MyMedR

Displaying publications 581 - 600 of 962 in total

Abstract:

Sort:

Fulltext Heterometrus Spinifer: An Untapped Source of Anti-Tumor Molecules

Soopramanien M, Khan NA, Ghimire A, Sagathevan K, Siddiqui R

Biology (Basel), 2020 Jul 02;9(7).
PMID: 32630812 DOI: 10.3390/biology9070150

Despite intensive research, cancer incidence and mortality continue to rise. Consequently, the necessity to develop effective anti-cancer therapy is apparent. We have recently shown that the gut bacteria of animals living in polluted environments, such as crocodiles, are a potential source of novel anti-tumor molecules. To extend this work to other resilient species, we investigated the anti-tumor effects of gut bacteria of Heterometrus spinifer (a scorpion). Bacteria from the feces and gut were isolated, identified and evaluated for their anti-tumor effects. Bacterial-conditioned media was prepared in Roswell Park Memorial Institute (RPMI) 1640 media, and cytotoxicity and growth inhibitory properties were examined against cervical (HeLa) cancer cells. Liquid chromatography-mass spectrometry (LC-MS) was conducted to establish the identity of the molecules. Eighteen bacteria species from the gut (HSG01-18) and ten bacteria species from feces (HSF01-10) were tested for anti-tumor effects. Bacterial-conditioned media from scorpion gut and feces exhibited significant growth inhibitory effects against HeLa cells of 66.9% and 83.8%, respectively. Microscopic analysis of cancer cells treated with conditioned media HSG12 and HSG16 revealed apoptosis-like effects. HSG12 was identified as Pseudomonas aeruginosa and HSG16 was identified as Bacillus subtilis. Both conditioned media exhibited 100% growth inhibitory effects versus a selection of cancer cells, comprising cervical, breast and prostate cancer cells. LC-MS indicated the presence of 72 and 38 compounds, detected from HSG12 and HSG16, respectively. Out of these compounds, 47 were successfully identified while the remainder were unidentified and are possibly novel. This study suggests that the fecal and gut microbiota of scorpions might possess molecules with anti-cancer properties, however, further intensive research is needed to assess these expectations.

Matched MeSH terms: Tandem Mass Spectrometry
Combination of light and oxygen accelerates formation of covalent protein-polyphenol bonding during chill storage of meat added 4-methyl catechol

Zainudin MAM, Jongberg S, Lund MN

Food Chem, 2021 Jan 01;334:127611.
PMID: 32712493 DOI: 10.1016/j.foodchem.2020.127611

Plant polyphenols applied as natural antioxidant ingredients, are known to bind to cysteine residues on meat proteins. The aim of this study was to examine the effect of light exposure on the formation of cysteine-phenol adduct in meat added 4-methylcatechol (4MC), a model polyphenol, during storage through quantitative LC-MS/MS-based analysis. Cysteine-4-methylcatechol adduct (Cys-4MC) formation in meat added 1500 ppm 4-MC increased significantly (by 50%) when stored under light in oxygen at 4 °C for 7 days as compared to storage in the dark. This was reflected by a significant decrease in thiol concentrations in the same sample. Gel electrophoresis showed loss in myosin heavy chain (MHC), and a resulting increase in cross-linked MHC (CL-MHC) and larger protein polymers in samples added 4MC. Protein blots stained with nitroblue tetrazolium (NBT) showed intensive protein-polyphenol binding in the meat samples added 4MC, but no major differences between storage conditions.

Matched MeSH terms: Tandem Mass Spectrometry
GC-MS metabolomics revealed protocatechuic acid as a cytotoxic and apoptosis-inducing compound from black rice brans

Yuliana ND, Tuarita MZ, Khatib A, Laila F, Sukarno S

Food Sci Biotechnol, 2020 Jun;29(6):825-835.
PMID: 32523792 DOI: 10.1007/s10068-019-00725-2

GC-MS metabolomics was used to discriminate the phytochemicals profile of Indonesian white, red, and black rice brans, and Japanese white rice brans. This technique was used for the first time to identify compounds in rice brans having cytotoxic activity against WiDr colon cancer cells. Orthogonal Projection to the Latent Structure (OPLS) analysis showed that protocatechuic acid (PA) was a discriminating factor found in black rice brans which strongly correlated with its cytotoxicity (IC50 8.53 ± 0.26 µM). Real time-PCR data demonstrated that PA cytotoxicity at different concentrations (1, 5, 10, 25 and 50 µg/mL) was mediated through different pathways. Bcl-2 expression was downregulated at all tested concentrations indicating apoptosis stimulation. At 1-10 ppm concentration, PA activated both intrinsic and extrinsic apoptosis pathways since the expression of p53, Bax, caspase-8, and caspase-9 were upregulated. At a higher dose (25 and 50 µg/mL), PA possibly involved in pyroptosis-mediated pro-inflammatory cell death by upregulating the expression of caspase-1 and caspase-7.

Matched MeSH terms: Gas Chromatography-Mass Spectrometry
Fulltext i-Propylammonium Lead Chloride Based Perovskite Photocatalysts for Depolymerization of Lignin Under UV Light

Kausar S, Altaf AA, Hamayun M, Rasool N, Hadait M, Akhtar A, et al.

Molecules, 2020 Jul 31;25(15).
PMID: 32752133 DOI: 10.3390/molecules25153520

Lignin depolymerization for the purpose of synthesizing aromatic molecules is a growing focus of research to find alternative energy sources. In current studies, the photocatalytic depolymerization of lignin has been investigated by two new iso-propylamine-based lead chloride perovskite nanomaterials (SK9 and SK10), synthesized by the facile hydrothermal method. Characterization was done by Powder X-Ray Diffraction (PXRD), Scanning Electron Microscopy (SEM), UV-Visible (UV-Vis), Photoluminescence (PL), and Fourier-Transform Infrared (FTIR) Spectroscopy and was used for the photocatalytic depolymerization of lignin under UV light. Lignin depolymerization was monitored by taking absorption spectra and catalytic paths studied by applying kinetic models. The %depolymerization was calculated for factors such as catalyst dose variation, initial concentration of lignin, and varying temperatures. Pseudo-second order was the best suited kinetic model, exhibiting a mechanism for lignin depolymerization that was chemically rate controlled. The activation energy (Ea) for the depolymerization reaction was found to be 15 kJ/mol, which is remarkably less than conventional depolymerization of the lignin, i.e., 59.75 kJ/mol, exhibiting significant catalytic efficiencies of synthesized perovskites. Products of lignin depolymerization obtained after photocatalytic activity at room temperature (20 °C) and at 90 °C were characterized by GC-MS analysis, indicating an increase in catalytic lignin depolymerization structural subunits into small monomeric functionalities at higher temperatures. Specifically, 2-methoxy-4-methylphenol (39%), benzene (17%), phenol (10%) and catechol (7%) were detected by GC-MS analysis of lignin depolymerization products.

Matched MeSH terms: Gas Chromatography-Mass Spectrometry
Understanding of the Role of Pretreatment Methods on Rapeseed Oil from the Perspective of Phenolic Compounds

Xu YJ, Jiang F, Song J, Yang X, Shu N, Yuan L, et al.

J Agric Food Chem, 2020 Aug 19;68(33):8847-8854.
PMID: 32806128 DOI: 10.1021/acs.jafc.0c03539

The thermal pretreatment of oilseed prior to oil extraction could increase the oil yield and improve the oil quality. Phenolic compounds are important antioxidants in rapeseed oil. In this study, we investigated the impact of thermal pretreatment method on the rapeseed oil based on phenolic compound levels. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis showed that the phenolic compound contents in the microwave-pretreated oil were higher than those in the oven- and infrared-treated oils. Sinapic acid (SA) and canolol (CA), which are the top two phenolic compounds in rapeseed oil, exerted well 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity with IC50 values of 8.45 and 8.80 μmol/L. The cell experiment uncovered that SA and CA have significant biological activities related to rapeseed oil quality, including increase of antioxidant enzymes superoxide dismutase (SOD), alleviation of reactive oxygen species (ROS), and cytotoxicity of HepG2 cells after the intake of excessive oleic acid. Further investigation indicated that SA and CA reduced cell apoptosis rate through Bax-Bcl-2-caspase-3 and p53-Bax-Bcl-2-caspase-3, respectively. Taken together, our findings suggest that microwave pretreatment is the best method to improve the content of phenolic compounds in rapeseed oil compared with oven and infrared pretreatments.

Matched MeSH terms: Tandem Mass Spectrometry
Fulltext Purification and characterization of charantin from Momordica charantia Linn.

Oh, J. H., Lim, P. K., Hamzah, S., Tan, C. P., Chan, Sook Wah

Malaysian Journal of Medicine and Health Sciences, 2019;15(102):43-43.
MyJurnal

Introduction: Diabetes mellitus (DM) is one of the top diseases that lead public health concern in Malaysia. It was believed to rise in number up to 4.5 million on cases by year 2020 based on the current figure. Momordica charantia Linn (MC), a climber belonging to family Cucurbitaceae, is well known in treating diabetic-related conditions. In earlier studies related to the hypoglycemic properties of MC mainly utilized the crude extract, which contain a mixture of bioactives (charantins, insulin-like peptides and alkaloids). Till now, there is no conclusive result on the major bioactives that play role in the hypoglycemic effect of MC and research regarding the charantin purification was not well established. Hence, the objectives of this study were to purify the charantin from MC and to characterize the purified charantin before further subjected to in vivo hypoglycemic study. Methods: The crude was first extracted from MC using ethanol as solvent via Soxhlet extraction following by a series of purification steps via washing, centrifugation, and C-18 cartridges. Results: The HPLC analysis showed that the charantin of purified extract after passing out from the cartridge exuded at 12.50 min with a concentration of 500 ppm, which is relatively 20 times higher than the crude extract (25 ppm). The structural properties of purified charantin were studied using FTIR and it showed strong peaks of carboxylic acids (2884 nm), alcohols (1023 nm) and diethyl ether (1114 nm) as compared
with the standard. The compound was reconfirmed in LC-MS analysis. The result displayed mass spectrum in positive mode indicates the presence of similar compound in the purified extract and standard charantin, as presented by ion m/z = 300. Conclusion: The charantin was successfully purified from MC and can act as a potent plant-based hypoglycemic agent for diabetes.

Matched MeSH terms: Tandem Mass Spectrometry
Fulltext Cytotoxic and anticancer properties of the Malaysian mangrove pit viper (Trimeresurus purpureomaculatus) venom and its disintegrin (purpureomaculin)

Tan CH, Liew JL, Navanesan S, Sim KS, Tan NH, Tan KY

J Venom Anim Toxins Incl Trop Dis, 2020;26:e20200013.
PMID: 32742279 DOI: 10.1590/1678-9199-JVATITD-2020-0013

Background: The Asiatic pit vipers from the Trimeresurus complex are medically important venomous snakes. These pit vipers are often associated with snakebite that leads to fatal coagulopathy and tissue necrosis. The cytotoxic venoms of Trimeresurus spp.; however, hold great potential for the development of peptide-based anticancer drugs.
Methods: This study investigated the cytotoxic effect of the venom from Trimeresurus purpureomaculatus, the mangrove pit viper (also known as shore pit viper) which is native in Malaysia, across a panel of human cancer cell lines from breast, lung, colon and prostate as well as the corresponding normal cell lines of each tissue.
Results: The venom exhibited dose-dependent cytotoxic activities on all cell lines tested, with median inhibition concentrations (IC50) ranging from 0.42 to 6.98 µg/mL. The venom has a high selectivity index (SI = 14.54) on breast cancer cell line (MCF7), indicating that it is significantly more cytotoxic toward the cancer than to normal cell lines. Furthermore, the venom was fractionated using C18 reversed-phase high-performance liquid chromatography and the anticancer effect of each protein fraction was examined. Fraction 1 that contains a hydrophilic low molecular weight (approximately 7.5 kDa) protein was found to be the most cytotoxic and selective toward the breast cancer cell line (MCF7). The protein was identified using liquid chromatography-tandem mass spectrometry as a venom disintegrin, termed purpureomaculin in this study.
Conclusion: Taken together, the findings revealed the potent and selective cytotoxicity of a disintegrin protein isolated from the Malaysian T. purpureomaculatus venom and suggested its anticancer potential in drug discovery.

Matched MeSH terms: Tandem Mass Spectrometry
Fulltext Identification of Haptoglobin as a Potential Biomarker in Young Adults with Acute Myocardial Infarction by Proteomic Analysis

Mohamed Bakrim N, Mohd Shah ANS, Talib NA, Ab Rahman J, Abdullah A

Malays J Med Sci, 2020 Mar;27(2):64-76.
PMID: 32788843 MyJurnal DOI: 10.21315/mjms2020.27.2.8

Background: Acute myocardial infarction (AMI) molecular research in young adults is still limited. The aim of this study is to identify AMI proteomic biomarker(s) in young adults.
Methods: This study comprised of two phases namely discovery and verification. In the discovery phase, proteins in the pooled plasma samples from young male adults between 18 and 45 years (10 AMI patients and 10 controls) were separated using two-dimensional electrophoresis. The protein spots that were expressed differently in the AMI patients were identified via matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. The plasma concentrations of these proteins were quantified using enzyme-linked immunosorbent assay during the verification phase (40 AMI patients and 80 controls).
Results: Haptoglobin (Hp), apolipoprotein AI (Apo AI) and apolipoprotein AIV (Apo AIV) were up-regulated in the discovery phase. In the verification phase, the plasma concentration of Hp was significantly higher in AMI patients than the controls (P < 0.001). Logistic regression showed an association between Hp and AMI in young adults (odds ratio [OR] = 1.016, 95% CI: 1.002-1.030, P = 0.025) independent of other AMI risk factors. Hp was significantly correlated with high sensitivity C-reactive protein (hs-CRP) (r = 0.424, P < 0.001).
Conclusion: In young adults with AMI, plasma Hp concentrations were elevated and it is independently associated with AMI. A positive correlation with hs-CRP suggests Hp could be a potential biomarker of AMI in young adults.

Matched MeSH terms: Mass Spectrometry
Fulltext Fatty Acid Composition of Hepatopancreas and Gonads in Both Sexes of Orange Mud Crab, Scylla olivacea Cultured at Various Water Flow Velocities

Taufik M, Shahrul I, Mohd Nordin AR, Ikhwanuddin M, Abol-Munafi AB

Trop Life Sci Res, 2020 Jul;31(2):79-105.
PMID: 32922670 DOI: 10.21315/tlsr2020.31.2.5

Nutritional quality of the hepatopancreas and gonads of orange portunid mud crab, Scylla olivacea was evaluated for each gender under four treatment of different water velocities (0, 20, 40 and 60 cm s-1), in terms of nutrient reserve and nutrient for reproduction. About 56 crabs were used in this study in which fatty acids composition was analysed using gas chromatography mass-spectrometry (GC-MS). For hepatopancreas analysis, monounsaturated fatty acids (MUFAs) were present in the highest fatty acids concentration, followed by polyunsaturated fatty acids (PUFAs) and, saturated fatty acids (SFAs). However, long-chain polyunsaturated fatty acids (LC-PUFAs) were displayed in low concentration in the hepatopancreas. Total fatty acid (TFAs) composition was significantly higher at moderate velocity of 20 cm s-1 compared to other water velocity treatments. For gonad analysis, 20 cm s-1 showed the highest TFA concentration of 93.34 mg g-1 while, the lowest concentration of 3.90 mg g-1 occurred at 0 cm s-1. There were significant differences in male and female crab's fatty acids contents of gonads at all flow velocities challenged (p < 0.05). PUFAs and MUFAs were dominant while, SFAs were observed at low concentration. This study revealed that, concentration of PUFAs increased as gonad maturation increased. The decreasing concentration of hepatopancreas fatty acids over the culture period indicated that nutrient was shifted from the hepatopancreas, to be used as energy reserved to gonads for further growth of eggs and offspring. The linkages between water flow strength, hepatopancreas, and gonad fatty acids concentrations, is fundamental knowledge useful in establishing efficient habitat velocities selection which will improve aquaculture production of mud crabs with high quality broodstock.

Matched MeSH terms: Gas Chromatography-Mass Spectrometry
Fulltext Cytotoxicity of Snake Venoms and Cytotoxins From Two Southeast Asian Cobras (Naja sumatrana, Naja kaouthia): Exploration of Anticancer Potential, Selectivity, and Cell Death Mechanism

Chong HP, Tan KY, Tan CH

Front Mol Biosci, 2020;7:583587.
PMID: 33263003 DOI: 10.3389/fmolb.2020.583587

Venoms of cobras (Naja spp.) contain high abundances of cytotoxins, which contribute to tissue necrosis in cobra envenomation. The tissue-necrotizing activity of cobra cytotoxins, nevertheless, indicates anticancer potentials. This study set to explore the anticancer properties of the venoms and cytotoxins from Naja sumatrana (equatorial spitting cobra) and Naja kaouthia (monocled cobra), two highly venomous species in Southeast Asia. The cytotoxicity, selectivity, and cell death mechanisms of their venoms and cytotoxins (NS-CTX from N. sumatrana: NS-CTX; N. kaouthia: NK-CTX) were elucidated in human lung (A549), prostate (PC-3), and breast (MCF-7) cancer cell lines. Cytotoxins were purified through a sequential fractionation approach using cation-exchange chromatography, followed by C18 reverse-phase high-performance liquid chromatography (HPLC) to homogeneity validated with sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and identified by liquid chromatography-tandem mass spectrometry (LCMS/MS). The cobra venoms and their respective cytotoxins exhibited concentration-dependent growth inhibitory effects in all cell lines tested, with the cytotoxins being more potent compared to the corresponding whole venoms. NS-CTX and NK-CTX are, respectively, P-type and S-type isoforms of cytotoxin, based on the amino acid sequences as per LCMS/MS analysis. Both cytotoxins exhibited differential cytotoxic effects in the cell lines tested, with NS-CTX (P-type cytotoxin) being significantly more potent in inhibiting the growth of the cancer cells. Both cytotoxins demonstrated promising selectivity only for the A549 lung cancer cell line (selectivity index = 2.17 and 2.26, respectively) but not in prostate (PC-3) and breast (MCF-7) cancer cell lines (selectivity index < 1). Flow cytometry revealed that the A549 lung cancer cells treated with NS-CTX and NK-CTX underwent necrosis predominantly. Meanwhile, the cytotoxins induced mainly caspase-independent late apoptosis in the prostate (PC-3) and breast (MCF-7) cancer cells lines but lacked selectivity. The findings revealed the limitations and challenges that could be faced during the development of new cancer therapy from cobra cytotoxins, notwithstanding their potent anticancer effects. Further studies should aim to overcome these impediments to unleash the anticancer potentials of the cytotoxins.

Matched MeSH terms: Tandem Mass Spectrometry
In-house validation of accelerated solvent extraction-gas chromatography-mass spectrometry for the determination of bound 3- and 2-monochloropropanediols (MCPD) and glycidol in food products

Shaari NA, Ahmad Tarmizi AH, Md Sikin A

Food Addit Contam Part A Chem Anal Control Expo Risk Assess, 2021 Feb;38(2):223-236.
PMID: 33332229 DOI: 10.1080/19440049.2020.1845400

The study aimed to establish the detection method for bound 3-, 2-MCPD, and glycidol using accelerated solvent extraction (ASE) and gas chromatography mass spectrometry (GC-MS). The ASE was modified for reduced solvent volume and process time to extract lipid from the chocolate spread, infant formula, potato chips, and sweetened creamer. The solvent selected for ASE was a mixture of iso-hexane and acetone at 100°C with the lipid and analyte recovery ranging from 96.9% to 98.6% and 84.1% to 107.5%, respectively. The derivatisation of analytes was adopted from the AOCS method Cd29a-13 for GC-MS analysis. The results showed that the coefficient of determination (R2) of all analytes was >0.99. The limit of detection (LOD) was 0.1 mg kg-1 expressed in lipid basis for both bound 3- and 2-MCPD and 0.2 mg kg-1 expressed in lipid basis for bound glycidol. The limit of quantitation (LOQ) was 0.3 mg kg-1 expressed in lipid basis for both bound 3- and 2-MCPD and 0.6 mg kg-1 expressed in lipid basis for bound glycidol. A blank spiked with 3-monochloropropanediols fatty acid esters (MCPDE) and 2-MCPDE (0.3, 2.1, and 7.2 mg kg-1) and glycidol esters (0.6, 4.7, and 16.6 mg kg-1) were chosen for accuracy and precision tests. The recoveries were 91.7% to 105.9%. Both repeatability and within-laboratory reproducibility of the analysis were within the acceptable level of precision ranging from 1.7% to 16%. This is the first time that a full validation procedure extending to both accuracy and precision tests has been carried out for sweetened creamer and chocolate spread. Overall, the combined protocol of ASE and AOCS Cd29a-13 was successfully validated for both solid and liquid food samples with lipid content from 10% to 30%.

Matched MeSH terms: Gas Chromatography-Mass Spectrometry
Fulltext Global metabolomics profiling of colorectal cancer in Malaysian patients

Amir Hashim NA, Ab-Rahim S, Wan Ngah WZ, Nathan S, Ab Mutalib NS, Sagap I, et al.

Bioimpacts, 2021;11(1):33-43.
PMID: 33469506 DOI: 10.34172/bi.2021.05

Introduction:
The serum metabolomics approach has been used to identify metabolite biomarkers that can diagnose colorectal cancer (CRC) accurately and specifically. However, the biomarkers identified differ between studies suggesting that more studies need to be performed to understand the influence of genetic and environmental factors. Therefore, this study aimed to identify biomarkers and affected metabolic pathways in Malaysian CRC patients.
Methods:
Serum from 50 healthy controls and 50 CRC patients were collected at UKM Medical Centre. The samples were deproteinized with acetonitrile and untargeted metabolomics profile determined using liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOFMS, Agilent USA). The data were analysed using Mass Profiler Professional (Agilent, USA) software. The panel of biomarkers determined were then used to identify CRC from a new set of 20 matched samples.
Results:
Eleven differential metabolites were identified whose levels were significantly different between CRC patients compared to normal controls. Based on the analysis of the area under the curve, 7 of these metabolites showed high sensitivity and specificity as biomarkers. The use of the 11 metabolites on a new set of samples was able to differentiate CRC from normal samples with 80% accuracy. These metabolites were hypoxanthine, acetylcarnitine, xanthine, uric acid, tyrosine, methionine, lysoPC, lysoPE, citric acid, 5-oxoproline, and pipercolic acid. The data also showed that the most perturbed pathways in CRC were purine, catecholamine, and amino acid metabolisms.
Conclusion:
Serum metabolomics profiling can be used to identify distinguishing biomarkers for CRC as well as to further our knowledge of its pathophysiological mechanisms.

Matched MeSH terms: Mass Spectrometry
Fulltext Flaxseed (linum usitatissimum) extract activity on human oral fibroblasts (HOrF) cell line

Sama Naziyah Shaban, Abdelkader Elbadawy Abbas Ashour, Khairani Idah Mokhtar, Solachuddin JauhariAriefIchwan, Basma Ezzat Mustafa Alahmad

Ann Dent, 2020;27(1):50-54.
MyJurnal

Natural products have demonstrated various activities beneficial to general health. Flaxseed (Linum usitatissimum) has been reported in many studies for its antimicrobial, antioxidant, and anti-inflammatory effects. Additionally, flaxseed extracts have skin wound healing activity and potential for treating oral ulcers.L. usitatissimum was extracted using 70% ethanol via soxhlet method and gas chromatography mass spectrum (GC-MS) was used to analyze the components of L. usitatissimum extract. The crude flaxseed oil were applied to human oral fibroblasts (HOrF), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to assess the cell viability after 24, 48 and 72 hours. Scratched HOrF cells were treated with crude flaxseed oil and healing was monitored per wound healing assay. GC-MS indicate that the major components present in L. usitatissimum oil extract are linolic, palmitic and oleic acid. L. usitatissimum crude oil extract showed high proliferation effect on HOrF cells at 24 and 48 hours, while the highest proliferation effect was recorded at 72 hours post-treatment. The wound healing assay results showed that healing activity of HOrF cells occurred as soon as 18 hours post-treatment when treated with L. usitatissimum crude oil extract. L. usitatissimum crude oil extract has proliferating and healing effects on HOrF cell line. Therefore, it can be considered as a potential promising oral wound healing agent.

Matched MeSH terms: Gas Chromatography-Mass Spectrometry
Shotgun Proteomics and Mass Spectrometry as a Tool for Protein Identification and Profiling of Bio-Carrier-Based Therapeutics on Human Cancer Cells

Abidin SAZ, Othman I, Naidu R

Methods Mol Biol, 2021;2211:233-240.
PMID: 33336281 DOI: 10.1007/978-1-0716-0943-9_16

Shotgun proteomics has been widely applied to study proteins in complex biological samples. Combination of high-performance liquid chromatography with mass spectrometry has allowed for comprehensive protein analysis with high resolution, sensitivity, and mass accuracy. Prior to mass spectrometry analysis, proteins are extracted from biological samples and subjected to in-solution trypsin digestion. The digested proteins are subjected for clean-up and injected into the liquid chromatography-mass spectrometry system for peptide mass identification. Protein identification is performed by analyzing the mass spectrometry data on a protein search engine software such as PEAKS studio loaded with protein database for the species of interest. Results such as protein score, protein coverage, number of peptides, and unique peptides identified will be obtained and can be used to determine proteins identified with high confidence. This method can be applied to understand the proteomic changes or profile brought by bio-carrier-based therapeutics in vitro. In this chapter, we describe methods in which proteins can be extracted for proteomic analysis using a shotgun approach. The chapter outlines important in vitro techniques and data analysis that can be applied to investigate the proteome dynamics.

Matched MeSH terms: Tandem Mass Spectrometry
Fulltext Secondary Metabolites, Antioxidant, and Antiproliferative Activities of Dioscorea bulbifera Leaf Collected from Endau Rompin, Johor, Malaysia

Mainasara MM, Abu Bakar MF, Md Akim A, Linatoc AC, Abu Bakar FI, Ranneh YKH

Evid Based Complement Alternat Med, 2021;2021:8826986.
PMID: 33505506 DOI: 10.1155/2021/8826986

Breast cancer is among the most commonly diagnosed cancer and the leading cause of cancer-related death among women globally. Malaysia is a country that is rich in medicinal plant species. Hence, this research aims to explore the secondary metabolites, antioxidant, and antiproliferative activities of Dioscorea bulbifera leaf collected from Endau Rompin, Johor, Malaysia. Antioxidant activity was assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assays, while the cytotoxicity of D. bulbifera on MDA-MB-231 and MCF-7 breast cancer cell lines was tested using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Cell cycle analysis and apoptosis were assessed using flow cytometry analysis. Phytochemical profiling was conducted using gas chromatography-mass spectrometry (GC-MS). Results showed that methanol extract had the highest antioxidant activity in DPPH, FRAP, and ABTS assays, followed by ethyl acetate and hexane extracts. D. bulbifera tested against MDA-MB-231 and MCF-7 cell lines showed a pronounced cytotoxic effect with IC50 values of 8.96 μg/mL, 6.88 μg/mL, and 3.27 μg/mL in MCF-7 and 14.29 μg/mL, 11.86 μg/mL, and 7.23 μg/mL in MDA-MB-231, respectively. Cell cycle analysis also indicated that D. bulbifera prompted apoptosis at various stages, and a significant decrease in viable cells was detected within 24 h and substantially improved after 48 h and 72 h of treatment. Phytochemical profiling of methanol extract revealed the presence of 39 metabolites such as acetic acid, n-hexadecanoic acid, acetin, hexadecanoate, 7-tetradecenal, phytol, octadecanoic acid, cholesterol, palmitic acid, and linolenate. Hence, these findings concluded that D. bulbifera extract has promising anticancer and natural antioxidant agents. However, further study is needed to isolate the bioactive compounds and validate the effectiveness of this extract in the In in vivo model.

Matched MeSH terms: Gas Chromatography-Mass Spectrometry
Pregnenolone metabolites in rat testis: endogenous concentrations, and intracellular distribution in whole testes during incubation in vitro

Pertiwi AK, Kwan TK, Gower DB

J Steroid Biochem Mol Biol, 2002 Aug;81(4-5):363-7.
PMID: 12361726

The intracellular movements of pregnenolone in rat testes were investigated. Whole testes were incubated in the presence or absence of pregnenolone (2.5mM) in the medium for 120 min (in some studies 30, 60, and 90 min). The testes were homogenised, subcellular fractions prepared and analysed in quadruplicate for steroid content by gas chromatography-mass spectrometry with selected ion monitoring. Quantification of pregnenolone and 11 of its metabolites, obtained from non-incubated whole testes, provided values for endogenous amounts. Pregnenolone was the only steroid of quantitative importance found initially in the mitochondrial fraction but was subsequently found in the microsomal fraction, where metabolism occurred. Identification and quantification of metabolites indicated that both classical pathways for testosterone production were operating, with the 4-en-3-oxosteroid pathway predominating. By 120 min, virtually all pregnenolone metabolites, including pregnenolone itself, were found in the cytosol, consistent with an overall movement from mitochondria to endoplasmic reticulum to cytosol.

Matched MeSH terms: Mass Spectrometry
New bisindole alkaloids from Tabernaemontana corymbosa

Kam TS, Sim KM, Pang HS

J Nat Prod, 2003 Jan;66(1):11-6.
PMID: 12542336

Ten new bisindole alkaloids of the vobasinyl-ibogan type, viz., conodiparines A-F (1-6), conodutarines A and B (7, 8), and cononitarines A and B (9, 10), were obtained from the leaf extract of the Malayan species Tabernaemontana corymbosa. The structures were determined using NMR and MS analysis.

Matched MeSH terms: Mass Spectrometry
Clean-up, detection and determination of salbutamol in human urine and serum

Saleh MI, Koh YM, Tan SC, Aishah AL

Analyst, 2000 Sep;125(9):1569-72.
PMID: 11064937

Salbutamol ¿2-(tert-butylamino)-1-[4-hydroxy-3- (hydroxymethyl)phenyl]ethanol¿, also known as albuterol, is clinically the most widely used beta 2-adrenoceptor agonist in the treatment of bronchial asthma. During this study, we evaluated liquid-liquid extraction (LLE) and solid-phase extraction (SPE) in order to develop a reliable extraction method followed by analysis using liquid chromatography and gas chromatography. An assay is described which involves SPE as the clean-up method followed by gas chromatography-mass spectrometry to determine salbutamol levels in human serum after oral administration. The SPE method requires the use of a hyper-cross-linked styrene-divinylbenzene bonded phase (ENV+) without involving any sample pre-treatment to obtain 60-65% recoveries for salbutamol and terbutaline as the internal standard. Distilled water and 1% trifluoroacetic acid in methanol were found to be the most suitable washing solvent and eluting solvent, respectively. A detection limit of 2 ng mL-1 was achieved by derivatization with N-methyl-N-trimethylsilyltrifluoroacetamide to form trimethylsilyl (TMS)-salbutamol (m/z 369) and TMS-terbutaline (m/z 356). The relationship between the ratio of the peak area of salbutamol to that of the internal standard and concentration was linear for the range tested (2-200 ng mL-1) and the correlation of coefficient was 0.9999 with a y-intercept not significantly different from zero. The inter-day relative standard deviation (RSD) was < 10% for all three concentrations. The intra-day RSD was 14% for 2 ng mL-1. This assay was then successfully applied to human serum samples obtained from clinical trials after oral administration of salbutamol.

Matched MeSH terms: Gas Chromatography-Mass Spectrometry
Four new clerodane diterpenoids from Callicarpa pentandra

Xu J, Harrison LJ, Vittal JJ, Xu YJ, Goh SH

J Nat Prod, 2000 Aug;63(8):1062-5.
PMID: 10978198

Leaf extracts of Callicarpa pentandra provided four new clerodane-type diterpenoids (1-4), of which 1, 2, and 4 have ring-A-contracted structures. Their structures and stereochemistry were established by spectral data interpretation, and for 3 also by single-crystal X-ray diffraction.

Matched MeSH terms: Gas Chromatography-Mass Spectrometry
Epi-sarsasapogenin and epi-smilagenin: two sapogenins isolated from the rumen content of sheep intoxicated by Brachiaria decumbens

Lajis NH, Abdullah AS, Salim SJ, Bremner JB, Khan MN

Steroids, 1993 Aug;58(8):387-9.
PMID: 8212090

Spectroscopic examination of purified extracts of the rumen content of sheep intoxicated by Brachiaria decumbens revealed the presence of two spirostanes, identified as epi-sarsasapogenin and epi-smilagenin. Sarsasapogenone was obtained by the oxidation of sarsasapogenin. The reduction of sarsasapogenone using lithium aluminum hydride yielded isomeric products, sarsasapogenin (20%) and epi-sarsasapogenin (80%).

Matched MeSH terms: Mass Spectrometry

Filters

Please provide feedback to Administrator (afdal@afpm.org.my)

External Links