Polycyclic aromatic hydrocarbons (PAHs) are hazardous and persistent organic pollutants that usually exist at low concentrations in the environment. In this study, dispersive liquid-liquid microextraction (DLLME) technique coupled with high performance liquid chromatography-fluorescence detection (HPLC-FD) was optimized for the analysis of selected PAHs, namely phenanthrene (PHE), fluoranthene (FLA) and benzo[a]pyrene (BaP) in apple juice. Under the optimal extraction conditions (the mixture of 200 µL of acetone and 50 µL of 1-octanol was applied to extract the selected PAHs for 1 min), the DLLME-HPLC-FD showed excellent linearity over the concentration range of 5 to 200 µg/L for both PHE and FLA, and 0.01 to 5 µg/L for BaP with correlation coefficients, r ≥ 0.9956. The method offered ultra-trace detection of selected PAHs in the range of 0.002 to 0.5 µg/L, and negligible matrix effects in determining selected PAHs with relative recovery average within the range of 92.6 to 109.6% in apple juice. The advantages of applying this method for the extraction of PAHs include rapidity, simple operation, as well as small consumption of organic extraction solvent, which is beneficial for routine analysis.
Exploiting natural resources for bioactive compounds is an attractive drug discovery strategy in search for new anti-malarial drugs with novel modes of action. Initial screening efforts in our laboratory revealed two preparations of soil-derived actinomycetes (H11809 and FH025) with potent anti-malarial activities. Both crude extracts showed glycogen synthase kinase 3β (GSK3β)-inhibitory activities in a yeast-based kinase assay. We have previously shown that the GSK3 inhibitor, lithium chloride (LiCl), was able to suppress parasitaemia development in a rodent model of malarial infection. The present study aims to evaluate whether anti-malarial activities of H11809 and FH025 involve the inhibition of GSK3β. The acetone crude extracts of H11809 and FH025 each exerted strong inhibition on the growth of Plasmodium falciparum 3D7 in vitro with 50% inhibitory concentration (IC50) values of 0.57 ± 0.09 and 1.28 ± 0.11 µg/mL, respectively. The tested extracts exhibited Selectivity Index (SI) values exceeding 10 for the 3D7 strain. Both H11809 and FH025 showed dosage-dependent chemo-suppressive activities in vivo and improved animal survivability compared to non-treated infected mice. Western analysis revealed increased phosphorylation of serine (Ser 9) GSK3β (by 6.79 to 6.83-fold) in liver samples from infected mice treated with H11809 or FH025 compared to samples from non-infected or non-treated infected mice. A compound already identified in H11809 (data not shown), dibutyl phthalate (DBP) showed active anti-plasmodial activity against 3D7 (IC50 4.87 ± 1.26 µg/mL which is equivalent to 17.50 µM) and good chemo-suppressive activity in vivo (60.80% chemo-suppression at 300 mg/kg body weight [bw] dosage). DBP administration also resulted in increased phosphorylation of Ser 9 GSK3β compared to controls. Findings from the present study demonstrate that the potent anti-malarial activities of H11809 and FH025 were mediated via inhibition of host GSK3β. In addition, our study suggests that DBP is in part the bioactive component contributing to the anti-malarial activity displayed by H11809 acting through the inhibition of GSK3β.
The reduced efficacy of the mainstay antimalarial drugs due to the widespread of drugresistant Plasmodium falciparum has necessitated efforts to discover new antimalarial drugs with new targets. Quercus infectoria (Olivier) has long been used to treat various ailments including fever. The acetone extract of the plant galls has recently been reported to have a promising antimalarial activity in vitro. This study was aimed to determine the effect of the Q. infectoria gall acetone crude extract on pH of the digestive vacuole of Plasmodium falciparum. A ratiometric fluorescent probe, fluorescein isothiocyanate-dextran (FITC-dextran) was used to facilitate a quantitative measurement of the digestive vacuole pH by flow cytometry. Mid trophozoite stage malaria parasites grown in resealed erythrocytes containing FITC-dextran were treated with different concentrations of the acetone extract based on the 50% inhibitory concentration (IC50). Saponin-permeabilized parasites were analyzed to obtain the ratio of green/yellow fluorescence intensity (Rgy) plotted as a function of pH in a pH calibration curve of FITC-dextran. Based on the pH calibration curve, the pH of the digestive vacuole of the acetone extract-treated parasites was significantly altered (pH values ranged from 6.35- 6.71) in a concentration-dependent manner compared to the untreated parasites (pH = 5.32) (p < 0.001). This study provides a valuable insight into the potential of the Q. infectoria galls as a promising antimalarial candidate with a novel mechanism of action.
The present study has been investigated the role of gallic acid (GA) in paclitaxel-induced neuropathic pain. The neuropathic pain was developed with paclitaxel (PT: 2 mg/kg, i.p.) administration in mice. GA (20 and 40 mg/kg) and pregabalin (PreG: 5 mg/kg) were administered intravenously for 10 consecutive days. The neuralgic sensations were investigated by assessing various pain tests like acetone drop, pinprick, plantar, tail flick, and tail pinch test. Mice pain behaviors were evaluated on 0, 4th, 8th, 12th and 16th days. The levels of sciatic nerve thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH), superoxide anion, calcium, myeloperoxidase (MPO), and TNF-α were estimated. Treatment of GA and PreG attenuate PT induced thermal &mechanical hyperalgesia and allodynia symptoms along with the reduction of TBARS, total calcium, TNF-α, superoxide anion, and MPO activity levels; and decreased GSH level. Therefore, it has been concluded that GA has potential neuroprotective actions against PT induced neuropathic pain due to it's anti-oxidant, anti-inflammation and regulation of intracellular calcium ion concentration.
Eupatorium odoratum (EO) contains many biologically active compounds, the anticancer effects of which are not well documented. This study evaluates the cytotoxic effects and mechanism of action of EO extracts on MCF-7 and Vero cell lines. Evaluation of the cytotoxic activity using MTT assay, morphological alterations, and apoptosis were carried out. Autophagy was evaluated by LC3-A protein expression. Cytotoxic activity, membrane blebbing and ballooning at 24 hours, replacement by mass vacuolation, and double membrane vesicles mimicking autophagy and cell death were observed in the cancer cells. No apoptosis was observed by DNA fragmentation assay. Overexpression of LC3-A protein indicated autophagic cell death. Cell cycle analysis showed G0 and G2/M arrest. The Vero cells did not show significant cell death at concentrations <100 μg/mL. These results thus suggest that acetone and ethyl acetate extracts of EO induce cell death through induction of autophagy and hold potential for development as potential anticancer drugs.
Oil palm (Elaeis guineensis) is an important economic crop cultivated for its nutritional palm oil. A significant amount of effort has been undertaken to understand oil palm growth and physiology at the molecular level, particularly in genomics and transcriptomics. Recently, proteomics studies have begun to garner interest. However, this effort is impeded by technical challenges. Plant sample preparation for proteomics analysis is plagued with technical challenges due to the presence of polysaccharides, secondary metabolites and other interfering compounds. Although protein extraction methods for plant tissues exist, none work universally on all sample types. Therefore, this study aims to compare and optimize different protein extraction protocols for use with two-dimensional gel electrophoresis of young and mature leaves from the oil palm. Four protein extraction methods were evaluated: phenol-guanidine isothiocyanate, trichloroacetic acid-acetone precipitation, sucrose and trichloroacetic acid-acetone-phenol. Of these four protocols, the trichloroacetic acid-acetone-phenol method was found to give the highest resolution and most reproducible gel. The results from this study can be used in sample preparations of oil palm tissue for proteomics work.
Superglue in the ear as a foreign body is an uncommon presentation. We report the case of a lady who accidentally instilled superglue directly onto her tympanic membrane and presented five days later. We successfully removed the glue with acetone and managed to preserve the integrity of the tympanic membrane.
Continuous fermentation of dilute acid-pretreated de-oiled rice bran (DRB) to butanol by the Clostridium acetobutylicum YM1 strain was investigated. Pretreatment of DRB with dilute sulfuric acid (1%) resulted in the production of 42.12 g/L total sugars, including 25.57 g/L glucose, 15.1 g/L xylose and 1.46 g/L cellobiose. Pretreated-DRB (SADRB) was used as a fermentation medium at various dilution rates, and a dilution rate of 0.02 h-1 was optimal for solvent production, in which 11.18 g/L of total solvent was produced (acetone 4.37 g/L, butanol 5.89 g/L and ethanol 0.92 g/L). Detoxification of SADRB with activated charcoal resulted in the high removal of fermentation inhibitory compounds. Fermentation of detoxified-SADRB in continuous fermentation with a dilution rate of 0.02 h-1 achieved higher concentrations of solvent (12.42 g/L) and butanol (6.87 g/L), respectively, with a solvent productivity of 0.248 g/L.h. This study showed that the solvent concentration and productivity in continuous fermentation from SADRB was higher than that obtained from batch culture fermentation. This study also provides an economic assessment for butanol production in continuous fermentation process from DRB to validate the commercial viability of this process.
Protease was extracted from two maturity stages of noni fruits (Morinda citrifolia L.), unripe (stage 1) and ripe (stage 5). The crude extract was partially purified by acetone precipitation method followed by dialysis, gel filtration chromatography and freeze drying. Protein concentrations, proteolytic activity, molecular weight distribution, pH stability, temperature stability and storage efficiency of the resulting protease were evaluated. The unripe and ripe noni fruit contains 0.65 and 0.35% protein, respectively. Molecular weight of the proteases from both stages ranged approximately between 3 to 28 kDa based on the SDS-PAGE results. The optimum activity were at pH 7s and 6, temperatures of 40 and 50°C, respectively for proteases obtained from the unripe and ripe fruit. Analysis from the freeze dried protease indicated that protease from ripe noni fruits had higher protein concentration and specific activity compared to those from unripe fruit. However, it is more sensitive to pH and temperature and less stable during storage as it shows lower proteolytic activity compared to protease from unripe fruit. Based on its high proteolytic activity reaching up to 70.31 U/mg and storage stability (30% lost of activity), noni fruit could be an alternative source of plant protease.
Proteases were extracted from starfruit at maturity Index 2 (unripe, light green) and Index 7 (very ripe, orange) and partially purified using acetone and 40% ammonium sulfate precipitations. Higher yield and proteolytic activity were observed for proteases purified using acetone than 40% ammonium sulfate. As for maturity index, yield and protein concentration of proteases from Index 2 were higher than those from Index 7. SDS-PAGE result showed intense bands for acetone proteases while a distinct band at 50 kDa was observed in all the proteases. Enzyme activity decreased during the seven days storage at 4°C with minimum relative activity of 70% achieved for acetone proteases at day seven. This study suggested that acetone precipitation is more effective method for purifying starfruit protease based on the yield and proteolytic activity compared to using 40% ammonium sulphate precipitation. In order to obtain higher protein concentration and proteolytic activity, starfruit at the unripe stage, Index 2 is a better raw material than Index 7 to be used for protease production.
Acacia species are multipurpose trees, widely used in the traditional systems of medicine to treat various ailments. The major objective of the present study was to determine the gene expression of enzymatic antioxidants by acetone extract from the stem bark of three Acacia species (Acacia dealbata, Acacia ferruginea and Acacia leucophloea) in hydrogen peroxide (H2O2)-induced human hepatoma (HepG2) cells. The expression of antioxidant enzymes such as superoxide dismutase containing copper-zinc (CuZnSOD)/manganese (MnSOD), catalase (CAT) and glutathione peroxidase (GPx) in HepG2 cells was evaluated by real-time PCR. The results of antioxidant enzyme expression in real-time PCR study revealed that the H2O2 (200 μM) challenged HepG2 cells reduced the expression of enzymes such as SOD, GPx and CAT. However, the cells pre-treated with acetone extracts of all the three Acacia species significantly (P > 0.05) up-regulated the expression of antioxidant enzymes in a concentration dependent manner (25, 50 and 75 μg/mL). In conclusion, the findings of our study demonstrated that the acetone extract of Acacia species effectively inhibited H2O2 mediated oxidative stress and may be useful as a therapeutic agent in preventing oxidative stress mediated diseases.
This research explores the possibility of using fluorescence technique to detect the presence of volatile organic compounds based on a single sensing material. The material used was TiO2 nanoparticles coated with porphyrin dye. The TiO2 nanoparticles colloid is in a sol-gel form synthesized from titanium (IV) ethoxide in ethanol with addition of kalium chloride (KCl) as stabilizer. TiO2 nanoparticles were then coated with porphyrin dye, Manganase (III) 5,10,15,20 tetra (4-pyridyl)-21H, 23H porphine chloride tetrakis (metachloride). The coated nanoparticles were deposited on quartz substrate using self-assembly through dip coating technique. The sensing properties of the thin film toward volatile organic compounds; ethanol, acetone, cyclohexane and 2-propanol were studied using luminescence spectrometer. It was found that the thin film produced different emission spectra peaks for different volatile organic compounds (VOCs). Hence, it eases chemical identification process and potentially be use as fluorescence gas sensor.
Acetone-butanol-ethanol (ABE) fermentation from Palm Oil Mill Effluent (POME) by C. acetobutylicum NCIMB 13357 in an oscillatory flow bioreactor was investigated. Experimental works were conducted in a U-shaped stainless steel oscillatory flow bioreactor at oscillation frequency between 0.45-0.78 Hz and a constant amplitude of 12.5 mm. Fermentations were carried out for 72 hr at 35oC using palm oil mill effluent and reinforced clostridia medium as a growth medium in batch culture. Result of this investigation showed that POME is a viable media for ABE fermentation and oscillatory flow bioreactor has an excellent potential as an alternative fermentation device.
The effect of solvent type in antioxidant compounds extraction from banana tissues was studied. The solvent system used was pure methanol, ethanol, acetone and their aqueous solution at 50% and 70% concentrations. Comparison among three common cultivars of banana in Malaysia (Berangan, Mas and Raja) had been done and their antioxidant activities were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging system, ferric reducing ability in plasma (FRAP)
assays and total phenolic content (TPC) assays. Acetone 70% had the strongest antioxidant compounds extraction power as compared to other solvent. All banana samples were found to be low in primary antioxidant but powerful secondary antioxidant source of fruit. The ascending order of banana cultivars in term of their antioxidant activities in all antioxidant assays carried out were Berangan < Mas < Raja. FRAP-TPC assays were highly correlated (R2>0.70) than FRAP-DPPH
and TPC-DPPH assays due to the same mechanism that occurred in the reaction of FRAP and TPC assays.
Palm oil mill effluent (POME) can be utilised directly as the sole substrate in the anaerobic fermentation of acetone-butanol-ethanol (ABE) and hydrogen by Clostridium acetobutylicum NClMB13357 in a submerged batch system. Effects of sedimented POME concentration and the initial culture pH on the production of ABE/H were studied. Sedimented POME with 90% v/v (POME90) at pH 5.8 is capable of producing 4.01 g/L ABE with acetone concentration at 1.97 g/L; butanol 1.74 g/L and ethanol 0.3 g/L. The highest concentration of butanol (1.86 g/L) was produced from a culture with initial pH 6.0. The production of hydrogen gas was proportioned to the concentration of POME. The highest hydrogen gas production was at pH 5.5 (31 mL). More than 50% (v/v) of hydrogen gas was produced at different pH except pH 4.5, when only 16% (v/v) or 5 mL of hydrogen was produced.
Titanium dioxide (TiO2), porphyrin and TiO2 coated with dye porphyrin thin films were prepared on Quartz Crystal Microbalance (QCM) using sol-gel dip coating method and were tested for sensing of volatile organic compounds (VOCs). The porphyrin used was 5,10,15,20-tetraphenyl-21H,23H-porphine manganese (III) chloride (MnTPPCl). The sensing sensitivity was based on the change in the fundamental frequency of the QCM upon exposure towards six vapor samples, namely ethanol, acetone, cyclohexane, toluene, o-xylene and 2-propanol. It was found that all the thin films were sensitive towards all the vapors. However, the TiO2 coated MnTPPCl thin film exhibit the most sensitive and has good selectivity property.
Plutella xylostella (L.) (Lepidoptera: Plutellidae), the major insect pest of cruciferous crops worldwide shows significant
resistance to almost all classes of insecticides. In order to effectively prevent and manage the insecticidal resistance,
it is crucial to understand the physiological adaptation of insects against insecticides. Identification of insect protein
that interacting with insecticides and characterization of their modification in resistant strains can be done by using
differential proteomics approach. This study focuses on optimizing a sensitive and rapid method for the extraction of
high quality protein of both larva and adult tissues of P. xylostella to be used in two-dimensional gel electrophoresis.
Five extraction methods were evaluated for protein concentration, yields and resolving patterns of one-dimensional
and two-dimensional electrophoresis. The results showed that trichloroacetic acid/acetone extraction methods with
two different concentrations of 2-mercaptoethanol produced the highest protein concentration and yield for both adult
and larva tissues, respectively. Meanwhile, trichloroacetic acid/acetone with dithiothreitol extraction method gave
better separation of spots and intensity for both larva and adult tissues compared to other methods tested. As such, we
concluded that trichloroacetic acid/acetone with dithiothreitol successfully yielded high total protein concentration and
good separation of two-dimensional electrophoresis gel spots in both adult and larva P. xylostella.
[BMIM]OTf and alcohol-based DES combination with a selected organic solvent (acetone and acetonitrile) have
been proven to efficiently extracting rotenone (isoflavonoid biopesticide) compound compared to individual organic
solvents. Their efficiency builds up interest to study the solvent-solute interaction that occurs between both selected
solvent systems with rotenone. The interaction study was analyzed using FTIR, 1D-NMR and 2D- NMR (NOESY, HMBC).
Correlation portrayed by NOESY and HMBC of [BMIM]OTf - standard rotenone mixture predicted probable hydrogen
bonding between the oxygen of rotenone with acidic proton C2-H of [BMIM]OTf. While for the alcohol-based DESrotenone
mixture, the correlation shows probable interaction to occur between methyl and methoxy group rotenone
with the hydroxyl group of 1,4-butanediol. In conclusion, potential hydrogen bonding that occurs between solvent
and solute aid towards the solvent efficiency in extracting rotenone compound while emphasizing on the low cost and
green mediated solvent systems.
Betta rubra is an ornamental freshwater fish endemic to northern Sumatra, Indonesia. The B. rubra population has decreased in recent decades, and is classified as an endangered species in the IUCN Red List. This study aims to report for the first time infection by L. cyprinacea in B. rubra harvested from the Aceh Besar region of Indonesia. The fish samples were obtained from the Cot Bira tributaries, Aceh Besar District, Indonesia from January to December 2020. The results showed that the parasite infected 6 out of 499 samples in August and September, with a prevalence and intensity rate of 1% and 2 parasites/fish, respectively. The eyes and pectoral fins were the common infection sites. Despite B. rubra is not an optimal host (small size) for the parasite, this parasite might serve as additional threatening factors for the endangered B. rubra fish population.
Date fruits are well known to be very nutritious. Nevertheless, the protein contents of the fruit, particularly the seed and flesh, are still understudied, largely due to their difficult physical characteristics. This study was conducted to compare three different protein extraction methods which were the trichloroacetic acid (TCA)-acetone (TCA-A), phenol (Phe), and TCA-acetone-phenol (TCA-A-Phe), and to perform proteomic analysis on date palm seed and flesh. Phe extraction method showed the highest protein yields for both seed (8.26 mg/g) and flesh (1.57 mg/g). Through sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Phe, and TCA-A-Phe extraction methods were shown to be efficient in removing interfering compounds and gave well-resolved bands over a wide range of molecular weights. Following liquid chromatography-tandem mass spectrometry analysis, about 50-64% of extracted proteins were identified with known functions including those involved in glycolysis, Krebs cycle, defense, and storage. Phe protein extraction method was proven to be the optimal method for date flesh and seed.