Environmental sustainability assessment of biodiesel production from Jatropha curcas L. seeds oil in Pakistan.

According to IPCC Annual Report (AR-5), environmental impact assessment of any product prototype is recommended before its large-scale commercialization; however, no environmental profile analysis of any biodiesel prototype has been conducted in Pakistan. Therefore, objective of this study was to conduct a comprehensive life cycle assessment (LCA), water footprint and cumulative energy demand (CED) of biodiesel production from Jatropha curcas L. (JC) seeds oil in Pakistan. A cradle-to-gate LCA approach was applied for 400 liter (L) JC biodiesel produced in Pakistan. JC biodiesel production chain was divided into three stages i.e., 1). cultivation of JC crop 2). crude oil extraction from JC seeds and 3). crude oil conversion to biodiesel. Primary data for all the stages were acquired through questionnaire surveys, field visits and measurements in the field. Potential environmental impacts were calculated in SimaPro v.9.2 software using Eco-indicator 99 methodology. Results showed that crude oil extraction stage accounted for highest emissions (77%) to the overall environmental impact categories evaluated, followed by oil conversion stage (21%) and JC cultivation stage (02%), respectively. The three stages of JC biodiesel production chain are major contributor to ecotoxicity with a contribution of 57% to this impact category. Higher contribution to ecotoxicity was due to agrochemicals used in the JC cultivation. Similarly, fossil fuels impact category was responsible for 38% of overall environmental impacts. In addition, water footprint of JC biodiesel production chain was 2632.54 m3/reference unit. Cumulative energy required for 400L JC biodiesel production chain was 46745.70 MJ in Pakistan. Fossil diesel consumption, synthetic fertilizers use and purchased electricity were major hotspot sources to environmental burdens caused by JC biodiesel production in Pakistan. By performing sensitivity analysis at 20% reduction of the baseline values of fossil diesel used, synthetic fertilizers and purchased electricity, a marked decrease in environmental footprint was observed. It is highly recommended that use of renewable energy instead of fossil energy would provide environmental benefits such as lower greenhouse gases and other toxic emissions as compared to conventional petroleum fuels. It is also recommended that JC as a biofuel plant, has been reported to have many desired characteristics such as quick growth, easy cultivation, drought resistance, pest and insect resistance, and mainly great oil content in JC seeds (27-40%). Therefore, JC plant is highly recommended to Billion Tree Afforestation Project (BTAP) for plantation on wasteland because it has multipurpose benefits.

Structural Distinctive 26SK, a Ribosome-Inactivating Protein from Jatropha curcas and Its Biological Activities.

Ribosome-inactivating proteins (RIPs) are a group of proteins exhibiting N-glycosidase activity leading to an inactivation of protein synthesis. Thirteen predicted Jatropha curcas RIP sequences could be grouped into RIP types 1 or 2. The expression of the RIP genes was detected in seed kernels, seed coats, and leaves. The full-length cDNA of two RIP genes (26SK and 34.7(A)SK) were cloned and studied. The 34.7(A)SK protein was successfully expressed in the host cells while it was difficult to produce even only a small amount of the 26SK protein. Therefore, the crude proteins were used from E. coli expressing 26SK and 34.7(A)SK constructs and they showed RIP activity. Only the cell lysate from 26SK could inhibit the growth of E. coli. In addition, the crude protein extracted from 26SK expressing cells displayed the effect on the growth of MDA-MB-231, a human breast cancer cell line. Based on in silico analysis, all 13 J. curcas RIPs contained RNA and ribosomal P2 stalk protein binding sites; however, the C-terminal region of the P2 stalk binding site was lacking in the 26SK structure. In addition, an amphipathic distribution between positive and negative potential was observed only in the 26SK protein, similar to that found in the anti-microbial peptide. These findings suggested that this 26SK protein structure might have contributed to its toxicity, suggesting potential uses against pathogenic bacteria in the future.

A comparative appraisal of three important oil yielding plants for their biodiesel potential.

In the present scenario, alternative energy sources are required to achieve the future economic prosperity where shortage of fossil fuels will be a limiting factor and hamper the global economic growth. Therefore, interest in biofuel is increasing continuously. The best way of sustainable development is fossil fuel supplementation with biodiesel to reduce the fossil fuel demand. Biodiesel is a clean burning, ester-based, oxygenated fuel derived from natural and renewable sources. Till now, majority of the people have worked on the biodiesel derived from edible oil. Instead of using edible oil, non-edible oil needs to be explored as feedstock for biofuel because half of the world's population is unable to afford the food oil as feedstock for fuel production. Looking at the significance of biodiesel and the resources of biofuel, in this paper, a comparative exhaustive study has been reported with for three important plants, namely Jatropha curcas, Pongemia pinnata and Balanites aegyptiaca. These plants were selected based on their biodiesel potential, availability, cultivation practices and general information available. The present study involves scientometric publications, comparison of fatty acid composition and biodiesel parameters. We have also compared climatic conditions for the growth of the plants, economic feasibility of biodiesel production and other ecological services. The study paves a way for sustainable solution to policy makers and foresters looking for selection of plant species as bioenergy resource.

Phytochemical prospecting, isolation, and protective effect of the ethanolic extract of the leaves of Jatropha mollissima (Pohl) Baill.

Jatropha mollissima is used in folk medicine as antimicrobial, antiparasitic, and larvicidal. However, few toxicogenetic studies have been carried out. Therefore, the aim of this study was to determine the phytochemical profile of ethanolic leaf extract of J. mollissima (EEJM) as well as potential cytotoxic, mutagenic, and antimutagenic properties. The EEJM was subjected to successive fractionation for the isolation of secondary metabolites, and five concentrations (0.01; 0.1; 1; 10 and 100 mg/ml) of extract were investigated using Allium cepa assay and the Somatic Mutation and Recombination (SMART) test. The mitotic index and % damage reduction were analyzed for A. cepa and the frequency of mutant hair for SMART. The presence of coumarins, alkaloids, flavonoids, saponins, and tannins was detected, while spinasterol and n-triacontane were the isolates identified for the first time for this species. EEJM did not exhibit cytotoxicity and was not mutagenic at 1 or 10 mg/ml using A. cepa and all concentrations of EEJM were not mutagenic in the SMART test. A cytoprotective effect was found at all concentrations. At 1 or 10 mg/ml EEJM exhibited antimutagenicity in A. cepa. In SMART, the protective effect was observed at 0.1 to 100 mg/ml EEJM. Our results demonstrate the important chemopreventive activity of EEJM, a desired quality in the search for natural anticarcinogenic compounds.

Evaluation of tissue repair using phytotherapeutic gel from Plectranthus neochilus, Schlechter (boldo-gambá) and Cnidoscolus quercifolius Pohl (favela) in Wistar rats / [Avaliação da reparação tecidual do gel fitoterápico de Plectranthus neochilus, Schlechter (Boldo-gambá) e Cnidoscolus quercifolius Pohl (Favela) em ratos Wistar]

The objective of this study was to investigate the effects of a gel formulation from the association of Plectranthus neochilus and Cnidoscolus quercifolius on tissue repair in cutaneous wounds in rats. A surgical wound was induced in 35 Wistar rats and treated according to group: G1 - commercial phytotherapeutic gel; G2 - Carbopol gel 1%; G3, 4, 5 - gel formulation from Boldo-gambá and Favela (FGBF) at 2.5%, 5%, and 10%, respectively. 1ml of the product was applied topically daily, for 14 days. Macroscopic evaluation of the wound showed inflammation, granulation, and epithelization in all groups. The FGBF 2.5% group showed greater angiogenic potential. There was a significant difference between the surgical area of the wounds treated with FGBF 2.5%, 5%, or 10% compared to the group with the commercial phytotherapeutic gel. On histomorphometry of the skin, there were reepithelization of the epidermis and superficial dermis, longitudinal collagen fibers, fibroblasts, and blood vessels, and in the deeper dermis, fibroblasts, transverse and longitudinal collagen fibers, blood vessels, and inflammatory cells. The 2.5% formulation had the greatest increase in fibroblast proliferation and most intense collagenization on day 14 of treatment.(AU)

Este trabalho objetivou investigar os efeitos de uma formulação em gel da associação da Plectranthus neochilus e da Cnidoscolus quercifolius no processo de reparação tecidual em feridas cutâneas de ratos. Foi induzida uma ferida cirúrgica em 35 ratos Wistar, sendo tratadas de acordo com os grupos: G1 - fitoterápico comercial; G2 - gel de carbopol 1%; G3, G4 e G5 - formulação gel boldo-gambá mais favela (FGBF) 2,5%, 5% e 10%, respectivamente. Aplicou-se 1mL do produto, via tópica, diariamente, durante 14 dias. Na avaliação macroscópica das feridas, verificou-se inflamação, granulação e epitelização em todos os grupos. O grupo FGBF 2,5% apresentou maior potencial angiogênico. Houve diferença significativa entre as áreas cirúrgicas das feridas tratadas com os FGBF 2,5%, 5% ou 10%, comparados ao grupo com o gel fitoterápico comercial. Na histomorfometria da pele, observou-se reepitelização da epiderme e da derme superficial, fibras colágenas longitudinais, fibroblastos e vasos sanguíneos e, na derme profunda, fibroblastos, fibras colágenas transversais e longitudinais, vasos sanguíneos e células inflamatórias. A formulação a 2,5% teve o maior aumento na proliferação de fibroblastos e mais intensa colagenização no dia 14 de tratamento.(AU)

Triterpenoids isolated from Jatropha macrantha (Müll. Arg.) inhibit the NF-κB and HIF-1α pathways in tumour cells.

Activity-guided fractionations of Jatropha macrantha Müll. Arg. led to the isolation of pomolic acid (1) and euscaphic acid (2). The potential for inhibition against NF-κB and HIF-1α production of these two compounds was tested in different tumour cell lines. Compounds 1 and 2 showed an inhibitory activity of HIF-1α in the SK-MEL-28 (IC50=3.01 ± 0.02 µM and 3.78 ± 0.02 µM), A549 (IC50=9.97 ± 0.01 µM and 10.25 ± 0.01 µM) and U-373 MG (IC50=6.34 ± 0.02 µM and 8.85 ± 0.02 µM) cell lines. In addition, compounds 1 and 2 showed an inhibitory activity on NF-κB in SK-MEL-28 (IC50=1.05 ± 0.02 µM and 2.71 ± 0.01 µM), A549 (IC50=3.63 ± 0.01 µM and 3.73 ± 0.02 µM) and U-373 MG (IC50=2.55 ± 0.02 µM and 3.39 ± 0.01 µM) cell lines. This is the first report that isolates these compounds from J. macrantha and tests their antitumor potential.

Curcusone C induces apoptosis in endometrial cancer cells via mitochondria-dependent apoptotic and ERK pathway.

OBJECTIVE: Jatropha curcashas been used in traditional medicine in Africa to treat cancer for thousands of years. This study aimed to examine the anti-endometrial cancer effect of Curcusone C, a naturally occurring rhamnofolane diterpene, isolated from J. curcas and reveal its molecular mechanism of action. RESULTS: Curcusone C treatment caused significant anti-proliferative and apoptotic effects in human endometrial cancer (EC) Ishikawa and HEC-1A cells in a dose-dependent manner. Exposure of EC cells to Curcusone C resulted in apoptosis, which was associated with cytochrome c release, caspase-3 and caspase-9 activation, Bcl-xL/Bax dysregulation, and decreased expression of inhibitors of apoptosis proteins, such as XIAP and survivin. The inhibitory effect induced by Curcusone C was greatly impaired by the overexpression of survivin or Bax-/- MEFs or the knockdown of Bim expression. Moreover, Curcusone C activated mitogen-activated protein kinases, and the ERK inhibitor U0126 significantly attenuated the growth-inhibitory and apoptotic effects of Curcusone C in Ishikawa cells. CONCLUSION: Taken together, the results demonstrate the anti-endometrial cancer potential of Curcusone C for the treatment of endometrial cancer.

Towards Thermally Reversible Networks Based on Furan-Functionalization of Jatropha Oil.

A novel biobased monomer for the preparation of thermally reversible networks based on the Diels-Alder reaction was synthesized from jatropha oil. The oil was epoxidized and subsequently reacted with furfurylamine to attach furan groups via an epoxide ring opening reaction. However, furfurylamine also reacted with the ester groups of the triglycerides via aminolysis, thus resulting in short-chain molecules that ultimately yielded brittle thermally reversible polymers upon cross-linking via a Diels-Alder reaction. A full-factorial experimental design was used in finding the optimum conditions to minimize ester aminolysis and to maximize the epoxide ring opening reaction as well as the number of furans attached to the modified oil. The optimum conditions were determined experimentally and were found to be 80 °C, 24 h, 1:1 molar ratio, with 50 mol % of LiBr with respect to the modified oil, resulting in 35% of ester conversion, 99% of epoxide conversion, and an average of 1.32 furans/triglyceride. Ultimately, further optimization by a statistical approach led to an average of 2.19 furans per triglyceride, which eventually yielded a flexible network upon cross-linking via a Diels-Alder reaction instead of the brittle one obtained when the furan-functionalization reaction was not optimized.

Chemical profile and gastroprotective effect of Jatropha elliptica (Pohl) Oken roots.

Jatropha elliptica (Pohl) Oken (Euphorbiaceae) roots are used in folk medicine to treat gastric ulcers. The purpose of this work was to evaluate the gastroprotective activity of ethanol extract (JER) and hexane fraction (ERH) of J. elliptica roots in mice, as well as to analyze the acute toxicity of the extract and identify the potential active compounds. No signs of toxicity were observed in JER. In both acidified ethanol and indometacin-induced gastric ulcer models, all doses tested of JER and ERH significantly reduced gastric lesions. Dereplication of JER was performed by HPLC-DAD-ESI-MS/MS and resulted in the annotation of compounds fraxetin, propacin, jatrophone and jatropholones A and B. GC-MS analysis of ERH revealed the diterpenes jatrophone, jatropholone A and jatropholone B as the major components. The chemical study of this fraction has led to the isolation of these compounds, in addition to the sequiterpene cyperenoic acid and the diterpene 2ß-hydroxyjatrophone, both reported for the first time in J. elliptica. The isolated compounds were tested against L929 cells and only cyperenoic acid and the mixture of jatropholones A and B did not show toxicity, being then selected as good candidates for bioassays using acidified ethanol-induced gastric ulcer model. Cyperenoic acid significantly decreased gastric lesions and preserved gastric mucus layer. The mixture of jatropholones A and B caused a smaller reduction of gastric lesions, without preservation of the gastric mucus layer. The study showed that J. elliptica roots present gastroprotective activity in mice, without causing acute toxic effects. The activity is related, at least in part, to the occurrence of terpenes, mainly the sesquiterpene cyperenoic acid.

Hydroxy-octadecenoic acids instead of phorbol esters are responsible for the Jatropha curcas kernel cake's toxicity.

The toxic kernel cake of Jatropha curcas (KCakeJ) is an emerging health and environmental concern. Although phorbol esters are widely recognized as the major toxin of KCakeJ, convincing evidence is absent. Here, we show that rather than phorbol esters an isomeric mixture of 11-hydroxy-9E-octadecenoic acid, 12-hydroxy-10E-octadecenoic acid and 12-hydroxy-10Z-octadecenoic acid (hydroxy-octadecenoic acids, molecular formula C18H34O3) is the major toxic component. The toxicities of hydroxy-octadecenoic acids on experimental animals, e.g. acute lethality, causing inflammation, pulmonary hemorrhage and thrombi, allergies, diarrhea and abortion, are consistent with those on human/animals caused by Jatropha seed and/or KCakeJ. The hydroxyl group and the double bond are essential for hydroxy-octadecenoic acids' toxicity. The main pathway of the toxicity mechanism includes down-regulating UCP3 gene expression, promoting ROS production, thus activating CD62P expression (platelet activation) and mast cell degranulation. The identification of the major toxin of KCakeJ lays a foundation for establishing an environmentally friendly Jatropha biofuel industry.

HPLC Method Validation for Jatropha dioica Extracts Analysis.

Jatropha dioica is a popular plant used in Mexican herbal medicine to treat several diseases. Cytotoxicity, antimicrobial and antiviral activities have been reported for root extracts, while riolozatrione, 6-epi-riolozatrione, citlalitrione and jatrophatrione, among others, have been identified as the principal components. In this work, an HPLC/DAD method for the analysis of riolozatrione and other major compounds in extracts of different polarities was validated. The analysis was carried out on an AccQ-Tag column with a water-acetonitrile mixture as mobile phase. Flow rate was 0.2 mL/min, and the separation was carried out in gradient mode with UV detection set at 254 nm. The resulting method showed good reproducibility in both retention times and peak areas of riolozatrione, 6-epi-riolozatrione, citlalitrione and jatrophatrione, with relative standard variations lower than 4.5 and 10.5% respectively. In addition, this method provides a good performance for riolozatrione quantitation, with recoveries between 102 and 108% and RSDs lower than 2.5%. The polarity of the extracting solvent did not affect the performance of the chromatographic method. The developed method was applied for the analysis and quantification of riolozatrione in extracts of Jatropha dioica collected in several seasonal stages and years (2014-2017).

Screening and characterization of medicinal plants extracts with bactericidal activity against Streptococcus mutans.

The aim of the present study was to perform a screening of extracts obtained from 15 medicinal plants using water (at 25 and 90 °C) or ethanol (at 25 °C), to bactericidal activity against cariogenic S. mutans ATCC 25175, as well as to carry out the preliminary phytochemical characterization of the extracts and HPLC/MS assay for selected extracts. The extractions were carried out for 5 h at 400 rpm. Only five from 45 tested extracts were selected based on their antibacterial activity. The IC50 varied from of 28 ppm for Quercus ilex up to 250 ppm for Jatropha dioica. Different polyphenolic and quinic acids, flavonoids, anthocyanin or tyrosol were identified by HPLC/MS in selected extracts from Rosa gallica L., Jatropha dioica Sessé, Mimosa tenuiflora (Willd.) Poir, Quercus ilex L., and Solanum nigrum. The obtained results confirm that selected extracts are good candidates to be used for cariogenic bacteria control.

Jatrofolianes A and B: Two Highly Modified Lathyrane Diterpenoids from Jatropha gossypiifolia.

Jatrofolianes A (1) and B (2), which are two highly modified lathyrane diterpenoids, were isolated from Jatropha gossypiifolia. 1 incorporates an unusual transannular 1,3-dioxolane moiety, forming a unique 5/6/5/8/3 ring system, while 2 possesses a new 10,11:13,14-diseco-lathyrane skeleton with a 12-membered macrocyclic lactone ring. Their structures were determined by spectroscopic analysis, quantum chemical calculations, and single-crystal X-ray diffraction. 2 showed significant multidrug resistance (MDR) reversal activity to cancer cells HepG2/ADR and HCT-15/5-FU at 10 µM.

Contenido de oligoelementos y factores antinutricionales de hojas comestibles nativas de Mesoamérica / Content of trace elements and antinutritional factors in edible leaves native to Mesoamerica

Los oligoelementos son importantes constituyentes nutricionales de las hierbas comestibles. Se colectaron 11 especies, nueve nativas (Amaranthus hybridus, Cnidoscolus aconitifolius, Crotalaria longirostrata, Dysphania ambrosioides, Lycianthes synanthera, Sechium edule, Solanum americanum, S. nigrescens, S. wendlandii) y dos introducidas (Moringa oleifera, Spinacea oleracea) en dos regiones de Guatemala. Se prepararon muestras de la hierba seca, cocida y de caldo de hierba fresca. Se cuantificaron por espectrofotometría de absorción atómica los macro (N, P, K) y oligoelementos (Ca, Mg, Na, Cu, Zn, Mn), taninos por espectrofotometría y oxalatos por permanganimetría. El contenido de oligoelementos es diverso, para Zn, la hierba control S. oleracea contienen buena cantidad (90-140 ppm); de las nativas D. ambrosioides (130-160 ppm) y A. hybridus (70-80 ppm) tienen la mayor cantidad. Respecto a Fe las hierbas control tienen buena composición (S. oleracea, 220-280 ppm y M. oleifera, 105-135 ppm); de las nativas A. hybridus (90-240 ppm), C. aconitifolius (75-185 ppm) y L. synanthera (75-140 ppm) tienen buenas concentraciones. Se encontraron niveles elevados de oxalatos en S. oleracea (67.30 (5.51) mg/g), L. synanthera (56.30 (9.67) mg/g) y S. nigrescens (33.6 (5.48) mg/g); en las demás hierbas se encontraron niveles menores. Los niveles de taninos fueron bajos (0.1-0.8 mg/g) para todas las especies. Se demuestra que cuatro especies nativas tienen un buen contenido de oligoelementos y presentan valores menores de antinutricionales que los controles.

Trace elements are important nutritional constituents from edible herbs. Eleven species were collected in two regions of Guatemala, nine native (Amaranthus hybridus, Cnidoscolus aconitifolius, Crotalaria longirostrata, Dysphania ambrosioides, Lycianthes synanthera, Sechium edule, Solanum americanum, S. nigrescens, S. wendlandii) and two introduced (Moringa oleifera, Spinacea oleracea). Dry, cooked and broth samples were prepared. By atomic absorption spectrometry, macro (N, P, K) and trace elements (Ca, Mg, Na, Cu, Zn, Mn) were quantified, tannins by spectrophotometry, and oxalates by permanganometry. Trace elements content is diverse, for Zn, control herb S. oleracea contained high quantity (90-140 ppm); from the natives D. ambrosioides (130-160 ppm) and A. hybridus (70-80 ppm) contained high amounts. For Fe, control herbs had high composition (S. oleracea, 220-280 ppm, M. oleifera, 105-135 ppm); from the natives A. hybridus (90-240 ppm), C. aconitifolius (75-185 ppm) and L. synanthera (75-140 ppm) had the highest amounts. High levels of oxalates were demonstrated in S. oleracea (67.30 (5.48) mg/g), L. synanthera (56.30 (9.67) mg/g), and S. nigrescens (33.6 (5.48) mg/g); from the others levels.

A sensitive refining of in vitro and in vivo toxicological behavior of green synthesized ZnO nanoparticles from the shells of Jatropha curcas for multifunctional biomaterials development.

ZnO nanoparticles (NPs) possess a wide range of biological functions in pharmaceutical and cosmetic applications due to their excellent antimicrobial, optical and UV protective properties. This study first reports the toxicological assessment of ZnO NPs green synthesized from Jatropha curcas shells for multifunctional biomedical applications. The hot water extract of J.curcas shells is utilized as a chelating agent for the reduction of zinc acetate and then, the prepared ZnO NPs are broadly characterized using X-ray spectroscopic and electron microscopic observations. The prepared ZnO NPs acquire high purity (100%) wurtzite crystal with hexagonal structure with the average particle size of 53 nm. In vitro and in vivo toxicity evaluation against human tumor cell lines and zebrafish embryos have ascertained the purpose of ZnO NPs in clinical research. Toxic effects of ZnO NPs were observed by a dose-dependent reduction of bacterial growth at ≥1   µg ml-1, by teratogenicity and genotoxicity in zebrafish embryos (from 3 to 90 µg ml-1) and by a significant nanoparticle uptake (0.5 ng µl-1) by a fish serum. In contrast, ZnO NPs fail to reduce the proliferation of human bladder tumor cells (UC6) and cell viability of A549 cells in vitro up to 500 µg ml-1. All these observations limit the unobstructed application of ZnO NPs at higher concentrations. Thus, abundantly used metal oxide nanoparticles like ZnO NPs examined in our present study in different animal models under in vitro and in vivo conditions will be the significant screening strategy to determine the nanotoxicity.

Lab-made solid phase microextraction phases for off line extraction and direct mass spectrometry analysis: Evaluating the extraction parameters.

The analyses of drugs and metabolites in complex matrices have been widely studied in recent years. However, due to high levels endogenous compounds and matrix complexity, these analyses require a sample pre-treatment step. To this aim, two lab-made extractive phases were integrated to probe electrospray ionization mass spectrometry (PESI-MS) technique for direct analysis of illicit drugs in biological fluids and phorbol esters in Jatropha curcas extract. The polypyrrole (PPy) phase was electropolymerized onto a platinum wire surface by cyclic voltammetry. The molecularly imprinted polymer (MIP) was synthesized and adhered onto a stainless-steel needle with epoxy resin. The PPy-PESI-MS method showed to be linear in a concentration range from 1 to 500 µg L-1, with accuracy values between -2.1 and 14%, and precision values between 0.8 and 10.8%. The MIP-PESI-MS method showed to be linear in a concentration range from 0.9 to 30 mg L-1, with accuracy values between -1.6 and -15.3%, and precision values between 4.1 and 13.5%.

Studies of Jatrogossone A as a Reactive Oxygen Species Inducer in Cancer Cellular Models.

Natural products continue to provide a platform to study biological systems. A bioguided study of cancer cell models led us to a new member of the jatrophane natural products from Jatropha gossypiifolia, which was independently identified and characterized as jatrogossone A (1). Purification and structure elucidation was performed by column chromatography and high-performance liquid chromatography-mass spectrometry and NMR techniques, and the structure was confirmed via X-ray crystallography. The unique molecular scaffold of jatrogossone A prompted an evaluation of its mode of action. Cytotoxicity assays demonstrated that jatrogossone A displays selective antiproliferative activity against cancer cell models in the low micromolar range with a therapeutic window. Jatrogossone A (1) affects mitochondrial membrane potential (ΔΨm) in a time- and dose-dependent manner. This natural product induces radical oxygen species (ROS) selectively in cancer cellular models, with minimal ROS induction in noncancerous cells. Compound 1 induces ROS in the mitochondria, as determined by colocalization studies, and it induces mitophagy. It promotes also in vitro cell death by causing cell arrest at the G2/M stage, caspase (3/7) activation, and PARP-1 cleavage. The combined findings provide a potential mechanism by which 1 relies on upregulation of mitochondrial ROS to potentiate cytotoxic effects through intracellular signaling.

Anti-melanin deposition activity and active constituents of Jatropha multifida stems.

Jatropha multifida is a medicinal plant that belongs to the Euphorbiaceae family. Our investigation revealed that the chloroform extract of J. multifida stems showed anti-melanin deposition activity against α-melanocyte stimulating hormone (α-MSH)- and 3-isobutyl-1-methylxanthine (IBMX)-induced melanogenesis in the mouse melanoma cell line (B16-F10). Further fractionation and purification of the major constituents led to the isolation of two coumarins (1 and 2) and seven known lignoids (3-9). All isolated compounds exhibited anti-melanin deposition activities against the mouse melanoma cell line (B16-F10) with IC50 values ranging from 37.5 to 560.1 µM, without any cytotoxicity even at high concentrations, except for 8. Further mechanistic studies suggested that 9 downregulated tyrosinase mRNA expression, while the anti-melanin deposition activities of 4 and 8 appeared to be unrelated to tyrosinase inhibition and the downregulated expression of the key melanogenesis-associated mRNAs. These results suggested that J. multifida could possess potent skin whitening ingredients.

Jatrogricaine A: a new diterpenoid with a 5/6/6/4 carbon ring system from the stems of Jatropha podagrica.

Jatrogricaine A (1), a new diterpenoid possessing a 5/6/6/4 carbon ring system, together with eight known diterpenoids (2-9) were isolated from the stems of Jatropha podagrica. Their structures were elucidated by extensive spectroscopic methods and the absolute configuration of 1 was determined by single crystal X-ray diffraction analysis. All compounds were evaluated for their anti-inflammatory activities in vitro, and compound 3 showed significant inhibitory effects against nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW264.7 macrophage cells with an IC50 of 13.44 ± 0.28 µmol·L-1, being comparable to the positive control, quercetin (IC50 17.00 ± 2.10 µmol·L-1).

Plant latex thrombin-like cysteine proteases alleviates bleeding by bypassing factor VIII in murine model.

Hemostasis is a tightly regulated process which maintains a fluid state of blood within the vasculature and provides thrombotic response upon tissue injury. Various scientific studies have implicated the role of plant latex proteases in hemostasis using in vitro experiments. However, in vivo models substantiate their role in hemostasis. Therefore, in the present study, the effect of plant latex thrombin-like proteases (PTLPs) on hemostasis was investigated systematically using mice tail bleeding as a preclinical model. In this direction, latex protease fractions (LPFs), which showed potent thrombin-like activity, were selected as they act directly on fibrinogen to form clot and quickly stop bleeding. Thrombin-like activity was exhibited mainly by cysteine proteases. Calotropis gigantea, Carica papaya, Jatropha curcas, Oxystelma esculentum, Tabernaemontana divaricata, and Vallaris solanacea LPFs and papain from C. papaya latex significantly reduced bleeding on a topical application in normal and aspirin administered mice. In addition, PTLPs accelerated the clotting of factor VIII deficient plasma, while, papain brought back the clotting time to normal levels acting like a bypassing agent. Further, papain failed to show activity in the presence of specific cysteine protease inhibitor iodoacetic acid; confirming protease role in all the activities exhibited. At the tested dose, PTLPs except C. gigantea did not show toxicity. Further, structural and sequence comparison between PTLPs and human thrombin revealed structural and sequence dissimilarity indicating their unique nature. The findings of the present study may open up a new avenue for considering PTLPs including papain in the treatment of bleeding wounds.