants not as taxonomic entities, but as sources of nutrients, primary metabolites, and mixtures of attractant and repellant chemicals. The present research examines herbivore responses to phytochemical diversity measured as volatile similarity in the tropical genus Piper. We quantified associations between naturally occurring volatile variation and herbivory by specialist and generalist insects. Intraspecific similarity of volatile compounds across individuals was associated with greater overall herbivory. A structural equation model supported the hypothesis that plot level volatile similarity caused greater herbivory by generalists, but not specialists, which led to increased understory plant richness. These results demonstrate that using volatiles as a functional diversity metric is informative for understanding tropical forest diversity and indicate that generalist herbivores contribute to the maintenance of diversity.
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http://dx.doi.org/10.1002/ecy.1875DOI Listing
July 2017

The Combined Use of Proteomics and Transcriptomics Reveals a Complex Secondary Metabolite Network in Peperomia obtusifolia.

J Nat Prod 2017 05 19;80(5):1275-1286. Epub 2017 Apr 19.

Instituto de Química, Universidade Estadual Paulista (Unesp) , Araraquara, SP 14800-060, Brazil.

Peperomia obtusifolia, an ornamental plant from the Piperaceae family, accumulates a series of secondary metabolites with interesting biological properties. From a biosynthesis standpoint, this species produces several benzopyrans derived from orsellinic acid, which is a polyketide typically found in fungi. Additionally, the chiral benzopyrans were reported as racemic and/or as diastereomeric mixtures, which raises questions about the level of enzymatic control in the cyclization step for the formation of the 3,4-dihydro-2H-pyran moiety. Therefore, this article describes the use of shotgun proteomic and transcriptome studies as well as phytochemical profiling for the characterization of the main biosynthesis pathways active in P. obtusifolia. This combined approach resulted in the identification of a series of proteins involved in its secondary metabolism, including tocopherol cyclase and prenyltransferases. The activity of these enzymes was supported by the phytochemical profiling performed in different organs of P. obtusifolia. However, the polyketide synthases possibly involved in the production of orsellinic acid could not be identified, suggesting that orsellinic acid may be produced by endophytes intimately associated with the plant.
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http://dx.doi.org/10.1021/acs.jnatprod.6b00827DOI Listing
May 2017

Synergistic effects of in vitro combinations of piplartine, epiisopiloturine and praziquantel against Schistosoma mansoni.

Biomed Pharmacother 2017 Apr 23;88:488-499. Epub 2017 Jan 23.

Biodiversity and Biotechnology Research Center, Biotec, Federal University of Piauí, UFPI, Parnaíba, PI, 64202020, Brazil; Faculty of Medicine, University of Brasilia, UnB, Brasília, DF, Brazil. Electronic address:

Schistosomiasis is a world health problem, and praziquantel is the only drug currently used for the treatment. There is some evidence that extensive monotherapy of praziquantel may be leading to drug resistance in the parasite. In order to find alternative treatments, the effects of the combination of epiisopiloturine (EPI), piplartine (PPT) and praziquantel (PZQ) were evaluated. Similarity analysis of these compounds was performed using optimized molecular structures to compare the shape and the charge modeling of combinations between PZQ and EPI or PPT. Supported by this data, in vitro association of PZQ-PPT, PZQ-EPI, and EPI-PPT was carried out, and the activity of these combinations against Schistosoma mansoni was assessed. The results showed synergistic activity with a combination index (CI) of 0.42 for the treatment with PZQ-PPT. Both PZQ-EPI and EPI-PPT combinations also showed synergistic effects, with CI values of 0.86 and 0.61, respectively. Surface alterations in the tegument of adult schistosomes after the treatments were observed using laser confocal microscopy and scanning electron microscopy. Additionally, the association of EPI-PPT decreased the cytotoxicity when compared with both isolated compounds in three different lines of mammalian cells. Thus, synergistic combinations of PZQ-PPT, PZQ-EPI, and EPI-PPT create the possibility of reduced doses to be used against Schistosoma mansoni.
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http://dx.doi.org/10.1016/j.biopha.2016.12.057DOI Listing
April 2017

Stabilization and detection of hydrophylloquinone as di-O-methyl derivative.

J Chromatogr B Analyt Technol Biomed Life Sci 2016 Oct 8;1033-1034:368-371. Epub 2016 Sep 8.

Department of Fundamental Chemistry, Institute of Chemistry, University of São Paulo, Av. Prof. Lineu Prestes, 748, São Paulo, SP, 05508-000, Brazil. Electronic address:

Phylloquinone is a redox active naphthoquinone involved in electron transport in plants. The function of this reduced form remains unclear due to its instability, which has precluded detection. Herein, a simple method that permits the stabilization of the reduced form of phylloquinone by di-O-methylation and HPLC detection is described.
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http://dx.doi.org/10.1016/j.jchromb.2016.09.011DOI Listing
October 2016

Bioactivity-guided isolation of laevicarpin, an antitrypanosomal and anticryptococcal lactam from Piper laevicarpu (Piperaceae).

Fitoterapia 2016 Jun 12;111:24-8. Epub 2016 Apr 12.

Instituto de Ciências Ambientais, Químicas e Farmacêuticas, Universidade Federal de São Paulo, 09972-270, São Paulo, SP, Brazil. Electronic address:

Crude CH2Cl2 extract from leaves of Piper laevicarpu (Piperaceae) displayed antitrypanosomal activity against trypomastigote forms of Trypanosoma cruzi (Y strain) and antimicrobial potential against Cryptococcus gattii (strain-type WM 178). Bioactivity-guided fractionation of crude extract afforded one new natural bioactive lactam derivative, named laevicarpin. The structure of isolated compound, which displayed a very rare ring system, was elucidated based on NMR, IR and MS spectral analysis. Using MTT assay, the trypomastigotes of T. cruzi demonstrated susceptibility to laevicarpin displaying IC50 value of 14.7μg/mL (49.6μM), about 10-fold more potent than the standard drug benznidazole. The mammalian cytotoxicity of laevicarpin was verified against murine fibroblasts (NCTC cells) and demonstrated a CC50 value of 100.3μg/mL (337.7μM-SI=7). When tested against Cryptococcus gattii, laevicarpin showed an IC50 value of 2.3μg/mL (7.9μM) and a MIC value of 7.4μg/mL (25μM). Based in the obtained results, laevicarpin could be used as a scaffold for future drug design studies against the Chagas disease and anti-cryptococosis agents.
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http://dx.doi.org/10.1016/j.fitote.2016.04.005DOI Listing
June 2016

UV-resistant yeasts isolated from a high-altitude volcanic area on the Atacama Desert as eukaryotic models for astrobiology.

Microbiologyopen 2015 Aug 4;4(4):574-88. Epub 2015 Jul 4.

Interunities Graduate Program in Biotechnology, Universidade de São Paulo, São Paulo, Brazil.

The Sairecabur volcano (5971 m), in the Atacama Desert, is a high-altitude extreme environment with high daily temperature variations, acidic soils, intense UV radiation, and low availability of water. Four different species of yeasts were isolated from this region using oligotrophic media, identified and characterized for their tolerance to extreme conditions. rRNA sequencing revealed high identity (>98%) to Cryptococcus friedmannii, Exophiala sp., Holtermanniella watticus, and Rhodosporidium toruloides. To our knowledge, this is the first report of these yeasts in the Atacama Desert. All isolates showed high resistance to UV-C, UV-B and environmental-UV radiation, capacity to grow at moderate saline media (0.75-2.25 mol/L NaCl) and at moderate to cold temperatures, being C. friedmannii and H. watticus able to grow in temperatures down to -6.5°C. The presence of pigments, analyzed by Raman spectroscopy, correlated with UV resistance in some cases, but there is evidence that, on the natural environment, other molecular mechanisms may be as important as pigmentation, which has implications for the search of spectroscopic biosignatures on planetary surfaces. Due to the extreme tolerances of the isolated yeasts, these organisms represent interesting eukaryotic models for astrobiological purposes.
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http://dx.doi.org/10.1002/mbo3.262DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4554453PMC
August 2015

Nitrogen acquisition in Agave tequilana from degradation of endophytic bacteria.

Sci Rep 2014 Nov 6;4:6938. Epub 2014 Nov 6.

Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, 05508-000, São Paulo, SP, Brazil.

Plants form symbiotic associations with endophytic bacteria within tissues of leaves, stems, and roots. It is unclear whether or how plants obtain nitrogen from these endophytic bacteria. Here we present evidence showing nitrogen flow from endophytic bacteria to plants in a process that appears to involve oxidative degradation of bacteria. In our experiments we employed Agave tequilana and its seed-transmitted endophyte Bacillus tequilensis to elucidate organic nitrogen transfer from (15)N-labeled bacteria to plants. Bacillus tequilensis cells grown in a minimal medium with (15)NH4Cl as the nitrogen source were watered onto plants growing in sand. We traced incorporation of (15)N into tryptophan, deoxynucleosides and pheophytin derived from chlorophyll a. Probes for hydrogen peroxide show its presence during degradation of bacteria in plant tissues, supporting involvement of reactive oxygen in the degradation process. In another experiment to assess nitrogen absorbed as a result of endophytic colonization of plants we demonstrated that endophytic bacteria potentially transfer more nitrogen to plants and stimulate greater biomass in plants than heat-killed bacteria that do not colonize plants but instead degrade in the soil. Findings presented here support the hypothesis that some plants under nutrient limitation may degrade and obtain nitrogen from endophytic microbes.
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http://dx.doi.org/10.1038/srep06938DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4221784PMC
November 2014

Isolation and characterization of nine polymorphic microsatellite loci in Piper solmsianum (piperaceae).

Appl Plant Sci 2014 Apr 4;2(4). Epub 2014 Apr 4.

Centro de Pesquisa e Desenvolvimento de Recursos Genéticos Vegetais, Instituto Agronômico de Campinas (IAC), C.P. 28, 13075-630 Campinas, São Paulo, Brazil.

Unlabelled:

Premise Of The Study: Nine microsatellite (simple sequence repeat [SSR]) loci were characterized for natural populations of Piper solmsianum, a potential source of bioactive secondary metabolites, and analyzed to assess the levels of genetic diversity in this species. •

Methods And Results: Based on an enriched library using the oligonucleotides (CT)8 and (GT)8, a total of 19 pairs of SSR primers were designed and nine of them were highly polymorphic after screening of 37 specimens from two populations. The number of alleles per locus ranged from one to six while the observed heterozygosity for polymorphic loci ranged from 0.000 to 0.875. •

Conclusions: The SSR regions characterized were informative, and the genetic markers will be useful to assess the genetic diversity and gene flow in populations of P. solmsianum.
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http://dx.doi.org/10.3732/apps.1300092DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4103135PMC
April 2014

Age-dependent changes from allylphenol to prenylated benzoic acid production in Piper gaudichaudianum Kunth.

Phytochemistry 2014 Oct 17;106:86-93. Epub 2014 Jul 17.

Research Support Center in Molecular Diversity of Natural Products, Institute of Chemistry, University of São Paulo, Av. Prof. Lineu Prestes, 748, 05508-000 São Paulo, SP, Brazil. Electronic address:

HPLC-DAD and principal component analysis (PCA) of the (1)H NMR spectrum of crude plant extracts showed high chemical variability among seedlings and adult organs of Piper gaudichaudianum. While gaudichaudianic acid was the major compound in the adult leaves, apiole and dillapiole were the major compounds in their seedling leaves. By the 15th month of seedling growth, the levels of apiole and dillapiole decreased and gaudichaudianic acid appeared along with two compounds, biosynthetically related to gaudichaudianic acid.
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http://dx.doi.org/10.1016/j.phytochem.2014.06.013DOI Listing
October 2014

Antifungal and cytotoxic 2-acylcyclohexane-1,3-diones from Peperomia alata and P. trineura.

J Nat Prod 2014 Jun 6;77(6):1377-82. Epub 2014 Jun 6.

Research Support Center in Molecular Diversity of Natural Products, Institute of Chemistry, University of São Paulo , 05508-000, São Paulo, SP, Brazil.

Bioactivity-guided fractionation of the separate CH2Cl2 extracts from the aerial parts of Peperomia alata and P. trineura yielded seven polyketides: alatanone A [3-hydroxy-2-(5'-phenylpent-4'E-enoyl)cyclohex-2-en-1-one, 1a] and alatanone B [3-hydroxy-2-(3'-phenyl-6'-methylenedioxypropanoyl)cyclohex-2-en-1-one, 2a] from P. alata and trineurone A [3-hydroxy-2-(11'-phenylundec-10'E-enoyl)cyclohex-2-en-1-one, 1b], trineurone B [3-hydroxy-2-(15'-phenyl-18'-methylenedioxypentadecanoyl)cyclohex-2-en-1-one, 2b], trineurone C [3-hydroxy-2-(17'-phenyl-20'-methylenedioxyheptadecanoyl)cyclohex-2-en-1-one, 2c], trineurone D [3-hydroxy-2-(hexadec-10'Z-enoyl)cyclohex-2-en-1-one, 3a], and trineurone E [(6R)-(+)-3,6-dihydroxy-2-(hexadec-10'Z-enoyl)cyclohex-2-en-1-one, 3b] from P. trineura. The isolated compounds were evaluated for antifungal activity against Cladosporium cladosporioides and C. sphaeospermum and for cytotoxicity against the K562 and Nalm-6 leukemia cell lines.
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http://dx.doi.org/10.1021/np500130xDOI Listing
June 2014

A benzoic acid derivative and flavokawains from Piper species as schistosomiasis vector controls.

Molecules 2014 Apr 23;19(4):5205-18. Epub 2014 Apr 23.

Research Support Center in Diversity of Natural Products, Instituto de Química, Universidade de São Paulo, Av. Prof. Lineu Prestes, 748, sala 1124, São Paulo, SP, CEP 05508-000, Brazil.

The search of alternative compounds to control tropical diseases such as schistosomiasis has pointed to secondary metabolites derived from natural sources. Piper species are candidates in strategies to control the transmission of schistosomiasis due to their production of molluscicidal compounds. A new benzoic acid derivative and three flavokawains from Piper diospyrifolium, P. cumanense and P. gaudichaudianum displayed significant activities against Biomphalaria glabrata snails. Additionally, "in silico" studies were performed using docking assays and Molecular Interaction Fields to evaluate the physical-chemical differences among the compounds in order to characterize the observed activities of the test compounds against Biomphalaria glabrata snails.
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http://dx.doi.org/10.3390/molecules19045205DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6271750PMC
April 2014

Antiherbivore prenylated benzoic acid derivatives from Piper kelleyi.

J Nat Prod 2014 Jan 14;77(1):148-53. Epub 2014 Jan 14.

Department of Chemistry, University of Nevada, Reno , Reno, Nevada 89557, United States.

The known prenylated benzoic acid derivative 3-geranyl-4-hydroxy-5-(3″,3″-dimethylallyl)benzoic acid (1) and two new chromane natural products were isolated from the methanolic extract of the leaves of Piper kelleyi Tepe (Piperaceae), a midcanopy tropical shrub that grows in lower montane rain forests in Ecuador and Peru. Structure determination using 1D and 2D NMR analysis led to the structure of the chromene 2 and to the reassignment of the structure of cumanensic acid as 4, an isomeric chromene previously isolated from Piper gaudichaudianum. The structure and relative configuration of new chromane 3 was determined using 1D and 2D NMR spectroscopic analysis and was found to be racemic by ECD spectropolarimetry. The biological activity of 1-3 was evaluated against a lab colony of the generalist caterpillar Spodoptera exigua (Noctuidae), and low concentrations of 2 and 3 were found to significantly reduce fitness. Further consideration of the biosynthetic relationship of the three compounds led to the proposal that 1 is converted to 2 via an oxidative process, whereas 3 is produced through hetero-[4+2] dimerization of a quinone methide derived from the chromene 2.
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http://dx.doi.org/10.1021/np400886sDOI Listing
January 2014

Cytotoxic non-aromatic B-ring flavanones from Piper carniconnectivum C. DC.

Phytochemistry 2014 Jan 16;97:81-7. Epub 2013 Nov 16.

Research Support Center in Molecular Diversity of Natural Products, Instituto de Química, Universidade de São Paulo, CP 26077, 05599-970 São Paulo, SP, Brazil. Electronic address:

The EtOAc extract from the leaves of Piper carniconnectivum C. DC. was subjected to chromatographic separation to afford two non-aromatic B-ring flavanone compounds: 5-hydroxy-2-(1'-hydroxy-4'-oxo-cyclohex-2'-en-1'-yl)-6,7-dimethoxy-2,3-dihydro-4H-chromen-4-one (1) and 5-hydroxy-2-(1',2'-dihydroxy-4'-oxo-cyclohexyl)-6,7-dimethoxy-2,3-dihydro-4H-chromen-4-one (2). The absolute configuration of (+)-1 was unambiguously determined as 2S,1'R by electronic circular dichroism (ECD) spectroscopy and comparison to simulated spectra that were calculated using time-dependent density functional theory (TDDFT). This methodology allowed the assignment of the absolute configuration of (+)-2 also as 2S,1'R, except for the stereogenic center at C-2', which was assigned as R because of the evidence drawn from high resolution NMR experiments. The cytotoxic activity of both compounds and 3 (hydrogenated B-ring derivative of 1) was evaluated on twelve human leukemia cell lines, and the IC50 values (<10 μM) indicated the activity of 1 against seven cell lines.
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http://dx.doi.org/10.1016/j.phytochem.2013.10.012DOI Listing
January 2014

Leishmanicidal activity of an alkenylphenol from Piper malacophyllum is related to plasma membrane disruption.

Exp Parasitol 2012 Nov 5;132(3):383-7. Epub 2012 Sep 5.

Instituto de Química, Universidade Federal de Uberlândia, Uberlândia, MG, Brazil.

Leishmaniasis and Chagas disease are parasitic protozoan infections that affect the poorest population in the world, causing high mortality and morbidity. As a result of highly toxic and long-duration treatments, novel, safe and more efficacious drugs are essential. In this work, the methanol (MeOH) extract from the leaves of Piper malacophyllum (Piperaceae) was fractioned to afford one alkenylphenol, which was characterized as 4-[(3'E)-decenyl]phenol (gibbilimbol B) by spectroscopic methods. Anti-protozoan in vitro assays demonstrated for the first time that Leishmania (L.) infantum chagasi was susceptible to gibbilimbol B, with an in vitro EC(50) of 23 μg/mL against axenic promastigotes and an EC(50) of 22 μg/mL against intracellular amastigotes. Gibbilimbol B was also tested for anti-trypanosomal activity (Trypanosoma cruzi) and showed an EC(50) value of 17 μg/mL against trypomastigotes. To evaluate the cytotoxic parameters, this alkenylphenol was tested in vitro against NCTC cells, showing a CC(50) of 59 μg/mL and absent hemolytic activity at the highest concentration of 75 μg/mL. Using the fluorescent probe SYTOX Green suggested that the alkenylphenol disrupted the Leishmania plasma membrane upon initial incubation. Further drug design studies aiming at derivatives could be a promising tool for the development of new therapeutic agents for leishmaniasis and Chagas disease.
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http://dx.doi.org/10.1016/j.exppara.2012.08.019DOI Listing
November 2012

Electrophysiological responses of the Naupactus bipes beetle to essential oils from Piperaceae species.

Nat Prod Commun 2012 Aug;7(8):1103-6

Departamento de Ciências Moleculares da Universidade Federal Rural de Pernambuco, 52.171-900 Recife, PE, Brazil.

Electrophysiological responses based on electroantennographic detection (GC-EAD) and electroantennography (EAG) analysis of Naupactus bipes beetles (Germar, 1824) (Coleoptera: Curculionidae: Brachycerinae) were used to test volatile oils of Piper gaudichaudianum, P. regnellii and P. hispidum. In the EAG experiments, female and male beetles showed significant EAG response to the three volatile oils of Piper species, with the females' responses slightly higher than the males'. The experiments with GC-EAD revealed that some terpenoids (namely, alpha-pinene, beta-pinene, myrcene, alpha-copaene and germacrene) present in the leaf essential oils of the Piper species are perceptible to female and male beetles.
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August 2012

Schistosoma mansoni: in vitro schistosomicidal activity and tegumental alterations induced by piplartine on schistosomula.

Exp Parasitol 2012 Oct 14;132(2):222-7. Epub 2012 Jul 14.

Laboratório de Parasitologia, Instituto Butantan, São Paulo, SP, Brazil.

Schistosomiasis is one of the most important parasitic infections in humans that occur in many tropical and subtropical countries. Currently, the control of schistosomiasis rests with a single drug, praziquantel, which is effective against adult worms but not the larval stages. Recent studies have shown that piplartine, an amide isolated from plants of the genus Piper (Piperaceae), reveals interesting antischistosomal properties against Schistosoma mansoni adult worms. Here, we report the in vitro antischistosomal activity of piplartine on S. mansoni schistosomula of different ages (3 h old and 1, 3, 5, and 7 days old), and examine alterations on the tegumental surface of worms by means of confocal laser scanning microscopy. Piplartine at a concentration of 7.5 μM caused the death of all schistosomula within 120 h. The lethal effect occurred in a dose-dependent manner and was also dependent on the age of the parasite. Microscopy observation revealed extensive tegumental destruction, including blebbing, granularity, and a shorter body length. This report provides the first evidence that piplartine is able to kill schistosomula of different ages and reinforce that piplartine is a promising compound that could be used for the development of new schistosomicidal agent.
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http://dx.doi.org/10.1016/j.exppara.2012.07.004DOI Listing
October 2012

VCD to determine absolute configuration of natural product molecules: secolignans from Peperomia blanda.

Org Biomol Chem 2012 Jun 30;10(21):4208-14. Epub 2012 Apr 30.

Organic Chemistry Department, Institute of Chemistry, São Paulo State University - UNESP, Araraquara, SP 14800-900, Brazil.

The absolute configuration and solution-state conformers of three peperomin-type secolignans isolated from Peperomia blanda (Piperaceae) are unambiguously determined by using vibrational circular dichroism (VCD) spectroscopy associated with density functional theory (DFT) calculations. Advantages of VCD over the electronic form of CD for the analysis of diastereomers are also discussed. This work extends our growing knowledge about secondary metabolites within the Piperaceae family species while providing a definitive and straightforward method to assess the absolute stereochemistry of secolignans.
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http://dx.doi.org/10.1039/c2ob25109dDOI Listing
June 2012

Secondary metabolites from the phloem of Piper solmsianum (Piperaceae) in the honeydew of Edessa meditabunda.

Phytochem Anal 2012 Nov-Dec;23(6):604-6. Epub 2012 Mar 7.

Departamento de Ciências Moleculares, Universidade Federal Rural de Pernambuco, CP 52.171-030, Recife, PE, Brazil.

Introduction: The phytochemistry of species of the genus Piper has been studied extensively, including Piper solmsianum. However, no studies have addressed the phytochemistry of the sap content of Piper species.

Objective: To evaluate the transferring of secondary compounds from the saps of P. solmsianum to the honeydew of Edessa meditabunda.

Methodology: The honeydew of E. meditabunda and saps of P. solmsianum were analysed by GC-MS, (1) H-NMR and LC-MS.

Results: The lignan (-)-grandisin and the phenylpropanoid (E)-isoelemicin were detected in both saps of P. solmsianum and honeydew of E. meditabunda.

Conclusion: Analysis of honeydew secreted by the sap-sucking insect E. meditabunda indicated that (-)-grandisin and (E)-isoelemicin are absorbed from the phloem of Piper solmsianum.
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http://dx.doi.org/10.1002/pca.2361DOI Listing
April 2013

Secondary metabolites isolated from Piper chimonantifolium and their antifungal activity.

Nat Prod Res 2012 24;26(8):770-3. Epub 2011 Oct 24.

Departamento de Ciências Exatas e da Terra, Universidade Federal de São Paulo, 09972-270, Diadema, SP, Brazil.

In this study, the CH(2)Cl(2) extract from leaves of Piper chimonantifolium was subjected to several chromatographic separation procedures to afford one chromene (gaudichaudianic acid) as a major compound as well as two flavonoids (dihydrooroxylin and pinocembrin) and three steroids (sitosterol, sitosteryl palmitate and stigmasterol). The structures of all determined compounds were characterised by spectrometric analysis, mainly mass spectrometry and NMR, as well as their optical properties. This article describes the first phytochemical study of the leaves of P. chimonantifolium and an evaluation of the antifungal activity of its major compounds.
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http://dx.doi.org/10.1080/14786419.2011.561435DOI Listing
August 2012

Antioxidant capacity and larvicidal and antifungal activities of essential oils and extracts from Piper krukoffii.

Nat Prod Commun 2011 Sep;6(9):1361-6

Programa de Pós-Graduação em Química, Universidade Federal do Pará, 66075-900 Belém, PA, Brazil.

The leaves and twigs of Piper krukoffii, collected in the Carajás National Forest, north Brazil, yielded essential oils (2.0% and 0.8%), the main constituents of which were myristicin (40.3% and 26.7%), apiole (25.4% and 34.1%) and elemicin (2.8% and 3.0%). The antioxidant activities of the oils, methanol extract and its sub-fractions were evaluated. The DPPH EC50 values varied from the ethyl acetate sub-fraction (73.4 +/- 3.7 microg/mL) to the methanol extract (24.9 +/- 0.8 microg/mL), and the ABTS TEAC values ranged in the same order from 265.7 to 349.2 microMol TE/g. These results indicated a significant antioxidant activity for the plant. The lignans (-)-kusunokin, yatein, (-)-hinokin and cubebin were identified in the methanol extract. The hydro-methanolic sub-fraction showed a high value for total phenol content (106.5 +/- 0.7 mg GAE/g), as well as 1H NMR signals for sugar moieties. Crude extracts and sub-fractions were also able to inhibit beta-carotene bleaching, varying from 22.4 to 47.1%. The oils from the leaves and twigs showed strong larvicidal (21.4 and 3.6 microg/mL) and fungicide (0.5 and 0.1 microg/mL) activities.
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September 2011