Crude Plant Extracts Research Articles

Terminalia arjuna Leaf Gall: The Possible Treatment for Sickle Cell Anaemia

Objective: To check the presence of various secondary metabolites produced in the plant under study Terminalia arjuna (Roxb.) Wight and Arn. post insect attack and gall formation.
 Study Design: Terminalia arjuna (Roxb.) Wight and Arn, a well known plant involved in treating heart ailments in the Ayurvedic system of medicine for centuries. The plant’s astringent bark exhibits cardioprotective properties and is commonly used in the treatment of hypertension, angina and coronary artery disease. T. arjuna plant suffers from gall development in its leaves due to the attack of an hemipteran insect Trioza fletcheri minor. Insect attack acts as a stress stimulus for the plant to produce various bio-active compounds as a part of their defence mechanism. The present study was intended for the identification of various compounds present in the crude extract obtained from the healthy and galled leaves of Terminalia arjuna.
 Place and Duration of Study: Experiments performed in the Plant pathology and tissue culture laboratory, Department of Botany University of Rajasthan, Jaipur, between April 2018 to March 2019.
 Methodology: Various methods are employed for the identification of possible bio-active compounds present in the plant crude extract, GC-MS (Gas Chromatography- Mass Spectroscopy) is one such method. To check the presence of phytochemicals produced post insect attack and infection in T. arjuna normal and galled leaves, Gas Chromatography-Mass Spectroscopy (GC-MS) of the methanolic extract was conducted. The normal healthy leaves and the galled leaves were collected, shade dried, pulverized and then soxhlet extracted for GC-MS analysis.
 Results: GC-MS analysis of methanolic extract of Terminalia arjuna leaves and galls confess the presence of 57 compounds in the gall extract while 21 compounds in normal (uninfected) leaf extract. The gall extract revealed the presence of various novel and medicinally important bio-active compounds, one of them was 5-Hydroxy Methyl Furfural (5-HMF). 
 Conclusion: 5-HMF, an aromatic aldehyde is an economically important chemical compound and well-known for its wide application in various fields. It is an efficient anti-sickling agent and has undergone pre-clinical testing as a potential treatment for the fatal sickle cell disease. Qualitative phytochemical screening revealed the presence of various bio-active compounds both in leaf and gall methanolic extract of Terminalia arjuna, thus proving the plant’s pharmaceutical importance. This study result will make a way for further research in plant’s pharmacological research.

Anthelmintic Activities of Polypodium Decumanum Leaf-Extracts

Various herbal plants used in traditional medicines contain a vast array of substances that can be used to treat various disease and complications, the present study deals with anthelmintic activity of different extracts of leaves of Polypodium decumanum, a potential medicinal plant. The crude plant extracts of leaf part exhibited anthelmintic activity. Two concentrations (30mg/ml and 40mg/ml) of each extract were studied, which involved for the determination of time of paralysis and time of death of the test worms. It was found that ethyl acetate extracts exhibited significant anthelmintic activity followed by ethanolic extract and aqueous extract in causing paralysis and death of earth worms.

Preparation of a pinoresinol diglucoside imprinted polymer using metal organic frame as the matrix for extracting target compound from Eucommia ulmoides

ABSTRACT Fabrication of a post-synthesis imprint modification method for metal-organic frameworks in order to improve its selectivity in the field of separation and analysis is of great interest. For this purpose, a pinoresinol diglucoside (PDG) imprinted polymer was attempted to graft on the surface of metal-organic frameworks MIL-101 to produce a new composite material named MIPs@ED-MIL-101. This modification procedure was monitored by using X-Ray Diffraction, fourier transforms infrared spectroscopy and scanning electron microscopy in the crystal structure, chemical groups and particles topography, respectively. Adsorption kinetics and thermodynamics behavior, rebinding selectivity and applicability in solid-phase extraction for this new material obtained were evaluated. The kinetics test indicated this composite material improved adsorption dynamics relative to the MIL-101, with a 28.6% decrease in adsorption equilibrium time. The adsorption capacity for the MIPs@ED-MIL-101 toward PDG was shown higher than that for the MIL-101 and nonimprint-modified material NIPs@ED-MIL-101 at the same substrate concentration when adsorption was carried out at 298 K in acetonitrile-H2O mixed solution (1:1, V/V). Freundlich model can best fit the isotherm data for the MIPs@ED-MIL-101, indicating site heterogeneity in this composite material and mean adsorption energy of 19.60 kJ mol-1 obtained by Dubinin–Radushkevich model fitting might imply chemical adsorption the main rebinding process. In addition, this imprint composite material could selectively bind template and be utilized as the adsorbent in solid-phase extraction to capture target compound from plant crude extract. This new material can also be reused many times with a slight decrease in adsorption capacity.

Biologically Active Constituents from Plants of the Genus Desmos.

The combination of traditional knowledge of medicinal plants with scientific rationale has yielded positive results in recent years. Bioactive compounds isolated from herbaceous plants have long been used as drugs that benefit human health, as well as providing useful compounds for drug development lead compound optimization. This chapter aims to provide a systematic overview of the structural types of Desmos secondary metabolites, along with their biological potential. Various chromatographic and spectroscopic methods have been utilized for isolating, purifying, and elucidating the structures of compounds from Desmos species. From 1982 to the present time, more than 200 metabolites have been isolated from members of this genus. Desmos spp. constituents include terpenoids, phytosterols, polyoxygenated cyclohexanes and cyclohexenes, oxepinones, fatty acids, with flavonoids, alkaloids, and miscellaneous phenols being the predominant compounds. The essential oils of Desmos species have also been investigated. Both crude plant extracts and isolated compounds from this genus have been evaluated for their biological activities. Desmos constituents have been found to exhibit cytotoxic, antimicrobial, antioxidative, anti-inflammatory, and aromatase and NFAT transcriptive inhibition effects.

Non-radioactive Assay to Determine Product Profile of Short-chain Isoprenyl Diphosphate Synthases

Isoprenoids represent the largest class of metabolites with amazing diversities in structure and function. They are involved in protecting plants against pathogens or herbivores or involved in attracting pollinators. Isoprenoids are derived from geranyl diphosphate (GPP; C10), farnesyl diphosphate (FPP; C15), geranylgeranyl diphosphate (GGPP; C20), and geranylfarnesyl diphosphate (GFPP; C25) that are in turn formed by sequential condensations of isopentenyl diphosphate (IPP; C5) with an allylic acceptor such as dimethylallyl diphosphate (DMAPP; C5), GPP, FPP, or GGPP in a reaction catalyzed by isoprenyl diphosphate synthases (IDSs). IDS enzyme assay for determination of prenyl diphosphate products is generally performed using radiolabelled substrates, and the products formed are identified by employing expensive instruments such as phosphor imager, radio-GC, or radio-HPLC. Though a non-radioactive assay for measuring IDS activity in crude plant extract has been reported, it requires a complex methodology utilizing chromatography coupled with tandem mass spectrometry (LC/MS-MS). Here, we describe a non-radioactive and simple inexpensive assay for determining the IDS assay products using non-radiolabeled IPP and its co-allylic substrates DMAPP, GPP, and FPP. The detection of prenyl diphosphate products generated in the assay was highly efficient and spots corresponding to prenyl alcohols were visible at >40 µM concentrations of IPP and DMAPP/GPP/FPP substrates. The protocol described here is sensitive, reliable, and technically simple, which could be used for functional characterization of IDS candidates.

Label-Free LC/HRMS-Based Enzymatic Activity Assay for the Detection of DDC, MAO and COMT Inhibitors in Natural Extracts

Pharmacological treatment of Parkinson’s disease consists of a combined chemotherapy that mostly relies on levodopa (L-DOPA) administration together with inhibitors of dopa-decarboxylase (DDC), monoamine oxidase (MAO) and catechol-methyltransferase (COMT). Identification of inhibitors specifically targeting these enzymes is still a significative part of the development of new alternative antiparkinsonian drugs. Most of the available methods use measurement of enzymatic reactions through radioactive labeling, antibody-recognized products or coupled enzymatic assays. Mass spectrometry (MS) represents an interesting alternative approach as it allows direct and specific detection and quantification of enzymatic reactions. We describe the development of a simple, reliable, label-free assay based on high-resolution mass spectrometry (HRMS) for the detection and relative quantification of three different enzymatic reactions using non-isolated enzymes. The assay was applied both to reference drugs and plant crude extracts. This method can be used to detect hits in extracts libraries as well as determine relative IC50 of inhibitors.

Effects of varying levels of nitrogen on plant growth, anti-Fusarium oxysporum activity and 3,5-dicaffeoylquinic acid concentration of extracts from hydroponically cultivated Helichrysum cymosum (L.)

Experiments were conducted to assess the effects of varied nitrogen (N) concentrations on plant growth, tissue nutrient content, and antimicrobial activity of acetone extract of aerial parts of Helichrysum cymosum. Six weeks old rooted cuttings were treated with 52.5, 105.0, 210.0, and 420.0 mg N/L by varying the nutrient N level in plant growth medium. Plant growth parameters, leaf tissue nutrient content, anti-Fusarium oxysporum activity, and a well-known bioactive secondary metabolite (3,5-dicaffeoylquinic acid) concentration in the plant extracts were analysed. Leaf number increased with increasing N level, while height increase was highest at the lowest N treatment. Tissue analysis of N content and LC-MS analysis of the acetone extracts of aerial parts indicated that lower N treatment corresponded to lower tissue N and higher accumulation of 3,5dicaffeoylquinic acid in the extracts, respectively. In conclusion, lower nutrient N level significantly correlated with increased antimicrobial activity and yield of the targeted compound of the crude acetone extract of H. cymosum. This study suggests that nitrogen fertilization may be manipulated to optimize bioactivities of crude extracts of medicinal plants.

Evaluation of the wound healing activity of the crude extract of root bark of Brucea antidysentrica, the leaves of Dodonaea angustifolia and Rhamnus prinoides in mice

BackgroundWounds are major problems of developing countries that can be managed alternatively using traditional medicinal plants. Since majority of currently available drugs for wound management are expensive and pose problems such as allergy and drug resistance, it is pivotal for the world to have intensified inquiries on the claimed medicinal plants to come up with wound healing chemicals being affordable, effective and safe. Ethiopian traditional healers recruit a wide range of medicinal plants with wound healing activities. Root bark juice of Brucea antidysentrica, the leaves of Rhamnus prinoides and Dodonaea angustifolia are claimed among others in the folklore medicine. Therefore, the aim of this study was to evaluate the in vivo wound healing activities of the root bark juice of Brucea antidysentrica, the leaves of Rhamnus prinoides and Dodonaea angustifolia in mice. MethodThe root bark juice of Brucea antidysentrica, the leaves of Rhamnus prinoides and Dodonaea angustifolia were collected, dried, ground to coarse powders. Then the crude extract was obtained by macerating with 80% methanol. The filtrate was dried, reconstituted in appropriate solvent and the wound healing activity was evaluated using excision and incision wound models. ResultsOn the last day of treatment, 80% methanol extracts from the selected medicinal plants showed a significant wound healing activity against control as supported by an increase in % wound contraction and a decrease in Epithelialization period. Ten percent of Rhamnus prinoides showed significant wound contraction against the control (P < 0.05) on days 2 and 4. But on day 6; except 5% extracts of Brucea antidysentrica and Rhamnus prinoides; all doses of extracts contracted the wound significantly (P < 0.05). Extracts of Dodonea angustifolia (5% & 10%) and 10% of Rhamnus prinoides and Bruceaantidysentrica increases wound contraction rate with increasing significant level on days 8 & 10 (P < 0.01) and 12 &14 (P < 0.001). Among the extracts, 10% of Dodonea angustifolia showed maximum percent (99.9%; P < 0.001) of wound contraction followed by 5% Dodonea angustifolia (99.15%; P < 0.001) and 10% Rhamnus prinoides (99.00%; P < 0.001) on the last day of treatment. In addition; significantly shorter healing time was attained by 5% &10% leaves of Dodonea angustifolia (P < 0.01), 10% leaves Rhamnus prinoides & root barks Brucea antidysentrica (P < 0.05). Ten percent of Rhamnus prinoides &Brucea antidysentrica (P < 0.05) and both doses (5% & 10%) of Dodonea angustifolia (P < 0.01) significantly increased the tensile strength by 54.10%, 56.58%, 63.04%, and 79.19%, respectively against the control. ConclusionThe 80% methanol crude extracts of the study plants support the traditional claims for healing of wounds as evidenced by an increase in wound contraction rate and tensile strength, decrease in Epithelialization period.

Inhibition of metastasis and suppression of pERK1/2 and pFAK expression by Solanum xanthocarpum crude extracts in human lung cancer cell line A549 in vitro

Solanum xanthocarpum is an important part of natural remedies for lung in Ayurveda. Lung cancer metastasis is a major challenge in the present time. Expressions and activation of focal adhesion kinase (FAK) and extracellular receptor kinases 1 and 2 (ERK1/2) are key checkpoints in cancer which lead to an invasion, epithelial-mesenchymal transition (EMT) and metastatic progression. In this study, the efficacy of the whole plant and root crude extracts prepared in ethyl acetate and chloroform on highly metastatic lung cancer cell line A549 was evaluated. The effect of crude extracts on cell migration was scrutinized using transwell migration assessment. The phosphorylated forms of proteins, pFAK and pERK1/2 levels, were estimated using ELISA. Zymography technique was utilized to evaluate the activities of MMP-2 and MMP-9 proteins. The EMT markers, vimentin and N-cadherin gene expressions were analyzed using semi-quantitative RT-PCR. The extracts significantly reduced single-cell migration. The expression levels of pFAK and pERK1/2 notably decreased and FAK and ERK1/2 remained unchanged. Moreover, gelatinase activity, vimentin and N-cadherin expression were also significantly reduced in treated cells compared to untreated cells. The crude extracts of S. xanthocarpum impaired the metastatic progression of A549 cells in vitro.

Appropriate Activity Assays Are Crucial for the Specific Determination of Proline Dehydrogenase and Pyrroline-5-Carboxylate Reductase Activities.

Accumulation of proline is a widespread plant response to a broad range of environmental stress conditions including salt and osmotic stress. Proline accumulation is achieved mainly by upregulation of proline biosynthesis in the cytosol and by inhibition of proline degradation in mitochondria. Changes in gene expression or activity levels of the two enzymes catalyzing the first reactions in these two pathways, namely pyrroline-5-carboxylate (P5C) synthetase and proline dehydrogenase (ProDH), are often used to assess the stress response of plants. The difficulty to isolate ProDH in active form has led several researchers to erroneously report proline-dependent NAD+ reduction at pH 10 as ProDH activity. We demonstrate that this activity is due to P5C reductase (P5CR), the second and last enzyme in proline biosynthesis, which works in the reverse direction at unphysiologically high pH. ProDH does not use NAD+ as electron acceptor but can be assayed with the artificial electron acceptor 2,6-dichlorophenolindophenol (DCPIP) after detergent-mediated solubilization or enrichment of mitochondria. Seemingly counter-intuitive results from previous publications can be explained in this way and our data highlight the importance of appropriate and specific assays for the detection of ProDH and P5CR activities in crude plant extracts.

“Phytochemical and antimicrobial investigation of methanolic extract/fraction of Ocimum basilicum L.”

Abstract The crude solvent extracts of medicinal plants may be exploited as medicines for the betterment of human health. The isolation, purification and characterization of active phytochemical components and their mechanism of action is of predominant grandness. Keeping in view the importance of Ocimum basilicum, the present study was designed to document the phytochemical and antimicrobial activity of methanolic extract of O. basilicum stem. For this purpose dried powdered stem was extracted by successive solvent extraction using petroleum ether (for non-polar Fractions), chloroform (for constituents of intermediate polarities) and methanol (for polar constituents). Six major fractions Ob-1 to Ob-6 were separated from the methanolic extract of O. basilicum stem by column chromatography and purified by thin layer chromatography. All biological active and purified fractions were analyzed by Ultra Violet (UV) and Fourier Transform Infrared (FTIR) spectroscopy. These extracts were initially subjected to screening of antimicrobial activities. The results of the initial screening indicated that methanolic extract was more active than other solvent extracts. Antimicrobial potential of six major fractions Ob-1 to Ob-6 was determined against three species of gram-positive (Clostridium defficile, Bacillus subtilis and Staphylococcus aureus) bacteria, three species of gram-negative (Escherichia coli, Salmonella typhi and Klebsiella pneumoniae) bacteria and three species of fungi (Aspergillus flavus, Aspergillus niger and Candida albicans). All the six isolated fractions displayed well-marked antimicrobial potential against nine microbes. Ob-4, Ob-5 and Ob-6 appeared to be most potent than other fractions, but lesser than the standard antibiotics.

Monoclonal Antibodies B38 and H4 Produced in Nicotiana benthamiana Neutralize SARS-CoV-2 in vitro

The ongoing coronavirus disease 2019 (COVID-19) outbreak caused by novel zoonotic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was initially reported in Wuhan city, Hubei Province of China, in late December 2019. The rapid global spread of the virus calls for the urgent development of vaccines or therapeutics for human applications to combat the coronavirus infection. Monoclonal antibodies (mAbs) have been utilized as effective therapeutics for treating various infectious diseases. In the present study, we evaluated the feasibility of plant expression system for the rapid production of recently identified therapeutically suitable human anti-SARS-CoV-2 mAbs B38 and H4. Transient co-expression of heavy-chain and light-chain sequences of both the antibodies by using plant expression geminiviral vector resulted in rapid accumulation of assembled mAbs in Nicotiana benthamiana leaves within 4 days post-infiltration. Furthermore, both the mAbs were purified from the plant crude extracts with single-step protein A affinity column chromatography. The expression level of mAb B38 and H4 was estimated to be 4 and 35 μg/g leaf fresh weight, respectively. Both plant-produced mAbs demonstrated specific binding to receptor binding domain (RBD) of SARS-CoV-2 and exhibited efficient virus neutralization activity in vitro. To the best of our knowledge, this is the first report of functional anti-SARS-CoV-2 mAbs produced in plants, which demonstrates the ability of using a plant expression system as a suitable platform for the production of effective, safe, and affordable SARS-CoV-2 mAbs to fight against the spread of this highly infectious pathogen.

Determination of the activity and kinetics parameters of proteases in the crude plant extracts of Mentha piperita L. and Thymus capitatus L.

Determination of the activity and kinetics parameters of proteases in the crude plant extracts of Mentha piperita L. and Thymus capitatus L.

Insecticidal potential of some plant extracts in nano and normal form on immatures stages of red palm weevil Rhynchophorus ferrugineus

Red palm weevil, R. ferrugineus (Coleoptera: Dryophthoridae) is the principal insect pest attacks date palm all over Egypt, Current work aims to take advantage of effective, ecofriendly, and safer plant sources in pest control. Toxicity of some plant crude extracts on both normal and nano form against eggs and larvae stags of RPW was assessed in laboratory exposure experiments. The treatments comprised three different concentration of plant extracts (5, 10 and 15 mg/100 ml) for both normal and nano extracts of selected plants using aquatic solutions. Data showed that all the evaluated extracts showed potential ovicidal and larvicidal activity against RPW, nano extracts recorded the highest ovicidal and larvicidal activities compared to normal extract, as ovicidal potential eucalyptus nano extract at 15% was the most toxic which give (80.00±1.00%) mortality, (LC50=9.011) followed by oleander nano extract at 15% 76.67±0.58% (LC50=9.48) and Datura extract at 15 (70.0±1.0 %) (LC50=9.652), while, in Larvicidal activity, nano eucalyptus was the highest effect among all extraction (LC50=5.228) followed by Oleander(LC50=6.042).

Plant-Produced Basic Fibroblast Growth Factor (bFGF) Promotes Cell Proliferation and Collagen Production

AbstractHuman fibroblast growth factor regulates a broad spectrum of biological functions, including cell proliferation and tissue differentiation, and has a wider application in tissue engineering. Here, we described the production of human basic fibroblast growth factor in plants by using a geminiviral vector system. In this study, we transiently expressed basic fibroblast growth factor containing a C-terminus 8X-Histidine with and without a barley alpha amylase signal peptide in Nicotiana benthamiana. The expression level of basic fibroblast growth factor without the signal peptide was found to be higher than the basic fibroblast growth factor with the signal peptide. Further, the recombinant basic fibroblast growth factor was purified from the plant crude extract by two-step purification viz., ammonium sulfate precipitation and Ni-affinity chromatography. Our results demonstrated that the purified plant-produced basic fibroblast growth factor was biologically active and promotes the proliferation of human periodontal ligament stem cells and human follicle dermal papilla cells in vitro. Moreover, the plant-produced basic fibroblast growth factor also induced collagen production in human dermal fibroblast cells. Our results suggest the potential use of plant-produced basic fibroblast growth factor as an antiaging and hair growth-promoting agent in the cosmetic industry.

Impact of Punica granatum-based green larvicide on the predation rate of Polypedates cruciger for the control of mosquito vectors, Anopheles stephensi and Culex quinquefasciatus (Diptera: Culicidae)

Mosquitoes can transmit the causal agent of various human diseases, resulting in millions of deaths every year. In the present study, four simple and inexpensive leaf extracts of Punica granatum, prepared with methanol solvents of increasing polarity, against various developmental stages of the malarial vector, Anopheles stephensi and filarial vector, Culex quinquefasciatus were assessed. The compound composition in each methanol extract was first identified using a gas chromatograph–mass spectrophotometer analysis and the main component was phenol, 2-methyl-5-(1-methylethyl); then it was analyzed by nuclear magnetic resonance (NMR) spectral analysis of 1H NMR. The LC50 and LC90 values of the methanol-derived extract against mosquito larvae were 125.78 and 225.98; 115.28 and 209.50 ppm, against both vectros. Under laboratory conditions, the predation rate of the Polypedates cruciger against An. stephensi and Cx. quinquefasciatus larvae was assessed. The mean number of mosquito prey per larval instars (I to IV) consumed daily by the tadpoles was 22.16 (I), 20.78 (II), 18.96 (III), 15.17 (IV) and 21.30 (I), 19.72 (II), 18.04 (III), 13.78 (IV) for An. stephensi and Cx. quinquefasciatus. Post-exposure to the methanol-derived leaf extract of P. granatum, the mean number of mosquito larvae prey consumed per tadpole per day increased which indicates that this natural mosquito larvicide on non-target tadpoles is compatible and can be used as part of a biological control program against these mosquitoes. This study suggests that the effective plant crude methanol-derived extracts has potential as an alternative eco-friendly approach for vector control of these mosquito larval pests.

Lantana trifolia: Phytochemical and elemental composition, proximate contents and gas chromatography–mass spectrometry profile

Background: With increasing concern over food insecurity, there is the need to incorporate wild edible plants in our meals as they can provide adequate level of nutrition when consumed as food. Aim: The objective of this study was to evaluate the proximate composition, elemental composition, total phenolic content, total flavonoid content and gas chromatography–mass spectrometry (GC-MS) profile of Lantana trifolia . Setting: This study was carried out in Juja, Kenya where the samples were collected, prepared and stored at the Department of Chemistry, Jomo Kenyatta University of Agriculture and Technology. Methods: The proximate and elemental compositions of the leaves, stalk and root samples were evaluated by using standard procedures, whilst the total phenolic and flavonoid contents were evaluated by using Folin–Ciocalteu and aluminium chloride method. The secondary metabolites present in the crude methanolic extracts of the whole plant were determined by using GC-MS. Results: The proximate and elemental analyses of the plant revealed that L. trifolia can be a good source of essential elements, proteins, crude fibre and carbohydrates. The protein, fat, crude fibre and carbohydrate contents in the leaves were found to be higher compared with the stalks and roots, whilst the ash and moisture contents were found to be higher in the roots. The concentrations of calcium, iron, magnesium and zinc in the leaves were found to be 8860.75 ± 565.27, 11 003.10 ± 143.24, 1520.25 ± 26.85 and 39.66 ± 15.68 mg/kg, respectively, compared with the roots and stalks, which were lower. Conclusion: The concentration of total phenolic and total flavonoid compounds and GC-MS profile of the methanolic extracts revealed that L. trifolia can be a good source of secondary metabolites, some of which have reported to be free radical scavengers. Hence, L. trifolia can not only be used as a source of important secondary metabolites, but its nutritional content suggests that the plant can be used to combat nutrient deficiency amongst many communities who lack adequate resources, because it thrives in the wild.

Phosphoenolpyruvate carboxykinase from T. cruzi magnetic beads affinity-based screening assays on crude plant extracts from Brazilian Cerrado

In T. cruzi, a causative agent of Chagas disease, phosphoenolpyruvate carboxykinase (TcPEPCK) is associated with carbohydrate catabolism. Due to its importance in the metabolism of the parasite, it has become a promising target for the development of new drugs against Chagas disease. Aiming to investigate different approaches for ligands screening, TcPEPCK was immobilized on amine-terminated magnetic beads (TcPEPCK-MB) and kinetically characterized by liquid chromatography tandem mass spectrometry activity assay with a KMapp value of 10 ± 1 μM to oxaloacetate as substrate. Natural products library affords highly diverse molecular frameworks through their secondary metabolites, herein a ligand fishing TcPEPCK-MB assay is described for prospecting ligands in four ethanolic extracts of Brazilian Cerrado plants: Qualea grandiflora (Vochysiaceae), Diospyros burchellii (Ebenaceae), Anadenanthera falcata (Fabaceae) and Byrsonima coccolobifolia (Malpighiaceae). The chemical characterization of eleven identified ligands was carried out by liquid chromatography tandem high-resolution mass spectrometry experiments. Senecic acid, syneilesinolide A, phytosphingosine and vanillic acid 4-glucopyranoside are herein reported for the first time for Q. grandiflora, D. burchellii, A. falcata, respectively. In addition, the specificity of the assay was observed since only catechin was fished out from the ethanolic extract of B. coccolobifolia leaves, despite the presence of epicatechin epimer.

Characterisation and performance of three Kenaf coagulation products under different operating conditions

The Sustainable Development Goal (SDG) 6.1, established by the United Nations General Assembly in 2015, targets universal and equitable access to safe and affordable drinking water for all by 2030. An essential factor in achieving this goal is the harnessing of “green” coagulants – naturally occurring, environmentally friendly materials which are effective coagulants for use in water treatment, with good availability in developing countries, inherent renewable properties and ease of biodegradation. In order to gain from these benefits, it is essential to fully understand how such coagulants may best be utilised, particularly concerning their practical application in developing countries. In this study, three different plant-based coagulation products (PCPs), namely Hexane (HxKP), saline (StKP) and crude (CrKP) extracts of Kenaf plant seed (Hibiscus cannabinus, a species of the Hibiscus plant), were applied to high (HTW), medium (MTW) and low (LTW) turbidity water in order to determine their performance and coagulation ability. The ability of the three Kenaf coagulant products (KCPs) to remove hydrophobic fractions of natural organic matter (NOM) was measured. The impact of KCPs on the treated water organic matter content (a known disinfection by-product (DBP) precursor) was examined using known surrogates of natural organic matter (NOM) i.e. the dissolved organic carbon (DOC), ultraviolet absorbance at 254 (UV254) and specific ultraviolet absorbance (SUVA254). Results obtained quantify the implications of using these coagulants during the water disinfection process. A parametric study, measuring the effect of different operating parameters, such as untreated water turbidity, pH, dosages, retention time, and KCP storage time, was completed. Turbidity removal performance for HxKP and StKP was very good with > 90% removal recorded for HTW and MTW, respectively, at pH seven within 2 hours retention time. Images obtained from scanning electron microscopy (SEM) analysis revealed a high likelihood of the coagulation mechanism of KCPs to be adsorption-interparticle bridging brought about by their flake-like structures and surfaces charges. Varying pH had no measurable influence on the coagulation performance of the KCPs. Comparing their efficiency with MoringaOleifera (MO, a previously researched PCP) and alum showed that HxKP had a negligibly different particle removal as MO. StKP turbidity removal performance was below HxKP by 1% for HTW and LTW and 2% for MTW but performed higher than the CrKP by 5% and 7% in HTW and MTW, respectively. The optimum dosage of HxKP and StKP reduced DBP surrogate values, indicating that its precursor is also minimized, although a slight shift from this optimum dosage showed a significant rise in their concentration thus signifying a potential increase in DBPs during disinfection.

Apoptosis in Cancer Cells Is Induced by Alternative Splicing of hnRNPA2/B1 Through Splicing of Bcl-x, a Mechanism that Can Be Stimulated by an Extract of the South African Medicinal Plant, Cotyledon orbiculata.

Alternative splicing is deregulated in cancer and alternatively spliced products can be linked to cancer hallmarks. Targeting alternative splicing could offer novel effective cancer treatments. We investigated the effects of the crude extract of a South African medicinal plant, Cotyledon orbiculata, on cell survival of colon (HCT116) and esophageal (OE33 and KYSE70) cancer cell lines. Using RNASeq, we discovered that the extract interfered with mRNA regulatory pathways. The extract caused hnRNPA2B1 to splice from the hnRNPB1 to the hnRNPA2 isoform, resulting in a switch in the BCL2L1 gene from Bcl-xL to Bcl-xS causing activation of caspase-3-cleavage and apoptosis. Similar splicing effects were induced by the known anti-cancer splicing modulator pladienolide B. Knockdown of hnRNPB1 using siRNA resulted in decreased cell viability and increased caspase-3-cleavage, and over-expression of hnRNPB1 prevented the effect of C. orbiculata extract on apoptosis and cell survival. The effect of the hnRNPA2/B1 splicing switch by the C. orbiculata extract increased hnRNPA2B1 binding to Bcl-xl/s, BCL2, MDM2, cMYC, CD44, CDK6, and cJUN mRNA. These findings suggest that apoptosis in HCT116, OE33, and KYSE cancer cells is controlled by switched splicing of hnRNPA2B1 and BCL2L1, providing evidence that hnRNPB1 regulates apoptosis. Inhibiting this splicing could have therapeutic potential for colon and esophageal cancers. Targeting hnRNPA2B1 splicing in colon cancer regulates splicing of BCL2L1 to induce apoptosis. This approach could be a useful therapeutic strategy to induce apoptosis and restrain cancer cell proliferation and tumor progression. Here, we found that the extract of Cotyledon orbiculata, a South African medicinal plant, had an anti-proliferative effect in cancer cells, mediated by apoptosis induced by alternative splicing of hnRNPA2B1 and BCL2L1.