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  1. IgA binding lectins isolated from distinct Artocarpus species demonstrate differential specificity
    Hashim OH, Ng CL, Gendeh S, Nik Jaafar MI
    Mol Immunol, 1991 4 1;28(4-5):393-8.
    PMID: 2062319
    The discovery of jacalin, a group of lectins from jackfruit seeds (Artocarpus heterophyllus), has attracted considerable attention due to its numerous interesting immunological properties as well as its usefulness in the isolation of various serum proteins. We have further identified a similar lectin from the seeds of Champedak (Artocarpus integer) which we refer to as lectin-C and performed comparative studies with two types of jacalin isolated from different batches of the Malaysian jackfruit seeds (jacalin-M1 and jacalin-M2). The three purified lectins demonstrated equivalent apparent Mr of about 52,500, each of which comprised of a combination of two types of non-covalently-linked subunits with apparent Mr of approximately 13,300 and 16,000. The lectins demonstrated equal haemagglutinating activity against human erythrocytes of blood groups A, B, AB and O. Our data also demonstrated that lectin-C, jacalin-M1 and jacalin-M2 are similar by selectively precipitating human serum IgA1 and colostral sIgA but not IgA2, IgD, IgG and IgM. When immunoelectrophoresis was performed on normal human sera and reacted with the lectins, single precipitin arcs corresponding to IgA immunoprecipitates were detected with lectin-C and jacalin-MI. Jacalin-M2, however, exhibited two closely associated precipitin arcs. The binding of these lectins with IgA was pronouncedly inhibited in the presence of p-nitrophenyl-beta-D-galactopyranoside, 1-o-methyl-alpha-D-galactopyranoside, D-melibiose, N-acetyl-D-galactosamine and D-galactose. The data therefore provide evidence on the differential specificity of IgA binding lectins isolated from seeds of similar as well as distinct Artocarpus species.
    Matched MeSH terms: Chromatography, Gel
  2. 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: Chromatography, Liquid
  3. Fulltext Pharmacokinetics of Free Oxytetracycline and Oxytetracycline Loaded Cockle Shell Calcium Carbonate-Based Nanoparticle in BALB/c Mice
    Idris SB, Abdul Kadir A, Abdullah JFF, Ramanoon SZ, Basit MA, Abubakar MZZA
    Front Vet Sci, 2020;7:270.
    PMID: 32613011 DOI: 10.3389/fvets.2020.00270
    The development and utilization of nano-antibiotics is currently gaining attention as a possible solution to antibiotic resistance. The aim of this study was therefore to determine the pharmacokinetics of free oxytetracycline (OTC) and oxytetracycline loaded cockle shell calcium carbonate-based nanoparticle (OTC-CNP) after a single dose of intraperitoneal (IP) administration in BALB/c mice. A total of 100 female BALB/c mice divided into two groups of equal number (n = 50) were administered with 10 mg/kg OTC and OTC-CNP, respectively. Blood samples were collected before and post-administration from both groups at time 0, 5, 10, 15, and 30 min and 1, 2, 6, 24, and 48 h, and OTC plasma concentration was quantified using a validated HPLC-UV method. The pharmacokinetic parameters were analyzed using a non-compartment model. The Cmax values of OTC in OTC-CNP and free OTC treated group were 64.99 and 23.53 μg/ml, respectively. OTC was detected up to 24 h in the OTC-CNP group as against 1 h in the free OTC group following intraperitoneal administration. In the OTC-CNP group, the plasma elimination rate of OTC was slower while the half-life, the area under the curve, and the volume of the distribution were increased. In conclusion, the pharmacokinetic profile of OTC in the OTC-CNP group differs significantly from that of free OTC. However, further studies are necessary to determine the antibacterial efficacy of OTC-CNP for the treatment of bacterial diseases.
    Matched MeSH terms: Chromatography, High Pressure Liquid
  4. 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: Chromatography, Liquid
  5. Fulltext Proteins from Lignosus tigris with selective apoptotic cytotoxicity towards MCF7 cell line and suppresses MCF7-xenograft tumor growth
    Kong BH, Teoh KH, Tan NH, Tan CS, Ng ST, Fung SY
    PeerJ, 2020;8:e9650.
    PMID: 32832273 DOI: 10.7717/peerj.9650
    Background: Lignosus tigris, a recently discovered species of the unique Lignosus family, has been traditionally used by the indigenous communities in Peninsular Malaysia to treat various ailments and as an alternative medicine for cancer treatment. The L. tigris cultivar sclerotia (Ligno TG-K) was found to contain numerous bioactive compounds with beneficial biomedicinal properties and the sclerotial extract exhibited potent antioxidant activity. However, the anticancer property of the Ligno TG-K including in vitro and in vivo antitumor effects as well as its anticancer active compounds and the mechanisms has yet to be investigated.

    Methods: The cytotoxicity of the Ligno TG-K against human breast (MCF7), prostate (PC3) and lung (A549) adenocarcinoma cell lines was evaluated using MTT cytotoxicity assay. The cytotoxic mechanisms of the active high molecular weight proteins (HMWp) fraction were investigated through detection of caspases activity and apoptotic-related proteins expression by Western blotting. The in vivo antitumor activity of the isolated HMWp was examined using MCF7 mouse xenograft model. Shotgun LC-MS/MS analysis was performed to identify the proteins in the HMWp.

    Results and Discussion: Cold water extract of the sclerotia inhibited proliferation of MCF7, A549 and PC3 cells with IC50 ranged from 28.9 to 95.0 µg/mL. Bioassay guided fractionation of the extract revealed that HMWp exhibited selective cytotoxicity against MCF7 cells via induction of cellular apoptosis by the activation of extrinsic and intrinsic signaling pathways. HMWp activated expression of caspase-8 and -9 enzymes, and pro-apoptotic Bax protein whilst inhibiting expression of tumor survivor protein, Bcl-2. HMWp induced tumor-cell apoptosis and suppressed growth of tumor in MCF-7 xenograft mice. Lectins, serine proteases, RNase Gf29 and a 230NA deoxyribonuclease are the major cytotoxic proteins that accounted for 55.93% of the HMWp.

    Conclusion: The findings from this study provided scientific evidences to support the traditional use of the L. tigris sclerotia for treatment of breast cancer. Several cytotoxic proteins with high abundance have been identified in the HMWp of the sclerotial extract and these proteins have potential to be developed into new anticancer agents or as adjunct cancer therapy.

    Matched MeSH terms: Chromatography, Liquid
  6. 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: Chromatography, Liquid
  7. Anti-proliferative and anti-invasive properties of a purified fraction from Streptomyces sp. H7372
    Yip WK, Cheenpracha S, Chang LC, Ho CC, Seow HF
    Int J Oncol, 2010 Nov;37(5):1229-41.
    PMID: 20878070
    Secondary metabolites from actinomycetes especially the genus Streptomyces may be one of the most important sources for novel anticancer agents. A purified fraction from a novel actinomycete strain, Streptomyces sp. H7372, was elucidated in breast cancer cells. We have isolated three purified fractions from a novel strain, Streptomyces sp. H7372. One of the fractions, designated as 31-2, exhibited the strongest growth-inhibitory effect and thereby was selected for further studies. 31-2 exerted a growth-inhibitory effect on a panel of 15 human cancer and 2 non-malignant cell lines. In MCF-7 and MDA-MB-231 breast cancer cells, 31-2 induced a cytostatic (anti-proliferative) effect without causing cytotoxicity (cell death). Our data suggest that the cytostasis resulted from cell cycle arrest at the G1 phase in MCF-7 cells and at the S phase in MDA-MB-231 cells. Western blot analysis demonstrated a modulation of phosphorylation of the Rb and CDC2 proteins and of CDK4, cyclin D1 and cyclin D3 in the 31-2-treated breast cancer cell lines. The protein levels of CDK2, CDK6, and PCNA were not affected by 31-2 treatment. 31-2 also exhibited an anti-invasive effect in MDA-MB-231 cells. However, this effect is not attributed to the modulation of proteolytic activity in MDA-MB-231 cells as the enzymatic degradation of type IV collagen was not affected by 31-2. The 31-2 is a potent cytostatic and anti-invasive agent and modulates the cell cycle pathway. Together, these results will have important implications in searching for novel approaches to treat cancer.
    Matched MeSH terms: Chromatography, High Pressure Liquid
  8. Purification and characterization of 31-kDa palm pollen glycoprotein (Ela g Bd 31 K), which is recognized by IgE from palm pollinosis patients
    Kimura Y, Maeda M, Kimupa M, Lai OM, Tan SH, Hon SM, et al.
    Biosci Biotechnol Biochem, 2002 Apr;66(4):820-7.
    PMID: 12036055
    A basic glycoprotein, which was recognized by IgE from oil palm pollinosis patients, has been purified from oil palm pollen (Elaeis guineensis Jacq.), which is a strong allergen and causes severe pollinosis in Malaysia and Singapore. Soluble proteins were extracted from defatted palm pollen with both Tris-HCl buffer (pH 7.8) and Na-acetate buffer (pH 4.0). The allergenic glycoprotein was purified from the total extract to homogeneity with 0.4% yield by a combination of DEAE- and CM-cellulose, SP-HPLC, and gel filtration. The purified oil palm pollen glycoprotein with molecular mass of 31 kDa was recognized by the beta1-2 xylose specific antibody, suggesting this basic glycoprotein bears plant complex type N-glycan(s). The palm pollen basic glycoprotein, designated Ela g Bd 31 K, was recognized by IgE of palm pollinosis patients, suggesting Ela g Bd 31 K should be one of the palm pollen allergens. The preliminary structural analysis of N-glycans linked to glycoproteins of palm pollens showed that the antigenic N-glycans having alpha1-3 fucose and alpha1-2 xylose residues (GlcNAc(2 to approximately 0)Man3Xyl1Fuc(1 to approximately 0)GlcNAc2) actually occur on the palm pollen glycoproteins, in addition to the high-mannose type structures (Man(9 to approximately 5)GlcNAc2).
    Matched MeSH terms: Chromatography, Gel
  9. Fulltext Trace Level Detection of Bisphenol A Analogues and Parabens by LC-MS/MS in Human Plasma from Malaysians
    Wiraagni IA, Mohd MA, Rashid RA, Haron DEBM
    Biomed Res Int, 2020;2020:2581287.
    PMID: 32420332 DOI: 10.1155/2020/2581287
    In this study, a novel LC-MS/MS method was designed using a simple extraction procedure that was scientifically developed to capture the most relevant bisphenol A (BPA) analogues (BPB, BPF, BPS, and BPAF) and parabens (propylparaben, ethylparaben, butylparaben, and methylparaben) in human plasma. The LC-MS/MS method was validated using US FDA guidelines, and all validation requirements were satisfactory. This is the method that allows for the detection of plasma bisphenols and parabens in one run and is also the fastest BPA analogue and paraben detection technique for human plasma. The method was used to analyze samples from 150 healthy volunteers from Malaysia who enrolled in the study. No BPB was detected in any of the volunteers; however, 99.3% were positive for BPF. Only 24% and 10.7% of volunteers were positive for BPAF and BPS, respectively. A high percentage of volunteers were negative for propylparaben, ethylparaben, butylparaben, and methylparaben (56%, 68%, 86.7%, and 83.3%, respectively). These results suggest that persons in Malaysia are exposed to different BPA analogues and parabens, from both the daily use of products (cosmetic and plastic products) and the environment.
    Matched MeSH terms: Chromatography, Liquid
  10. Fulltext Nutrient and Sensory Metabolites Profiling of Averrhoa Carambola L. (Starfruit) in the Context of Its Origin and Ripening Stage by GC/MS and Chemometric Analysis
    Ramadan NS, Wessjohann LA, Mocan A, Vodnar DC, El-Sayed NH, El-Toumy SA, et al.
    Molecules, 2020 May 22;25(10).
    PMID: 32455938 DOI: 10.3390/molecules25102423
    Averrhoa carambola L. is a tropical tree with edible fruit that grows at different climatic conditions. Despite its nutritive value and reported health benefits, it is a controversial fruit owing to its rich oxalate content. The present study aimed at investigating aroma and nutrient primary metabolites distribution in A. carambola fruits grown in Indonesia, Malaysia (its endemic origin) versus Egypt, and at different ripening stages. Two techniques were employed to assess volatile and non-volatile metabolites including headspace solid-phase micro-extraction (HS-SPME) joined with gas chromatography coupled with mass-spectrometry (GC-MS) and GC-MS post silylation, respectively. Twenty-four volatiles were detected, with esters amounting for the major class of volatiles in Egyptian fruit at ca. 66%, with methyl caproate as the major component, distinguishing it from other origins. In contrast, aldehydes predominated tropically grown fruits with the ether myristicin found exclusively in these. Primary metabolites profiling led to the identification of 117 metabolites viz. sugars, polyols and organic acids. Fructose (38-48%) and glucose (21-25%) predominated sugar compositions in ripe fruits, whereas sorbitol was the major sugar alcohol (2.4-10.5%) in ripe fruits as well. Oxalic acid, an anti-nutrient with potential health risks, was the major organic acid detected in all the studied fruits (1.7-2.7%), except the Malaysian one (0.07%). It increases upon fruit ripening, including considerable amounts of volatile oxalate esters detected via SPME, and which must not be omitted in total oxalate determinations for safety assessments.
    Matched MeSH terms: Gas Chromatography-Mass Spectrometry
  11. Molecular description of familial defective APOB-100 in Malaysia
    Al-Khateeb AR, Mohd MS, Yusof Z, Zilfalil BA
    Biochem Genet, 2013 Oct;51(9-10):811-23.
    PMID: 23775634 DOI: 10.1007/s10528-013-9609-6
    Familial ligand-defective apolipoprotein B-100 is characterized by elevated plasma low-density lipoprotein levels and premature heart disease. This study aims to determine apolipoprotein B gene mutations among Malaysians with clinical diagnoses of familial hypercholesterolemia and to compare the phenotype of patients with apolipoprotein B gene mutations to those with a low-density lipoprotein receptor gene mutation. A group of 164 patients with a clinical diagnosis of familial hypercholesterolemia was analyzed. Amplicons in exon 26 and exon 29 of the apolipoprotein B gene were screened for genetic variants using denaturing gradient high-performance liquid chromatography; 10 variants were identified. Five novel mutations were detected (p.Gln2485Arg, p.Thr3526Ala, p.Glu3666Lys, p.Tyr4343CysfsX221, and p.Arg4297His). Those with familial defective apolipoprotein had a less severe phenotype than those with familial hypercholesterolemia. An apolipoprotein gene defect is present among Malaysian familial hypercholesterolemics. Those with both mutations show a more severe phenotype than those with one gene defect.
    Matched MeSH terms: Chromatography, Liquid
  12. Fulltext LC-MS Data set on the Malayan Deer (Cervus timorensis) Antler Velvet and its antibiofilm activity against Candida species: LC-MS Data set on the Malayan Deer (Cervus timorensis) Antler Velvet and its antibiofilm properties against Candida species
    Arzmi MH, John A, Rismayuddin NAR, Kenali NM, Darnis DS
    Data Brief, 2021 Apr;35:106769.
    PMID: 33537383 DOI: 10.1016/j.dib.2021.106769
    Deer antler velvet (DAV) has been traditionally used in Chinese medicine, including treatment on toothache [1]. Due to its rapid and regenerative capacity, deer antlers were proposed to be the good model for bone remodelling in mammals [2]. The data presented in this work is on the liquid chromatography and mass spectrometry (LC-MS) profile and bioactive potential of Malayan deer antler velvet (DAV) on different Candida species that has clinical importance. Aqueous extraction of DAV samples was subjected to Liquid chromatography quadrupole time of flight mass spectrometry (LC-QTOF-MS) profiling. Reverse phase (RP) separation was used due to the process extraction using water as a solvent to separate polar compound. The data was interpreted using Profile Analysis 2.1V. The DAV samples were also tested for the effect on the biofilm formation of seven Candida species in a 96 well plate [3]. The biofilms were developed for 72 h in aerobic environment. Following that, the biofilms biomass was determined using crystal violet assay.
    Matched MeSH terms: Chromatography, Liquid
  13. Immunomodulatory effects of Tinospora crispa extract and its major compounds on the immune functions of RAW 264.7 macrophages
    Ahmad W, Jantan I, Kumolosasi E, Haque MA, Bukhari SNA
    Int Immunopharmacol, 2018 Jul;60:141-151.
    PMID: 29730557 DOI: 10.1016/j.intimp.2018.04.046
    The in vivo immunomodulatory activities of Tinospora crispa have been reported but its molecular mechanisms underlying its immunomodulatory properties remains obscure and the active constituents contributing to the activities have not been identified. The present study was aimed to investigate the immunomodulatory effects of T. crispa extract (TCE) and its chemical constituents on RAW 264.7 macrophages. Six known compounds including magnoflorine and syringin were isolated by various chromatographic techniques from TCE and their structures were determined spectroscopically. A validated HPLC method was used to quantify magnoflorine and syringin in the extract. The immunomodulatory effects of TCE and its isolated compounds on chemotaxis, phagocytosis, production of inflammatory mediators including reactive oxygen species (ROS), nitric oxide (NO), prostaglandin E2 (PGE2) and pro-inflammatory cytokines which include tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and monocyte chemoattractant protein-1 (MCP-1) on macrophages were assessed. TCE increased the chemotaxis and phagocytic activity of macrophages and significantly enhanced the production of ROS, NO and pro-inflammatory cytokines. All alkaloids isolated, specifically magnoflorine showed remarkable inducing effects on the chemotaxis, phagocytic activity, ROS and NO productions and the secretions of IL-1β, TNF-α, IL6, PGE2 and MCP-1. In contrast, syringin potently reduced the chemotaxis, phagocytic activity, ROS and NO productions and secretions of IL-1β, TNF-α, IL6, PGE2 and MCP-1. TCE showed strong immunostimulant effects on various components of the immune system and these activities were possibly contributed mainly by the alkaloids specifically magnoflorine. TCE has potential to be developed as an effective natural immunostimulant for improvement of immune-related disorders.
    Matched MeSH terms: Chromatography, High Pressure Liquid
  14. Fulltext Over-Expression of the Pikh Gene with a CaMV 35S Promoter Leads to Improved Blast Disease (Magnaporthe oryzae) Tolerance in Rice
    Azizi P, Rafii MY, Abdullah SN, Hanafi MM, Maziah M, Sahebi M, et al.
    Front Plant Sci, 2016;7:773.
    PMID: 27379107 DOI: 10.3389/fpls.2016.00773
    Magnaporthe oryzae is a rice blast fungus and plant pathogen that causes a serious rice disease and, therefore, poses a threat to the world's second most important food security crop. Plant transformation technology has become an adaptable system for cultivar improvement and to functionally analyze genes in plants. The objective of this study was to determine the effects (through over-expressing and using the CaMV 35S promoter) of Pikh on MR219 resistance because it is a rice variety that is susceptible to the blast fungus pathotype P7.2. Thus, a full DNA and coding DNA sequence (CDS) of the Pikh gene, 3172 bp, and 1206 bp in length, were obtained through amplifying the gDNA and cDNA template from a PH9-resistant rice variety using a specific primer. Agrobacterium-mediated transformation technology was also used to introduce the Pikh gene into the MR219 callus. Subsequently, transgenic plants were evaluated from the DNA to protein stages using polymerase chain reaction (PCR), semi-quantitative RT-PCR, real-time quantitative PCR and high performance liquid chromatography (HPLC). Transgenic plants were also compared with a control using a real-time quantification technique (to quantify the pathogen population), and transgenic and control plants were challenged with the local most virulent M. oryzae pathotype, P7.2. Based on the results, the Pikh gene encodes a hydrophilic protein with 18 sheets, 4 helixes, and 21 coils. This protein contains 401 amino acids, among which the amino acid sequence from 1 to 376 is a non-cytoplasmic region, that from 377 to 397 is a transmembrane region, and that from 398 to 401 is a cytoplasmic region with no identified disordered regions. The Pikh gene was up-regulated in the transgenic plants compared with the control plants. The quantity of the amino acid leucine in the transgenic rice plants increased significantly from 17.131 in the wild-type to 47.865 mg g(-1) in transgenic plants. The M. oryzae population was constant at 31, 48, and 72 h after inoculation in transgenic plants, while it was increased in the inoculated control plants. This study successfully clarified that over-expression of the Pikh gene in transgenic plants can improve their blast resistance against the M. oryzae pathotype P7.2.
    Matched MeSH terms: Chromatography, High Pressure Liquid
  15. 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
  16. Transcriptomic and Proteomic Analyses of Nepenthes ampullaria and Nepenthes rafflesiana Reveal Parental Molecular Expression in the Pitchers of Their Hybrid, Nepenthes × hookeriana
    Zulkapli MM, Ab Ghani NS, Ting TY, Aizat WM, Goh HH
    Front Plant Sci, 2020;11:625507.
    PMID: 33552113 DOI: 10.3389/fpls.2020.625507
    Nepenthes is a genus comprising carnivorous tropical pitcher plants that have evolved trapping organs at the tip of their leaves for nutrient acquisition from insect trapping. Recent studies have applied proteomics approaches to identify proteins in the pitcher fluids for better understanding the carnivory mechanism, but protein identification is hindered by limited species-specific transcriptomes for Nepenthes. In this study, the proteomics informed by transcriptomics (PIT) approach was utilized to identify and compare proteins in the pitcher fluids of Nepenthes ampullaria, Nepenthes rafflesiana, and their hybrid Nepenthes × hookeriana through PacBio isoform sequencing (Iso-Seq) and liquid chromatography-mass spectrometry (LC-MS) proteomic profiling. We generated full-length transcriptomes from all three species of 80,791 consensus isoforms with an average length of 1,692 bp as a reference for protein identification. The comparative analysis found that transcripts and proteins identified in the hybrid N. × hookeriana were more resembling N. rafflesiana, both of which are insectivorous compared with omnivorous N. ampullaria that can derive nutrients from leaf litters. Previously reported hydrolytic proteins were detected, including proteases, glucanases, chitinases, phosphatases, nucleases, peroxidases, lipid transfer protein, thaumatin-like protein, pathogenesis-related protein, and disease resistance proteins. Many new proteins with diverse predicted functions were also identified, such as amylase, invertase, catalase, kinases, ligases, synthases, esterases, transferases, transporters, and transcription factors. Despite the discovery of a few unique enzymes in N. ampullaria, we found no strong evidence of adaptive evolution to produce endogenous enzymes for the breakdown of leaf litter. A more complete picture of digestive fluid protein composition in this study provides important insights on the molecular physiology of pitchers and carnivory mechanism of Nepenthes species with distinct dietary habits.
    Matched MeSH terms: Chromatography, Liquid
  17. Isolation and characterization of the thrombin-like enzyme from Cryptelytrops purpureomaculatus venom
    Tan NH
    Comp Biochem Physiol C Toxicol Pharmacol, 2010 Jan;151(1):131-6.
    PMID: 19770070 DOI: 10.1016/j.cbpc.2009.09.002
    A thrombin-like enzyme, purpurase, was purified from the Cryptelytrops purpureomaculatus (mangrove pit viper) venom using high performance ion-exchange and gel filtration chromatography. The purified sample (termed purpurase) yielded a homogeneous band in SDS-polyacrylamide gel electrophoresis with a molecular weight of 35,000. The N-terminal sequence of purpurase was determined to be VVGGDECNINDHRSLVRIF and is homologous to many other venom thrombin-like enzymes. Purpurase exhibits both arginine ester hydrolase and amidase activities. Kinetic studies using tripeptide chromogenic anilide substrates showed that purpurase is not fastidious towards its substrate. The clotting times of fibrinogen by purpurase were concentration dependent, with optimum clotting activity at 3mg fibronogen/mL. The clotting activity by purpurase was in the following decreasing order: cat fibrinogen>human fibrinogen>dog fibrinogen>goat fibrinogen>rabbit fibrinogen. Reversed-phase HPLC analysis of the products of action of purpurase on bovine fibrinogen showed that only fibrinopeptide A was released. Indirect ELISA studies showed that anti-purpurase cross-reacted strongly with venoms of most crotalid venoms, indicating the snake venom thrombin-like enzymes generally possess similar epitopes. In the more specific double-sandwich ELISA, however, anti-purpurase cross-reacted only with venoms of certain species of the Trimeresurus complex, and the results support the recent proposed taxonomy changes concerning the Trimeresurus complex.
    Matched MeSH terms: Chromatography, Gel; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Chromatography, Reverse-Phase
  18. Fulltext Ergogenic, anti-diabetic and antioxidant attributes of selected Malaysian herbs: characterisation of flavonoids and correlation of functional activities
    Sallehuddin, N. A., Azizah Abdul Hamid, Salleh, S. Z., Nazia Abdul Majid, Hani Hafeeza Halim, Nurul Shazini Ramli, et al.
    International Food Research Journal, 2020;27(1):197-207.
    MyJurnal
    In the present work, aqueous ethanolic (60% ethanol) extracts from selected Malaysian herbs
    including Murraya koenigii L. Spreng, Lawsonia inermis L., Cosmos caudatus Kunth, Piper
    betle L., and P. sarmentosum Roxb. were evaluated for their ergogenic, anti-diabetic and
    antioxidant potentials. Results showed that the analysed herbs had ergogenic property and
    were able to activate 5'AMP-activated protein kinase (AMPK) in a concentration dependant
    manner. The highest AMPK activation was exhibited by M. koenigii extract which showed no
    significant (p > 0.05) difference with green tea (positive control). For anti-diabetic potential,
    the highest α-glucosidase inhibition was exhibited by M. koenigii extract with IC50 of 43.35
    ± 7.5 µg/mL, which was higher than acarbose (positive control). The determinations of free
    radical scavenging activity and total phenolics content (TPC) indicated that the analysed herbs
    had good antioxidant activity. However, C. caudatus extract showed superior antioxidant
    activity with IC50 against free radical and TPC of 21.12 ± 3.20 µg/mL and 221.61 ± 7.49 mg
    GAE/g, respectively. RP-HPLC analysis established the presence of flavonoids in the herbs
    wherein L. inermis contained the highest flavonoid (catechin, epicatechin, naringin and rutin)
    content (668.87 mg/kg of extract). Correlations between the analyses were conducted, and
    revealed incoherent trends. Overall, M. koenigii was noted to be the most potent herb for
    enhancement of AMPK activity and α-glucosidase inhibition but exhibited moderate antioxidant activity. These results revealed that the selected herbs could be potential sources of
    natural ergogenic and anti-diabetic/antioxidant agents due to their rich profile of phenolics.
    Further analysis in vivo should be carried out to further elucidate the mechanism of actions of
    these herbs as ergogenic aids and anti-diabetic/antioxidant agents.
    Matched MeSH terms: Chromatography, High Pressure Liquid
  19. Fulltext Characterization of phenolics and biological activities of different solvent extracts from Withania somnifera fruit
    Tabassam, Q., Mehmood, T., Anwar, F., Saari, N., Qadir, R.
    International Food Research Journal, 2020;27(5):915-924.
    MyJurnal
    The present work studies the profiling of phenolic bioactive and in vitro biological (anticancer, antioxidant, and antimicrobial) activities of different solvent extracts from Withania
    somnifera fruit. Anticancer activity was performed using potato-disc assay and Agrobacterium tumefaciens. While antibacterial and antifungal evaluation was done by using disc diffusion method against bacterial (Staphylococcus aureus, S. epidermidis, Escherichia coli, and
    Klebsiella pneumonia) and fungal (Aspergillus flavus and Fusarium oxysporum) strains.
    Among different extraction solvents used, n-hexane extract exhibited the highest inhibition of
    tumour initiation (64%), whereas ethyl acetate (15%) was the lowest by using potato-disc
    assay. Highest total phenolic and total flavonoid contents were noted for methanolic (69.10
    GAE mg/g DW%) and n-hexane (29.45 CE mg/g DW%) extracts, respectively. For antioxidant potential, 2,2,1-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging (IC50) and reducing power EC50 were noted to be superior (0.6 and 2.0 mg/mL, respectively) for n-hexane
    extract. All the tested extracts showed considerable antibacterial and antifungal activity with
    the highest growth inhibition zones for K. pneumoniae (31.70 mm) and A. flavus (27.09 mm)
    were shown by n-hexane extract. High Performance Liquid Chromatographic (HPLC) analysis of individual phenolics (gallic acid, 2,288.48 mg/kg) indicated the highest contents of these
    compounds in n-hexane extract, which might explain the potent biological activities of this
    extract. Our findings revealed that the bioactive present in the tested fruit had significant
    potential as anticancer, antibacterial, and antifungal agents. Further studies are needed to
    elucidate the mechanism of actions of isolated bioactive against specific diseases such as
    cancer, especially in the case of n-hexane fraction.
    Matched MeSH terms: Chromatography, High Pressure Liquid
  20. Fulltext Osmotic stress in banana is relieved by exogenous nitric oxide
    Mohd Amnan MA, Pua TL, Lau SE, Tan BC, Yamaguchi H, Hitachi K, et al.
    PeerJ, 2021;9:e10879.
    PMID: 33614294 DOI: 10.7717/peerj.10879
    Drought is one of the severe environmental stresses threatening agriculture around the globe. Nitric oxide plays diverse roles in plant growth and defensive responses. Despite a few studies supporting the role of nitric oxide in plants under drought responses, little is known about its pivotal molecular amendment in the regulation of stress signaling. In this study, a label-free nano-liquid chromatography-mass spectrometry approach was used to determine the effects of sodium nitroprusside (SNP) on polyethylene glycol (PEG)-induced osmotic stress in banana roots. Plant treatment with SNP improved plant growth and reduced the percentage of yellow leaves. A total of 30 and 90 proteins were differentially identified in PEG+SNP against PEG and PEG+SNP against the control, respectively. The majority of proteins differing between them were related to carbohydrate and energy metabolisms. Antioxidant enzyme activities, such as superoxide dismutase and ascorbate peroxidase, decreased in SNP-treated banana roots compared to PEG-treated banana. These results suggest that the nitric oxide-induced osmotic stress tolerance could be associated with improved carbohydrate and energy metabolism capability in higher plants.
    Matched MeSH terms: Chromatography
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