Differential biological activity of two extracts of ripe fruits of Carica candamarcensis Hook on human neutrophils / Actividad biologica diferencial de dos extractos de frutos maduros de Carica candamarcensis Hook en neutrófilos humanos

We evaluated the effect of the total macerate (TM) and seed oil (SO) of mature Carica candamarcensis fruits, on the release of Matrix metalloproteinase 9 (MMP9) and the phosphorylation of MAPK in neutrophils. The antioxidant capacity of these extracts was evaluated by ABTS assay. Neutrophils stimulated with different dilutions of TM or SO were analyzed for cytotoxicity, MMP9 release, and MAPK phosphorylation, using trypan blue exclusion assays, zymography, and immunoblotting, respectively. Both extracts show antioxidant activity, being higher in TM; none presented cytotoxic effect. The 5% and 2.5% dilutions of TM significantly reduced MMP9 release, and all decreased MAPK phosphorylation. SO significantly increased the release o f MMP9 and MAPK phosphorylation, the effect being greater when they were prestimulated with lipopolysaccharide.TM may have anti - inflammatory potential, while SO could have a priming effect that needs to be confirmed

Evaluamos el efecto del macerado total (MT) y aceite de semillas (AV) de frutos maduros de Carica candamarcensis , en la liberación de Matriz metaloproteinasa 9 (MMP9) y la fosfor ilación de MAPK en neutrófilos. La capacidad antioxidante de estos extractos se evaluó por ensayo ABTS. En neutrófilos estimulados con diferentes diluciones de MT o AV se analizó la citotoxicidad, liberación de MMP9 y fosforilación de MAPK, mediante ensayo s de exclusión con azul de tripano, zimografía e inmunotransferencia, respectivamente. Ambos extractos muestran actividad antioxidante, siendo mayor en MT; ninguno presentó efecto citotóxico. Las diluciones 5% y 2,5% de MT redujeron significativamente la l iberación de MMP9, y todas disminuyeron la fosforilación de MAPK. El AV incrementó significativamente la liberación de MMP9 y la fosforilación de MAPK, el efecto fue mayor cuando se preestimularon con lipopolisacárido. El MT puede tener potencial antiinfla matorio, mientras que el AV podría tener un efecto "priming" que necesita ser corroborado.

Biochemical characterization of Euphorbia resinifera floral cyathia.

Euphorbia resinifera O. Berg is a plant endemic to the Northern and Central regions of Morocco known since the ancient Roman and Greek times for secreting a poisonous latex containing resiniferatoxin. However, E. resinifera pseudo-inflorescences called cyathia are devoid of laticifers and, therefore, do not secrete latex. Instead, they exudate nectar that local honey bees collect and craft into honey. Honey and cyathium water extracts find a broad range of applications in the traditional medicine of Northern Africa as ointments and water decoctions. Moreover, E. resinifera monofloral honey has received the Protected Geographic Indication certification for its outstanding qualities. Given the relevance of E. resinifera cyathia for bee nutrition, honey production, and the health benefit of cyathium-derived products, this study aimed to screen metabolites synthesized and accumulated in its pseudo-inflorescences. Our analyses revealed that E. resinifera cyathia accumulate primary metabolites in considerable abundance, including hexoses, amino acids and vitamins that honey bees may collect from nectar and craft into honey. Cyathia also synthesize volatile organic compounds of the class of benzenoids and terpenes, which are emitted by flowers pollinated by honey bees and bumblebees. Many specialized metabolites, including carotenoids, flavonoids, and polyamines, were also detected, which, while protecting the reproductive organs against abiotic stresses, also confer antioxidant properties to water decoctions. In conclusion, our analyses revealed that E. resinifera cyathia are a great source of antioxidant molecules and a good food source for the local foraging honeybees, revealing the central role of the flowers from this species in mediating interactions with local pollinators and the conferral of medicinal properties to plant extracts.

Comparison of direct analysis in real-time mass spectrometry, atmospheric solids analysis probe-mass spectrometry, and ion mobility spectrometry for ensuring food safety by rapid screening of poppy seeds.

The opium poppy (Papaver somniferum) is a global commercial crop that has been historically valued for both medicinal and culinary purposes. Naturally occurring opium alkaloids including morphine, codeine, thebaine, noscapine, and papaverine are found primarily in the latex produced by the plant. If the plant is allowed to fully mature, poppy seeds that do not contain the opium alkaloids will form within the pods and may be used in the food industry. It is possible for the seeds to become contaminated with alkaloids by the latex during harvesting, posing a potential health risk for consumers. In the USA, there have been more than 600 reported adverse events including 19 fatalities that may be linked to the consumption of a contaminated poppy-containing product such as home-brewed poppy seed tea. Unwashed poppy seeds and pods may be purchased over the Internet and shipped worldwide. The Forensic Chemistry Center, US Food and Drug Administration (FDA) has evaluated several mass spectrometers (MS) capable of rapid screening to be used for high-throughput analysis of samples such as poppy seeds. These include a direct analysis in real-time (DART) ambient ionization source coupled to a single-quadrupole MS, an atmospheric solids analysis probe (ASAP) ionization source coupled to the same MS, and ion mobility spectrometers (IMS). These instruments have been used to analyze 17 poppy seed samples for the presence of alkaloids, and the results were compared to data obtained using liquid chromatography with mass spectral detection (LC-MS/MS). Results from the 17 poppy seed samples indicate that the DART-MS, ASAP-MS, and IMS devices detect many of the same alkaloids confirmed during the LC-MS/MS analyses, although both the false-positive and false-negative rates are higher, possibly due to the non-homogeneity of the samples and the lack of chromatographic separation.

Development of a Method to Extract Opium Poppy (Papaver somniferum L.) DNA from Heroin.

This study is the first to report the successful development of a method to extract opium poppy (Papaver somniferum L.) DNA from heroin samples. Determining of the source of an unknown heroin sample (forensic geosourcing) is vital to informing domestic and foreign policy related to counter-narcoterrorism. Current profiling methods focus on identifying process-related chemical impurities found in heroin samples. Changes to the geographically distinct processing methods may lead to difficulties in classifying and attributing heroin samples to a region/country. This study focuses on methods to optimize the DNA extraction and amplification of samples with low levels of degraded DNA and inhibiting compounds such as heroin. We compared modified commercial-off-the-shelf extraction methods such as the Qiagen Plant, Stool and the Promega Maxwell-16 RNA-LEV tissue kits for the ability to extract opium poppy DNA from latex, raw and cooked opium, white and brown powder heroin and black tar heroin. Opium poppy DNA was successfully detected in all poppy-derived samples, including heroin. The modified Qiagen stool method with post-extraction purification and a two-stage, dual DNA polymerase amplification procedure resulted in the highest DNA yield and minimized inhibition. This paper describes the initial phase in establishing a DNA-based signature method to characterize heroin.

Preliminary study to characterize plastic polymers using elemental analyser/isotope ratio mass spectrometry (EA/IRMS).

Plastic waste is a growing global environmental problem, particularly in the marine ecosystems, in consideration of its persistence. The monitoring of the plastic waste has become a global issue, as reported by several surveillance guidelines proposed by Regional Sea Conventions (OSPAR, UNEP) and appointed by the EU Marine Strategy Framework Directive. Policy responses to plastic waste vary at many levels, ranging from beach clean-up to bans on the commercialization of plastic bags and to Regional Plans for waste management and recycling. Moreover, in recent years, the production of plant-derived biodegradable plastic polymers has assumed increasing importance. This study reports the first preliminary characterization of carbon stable isotopes (δ13C) of different plastic polymers (petroleum- and plant-derived) in order to increase the dataset of isotopic values as a tool for further investigation in different fields of polymers research as well as in the marine environment surveillance. The δ13C values determined in different packaging for food uses reflect the plant origin of "BIO" materials, whereas the recycled plastic materials displayed a δ13C signatures between plant- and petroleum-derived polymers source. In a preliminary estimation, the different colours of plastic did not affect the variability of δ13C values, whereas the abiotic and biotic degradation processes that occurred in the plastic materials collected on beaches and in seawater, showed less negative δ13C values. A preliminary experimental field test confirmed these results. The advantages offered by isotope ratio mass spectrometry with respect to other analytical methods used to characterize the composition of plastic polymers are: high sensitivity, small amount of material required, rapidity of analysis, low cost and no limitation in black/dark samples compared with spectroscopic analysis.

Avaliação da qualidade de amostras comerciais de leite de janaguba (Himatanthus drasticus (Mart. ) Plumel) em Fortaleza – Ceará / Quality assessment of commercial samples of janaguba’s milk (Himatanthus drasticus (Mart. ) Plumel) in Fortaleza – Ceará.

RESUMO Os fitoterápicos à base de leite de janaguba (Himatanthus drasticus (Mart.) Plumel), usados para o tratamento de câncer, úlcera gástrica e outras doenças, são muito vendidos em mercados públicos de Fortaleza (CE). No entanto, registros mencionam que é comum a troca deste leite por látex de mangabeira (Hancornia speciosa Gomes). O trabalho objetivou avaliar a qualidade físico-química, química e microbiológica de amostras comerciais do leite de janaguba. Dez amostras comerciais foram adquiridas de um mercado de Fortaleza; quatro amostras autênticas de látex de janaguba foram obtidas da chapada do Araripe e uma amostra de látex de mangabeira foi obtida em Paracuru (CE). Foram determinados o aspecto geral, densidade, pH, resíduo seco, volume de sedimentação, perfil cromatográfico e qualidade microbiológica das amostras. Os resultados mostraram elevada contaminação microbiológica nas preparações comerciais e adulteração em seis destas amostras, o que aponta a urgente implantação de uma efetiva farmacovigilância dos fitoterápicos a fim de tornar seu consumo mais seguro e racional.

ABSTRACT The herbal medicines made from janaguba milk (Himatanthus drasticus (Mart.) Plumel), used to treat gastric ulcer, cancer and other diseases, and arewidely sold in the public markets of Fortaleza (CE). However, records mention that it is common to make fake copies of the milk by using mango tree latex (Hanconia speciosa Gomes). This study aimed to evaluate the chemical, physical-chemical and microbiological quality of commercial samples of janaguba milk. Ten commercial samples were purchased from a Fortaleza market; four authentic samples of janaguba latex were obtained from the Araripe plateau, and one mango tree latex sample was obtained in Paracuru (CE). We determined the general appearance, density, pH, dry residue, sedimentation volume, chromatographic profile and microbiological qualities. Results showed high microbiological contamination in the commercial preparations and evidence of tampering in six of these samples, which indicates the need for an urgent implementation of an effective pharmacovigilance strategy for herbal medicines in order to make its consumption more secure and rational.

Estudo etnofarmacológico e etnobotânico de Himatanthus drasticus (Mart. ) Plumel (janaguba) / Ethnopharmacological and ethnobotanical study of Himatanthus drasticus (Mart. ) Plumel (janaguba).

RESUMO Himatanthus drasticus (Mart.) Plumel, janaguba, produz um látex em seu caule que é usado para o tratamento de alguns tipos de câncer, vermes intestinais, febre, menstruações irregulares, infertilidade feminina e úlceras gástricas. Estudos fitoquímicos revelaram a presença de acetato de lupeol e proteínas; e estudos farmacológicos, os efeitos antiúlcera, antitumoral, imunomodulador, analgésico e anti-inflamatório. O presente trabalho objetivou realizar levantamento etnobotânico e etnofarmacológico sobre H. drasticus como contribuição ao conhecimento científico da referida espécie. Foi elaborado um questionário, que foi aplicado em entrevista a 21 raizeiros nos mercados das cidades de Crato, Fortaleza e Pacajus do estado do Ceará (CE). Observou-se que 38,1% dos raizeiros têm mais de 10 anos de serviço e 47,6%, estão na faixa etária de 40 a 59 anos, o que representa um fator positivo quanto à experiência com plantas medicinais. Aproximadamente 85,7% dos raizeiros relataram o uso do látex da H. drasticus para tratamento de úlcera; 66,7% para inflamação; 66,7% para câncer. Verificou-se que as principais indicações terapêuticas relatadas pelos raizeiros sobre esta espécie estão de acordo com as informações científicas. Os levantamentos etnobotânicos e etnofarmacológicos exercem um papel primordial no resgate do conhecimento tradicional, nos meios rurais e urbanos, tanto por seu valor histórico-cultural, como também pela necessidade de confirmação das indicações de uso.

ABSTRACT The Himatanthus drasticus (Mart.) Plumel, janaguba, produces a latex whose stem is used to treat some types of cancer, intestinal worms, fever, irregular menses, female infertility and gastric ulcers. Phytochemical studies revealed the presence of lupeol acetate and proteins in the latex; and pharmacological studies indicated antiulcer, anti-tumor, immunomodulating, analgesic and anti-inflammatory effects. This study aimed to conduct surveys on ethnopharmacological and ethnobotanical species H. drasticus as a contribution to the scientific knowledge about the species. A questionnaire and interviews were made with 21 healers in the markets of the cities of Crato, Fortaleza and Pacajús, in state of Ceara (CE). It was observed that 38.1% of the healers have more than 10 years of service, and 47.6% are between 40-59 years old, which is a positive factor about the experience with medicinal plants. Approximately 85.7% of the healers reported to apply latex H. drasticus for the treatment of ulcers, 66.7% for inflammation, and 66.7% for cancer. It was found that the main therapeutic indications mentioned by the healers about this species are consistent with the scientific information available. The ethnobotanical and ethnopharmacological surveys play an essential role on keeping the traditional knowledge in rural and urban areas, both for its historical and cultural value, but also for the need of confirmation of the intended use.

Análise sazonal da produção e da atividade enzimática de látex fresco coletado de frutos de plantas femininas e hermafroditas de mamão (Carica papaya L. ) / Seasonal analysis of yield and enzyme activity of fresh latex collected from fruits of female and hermaphrodite plants of papaya (Carica papaya L. )

RESUMO O látex obtido do fruto de Carica papaya L. (mamão) é de grande importância industrial e farmacêutica. Sua obtenção, através do cultivo dessa espécie, depende de vários fatores ainda pouco estudados. Nesse sentido, o presente trabalho tem por objetivo avaliar e comparar plantas femininas e hermafroditas de C. papaya em relação à produção de látex e sua atividade enzimática em coletas semanais e consecutivas durante o 12º mês de cultivo (verão) e no decorrer do segundo ano de desenvolvimento das plantas, e em mais três coletas abrangendo as demais estações do ano (outono, inverno e primavera). Os resultados mostraram que, em quatro coletas semanais e sucessivas durante o verão, a massa de látex da primeira coleta foi superior às demais para plantas femininas. Plantas hermafroditas tiveram comportamento oscilante para esta variável. Plantas femininas tiveram médias de produção superiores comparadas com as hermafroditas. As médias das atividades enzimáticas desse período se mostraram superiores para plantas hermafroditas. Nas coletas estacionais foi observado que plantas hermafroditas têm um comportamento mais oscilante em relação à produção de látex e atividade enzimática. Plantas femininas produzem, em média, maior massa de látex e são mais constantes na produção. Nas estações avaliadas, observaram-se semelhanças nas médias das atividades enzimáticas entre plantas femininas e hermafroditas.

ABSTRACT The latex obtained from the fruit of Carica papaya L. (papaya) is of great industrial and pharmaceutical importance. Its obtainment, through the cultivation of this specie, depends on several factors still poorly studied. In this sense, this study aims to evaluate and compare hermaphrodite and female plants of C. papaya for the production of latex and its enzymatic activity in weekly and consecutive collections of fruits during the 12th month of cultivation (summer) and during the second year of the plant´s development, in three collections covering the other seasons (autumn, winter and spring). The results showed that, in four successive weekly tapping during the summer, the latex mass of the first collection was superior to the others, for female plants. Hermaphrodite plants had oscillating behavior for this variable. Female plants had higher production averages, compared with the hermaphrodites ones. The averages of the enzymatic activities of this period were higher for hermaphrodite plants. The seasonal tapping showed that hermaphrodite plants had oscillating behavior in the production of latex and enzyme activity. Female plants produced, on average, greater mass and were constant in latex production. In the evaluated seasons, similarities in the averages of the enzymatic activities between female and hermaphrodite plants were observed.

Effects of Euphorbia milii latex on mitogen-induced lymphocyte proliferation / Efeito do latex de Euphorbia milii sobre a proliferação de linfócitos induzida por mitogénio

The crude latex of "Crown-of-Thorns" (Euphorbia milii var hislopii, syn E.splendens) is a potent plant molluscicide. For this reason, toxicological studies have been performed to evaluate the health risks posed by its use in schistosomiasis control programs. The present study is part of a more comprehensive immunotoxicological evaluation of this molluscicide. Here, we investigated the effects of E. milii latex on the proliferation of human lymphocytes in vitro. Lyophilized latex of E. milii (0, 0.5, 5, 25 and 50 µg/ml) was incubated with whole blood in the presence of proliferation stimulators, i.e. lectins (phytohemagglutinin, concanavalin A and pokeweed mitogen), as well as with human monoclonal antibody against CD3 and tetanus toxoid. Cell proliferation was measured by ³H-thymidine incorporation, and the effects of latex on mitogen-induced cell proliferation were compared to the effects of 10 ng/ml of 12-O-tetradecanoylphorbol-13-acetate (TPA). Results showed that mitogen-induced cell proliferation was markedly enhanced by E. milii latex. This synergistic effect of latex on mitogen-induced lymphocyte proliferation may be due to the presence of TPA-like phorbol esters and/or to mitogenic plant lectins.

O látexbrutoda "Coroa de Cristo" (Euphorbia miliivarhislopii, syn E.splendens) é um potente moluscicidavegetal. Neste sentido, são necessários estudos toxicológicosque visemavaliar possíveis riscos à saúdeassociados ao uso em larga escala desta espécie em áreas endêmicas para esquistossomose. O presente estudo é parte deuma avaliação mais abrangentesobre o potencial tóxico destemoluscicida. Foram investigados in vitro osefeitos dolátex da E.miliisobre a proliferação delinfócitoshumanos. O látexliofilizado (0; 0,5;5;25 e 50 µg/ml)foi incubado comsangue totalna presençade agentes mitogênicos, tais como lectinas(fitohemaglutinina, concanavalina Ae pokeweed), anticorpomonoclonalhumano anti-CD3etoxóide tetânico. A proliferação celularfoi quantificada atravésincorporaçãode ³H-timidina eos efeitos do látexnaproliferação celular induzida por agentes mitogênicosforam comparados comos efeitos de10 ng/mlde12-O-tetradecanoilforbol-13-acetato (TPA). Os resultados demonstram quea proliferação celular induzida poragentes mitogênicosfoimarcadamenteaumentada na presença do látex daE.milii.Oefeito sinérgico observado pode ser devidoà presença deésteres de forbol, como o TPA, e/oude lectinas com ação mitogênica presentes nesta espécie vegetal.

Evaluation of antioxidant and antimicrobial activities of essential oils from Carum copticum seed and Ferula assafoetida latex.

UNLABELLED: Carum copticum and Ferula assafoetida have several medicinal properties including antispasmodic, carminative, sedative, analgesic, and antiseptic. Reactive oxygen species (ROS), reactive nitrogen species (RNS), hydrogen peroxide (H(2) O(2) ), and thiobarbituric acid reactive substances (TBARS) scavenging activities of Carum and Ferula oils along with their antibacterial and antifungal activities were examined. Thymol (40.25%), γ-terpinene (38.7%) and p-cymene (15.8%) were detected as the main components of Carum oil while, ß-pinene (47.1%), α-pinene (21.36%), and 1, 2-dithiolane (18.6%) were the main components of Ferula oil. Inhibitory concentrations (IC50) for total radical scavenging were between 40 and 60 and 130 and 160 µg/mL of Carum and Ferula oil, respectively. Minimal inhibitory concentration (MIC) for Salmonella typhi, Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Aspergillus niger, and Candida albicans were 78 ± 8, 65 ± 7, 14 ± 3, 5 ± 2, 5.6 ± 1.3, and 8.8 ± 2.2 µg/mL of Carum oil, respectively. MIC for S. typhi, E. coli, S. aureus, B. subtilis, A. niger, and C. albicans were >200, >200, 125 ± 17, 80 ± 12, 85 ± 5, and 90 ± 11 µg/mL of Ferula oil, respectively. Accordingly, Carum and Ferula oils could be used as safe and effective natural antioxidants to improve the oxidative stability of fatty foods during storage and to preserve foods against food burn pathogens. PRACTICAL APPLICATION: This study clearly demonstrates the potential of Carum and Ferula oil especially Carum oil as natural antioxidant and antimicrobial agent. The chemical composition of essential oils was identified. Thus, identification of such compounds also helps to discover of new antioxidant, antibacterial and antifungal agents for potential applications in food safety and food preservation.

Identification of minor secondary metabolites from the latex of Croton lechleri (Muell-Arg) and evaluation of their antioxidant activity.

Dragon's blood (Sangre de drago), a viscous red sap derived from Croton lechleri Muell-Arg (Euphorbiaceae), is extensively used by indigenous cultures of the Amazonian basin for its wound healing properties. The aim of this study was to identify the minor secondary metabolites and test the antioxidant activity of this sustance. A bioguided fractionation of the n-hexane, chloroform, n-butanol, and aqueous extracts led to the isolation of 15 compounds: three megastigmanes, four flavan-3-ols, three phenylpropanoids, three lignans, a clerodane, and the alkaloid taspine. In addition to these known molecules, six compounds were isolated and identified for the first time in the latex: blumenol B, blumenol C, 4,5-dihydroblumenol A, erythro-guaiacyl-glyceryl-beta-O-4'- dihydroconiferyl ether, 2-[4-(3-hydroxypropyl)-2-methoxyphenoxy]-propane-1,3-diol and floribundic acid glucoside. Combinations of spectroscopic methods ((1)H-, (13)C- NMR and 2D-NMR experiments), ESI-MS, and literature comparisons were used for compound identification. In vitro antioxidant activities were assessed by DPPH, total antioxidant capacity and lipid peroxidation assays. Flavan-3-ols derivatives (as major phenolic compounds in the latex) exhibited the highest antioxidant activity.

Anti-inflammatory and analgesic activities of the latex containing triterpenes from Himatanthus sucuuba.

Some triterpenes and iridoids were previously isolated from the stem bark of Himatanthus sucuuba. The latex from Himatanthus sucuuba is used in popular amazonian medicine as an anti-inflammatory remedy. Fractions of the latex were pharmacologically evaluated with a view to verify this popular use in the carrageenan-induced rat paw edema and in the acetic acid-induced mouse constriction tests. The hexane fraction inhibited the edema formation by 35.9% at a dose of 200 mg/kg (p.o.) but no activity was observed at 100 mg/kg (p.o.). The triterpenes present in the hexane fraction were identified as lupeol acetate, alpha-amyrin and lupeol cinnamates. The fraction containing only cinnamates inhibited the edema and the abdominal constrictions by 50-40% and 57.9%, respectively, at 100 mg/kg (p.o.). Among all the fractions studied, the fraction containing only cinnamates showed the greatest anti-inflammatory activity which suggests that these compounds were responsible for the previously described activity of the crude extract.

Extraction and purification of curcain, a protease from the latex of Jatropha curcas Linn.

A proteolytic enzyme, curcain, has been extracted from the latex of Jatropha curcas Linn. The enzyme was purified by chromatography on carboxymethyl cellulose and gel filtration on Sephadex G-200. The homogeneity of protein associated with curcain was established by non-denatured polyacrylamide gel electrophoresis using a discontinuous buffer system. The molecular weight of curcain was estimated by Sephadex G-100 gel filtration using a calibration curve of standard proteins to be around 22,000 daltons.

Dopamine biosynthesis at different stages of plant development in Papaver somniferum.

Latex and cell-free extracts of various organs and stages of plant and capsule development in Papaver somniferum, the opium poppy, synthesized dopamine, an alkaloid precursor, from 14C-dopa. The 1000 g X 30 min supernatant from latex of the pedicel-capsule junction converted more dopa than latex supernatant from the upper capsule or lower pedicel regions, although there was more protein in the latex from the capsule. Percent conversion of pedicel-capsule latex into dopamine was maximum in unopened flower buds and decreased within 14 days after flowering. Dopamine biosynthesis in latex and cell-free extracts also varied with the stage of organ development. Extracts from capsule tissue converted more labeled dopa into dopamine than did extracts from pedicels, leaves from vegetative plants at the rosette stage, leaves from flowering plants, or pedicels connected to capsules.