Fragaria chiloensis fruit has a short postharvest life mainly due to its rapid softening. In order to improve its postharvest life, preharvest applications of methyl jasmonate (MeJA) and chitosan were evaluated during postharvest storage at room temperature. The quality and chemical parameters, and protection against decay were evaluated at 0, 24, 48 and 72 h of storage from fruits of two subsequent picks (termed as first harvest and second harvest). In general, fruits treated with MeJA and chitosan maintained higher levels of fruit firmness, anthocyanin, and showed significant delays in decay incidence compared to control fruit. MeJA-treated fruits exhibited a greater lignin content and SSC/TA ratio, and delayed decay incidences. Instead, chitosan-treated fruits presented higher antioxidant capacity and total phenol content. In short, both the elicitors were able to increase the shelf life of fruits as evidenced by the increased levels of lignin and anthocyanin, especially of the second harvest.
A novel microextraction method based on vortex- and CO2 -assisted liquid-liquid microextraction with salt addition for the isolation of furanic compounds (5-hydroxymethyl-2-furaldehyde, 5-methyl-2-furaldehyde, 2-furaldehyde, 3-furaldehyde, 2-furoic and 3-furoic acids) was developed. Purging the sample with CO2 was applied after vortexing to enhance the phase separation and mass transfer of the analytes. The optimum extraction conditions were: extraction solvent (volume), propyl acetate (125 μL); sample pH, 2.4; vortexing time, 45 s; salt concentration, 25% w/v and purging time, 5 min. The analytes were separated using an ODS Hypersil C18 column (250×4.6 mm i.d, 5 μm) under gradient flow. The proposed method showed good linearities (r(2) >0.999), low detection limits (0.08-1.9 μg/L) and good recoveries (80.7-122%). The validated method was successfully applied for the determination of the furanic compounds in concentrated juice (mango, date, orange, pomegranate, roselle, mangosteen and soursop) and dried fruit (prune, date and apricot paste) samples.
Actinomycete strain AUM 00500 was 99.5 % similar to Streptomyces sanglieri NBRC 100784(T) and was evaluated for antagonistic activity towards Ganoderma boninense, the causative fungus of basal stem rot of oil palm. The strain showed strong antifungal activity towards G. boninense in in vitro and SEM analysis showed various modes of inhibition of the fungus. Ethyl acetate extracts of single culture and inhibition zone of cross-plug culture by HPLC indicated that strain AUM 00500 produced two different antibiotics of the glutarimide group namely cycloheximide and actiphenol. In greenhouse trials, oil palm seed treated with spores of S. sanglieri strain AUM 00500 at 10(9) cfu/ml showed significant (P
Proteins of the Bcl-2 family are key targets in anticancer drug discovery. Disrupting the interaction between anti- and pro-apoptotic members of this protein family was the approach chosen in this study to restore apoptosis. Thus, a biological screening on the modulation of the Bcl-xL/Bak and Mcl-1/Bid interactions permitted the selection of Knema hookeriana for further phytochemical investigations. The ethyl acetate extract from the stem bark led to the isolation of six new compounds, three acetophenone derivatives (1-3) and three anacardic acid derivatives (4-6), along with four known anacardic acids (7-10) and two cardanols (11, 12). Their structures were elucidated by 1D and 2D NMR analysis in combination with HRMS experiments. The ability of these compounds to antagonize Bcl-xL/Bak and Mcl-1/Bid association was determined, using a protein-protein interaction assay, but only anacardic acid derivatives (4-10) exhibited significant binding properties, with Ki values ranging from 0.2 to 18 μM. Protein-ligand NMR experiments further revealed that anacardic acid 9, the most active compound, does not interact with the anti-apoptotic proteins Bcl-xL and Mcl-1 but instead interacts with pro-apoptotic protein Bid.
Lead, a chemical element in the carbon group with symbol Pb (from Latin: Plumbum, meaning "the liquid silver") and has an atomic number 82 in the periodic table. It was the first element that was characterized by its kind of toxicity. In animal systems, lead (Pb) has been incriminated in a wide spectrum of toxic effects and it is considered one of the persistent ubiquitous heavy metals. Being exposed to this metal could lead to the change of testicular functions in human beings as well as in the wildlife. The lead poising is a real threat to the public health, especially in the developing countries. Accordingly, great efforts on the part of the occupational and public health have been taken to curb the dangers of this metal. Hematopoietic, renal, reproductive, and central nervous system are among the parts of the human body and systems that are vulnerable toward the dangers following exposure to high level of Pb. In this review, we discussed the massive harmful impact that leads acetate toxicity has on the animals and the worrying fact that this harmful toxicant can be found quite easily in the environment and abundance. Highlighting its (Pb) effects on various organs in the biological systems, its economic, as well as scientific importance, with the view to educate the public/professionals who work in this area. In this study, we focus on the current studies and research related to lead toxicity in animals and also to a certain extent toward human as well.
In the present investigation, we prepared four different solvent fractions (chloroform, hexane, butanol, and ethyl acetate) of Moringa oleifera extract to evaluate its anti-inflammatory potential and cellular mechanism of action in lipopolysaccharide (LPS)-induced RAW264.7 cells. Cell cytotoxicity assay suggested that the solvent fractions were not cytotoxic to macrophages at concentrations up to 200 µg/mL. The ethyl acetate fraction suppressed LPS-induced production of nitric oxide and proinflammatory cytokines in macrophages in a concentration-dependent manner and was more effective than the other fractions. Immunoblot observations revealed that the ethyl acetate fraction effectively inhibited the expression of inflammatory mediators including cyclooxygenase-2, inducible nitric oxide synthase, and nuclear factor (NF)-κB p65 through suppression of the NF-κB signaling pathway. Furthermore, it upregulated the expression of the inhibitor of κB (IκBα) and blocked the nuclear translocation of NF-κB. These findings indicated that the ethyl acetate fraction of M. oleifera exhibited potent anti-inflammatory activity in LPS-stimulated macrophages via suppression of the NF-κB signaling pathway.
OBJECTIVE: Mimosa pudica Linn. (Mimosaceae) is traditionally used as a folk medicine to treat various ailments including convulsions, alopecia, diarrhea, dysentery, insomnia, tumor, wound healing, snake bite, etc., Here, the study was aimed to evaluate the antioxidant potential of M. pudica leaves extract against 2, 2-diphenyl-1-picrylhydrazyl (DPPH) (in vitro) and its modulatory effect on rat brain enzymes.
MATERIALS AND METHODS: Total phenolic, flavonoid contents, and in vitro antioxidant potential against DPPH radical were evaluated from various extracts of M. pudica leaves. In addition, ethyl acetate extract of Mimosa pudica leaves (EAMP) in doses of 100, 200, and 400 mg/kg/day were administered orally for 7 consecutive days to albino rats and evaluated for the oxidative stress markers as thiobarbituric acid reactive substances (TBARS), superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) from rat brain homogenate.
RESULTS: The ethyl acetate extract showed the highest total phenolic content and total flavonoid content among other extracts of M. pudica leaves. The percentage inhibition and IC50 value of all the extracts were followed dose-dependency and found significant (P < 0.01) as compared to standard (ascorbic acid). The oxidative stress markers as SOD, CAT, and GSH were increased significantly (P < 0.01) at 200 and 400 mg/kg of EAMP treated animals and decreased significantly the TBARS level at 400 mg/kg of EAMP as compared to control group.
CONCLUSION: These results revealed that the ethyl acetate extract of M. pudica exhibits both in vitro antioxidant activity against DPPH and in vivo antioxidant activity by modulating brain enzymes in the rat. This could be further correlated with its potential to neuroprotective activity due to the presence of flavonoids and phenolic contents in the extract.
SUMMARY: Total phenolic, flavonoid contents and in-vitro antioxidant potential were evaluated from various extracts of M. pudica leaves. Again, in-vivo antioxidant evaluation from brain homogenate on oxidative stress markers as TBARS, SOD, CAT and GSH from rat was investigated. Our findings revealed that M. pudica possesses both in-vitro and in-vivo antioxidant activity due to presence of phenolics and flavonoids.
KEYWORDS: 2; 2-diphenyl-1-picrylhydrazyl; Brain homogenate; Flavonoids; Mimosa pudica; Oxidative stress
Mixed culture anaerobic fermentation generates a wide range of products from simple sugars, and is potentially an effective process for producing renewable commodity chemicals. However it is difficult to predict product spectrum, and to control the process. One of the key control handles is pH, but the response is commonly dependent on culture history. In this work, we assess the impact of pH regulation mode on the product spectrum. Two regulation modes were applied: in the first, pH was adjusted from 4.5 to 8.5 in progressive steps of 0.5 and in the second, covered the same pH range, but the pH was reset to 5.5 before each change. Acetate, butyrate, and ethanol were produced throughout all pH ranges, but there was a shift from butyrate at pH < 6.5 to ethanol at pH > 6.5, as well as a strong and consistent shift from hydrogen to formate as pH increased. Microbial analysis indicated that progressive pH resulted in dominance by Klebsiella, while reset pH resulted in a bias towards Clostridium spp., particularly at low pH, with higher variance in community between different pH levels. Reset pH was more responsive to changes in pH, and analysis of Gibbs free energy indicated that the reset pH experiments operated closer to thermodynamic equilibrium, particularly with respect to the formate/hydrogen balance. This may indicate that periodically resetting pH conforms better to thermodynamic expectations.
The edible mushroom Pleurotus giganteus was tested for its effect on adipocyte differentiation and glucose uptake activity in 3T3-L1 cells. The basidiocarps of P. giganteus were soaked in methanol to obtain a crude methanol extract and then fractionated to obtain an ethyl acetate extract. In this study, cell proliferation was measured using an MTT assay, lipid accumulation using an Oil Red O assay, and glucose uptake using a fluorescence glucose uptake assay. Gene expression was measured via real-time polymerase chain reaction analysis with TaqMan primer. Ethyl acetate extract significantly enhanced adipogenic differentiation and glucose uptake in 3T3-L1 adipocytes via the expression of sterol regulatory element-binding protein, peroxisome proliferator-activated receptor γ, and phos-phatidylinositol 3-kinase/Akt. Glucose uptake was facilitated by the highly expressed glucose transporters Glut1 and Glut4. Taken together, these results suggest that P. giganteus ethyl acetate extract has an insulin-sensitizing effect on adipocytes and has potential as an adjuvant for the management of type 2 diabetes.
The study aimed to isolate and elucidate the chemical compounds that are found in banana
(Musa balbisiana cv. Saba) inflorescences. Banana inflorescence buds were extracted using
methanol and the resulted methanolic extract was partitioned using chloroform, ethyl acetate
and butanol against deionized water. The chloroform partition was further separated into
fractions using column chromatography assisted by thin layer chromatography. The structure
elucidation was performed using nuclear magnetic resonance spectrometry (NMR). Three
triterpenes were isolated namely 31-norcyclolaudenone (1), cycloartenol (2) and (24R)-4a,24-
trimethyl-5a-cholesta-8,25(27)-dien-3b-ol (3). This is the first report on the isolation of these
triterpenes from Musa balbisiana inflorescence. The discovery of new triterpenes from banana
inflorescence should be further explored to open a new perspective that banana by-products
might serve as new source of natural products for food and pharmaceutical applications.
This article discusses on the natural compounds from the ant plant (Myrmecodia species, family: Rubiaceae). The ethyl
acetate (EtOAc) extract from the tuber of M. platytyrea was fractionated by using medium pressure liquid chromatography,
giving eight fractions (F1-F8). Those fractions were evaluated using the 2, 2-diphenyl-1-picrylhydrazyl (DPPH)
assay. Fraction F5 was recorded as potent (EC50 = 21.57 ± 1.40 µg/mL). Then, it was purified by using column
chromatography (CC) (mobile phase = chloroform: EtOAc). From the CC, ten fractions (F5F1-F5F10) were obtained
and compound (1) was isolated from F5F3 via preparative thin layer chromatography (TLC). After spraying with
anisaldehyde-sulphuric reagent, compound (1) gave a green TLC spot (Rf
= 0.65, 100% CHCl3
, multiple development).
The 1
H-Nuclear Magnetic Resonance (NMR) spectroscopy (500 MHz, CDCl3
) was performed to determine the chemical
framework of (1). This compound was identified as morindolide, having an iridoid structure. Meanwhile, the mass
spectra for compounds (2) and (3) were analysed. The data presented the molecular ion at m/z 375 [M-H]- and 255,
suggesting the formulation of 2-(2-methylbutyryl)phloroglucinol glucoside and a flavanone, respectively. From the
literature, compound (1) was firstly isolated from a Chinese natural medicine, the dried root of Morinda officinalis
(family: Rubiaceae). The flavonoids are also included as the biologically active compounds from Myrmecodia. In
short, this is the first occurrence of morindolide from the ant plant.
Elephantopus scaber and Clinacanthus nutans are traditionally used as wound healing herb. The objective of the present study is to develop a new polyherbal formulation, by comparison, the herbal combination of Elephantopus scaber and Clinacanthus nutans as an in vitro antioxidant activity with their individual herbal activity followed by fractionation of polyherbal formulation for in vivo wound healing activities and identification of bioactive compounds from their active fractions. Antioxidant activity was performed in vitro by DPPH scavenging antioxidant activity followed by in vivo wound healing activities using excision wound model, incision wound model, and burn wound model. Toxicity of the fractions of the polyherbal formulation was performed by a dermal toxicity test. The result showed that Elephantopus scaber crude extract on the basis of EC50 performs a much faster action (15.67 μg/mL) but with less % inhibition (87.66%) as compared to the combination of the new polyherbal formulation of crude extract (30 μg/mL). The polyherbal formulation has the highest % inhibition (89.49%) at the same dose as compared to Elephantopus scaber (87.66%). In comparison among all crude and fractions of new polyherbal formulation, it was found that the ethyl acetate fraction of polyherbal formulation has the fastest activity (EC50 14.83 μg/mL) with % inhibition (89.28%). Furthermore, during evaluation of wound contraction on excision and incision wound model, ethyl acetate fraction possesses the highest activity with (P < 0.001) and (P < 0.0001), respectively. During burn wound model, aqueous fraction (P < 0.001) possesses the highest activity followed by an ethyl acetate fraction (P < 0.0001). LC-MS analysis discovered the presence of several flavonoid-based compounds that work synergistically with sesquiterpene lactone and other bioactive compounds. In conclusion, flavonoid increases the antioxidant activity that surges the rate of wound contraction and works synergistically with other bioactive compounds.
Papaya leaf methanolic extract (PLE) at concentrations of 0 (CON), 5 (LLE), 10 (MLE) and 15 (HLE) mg/250 mg dry matter (DM) with 30 mL buffered rumen fluid were incubated for 24 h to identify its effect on in vitro ruminal methanogenesis and ruminal biohydrogenation (BH). Total gas production was not affected (P > 0.05) by addition of PLE compared to the CON at 24 h of incubation. Methane (CH4 ) production (mL/250 mg DM) decreased (P
Swietenia macrophylla (SM) is a medicinally important plant found in tropical and subtropical regions of the world. The ethyl acetate fraction of the seeds of S. macrophylla (SMEAF) is reported to exhibit potent anticancer, antitumor, anti-inflammatory and antifeedant activities. Till date, there have been no studies reported on the acute oral toxicity profile of the ethyl acetate fraction of the seeds of SM. The objective of the present study was to determine the acute toxicity of SMEAF and evaluate the in-vitro neuroprotective activity of SMEAF using primary neuronal cell cultures. In acute oral toxicity study, the SMEAF did not produce any lethal signs of morbidity and mortality. Histo-pathological findings, support the safety of SMEAF, as there were no significant changes observed in any of the parameters studied. Based on the results obtained in MTT assay, we infer that SMEAF has a significant neuroprotective effect, as it increased the cell viability and exhibited protection to the neuronal cells against TBHP induced oxidative stress. Thus, SMEAF can be suggested for use in the development of herbal drug formulations with neuroprotective potential.
Phaleria macrocarpa (Boerl.) is a well-known medicinal plant and have been extensively used as traditional medicine for ages in treatment of various diseases. The purpose of this study was to determine the in situ cytotoxicity effect P. macrocarpa fruit ethyl acetate fraction (PMEAF) by using various conventional and modern microscopy techniques. The cytotoxicity of PMEAF treated MDA-MB-231 cells was determined through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay and CyQuant Cell Proliferation Assay after 24h of treatment. Both results were indicated that the PMEAF is a potential anticancer agent with the average IC50 values of 18.10μg/mL by inhibiting the MDA-MB-231 cell proliferation. Various conventional and modern microscopy techniques such as light microscopy, holographic microscopy, transmission (TEM) and scanning (SEM) electron microscope were used for the observation of morphological changes in PMEAF treated MDA-MB-231cells for 24h. The characteristic of apoptotic cell death includes cell shrinkage, membrane blebs, chromatin condensation and the formation of apoptotic bodies were observed. PMEAF might be the best candidate for developing more potent anticancer drugs or chemo-preventive supplements.
Background: Clinacanthus nutans (C.nutans) is a plant consumed as a cancer treatment in tropical Asia. Despite the availability of numerous anecdotal reports, evaluation of active anticancer effects has remained elusive. Therefore we here examined antiproliferative, reactive oxygen species (ROS)-inducing and apoptosis mechanisms of whole plant extracts in different cancer cell lines. Methods: Antiproliferative actions of five solvent extracts (hexane, chloroform, ethyl acetate, methanol and water) of C.nutans were tested on non-small cell lung cancer (A549), nasopharygeal cancer (CNE1) and liver cancer (HepG2) cells using MTT assay. The most potent anticancer extract was then assessed by flow cytometry to study cell cycle changes . Intracellular levels of ROS were quantified by DCFH-DA assay. Involvement of the caspase pathway in induction of apoptosis was assessed using caspase assay kits. GC-MS analysis was performed to identify phytoconstituents in the extracts. Results: Hexane and chloroform extracts were antiproliferative against all three cell lines, while the ethyl acetate extract, at 300 μg/mL, was antiproliferative in the CNE1 but not A549 and HepG2 cases. Methanol and water extracts did not inhibit cancer cell proliferation. The most potent anticancer hexane extract was selected for further testing. It induced apoptosis in all three cell lines as shown by an increase in the percentage of cell in sub-G1 phase. Dose-dependent increase in ROS levels in all three cell lines indicated apoptosis to be possibly modulated by oxidative stress. At high concentrations (>100 μg/mL), hexane extracts upregulated caspases 8, 9 and 3/7 across all three cell lines. GC-MS analysis of the hexane extract revealed abundance of 31 compounds. Conclusion : Among the five extracts of C.nutans, that with hexane extract demonstrated the highest antiproliferative activity against all three cancer cell lines tested. Action appeared to be via ion of intracellular ROS, and induction of apoptosis via intrinsic and extrinsic caspase pathways.
In earlier ex vivo studies, we reported the biphasic effect of a methanolic extract of unripe Morinda citrifolia fruit (MMC) on dopamine-induced contractility in isolated rat vas deferens preparations. The present in vivo study was designed and undertaken to further explore our earlier ex vivo findings. This study examined the effect of the ethyl acetate fraction of a methanolic extract of unripe Morinda citrifolia Linn. fruit (EA-MMC; 5-100 mg/kg, p.o.) on the dopaminergic system using mouse models of apomorphine-induced climbing time and climbing behavior, methamphetamine-induced stereotypy (sniffing, biting, gnawing, and licking) and haloperidol-induced catalepsy using the bar test. Acute treatment with EA-MMC at a low dose (25 mg/kg, p.o.) significantly attenuated the apomorphine-induced climbing time and climbing behavior in mice. Similarly, EA-MMC (5 and 10 mg/kg, p.o.) significantly inhibited methamphetamine-induced stereotyped behavior in mice. These results demonstrated that the antidopaminergic effect of EA-MMC was observed at relatively lower doses (<25 mg/kg, p.o.). On the other hand, EA-MMC showed dopaminergic agonistic activity at a high dose (3,000 mg/kg, p.o.), which was evident from alleviation of haloperidol (a dopamine D2 blocker)-induced catalepsy in mice. Therefore, it is concluded that EA-MMC might possess a biphasic effect on the dopaminergic system, i.e., an antagonistic effect at lower doses (<25 mg/kg, p.o.) and an agonistic effect at higher doses (>1,000 mg/kg, p.o.). However, further receptor-ligand binding assays are necessary to confirm the biphasic effects of M. citrifolia fruit on the dopaminergic system.
Red seaweed (Kappaphycus alvarezii) cultivated from Sabah (RSS) and Langkawi (RSL) were digested using in vitro mouth, gastric and duodenal model. The digested seaweed then fermented in a pH-controlled batch culture system inoculated with human faeces to mimic the distal colon. Bacterial enumeration were monitored using fluorescent in situ hybridisation, and the fermentation end products, the short chain fatty acids (SCFA), were analysed using HPLC. Both RSS and RSL showed significant increase of Bifidobacterium sp.; from log10 7.96 at 0 h to log10 8.72 at 24 h, and from log10 7.96 at 0 h to log10 8.60 at 24 h, respectively, and shows no significant difference when compared to the Bifidobacterium sp. count at 24 h of inulin fermentation. Both seaweeds also showed significant increase in total SCFA production, particularly acetate and propionate. Overall, this data suggested that K. alvarezii might have the potential as a prebiotic ingredient.
Previous studies have shown that enhanced biological phosphorus removal (EBPR) performance under continuous aerobic conditions always eventually deteriorates; however, the speed at which this happens depends on the carbon source supplied. The published data suggest that propionate is a better carbon source than acetate is for maintaining operational stability, although it is not clear why. A lab-scale sequencing batch reactor was run initially under conventional anaerobic/aerobic conditions with either acetate or propionate as the carbon source. Chemical and microbiological analyses revealed that both sources performed as expected for such systems. When continuous aerobic conditions were imposed on both these established communities, marked shifts of the "Candidatus Accumulibacter" clades were recorded for both carbon sources. Here, we discuss whether this shift could explain the prolonged EBPR stability observed with propionate.
DNA is widely used in plant genetic and molecular biology studies. In this chapter, we describe how to extract DNA from wheat tissues. The tissue samples are ground to disrupt the cell wall. Then cetyltrimethylammonium bromide (CTAB) or sodium dodecyl sulfate (SDS) is used to disrupt the cell and nuclear membranes to release the DNA into solution. A reducing agent, β-mercaptoethanol, is added to break the disulfide bonds between the cysteine residues and to help remove the tanins and polyphenols. A high concentration of salt is employed to remove polysaccharides. Ethylenediaminetetraacetic acid (EDTA) stops DNase activity by chelating the magnesium ions. The nucleic acid solution is extracted with chloroform-isoamyl alcohol (24:1) or 6 M ammonium acetate. The DNA in aqueous phase is precipated with ethanol or isopropanol, which makes DNA less hydrophilic in the presence of sodium ions (Na+).