Papers by Pardhasaradhi Mathi
Handling the Out of Specification (Oos) Results in Pharmaceutical/ Biopharmaceutical Industries for Compliance – a Holistic Review
Handling of written operational procedures is mandatory in pharma/biopharma industries in view of... more Handling of written operational procedures is mandatory in pharma/biopharma industries in view of, to carry out activities for compliance. Out of all these procedures, handling of Out Of Specification (OOS) results attracted the attention of pharma/biopharma industrialists. To investigate the OOS result, an operating procedure should be in place. If any result found out of specification that should be investigated. The source of the OOS result should be identified either as an aberration of the measurement process i.e., laboratory error or an aberration of the manufacturing process. Phase I investigation (laboratory investigation) should include an initial assessment of the accuracy of the laboratory's data which may occur from instrument, reagent, reference standard, environment condition, test method, analyst and calculation within laboratory. Phase II investigation (full-scale OOS investigation) consist of a production/ user department process review and/or additional laboratory investigation. The written procedure should give guidance on the number of retests/ resamples permitted. After impact assessment of OOS on product/ process, appropriate CAPA to be initiated to avoid reoccurrences. This is the first reported and ideal form of handling the out of specification results in different pharmaceutical/ biopharmaceutical industries across the world.
Analysis of Phytochemicals, Antibacterial and Antioxidant activities ofMoringa oleifera Lam. Leaf extract- an in vitro study
International Journal of Drug Development and Research, 2014
The leading important things about utilizing plant-derived medication are relatively less dangero... more The leading important things about utilizing plant-derived medication are relatively less dangerous than artificial drugs and provide deep restorative benefits. In this regard, Moringa oleifera plant was evaluated for its nutritional effects. The phytochemical study, antibacterial activity and the in vitro antioxidant activity of aqueous, chloroform and petroleum ether extracts of Moringa oleifera leaves were investigated. Phytochemical analysis revealed the presence of alkaloids, flavonoids, steroids, tannins, saponins and glycosides as major components. The Petroleum ether extracts of Moringa oleifera contains the high content of bioactive compounds such as phenol and flavonoids. The extracts were screened for in vitro antibacterial potential against enteric pathogenic bacteria such as Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella typhi, E. coli and Vibrio cholera by disc diffusion method. Moringa oleifera showed potent antibacterial activity against several gram negative and gram positive bacteria. Antioxidants are shown to play an important position in your body immune system next to Reactive oxygen species (ROS). The enzymatic and non-enzymatic activities of SOD, Catalase, Peroxidase, Vitamin C, Carotenoids, and reduced Glutathione were high in petroleum ether extracts of Moringa oleifera. It shows high scavenging activity against DPPH, Nitrous Oxide and Hydroxyl Radical scavenging assays. As a result, Moringa oleifera possesses a good antioxidant, it has a scavenging property against ROS and also it has good antibacterial properties. Thus, Moringa oleifera can be used to synthesize a new drug preparation against various diseases responsible for severe illness.
Journal of Molecular Graphics & Modelling, Jul 1, 2015
Staphylococcus aureus sortase A is an attractive target of Gram-positive bacteria that plays a cr... more Staphylococcus aureus sortase A is an attractive target of Gram-positive bacteria that plays a crucial role in anchoring of surface proteins to peptidoglycan present in bacterial cell wall. Inhibiting sortase A is an elementary and essential effort in preventing the pathogenesis. In this context, in silico virtual screening of in-house database was performed using ligand based pharmacophore model as a filter. The developed pharmacophore model AAHR 11 consists of two acceptors, one hydrophobic and one ring aromatic feature. Top ranked molecule KKR1 was docked into the active site of the target. After profound analysis, it was analyzed and optimized based on the observations from its binding pose orientation. Upgraded version of KKR1 was KKR2 and has improved docking score, binding interactions and best fit in the binding pocket. KKR1 along with KKR2 were further validated using 100 ns molecular dynamic studies. Both KKR1 and KKR2 contain Indole-thiazolidine moiety and were synthesized. The disk diffusion assay has good initial results (ZI of KKR1, KKR2 were 24, 38 mm at 10 g/mL and ZI of Ampicillin was 22 at 10 g/mL) and calculated MICs of the molecules (KKR1 5.56 ± 0.28 g/mL, KKR2 1.32 ± 0.12 g/mL, Ampicillin 8 ± 1.1 g/mL) were in good agreement with standard drug Ampicillin. KKR1 has shown IC 50 of 1.23 ± 0.14 M whereas the optimized lead molecule KKR2 show IC 50 of 0.008 ± 0.07 M. Results from in silico were validated by in vitro studies and proved that indole-thiazolidine molecules would be useful for future development as lead molecules against S. aureus sortase A.
Analysis of Phytochemicals, Antibacterial and Antioxidant activities ofMoringa oleifera Lam. Leaf extract- an in vitro study
International Journal of Drug Development and Research, 2014
The leading important things about utilizing plant-derived medication are relatively less dangero... more The leading important things about utilizing plant-derived medication are relatively less dangerous than artificial drugs and provide deep restorative benefits. In this regard, Moringa oleifera plant was evaluated for its nutritional effects. The phytochemical study, antibacterial activity and the in vitro antioxidant activity of aqueous, chloroform and petroleum ether extracts of Moringa oleifera leaves were investigated. Phytochemical analysis revealed the presence of alkaloids, flavonoids, steroids, tannins, saponins and glycosides as major components. The Petroleum ether extracts of Moringa oleifera contains the high content of bioactive compounds such as phenol and flavonoids. The extracts were screened for in vitro antibacterial potential against enteric pathogenic bacteria such as Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella typhi, E. coli and Vibrio cholera by disc diffusion method. Moringa oleifera showed potent antibacterial activity against several gram negat...
Entada pursaetha DC is a perennial woody climber belonging to family Fabaceae and widely used in ... more Entada pursaetha DC is a perennial woody climber belonging to family Fabaceae and widely used in traditional medicine especially in curing joint pains. The present study was designed to validate the antiinflammatory activity of the ethanol seed extract. E. pursaetha was validated for its antiinflammatory activity using macrophage RAW 264.7 cells, further bioassay guided fractionation for isolating and identifying the potent fraction against COX-2 as well as the biochemical compositions from the active fractions using GC-MS analysis. The results showed significant down regulation of nitric oxide (NO) production in RAW cells IC = 130 ± 1.54 µg/ml without altering the normal growth of cells. Fractionation yielded 25 fractions from which 50 Methanol (100%), Chloroform: Methanol (7:3), Ethyl acetate: Chloroform (9:1) and Hexane: Ethyl acetate (7:3) were active fractions against COX-2 enzyme. However, Chloroform: Methanol (7:3) has potent inhibition IC = 2.9 µg/ml was observed. A total of...
Sophora interrupta Bedd, was a common woody perennial herb native to India. Reports suggest that ... more Sophora interrupta Bedd, was a common woody perennial herb native to India. Reports suggest that the plant possesses numerous health promoting benefits such as anticancer, antiinflammatory, antimicrobial..
African Journal of Biotechnology, 2014
An attempt was made to study the somatic embryogenesis and plant regeneration from the in vitro l... more An attempt was made to study the somatic embryogenesis and plant regeneration from the in vitro leaf explants of Rumex vesicarius L. a renowned medicinal plant, which belongs to polygonaceae family. Effective in vitro regeneration of R. vesicarius was achieved via young leaf derived somatic embryo cultures. Embryogenic callus was induced from leaf explants on Schenk and Hildebrandt (SH) medium supplemented with various concentrations of 2,4-dichlorophenoxy acetic acid (2,4-D) (0.5 to 3.0 mg/l) along with Kinetin (Kn) (0.5 mg/l). High frequency of somatic embryogenesis was effective on SH medium with 2, 4-D (2.5 mg/l) + Kn (0.5 mg/l) from leaf explants. Secondary somatic embryogenesis was also observed when primary somatic embryos were subculture on the same somatic embryo induction medium. Well developed cotyledonary shaped embryos regenerate 80% of shoots on media containing 2,4-D 0.5 mg/l + 2.0 mg/l BA. The regenerated shoots transferred to rooting media containing Indole- 3- buty...
Pharmacognosy Magazine, 2019
Background: Biochanin-A and Piceatannol are phytochemical constituents extracted from Sophora int... more Background: Biochanin-A and Piceatannol are phytochemical constituents extracted from Sophora interrupta. Although both the compounds were isolated from a single plant, these compounds were not compared against anticancer activity. Objective: A systematic comparative analysis of biochanin-A, piceatannol, and resveratrol was performed to investigate cancer cell viability, motility, metabolic changes in Michigan Cancer Foundation-7 breast cancer cells, and structure compound interaction with the vascular endothelial growth factor (VEGF) receptors were studied. Materials and Methods: Cancer cell viability was studied using 3 (4, 5 dimethyl thiazol 2yl) 2, 5 diphenyltetrazo- lium bromide and acridine orange (AO)/ethidium bromide (EtBr) assay. The wound-healing assay was performed by measuring cell migration from the scratch area. Metabolic changes of the compounds in culture conditions were recorded using Fourier-transform infrared (FT-IR) spectroscopy. Molecular docking and dynamic simulations were performed using Schrödinger software. Results: Our results showed that the half-maximal growth inhibitory concentration for biochanin-A is 150 μM/ml and piceatannol and resveratrol showed 150 μM/ml, which is evident from the uptake of AO and EtBr dyes by live/dead cells. Moreover, drug-treated cells were unable to fill the cleared area from the scratch area, which suggests that all compounds effectively inhibit cell motility. FT-IR fingerprint showed a marked difference in the percentage of transition and dynamic structural changes between untreated and treated samples. Strong hydrogen-bond interaction with VEGF receptor-1 (VEGFR1) and VEGFR2 proteins and their interactions were stable throughout the simulation period. Moreover, these compounds inhibited sprouting of a new blood vessel from the chicken aorta and microvessels formation in the in ovo chorioallantoic membrane assay. Conclusion: Taken together, we conclude that anticancer and anti-angiogenic activity, structure-function relationship of biochanin-A is like well-known anticancer compound resveratrol and its metabolic product piceatannol in breast cancer cells.
Journal of Molecular Modeling, 2016
Sophora interrupta Bedd, (Fabaceae) is used in Indian folk medicine to treat cancer. Angiogenesis... more Sophora interrupta Bedd, (Fabaceae) is used in Indian folk medicine to treat cancer. Angiogenesis is one of the crucial characteristics of cancer metastasis and is regulated by vascular endothelial growth factor (VEGF). In this study, we examined the antiangiogenic properties of the root ethyl acetate extract of Sophora interrupta by various methods. In vitro antioxidant activity (100-600 μg/ml) of S. interrupta ethyl acetate (SEA) extract was evaluated by DPPH and ABTS, anti-inflammatory activity (50, 100 and 150 μg/ml) by estimating nitric oxide (NO) levels, anti-angiogenic activity (200 and 500 μg/ml) was validated by chorio allantoic membrane (CAM) assay and in silico molecular dynamic (MD) simulations analyses (25 ns) were performed to identify the anti-angiogenic compounds extracted from root extract. The antioxidative activity of SEA extract at IC 50 (200 ± 0.6 μg/mL) is equal to that of ascorbic acid at IC 50 (50 ± 0.6 μg/mL), and the anti-inflammatory activity of SEA extract at IC 50 (150 ± 0.2 μg/mL) was inhibited significantly by nitric oxide (NO) production. The SEA extract significantly reduced the sprouting of new blood vessels at ID 50 500 ± 0.13 μg/mL in the CAM assay. Gas chromatography-mass spectrometry analysis of the SEA extract detected 34 secondary metabolites, of which 6a,12a-dihydro-6H-(1,3)dioxolo(5,6)benzofuro(3,2-c)chromen-3-ol (maackiain) and funiculosin formed strong hydrogen bond interactions with Lys 920, Thr 916 and Cys 919 (2H), as well as Glu 917 of VEGFR2, and these interactions were similar to those of the anti-angiogenic compound axitinib. Significant findings in all the assays performed indicate that SEA extract has potential anti-angiogenic compounds that may interfere with VEGF-induced cancer malignancy.
Characterization of vitamin–cisplatin-loaded chitosan nano-particles for chemoprevention and cancer fatigue
Journal of Microencapsulation, 2015
Vitamins have been shown to reduce chemotherapy-related fatigue (CRF) by conserving energy loss b... more Vitamins have been shown to reduce chemotherapy-related fatigue (CRF) by conserving energy loss both during and after cancer treatment. However, it remains unknown whether this reduction of fatigue interferes with the cancer drugs or alters the effectiveness of these agents. The objective was to synthesize vitamin-cisplatin-loaded chitosan nano-particles for chemoprevention and cancer fatigue. Multi-vitamin (C, D3, and B12)-cisplatin composite nano-formulation called NanoCisVital (NCV) to overcome CRF. The interactions between vitamins and NCV were characterized using scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, and a particle size analyser. The chemo-preventive activity was performed by in vitro bio assays. SEM analysis showed spherical shape and the size is < 225 nm. NCV inhibited the proliferating yeast cells as well as denaturation of bovine serum albumin, and it also reduced the sprouting of new blood vessels in dose-dependent manner. Collectively, these results demonstrate that the NCV particles can be used to reduce CRF without much affecting the anti-cancer properties of cisplatin.
Asian Journal of Pharmaceutical and Clinical Research, Jan 7, 2015
Climatic conditions are fundamental to life on earth and their destruction or disturbance by dire... more Climatic conditions are fundamental to life on earth and their destruction or disturbance by direct or indirect human activities is the greatest threat to human health. Human life on earth is directly associated with environmental factors such as "air" and "water." Pollution of air by toxic substances by the activities of mankind has shown to cause serious health issues, including damage to the immune, respiratory, neurological, and reproductive systems, and other health problems like cancer. Water intended for human consumption should be free from microorganisms and toxic substances. The impact and drastic effects of chlorinated water and their impact on human health are poorly studied. Chlorination is an inexpensive and effective process for disinfecting water worldwide. During the disinfection, the chlorine generates hundreds of different by-products called chlorination by-products such as trihalomethanes and halo acetic acids (HAA's) at low levels. In this article we address the action of two HAA's, tri-and di-chloroacetic acid and their impact on the progression of cancer, respiratory disorder, and neurological anomalies.
Journal of Molecular Graphics and Modelling, 2015
Staphylococcus aureus sortase A is an attractive target of Gram-positive bacteria that plays a cr... more Staphylococcus aureus sortase A is an attractive target of Gram-positive bacteria that plays a crucial role in anchoring of surface proteins to peptidoglycan present in bacterial cell wall. Inhibiting sortase A is an elementary and essential effort in preventing the pathogenesis. In this context, in silico virtual screening of in-house database was performed using ligand based pharmacophore model as a filter. The developed pharmacophore model AAHR 11 consists of two acceptors, one hydrophobic and one ring aromatic feature. Top ranked molecule KKR1 was docked into the active site of the target. After profound analysis, it was analyzed and optimized based on the observations from its binding pose orientation. Upgraded version of KKR1 was KKR2 and has improved docking score, binding interactions and best fit in the binding pocket. KKR1 along with KKR2 were further validated using 100 ns molecular dynamic studies. Both KKR1 and KKR2 contain Indole-thiazolidine moiety and were synthesized. The disk diffusion assay has good initial results (ZI of KKR1, KKR2 were 24, 38 mm at 10 g/mL and ZI of Ampicillin was 22 at 10 g/mL) and calculated MICs of the molecules (KKR1 5.56 ± 0.28 g/mL, KKR2 1.32 ± 0.12 g/mL, Ampicillin 8 ± 1.1 g/mL) were in good agreement with standard drug Ampicillin. KKR1 has shown IC 50 of 1.23 ± 0.14 M whereas the optimized lead molecule KKR2 show IC 50 of 0.008 ± 0.07 M. Results from in silico were validated by in vitro studies and proved that indole-thiazolidine molecules would be useful for future development as lead molecules against S. aureus sortase A.
Bioinformation, 2013
Follicle stimulating hormone (FSH) is a glycoprotein secreted by gonadotrophs of the anterior pit... more Follicle stimulating hormone (FSH) is a glycoprotein secreted by gonadotrophs of the anterior pituitary gland that regulates reproduction in mammals. FSH targets its receptor (FSHR) expressed only on grannulosa cells and induce the maturation of ovarian follicles in females. The levels of both FSH and FSHR rise until the middle of estrus cycle and then falls on level at the time of ovulation. It is associated with stimulated sertoli cell proliferation in testes and supports spermatogenesis in males. The interaction between the polypeptide FSH hormone and its corresponding receptor is highly selective. Therefore, it is of interest to inhibit FSH in the context of infertility. The structure of FSH (PDB ID: 1XWD) is screened using molecular docking techniques against the ZINC database (a database of 2.7 million compounds) with reference to known standard compounds. This exercise identifies compounds with better binding and ADMET (Absorption, Digestion, Metabolism, Excretion and Toxicity) properties compared to known standard compounds. These observations find application for the consideration of such compounds for further validation towards inhibiting the FSH.
Bioinformation, 2014
Sophora interrupta belongs to the family of Fabaceae and the species in this genus have a diverse... more Sophora interrupta belongs to the family of Fabaceae and the species in this genus have a diverse medicinal importance as a folk medicine for preventing many ailments including cancer. In order to evaluate the anticancer activity of S.interrupta, we have performed in vitro anti-oxidant, anti-inflammatory, anti-proliferative, and cell based anticancer activity in MCF-7 and PC-3 cell lines. Secondary metabolites of S.interrupta were used to identify anticancer compounds using Open Eye software. The antioxidant activity of the S.interrupta root ethylacetate (SEA) extract at 100 μg/ml is equal to that of ascorbic acid at 50 μg/ml. The antiinflammatory activity of SEA is half of that of diclofenac at 50 μg/ml. Anticancer activity was detected by measuring the mitochondrial dehydrogenase activity (MTT assay). The half maximal inhibitory concentrations (IC50) for MCF-7 and PC-3 cell lines are 250 and 700 µg/ml respectively. This was supported by the morphological changes such as membrane blebbing, cell detachment and rounded cell morphology when compared to the parental cells. In addition, we observed few green cells (live) over red cells (dead) based on the uptake of acridine orange and ethidium bromide dyes. Kaempferol-3-O-b-D-glucopyranoside, a Secondary metabolite of S.interrupta form 6 hydrogen bond interactions with Arg 202, Gln 207, Gly 227, Gly 229, Thr 231 and Ala 232 human DEAD box RNA helicase, DDX3 protein and is equivalent to crystal structure of adenosine mono phosphate to DDX3. Overall, it suggests that the SEA extract has anticancer compounds, and it can be used to enhance death receptor mediated cancer cell death.
Bioinformation, 2014
Cyperus scariosus (R.Br) belongs to the family Cyperaceae and it has a diverse medicinal importan... more Cyperus scariosus (R.Br) belongs to the family Cyperaceae and it has a diverse medicinal importance. To identify human cyclooxegenase-2 (COX-2) inhibitors from C. scariosus, the rhizome powder was exhaustively extracted with various solvents based on the increasing polarity. Based on the presence and absence of secondary metabolites, we have selected the methanolic extract to evaluate the anti-oxidant and anti-inflammatory activity. The same extract was further subjected to gas chromatography-mass spectroscopy (GC-MS) analysis to identify the active compounds. Binding affinities of these compounds towards anti-inflammatory protein COX-2 were analyzed using molecular docking interaction studies. Phytochemical analysis showed that methanol extract is positive for all secondary metabolites. The antioxidant activity of the C. scariosus rhizomes methanolic extract (CSRME) is half to that of ascorbic acid at 50 μg/ml. The anti-inflammatory activity of CSRME is higher than that of diclofenac sodium salt at high concentration, which is evident from the dose dependent inhibition of bovine serum albumin denaturation at 40 μg/ml-5 mg/ml. GC-MS analysis showed the presence of nine compounds, among all N-methyl-1-adamantaneacetamide and 1,5,diphenyl-2H-1,2,4triazine form a hydrogen bond interactions with Ser-530 and Tyr-385 respectively and found similar interactions with crystal structure of diclofenac bound COX-2 protein. Benzene-1, 2-diol, 4-(4-bromo-3 chlorophenyl iminomethyl forms hydrogen bond interactions with Thr-199 and Thr-200 as similar to crystallized COX-2 protein with valdecoxib. Collectively our results suggest that CSRME contains medicinally important anti-inflammatory compounds and this justifies the use of this plant as a folklore medicine for preventing inflammation associated disorders.
Journal of Molecular Modeling, Sep 28, 2016
Sophora interrupta Bedd, (Fabaceae) is used in Indian folk medicine to treat cancer. Angiogenesis... more Sophora interrupta Bedd, (Fabaceae) is used in Indian folk medicine to treat cancer. Angiogenesis is one of the crucial characteristics of cancer metastasis and is regulated by vascular endothelial growth factor (VEGF). In this study, we examined the antiangiogenic properties of the root ethyl acetate extract of Sophora interrupta by various methods. In vitro antioxidant activity (100-600 μg/ml) of S. interrupta ethyl acetate (SEA) extract was evaluated by DPPH and ABTS, anti-inflammatory activity (50, 100 and 150 μg/ml) by estimating nitric oxide (NO) levels, anti-angiogenic activity (200 and 500 μg/ml) was validated by chorio allantoic membrane (CAM) assay and in silico molecular dynamic (MD) simulations analyses (25 ns) were performed to identify the anti-angiogenic compounds extracted from root extract. The antioxidative activity of SEA extract at IC 50 (200 ± 0.6 μg/mL) is equal to that of ascorbic acid at IC 50 (50 ± 0.6 μg/mL), and the anti-inflammatory activity of SEA extract at IC 50 (150 ± 0.2 μg/mL) was inhibited significantly by nitric oxide (NO) production. The SEA extract significantly reduced the sprouting of new blood vessels at ID 50 500 ± 0.13 μg/mL in the CAM assay. Gas chromatography-mass spectrometry analysis of the SEA extract detected 34 secondary metabolites, of which 6a,12a-dihydro-6H-(1,3)dioxolo(5,6)benzofuro(3,2-c)chromen-3-ol (maackiain) and funiculosin formed strong hydrogen bond interactions with Lys 920, Thr 916 and Cys 919 (2H), as well as Glu 917 of VEGFR2, and these interactions were similar to those of the anti-angiogenic compound axitinib. Significant findings in all the assays performed indicate that SEA extract has potential anti-angiogenic compounds that may interfere with VEGF-induced cancer malignancy.
Chemical Data Collections, Aug 1, 2018
Briefly describe the contentof thisdata article. If data is supplied with the data article, pleas... more Briefly describe the contentof thisdata article. If data is supplied with the data article, please explicitly state this or indicate the reference number and repository name for the data stored elsewhere. If this article is related to a published full research article, please also directly mention the article in the abstract] Novel molecules were designed against Staphylococcus aureus sortase A, an imperative and vital target involved in the bacterial virulence. Structure guided de novo designing was performed using different chemical fragments. These fragments were extended based on the requirements of the binding cavity. All the compounds in its binding orientation has formed good interaction network with amino acids HIS 120 and ARG 197. Molecules designed contain indole-chroman and indolin-chroman moieties and their docking scores were ranging from-6.92 to-5.67. Designed molecules were selected for synthesis and tested in vitro against Staphylococcus aureus sortase A using Fluorescence resonance energy transfer assay. The highest active molecule KK4 ((Z)-3-((5-nitro-1H-indol-3-yl) methylene) chroman-2,4-dione)
Identification of human cyclooxegenase-2 inhibitors from Cyperus scariosus (R.Br) rhizomes
Bioinformation, Oct 30, 2014
DOAJ (DOAJ: Directory of Open Access Journals), 2015
Background: Sophora belongs to the family of Fabaceae and the species in this genus are currently... more Background: Sophora belongs to the family of Fabaceae and the species in this genus are currently used as a folklore medicine for preventing a variety of ailments including cancer. Our aim was to identify and validate an anticancer compound from Sophora interrupta using multi-spectroscopic, anticancer screening, and molecular docking approach. Methods: The cytotoxicity of the various solvent extracts, petroleum ether, n-butanol, and ethyl acetate (EtOAc) of the S. interrupta root powder was evaluated in a breast cancer cell lines (MCF-7). The extract that had anticancer activity was subjected to column chromatography based on the polarity of the solvents. The anticancer activity of the elution fractions was validated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The isolated metabolite fraction with anticancer activity was run through a C18 column isocratic and gradient high-performance liquid chromatography (HPLC). The structure of the isolated compound was characterized using 1 H nuclear magnetic resonance (NMR), 13 C-NMR, Fourier transform infrared spectroscopy, and liquid chromatography-mass spectrometer methods. Results: The crude EtAOc extract effectively inhibited the proliferation of MCF-7 cells. The column eluted chloroform and EtOAc (4:6) fraction of the EtOAc extract showed significant anticancer activity in the MCF-7 cells compared with normal mesenchymal stem cells. This fraction showed three major peaks in the HPLC chromatogram and the first major peak with a retention time (RT) of 7.153 was purified using preparative-HPLC. The structure of the compound is a piceatannol, which is a metabolic product of resveratrol. Piceatannol formed direct two hydrogen bond interactions between Cys912 (2H), and Glu878 of vascular endothelial growth factor receptor 1 (VEGFR1) with a glide-score (G-score) of −10.193, and two hydrogen bond interactions between Cys919, and Asp1046 of VEGFR2, with a G-score of −8.359. The structure is similar to that of the crystallized protein for VEGFR1 and R2. Conclusions: Piceatannol is a secondary metabolite of S. interrupta that has anticancer activity. Moreover, piceatannol has been isolated for the first time from S. interrupta.
Competitive metal‐binding stoichiometry between calcium and strontium by cell wall proteins of <i>Neurospora crassa</i>
Journal of Basic Microbiology, Mar 29, 2022
Cellwall proteins from Neurospora crassa were isolated and evaluated to demonstrate their metal a... more Cellwall proteins from Neurospora crassa were isolated and evaluated to demonstrate their metal ability to bind Ca2+ /Sr2+ by loading the solubilized protein fraction on to IMAC column pre-equilibrated with Ca2+ /Sr2+ . The SDS-PAGE analysis IMAC elutant, revealed ~18 proteins with a similarity in the proteome pattern of Ca2+ /Sr2+ fractions. DEAE chromatography showed 5 proteins in common in binding to Ca2+ and Sr2+ , were subjected to N-terminal sequencing. The sequence analysis were studied for determination of metal binding site prediction by CHED software indicates that all the five were found to have high affinity towards Ca2+ . From these 5, two were randomly selected and denoted as CWP-A (possess 5 Ca binding sites of 6 metal binding sites) and CWP-B (possess 6 binding sites of 8 metal binding sites). They were selected for further characterization studies to determine their Ca2+ bound Sr2+ binding properties. Surprisingly, these proteins were able to bind Sr2+ ions (29 μ moles) with equal affinity as to Ca2+ ions (42 μmoles) by means of direct binding, and/or by displacing calcium as observed in metal dependent proteolytic protection, fluorescence based metal exchange assays and molecular simulation studies. From the results, we demonstrate for the first time, that there is a stoichiometry between Ca2+ (an essential macro elemental metal ion) and Sr2+ ions (a non-essential element for which no reported metabolic activity is reported) for the metal binding sites on cell wall proteins. This stoichiometry could be due to similar atomic dimensions and metal-protein structure stabilizing properties of Sr2+ compared to Ca2+ . This article is protected by copyright. All rights reserved.
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Papers by Pardhasaradhi Mathi