Publications by authors named "Rumana Ahmad"

31 Publications

Phytochemicals from Ajwa dates pulp extract induce apoptosis in human triple-negative breast cancer by inhibiting AKT/mTOR pathway and modulating Bcl-2 family proteins.

Sci Rep 2021 May 14;11(1):10322. Epub 2021 May 14.

Department of Biochemistry, Era's Lucknow Medical College and Hospital, Era University, Lucknow, 226003, India.

Ajwa dates (Phoenix dactylifera L.) have been described in traditional and alternative medicine to provide several health benefits, but their mechanism of apoptosis induction against human triple-negative breast cancer MDA-MB-231 cells remains to be investigated. In this study, we analyzed the phytoconstituents in ethanolic Ajwa Dates Pulp Extract (ADPE) by liquid chromatography-mass spectrometry (LC-MS) and investigated anticancer effects against MDA-MB-231 cells. LC-MS analysis revealed that ADPE contained phytocomponents belonging to classes such as carbohydrates, phenolics, flavonoids and terpenoids. MTT assay demonstrated statistically significant dose- and time-dependent inhibition of MDA-MB-231 cells with IC values of 17.45 and 16.67 mg/mL at 24 and 48 h, respectively. Hoechst 33342 dye and DNA fragmentation data showed apoptotic cell death while AO/PI and Annexin V-FITC data revealed cells in late apoptosis at higher doses of ADPE. More importantly, ADPE prompted reactive oxygen species (ROS) induced alterations in mitochondrial membrane potential (MMP) in ADPE treated MDA-MB-231 cells. Cell cycle analysis demonstrated that ADPE induced cell arrest in S and G2/M checkpoints. ADPE upregulated the p53, Bax and cleaved caspase-3, thereby leading to the downregulation of Bcl-2 and AKT/mTOR pathway. ADPE did not show any significant toxicity on normal human peripheral blood mononuclear cells which suggests its safe application to biological systems under study. Thus, ADPE has the potential to be used as an adjunct to the mainline of treatment against breast cancer.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41598-021-89420-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8121835PMC
May 2021

Flavonol morin targets host ACE2, IMP-α, PARP-1 and viral proteins of SARS-CoV-2, SARS-CoV and MERS-CoV critical for infection and survival: a computational analysis.

J Biomol Struct Dyn 2021 Feb 1:1-32. Epub 2021 Feb 1.

Department of Orthopedics, Era's Lucknow Medical College and Hospital, Era University, Lucknow, UP, India.

A sudden outbreak of a novel coronavirus SARS-CoV-2 in 2019 has now emerged as a pandemic threatening to efface the existence of mankind. In absence of any valid and appropriate vaccines to combat this newly evolved agent, there is need of novel resource molecules for treatment and prophylaxis. To this effect, flavonol morin which is found in fruits, vegetables and various medicinal herbs has been evaluated for its antiviral potential in the present study. PASS analysis of morin reference antiviral drugs baricitinib, remdesivir and hydroxychloroquine revealed that morin displayed no violations of Lipinski's rule of five and other druglikeness filters. Morin also displayed no tumorigenic, reproductive or irritant effects and exhibited good absorption and permeation through GI (clogP <5). In principal component analysis, morin appeared closest to baricitinib in 3D space. Morin displayed potent binding to spike glycoprotein, main protease 3CLPro and papain-like protease PLPro of SARS-CoV-2, SARS-CoV and MERS-CoV using molecular docking and significant binding to three viral-specific host proteins human ACE2, importin-α and poly (ADP-ribose) polymerase (PARP)-1, further lending support to its antiviral efficacy. Additionally, morin displayed potent binding to pro-inflammatory cytokines IL-6, 8 and 10 also supporting its anti-inflammatory activity. MD simulation of morin with SARS-CoV-2 3CLPro and PLPro displayed strong stability at 300 K. Both complexes exhibited constant RMSDs of protein side chains and Cα atoms throughout the simulation run time. In conclusion, morin might hold considerable therapeutic potential for the treatment and management of not only COVID-19, but also SARS and MERS if studied further. Communicated by Ramaswamy H. Sarma.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/07391102.2021.1871863DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7869441PMC
February 2021

Virtual screening of phytoconstituents from miracle herb targeting nucleocapsid protein and papain-like protease of SARS-CoV-2 for COVID-19 treatment.

J Biomol Struct Dyn 2020 Dec 8:1-21. Epub 2020 Dec 8.

Chancellor, Era's Lucknow Medical College and Hospital, Era University, Lucknow, Uttar Pradesh, India.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a novel etiological agent of coronavirus disease 2019 (COVID-19). , commonly known as black seed or black cumin, has been a historical and traditional plant since thousands of years. Based on their therapeutic efficacy, the chief components of terpenoids and flavonoids were selected from seeds and seed oil. This study was designed to check the antiviral efficacy of main phytoconstituents against five potential targets of SARS-CoV-2 using structure-based virtual screening approach. Out of twenty five phytocomponents, ten components showed best binding affinity against two viral proteins N-terminal RNA binding domain (NRBD; PDB ID: 6M3M) of nucleocapsid protein and papain-like protease (PL-; PDB ID: 6W9C) of SARS-CoV-2 using AutoDock 4.2.6, AutoDock Vina and iGEMDOCK. PASS analyses of all ten phytocomponents using Lipinski's Rule of five showed promising results. Further, druglikeness and toxicity assessment using OSIRIS Data Warrior v5.2.1 software exhibited the feasibility of phytocomponents as drug candidates with no predicted toxicity. Molecular dynamics simulation study of NRBD of SARS-CoV-2 nucleocapsid protein-alpha-spinasterol complex and PL--cycloeucalenol complex displayed strong stability at 300 K. Both these complexes exhibited constant root mean square deviation (RMSDs) of protein side chains and Cα atoms throughout the simulation run time. Interestingly, PL- and NRBD are key proteins in viral replication, host cell immune evasion and viral assembly. Thus, NRBD and PL- have the potential to serve as therapeutic targets for phytoconstituents in drug discovery process against COVID-19.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/07391102.2020.1852117DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7738213PMC
December 2020

Exploring nature's bounty: identification of as a promising source of therapeutic agents against COVID-19 by virtual screening and evaluation.

J Biomol Struct Dyn 2020 Nov 27:1-51. Epub 2020 Nov 27.

Department of Pathology, Era's Lucknow Medical College and Hospital, Era University, Lucknow, UP, India.

Coronaviruses are etiological agents of extreme human and animal infection resulting in abnormalities primarily in the respiratory tract. Presently, there is no defined COVID-19 intervention and clinical trials of prospective therapeutic agents are still in the nascent stage. (L.) Dunal (WS), is an important medicinal plant in Ayurveda. The present study aimed to evaluate the antiviral potential of selected WS phytoconstituents against the novel SARS-CoV-2 target proteins and human ACE2 receptor using methods. Most of the phytoconstituents displayed good absorption and transport kinetics and were also found to display no associated mutagenic or adverse effect(s). Molecular docking analyses revealed that most of the WS phytoconstituents exhibited potent binding to human ACE2 receptor, SAR-CoV and SARS-CoV-2 spike glycoproteins as well as the two main SARS-CoV-2 proteases. Most of the phytoconstituents were predicted to undergo Phase-I metabolism prior to excretion. All phytoconstituents had favorable bioactivity scores with respect to various receptor proteins and target enzymes. SAR analysis revealed that the number of oxygen atoms in the withanolide backbone and structural rearrangements were crucial for effective binding. Molecular simulation analyses of SARS-CoV-2 spike protein and papain-like protease with Withanolides A and B, respectively, displayed a stability profile at 300 K and constant RMSDs of protein side chains and Cα atoms throughout the simulation run time. In a nutshell, WS phytoconstituents warrant further investigations and to unravel their molecular mechanism(s) and modes of action for their future development as novel antiviral agents against COVID-19.Communicated by Ramaswamy H. Sarma.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/07391102.2020.1835725DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7755033PMC
November 2020

Assessment of micronuclei counts as tumour marker in cervical carcinogenesis: A follow-up study.

Cytopathology 2020 07;31(4):292-297

Department of Biochemistry, Era's Lucknow Medical College and Hospital, Era University, Lucknow, India.

Objective: Micronuclei counts were performed in cervical smears with low-grade squamous intraepithelial lesions of the cervix (LSIL) to assess its potentiality as tumour marker in cervical carcinogenesis.

Methods: The cases studied were from the ongoing rural cervical cancer screening in west Lucknow, India. Micronuclei counts were performed in the cervical smears of 100 LSIL cases, and the number of cells with micronuclei was defined as micronucleated cells (MNC) and the number of micronuclei per 1000 cells as MNC score. Human papillomavirus (HPV) DNA testing was also done in 100 LSIL cases by GeneNav qPCR test.

Results: A high MNC score was found in 20 of the 100 LSIL cases while the counts were low in the remaining 80. Persistence of LSIL was seen in 19 of the 20 LSIL cases with high MNC score while only six cases of the 80 cases with low MNC score showed persistence. The persistence of LSIL was very high in cases with high MNC score. The multiple high-risk HPV types such as 18, 31, 33 and 35 were seen in 12 of the 100 LSIL cases and a high positivity rate was seen in women with high MNC score. The persistence of LSIL was also higher with HPV positivity.

Conclusion: The study revealed correlation between high MNC score, persistence of LSIL and HPV positivity. Hence, MNC score can prove to be very useful in discriminating high-risk LSIL cases that are less likely to regress and possibly may progress to high-grade squamous intraepithelial lesions or carcinoma.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/cyt.12844DOI Listing
July 2020

Phenanthridine derivatives as promising new anticancer agents: synthesis, biological evaluation and binding studies.

Future Med Chem 2020 04 25;12(8):709-739. Epub 2020 Mar 25.

Department of Chemistry, Integral University, Dasauli, P.O. Bas-ha, Kursi Road, Lucknow, 226026, Uttar Pradesh, India.

Phenanthridines are an essential class of nitrogenous heterocycles with extensive applications in medicinal chemistry. The development of efficient and eco-friendly methods for the preparation of chirally pure dihydropyrrolo[1,2-]phenanthridines (), and their evaluation and modeling studies as potential anticancer, antioxidant and DNA cleavage agents is reported. Compounds were prepared through a facile one-pot synthesis and characterized by infrared, high resolution mass spectrometry, H and C nuclear magnetic resonance. The molecules were subjected to virtual screening and docking analysis against selected human molecular targets. Compound displayed good binding properties as well as significant anticancer and DNA cleavage activity. Compound has been identified as a potential lead candidate for further testing against additional cancer cell lines and animal models in future.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.4155/fmc-2019-0016DOI Listing
April 2020

Anticancer Potential of Dietary Natural Products: A Comprehensive Review.

Anticancer Agents Med Chem 2020 ;20(2):122-236

Department of Food and Nutrition, Era University, Sarfarazganj, Lucknow-226003, UP, India.

Nature is a rich source of natural drug-like compounds with minimal side effects. Phytochemicals better known as "Natural Products" are found abundantly in a number of plants. Since time immemorial, spices have been widely used in Indian cuisine as flavoring and coloring agents. Most of these spices and condiments are derived from various biodiversity hotspots in India (which contribute 75% of global spice production) and form the crux of India's multidiverse and multicultural cuisine. Apart from their aroma, flavor and taste, these spices and condiments are known to possess several medicinal properties also. Most of these spices are mentioned in the Ayurveda, the indigenous system of medicine. The antimicrobial, antioxidant, antiproliferative, antihypertensive and antidiabetic properties of several of these natural products are well documented in Ayurveda. These phytoconstituemts are known to act as functional immunoboosters, immunomodulators as well as anti-inflammatory agents. As anticancer agents, their mechanistic action involves cancer cell death via induction of apoptosis, necrosis and autophagy. The present review provides a comprehensive and collective update on the potential of 66 commonly used spices as well as their bioactive constituents as anticancer agents. The review also provides an in-depth update of all major in vitro, in vivo, clinical and pharmacological studies done on these spices with special emphasis on the potential of these spices and their bioactive constituents as potential functional foods for prevention, treatment and management of cancer.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.2174/1871520619666191015103712DOI Listing
January 2021

Mixed Ligand-metal Complexes of 2-(butan-2-ylidene) Hydrazinecarbothioamide- Synthesis, Characterization, Computer-Aided Drug Character Evaluation and in vitro Biological Activity Assessment.

Curr Comput Aided Drug Des 2021 ;17(1):107-122

Department of Chemistry, Integral University, Lucknow, UP 226026, India.

Background: Mixed ligand-metal complexes are efficient chelating agents because of their flexible donor ability. Mixed ligand complexes containing hetero atoms sulphur, nitrogen and oxygen have been probed for their biological significance.

Methods: Nine mixed ligand-metal complexes of 2-(butan-2-ylidene) hydrazinecarbothioamide (2- butanone thiosemicarbazone) with pyridine, bipyridine and 2-picoline as co-ligands were synthesized with Cu, Co and Zn salts. The complexes were tested against MDA-MB231 (MDA) and A549 cell lines. Antibacterial activity was tested against Staphylococcus aureus and Escherichia coli. The drug character of the complexes was evaluated on parameters viz. physicochemical properties, bioactivity scores, toxicity assessment and Absorption, Distribution, Metabolism, Excretion and Toxicity (ADMET) profile using various automated softwares. Molecular docking was performed against Ribonucleotide Reductase (RR) and topoisomerase II (topo II).

Results: The mixed ligand-metal complexes were synthesized by condensation reaction for 4-5 h. The characterization was done by elemental analysis, 1H-NMR, FT-IR, molar conductance and UV spectroscopic techniques. Molecular docking results showed that [Cu(CHNS)(py)2(CH3COO)2], [Zn(CHNS)(bpy)(SO)] and [Zn(CHNS)(2-pic)2(SO4)] displayed the lowest binding energies with respect to RR. Against topo II [Cu(CHNS)(py)2(CH3COO)2], [Cu(CHNS)(bpy)(CH3COO)2] and [Zn(CHNS)(2-pic)2(SO)] had the lowest energies. The druglikness assessment was done using Leadlikeness and Lipinski's rules. Not more than two violations were obtained in case of each filtering rule showing drug-like character of the mixed ligand complexes. Some of the complexes exhibited positive bioactivity scores and almost all the complexes were predicted to be safe with no hazardous effects as predicted by the toxicity assessment. Ames test predicted the non-mutagenic nature of the complexes.

Conclusion: In vitro activity evaluation showed that [Zn(CHNS)(py)2(SO)], [Co(CHNS(bpy) (Cl)2] and [Cu(CHNS)(2-pic)2(CH3COO)2] were active against MDA. Against A549 [Co(CHNS)(py)2(Cl)2], [Cu(CHNS)(py)2(CH3COO)2] and [Co(CHNS(bpy)(Cl)2] were active. Antibacterial evaluation showed that [Co(CHNS)(bpy)(Cl)2], [Zn(CHNS)(2-pic)2(SO)] and [Cu(CHNS)(2-pic)2(CH3COO)2] were active against S. aureus. Against E. coli, [Zn(CHNS)(2- pic)2(SO)] showed activity at 18-20 mg dose range.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.2174/1573409915666190926122103DOI Listing
January 2021

Clinicopathological correlation of cancer stem cell markers Oct-4 and CD133 expression as prognostic factor in malignant lesions of gallbladder: An immunohistochemical study.

Indian J Pathol Microbiol 2019 Jul-Sep;62(3):384-390

Department of Surgical Oncology, King George's Medical University, Lucknow, Uttar Pradesh, India.

Background: Gallbladder cancer (GBC) is the most frequent biliary tract cancer, with high morbidity and poor prognosis, and shows early metastasis and invasiveness. No reliable biomarkers are available for detection of GBC progression.

Aim: To investigate the immunohistochemical expression of Oct-4 and CD133 in malignant and nonneoplastic lesions of gallbladder and to analyze the clinical significance of the expressions related to clinicopathological parameters.

Settings And Design: This is a prospective case control study, conducted in medical college background.

Materials And Methods: A total of 103 cases of gallbladder were grouped into malignant lesions (n = 48) and nonneoplastic lesions (simple epithelial hyperplasia; n = 35 and chronic cholecystitis; n = 20). All tissue samples were evaluated for expression of Oct-4 and CD133 using immunohistochemistry in an effort to elucidate the correlation between their expressions with clinicopathological parameters.

Statistical Analysis: The final score was calculated by multiplying the intensity to the percentage of positive cells. The scores ≥2 were considered as positive.

Results: Significant positive correlation of higher expression levels of Oct-4 and CD133 were observed in malignant as compared to nonneoplastic lesions of gallbladder (P < 0.0001). High expression of Oct-4 and CD133 were significantly associated with tumor grading (Oct-4, P = 0.04; CD133, P = 0.02), staging (Oct-4, P = 0.03; CD133, P = 0.02), and liver metastasis (Oct-4, P = 0.01; CD133, P = 0.007). Significantly reduced survival was observed with high expression of Oct-4 (P = 0.002). No significant correction was observed between CD 133 and survival.

Conclusion: This study revealed that high expression level of Oct-4 may provide a new insight for the prognosis of the disease in terms of clinical staging and grade.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.4103/IJPM.IJPM_134_19DOI Listing
December 2019

Cytostatic and Anti-tumor Potential of Ajwa Date Pulp against Human Hepatocellular Carcinoma HepG2 Cells.

Sci Rep 2019 01 21;9(1):245. Epub 2019 Jan 21.

Department of Pathology, Era's Lucknow Medical College & Hospital, Era University, Lucknow, 226003, UP, India.

Ajwa dates (Phoenix dactylifera L.) are used by traditional therapeutic practitioners for several health benefits but most remain to be scientifically validated. In this study, we evaluated the apoptosis-inducing effect of ethanolic extract of Ajwa date pulp (ADP) on human hepatocellular carcinoma (HCC) HepG2 cells. High performance liquid chromatography analysis revealed the presence of polysaccharide β-D-glucan in ADP extract. Treated HCC cells revealed morphological characteristics of apoptosis under phase contrast microscopy. MTT assay demonstrated significant (p < 0.05) dose- and time-dependent inhibition of HCC cell growth. HCC cells were found to be in late apoptotic stage on treatment with higher doses of ADP extract as depicted by acridine orange/ethidium bromide and Annexin V-FITC/PI double stain. Importantly, ADP extract increased the reactive oxygen species level and decreased the mitochondrial membrane potential in treated HCC cells. Flow cytometry analysis demonstrated that ADP extract induced elevation of S and G2/M phases of cell cycle. Moreover, ADP extract induced apoptosis in HCC cells independent of tumor suppressor genes viz. CHEK2, ATM and TP53. Interestingly, ADP extract did not display any significant effect on normal cell line Vero. This study provides validation that ADP extract can be considered as a safe and natural potential drug candidate against human liver cancer.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41598-018-36475-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6341075PMC
January 2019

Steroidal glycoalkaloids from target cytoskeletal proteins: an analysis.

Authors:
Rumana Ahmad

PeerJ 2019 3;7:e6012. Epub 2019 Jan 3.

Department of Biochemisty, Era's Lucknow Medical College and Hospital, Era University, Lucknow, Uttar Pradesh, India.

Background: (black nightshade; ), a member of family Solanaceae, has been endowed with a heterogeneous array of secondary metabolites of which the steroidal glycoalkaloids (SGAs) and steroidal saponins (SS) have vast potential to serve as anticancer agents. Since there has been much controversy regarding safety of use of glycoalkaloids as anticancer agents, this area has remained more or less unexplored. Cytoskeletal proteins like actin play an important role in maintaining cell shape, synchronizing cell division, cell motility, etc. and along with their accessory proteins may also serve as important therapeutic targets for potential anticancer candidates. In the present study, glycoalkaloids and saponins from were screened for their interaction and binding affinity to cytoskeletal proteins, using molecular docking.

Methods: Bioactivity score and Prediction of Activity Spectra for Substances (PASS) analysis were performed using softwares Molinspiration and Osiris Data Explorer respectively, to assess the feasibility of selected phytoconstituents as potential drug candidates. The results were compared with two standard reference drugs doxorubicin hydrochloride (anticancer) and tetracycline (antibiotic). Multivariate data obtained were analyzed using principal component analysis (PCA).

Results: Docking analysis revealed that the binding affinities of the phytoconstituents towards the target cytoskeletal proteins decreased in the order coronin>villin>ezrin>vimentin>gelsolin>thymosin>cofilin. Glycoalkaloid solasonine displayed the greatest binding affinity towards the target proteins followed by alpha-solanine whereas amongst the saponins, nigrumnin-I showed maximum binding affinity. PASS Analysis of the selected phytoconstituents revealed 1 to 3 violations of Lipinski's parameters indicating the need for modification of their structure-activity relationship (SAR) for improvement of their bioactivity and bioavailability. Glycoalkaloids and saponins all had bioactivity scores between -5.0 and 0.0 with respect to various receptor proteins and target enzymes. Solanidine, solasodine and solamargine had positive values of druglikeness which indicated that these compounds have the potential for development into future anticancer drugs. Toxicity potential evaluation revealed that glycoalkaloids and saponins had no toxicity, tumorigenicity or irritant effect(s). SAR analysis revealed that the number, type and location of sugar or the substitution of hydroxyl group on alkaloid backbone had an effect on the activity and that the presence of α-L-rhamnopyranose sugar at C-2 was critical for a compound to exhibit anticancer activity.

Conclusion: The present study revealed some cytoskeletal target(s) for phytoconstituents by docking analysis that have not been previously reported and thus warrant further investigations both and .
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.7717/peerj.6012DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6321755PMC
January 2019

Computer-aided drug design and virtual screening of targeted combinatorial libraries of mixed-ligand transition metal complexes of 2-butanone thiosemicarbazone.

Comput Biol Chem 2018 Aug 8;75:178-195. Epub 2018 May 8.

Department of Chemistry, Integral University, Dasauli, P.O. Bas-ha, Kursi Road, Lucknow 226026, UP, India.

The present paper deals with in silico evaluation of 32 virtually designed transition metal complexes of 2-butanone thiosemicarbazone and N,S,O containing donor hetero-ligands namely py, bpy, furan, thiophene, 2-picoline, 1,10-phenanthroline, piperazine and liquid ammonia. The complexes were designed with a view to assess their potential anticancer, antioxidant and antibacterial activity. The absorption, distribution, metabolism, excretion and toxicity (ADMET) properties of the chosen ligands were calculated by admetSAR software. Metabolic sites of different ligands likely to undergo metabolism were predicted using Metaprint 2D. The proposed complexes were also evaluated for their drug-like quality based on Lipinski's, Veber, Ghose and leadlikeness filters. Druglikeness and toxicity potential were predicted by OSIRIS property explorer. The pharmacokinetic properties and bioactivity scores were calculated by Molinspiration tool. Bioactivity scores of the complexes were predicted for drug targets including enzymes, nuclear receptors, kinase inhibitors, G-protein coupled receptor ligands and ion channel modulators. Molecular docking of selected Fe(II) mixed-ligand complexes was performed using AutoDock version 4.2.6 and i-GEMDOCK version 2.1 with two target proteins namely Ribonucleotide reductase (RR) and Topoisomerase II (Topo II). The results were compared with three standard reference drugs viz. Doxorubicin HCl, Letrozole (anticancer) and Tetracycline (antibiotic). Multivariate data obtained were analyzed using principal component analysis (PCA) for visualization and projection as scatter and 3D plots. Positive results obtained for hetero-ligands using admetSAR version 1.0 indicated good absorption and transport kinetics of the hetero-ligand complexes through the human intestine and blood-brain barrier. The hetero-ligands were predicted to have no associated mutagenic effect(s) also. However, none of the hetero-ligands was predicted to be Caco-2 (human colon cancer cell line) permeable. Most of the hetero-ligands and the parent ligand (2-butanone thiosemicarbazone) were predicted to undergo Phase-I metabolism prior to excretion using MetaPrint2D software. Pharmacokinetic evaluation of the proposed complexes revealed that all complexes displayed drug-like character and were predicted to have no apparent toxic side-effects. All the proposed complexes showed moderate to good biological activity scores (-5.0 to 0.0). Mixed complexes with bpy, 2-picoline and 1,10-phenanthroline showed significant bioactivity scores (as enzyme inhibitors) in the range 0.02-0.13. Likewise, good docking scores were obtained for Fe (II) complexes with the same ligands. [FeL(2-pic)] displayed the lowest binding energy (-6.47 kcal/mol) with respect to Topo II followed by [FeL(py)] (-6.14 kcal/mol) as calculated by AutoDock version 4.2.6. With respect to binding with RR, [FeL(2--pic)] again displayed the lowest binding energy (-7.21 kcal/mol) followed by [FeL(py)] (-5.96 kcal/mol). On the basis of docking predictions and various other computational evaluations, four mixed-ligand complexes of Fe in +2 oxidation state with py, bpy, 2--picoline and 1,10-phenanthroline were synthesized with 2-butanone thiosemicarbazone. All the synthesized Fe complexes were characterized using various spectroscopic techniques and tested for their potential anticancer activity in vitro against human breast carcinoma cell line MDA-MB 231 and human lung carcinoma cell line A549 cell line using MTT assay. [FeL(2-pic)], [FeL(bpy)], and [FeL(py)] were found to exhibit significant antiproliferative activity with IC values in the range of 80-100 μM against breast and lung cancer cells. The synthesized Fe complexes also displayed mild antioxidant activities. The synthesized and studied Fe complexes have the potential for development into future anticancer agents if analyzed and modified further for improvement of their ADMET, solubility and permeability criteria set for potential drug-candidates.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.compbiolchem.2018.05.008DOI Listing
August 2018

Synthesis, characterization, computational studies and biological activity evaluation of Cu, Fe, Co and Zn complexes with 2-butanone thiosemicarbazone and 1,10-phenanthroline ligands as anticancer and antibacterial agents.

EXCLI J 2018 29;17:331-348. Epub 2018 Mar 29.

Department of Chemistry, Integral University, Dasauli, P.O. Bas-ha, Kursi Road, Lucknow-226026, UP, India.

Mixed-ligand metal (II) (M=Cu, Fe, Co and Zn) complexes containing 2-butanone thiosemicarbazone and 1, 10-phenanthroline have been synthesized and characterized by melting point, FT-IR, H-NMR, UV-spectrophotometry and molar conductance measurements. All the complexes were soluble in DMSO and DMF. They were thermally stable with high melting points. The computational studies of the complexes were also performed to assess toxicity potential, bioactivity score prediction and drug likeliness assessment based on various drug filters. All the complexes showed no Veber's violations whereas only Cu complex showed one Lipinski's violation. Almost all synthesized compounds were predicted to have no toxic effects. Some of them showed positive bioactivity as enzyme inhibitors. Molecular docking of the complexes was also performed against ribonucleotide diphosphate reductase (RR) and topoisomerase II (Topo II) for minimum binding energy (kcal/mol) calculations. Cu complex was found to have minimum binding energy (-101.13 kcal/mol) released on interaction with Topo II showing a high affinity towards the enzyme, whereas Fe complex had the lowest binding energy (-99.8349 kcal/mol) when docked with RR. The results were compared with two standard drugs i.e. doxorubicin HCl and tetracycline. The ligand was tested for its potential anticancer activity against MDA-MB-231 cell line using MTT assay. Antibacterial activity of the complexes was tested against and using the disc diffusion method. Cu (II) complex showed maximum activity against the MDA cells and also exhibited mild antibacterial activity against .
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.17179/excli2017-984DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5938540PMC
March 2018

Phytochemical characterization and biological activity evaluation of ethanolic extract of Cinnamomum zeylanicum.

J Ethnopharmacol 2018 Jun 22;219:110-116. Epub 2018 Mar 22.

Dept. of Biochemistry, Era's Lucknow Medical College & Hospital, Era University, Sarfarazganj, Hardoi Road, Lucknow 226003, UP, India. Electronic address:

Ethnopharmacological Relevance: India being a multicultural nation, every region of the country offers a distinct culinary flavor and taste. These flavors are attributed to spices and condiments which form the mainstay of Indian cuisine. Most of these spices and condiments are derived from various biodiversity hotspots in India and form the crux of India's multidiverse and multicultural cuisine. Apart from their varying aromas, flavors and tastes, these spices and condiments are known to possess several medicinal properties also. Most of these spices find considerable mention in Ayurveda, the indigenous system of medicine, as panaceas for several aliments. Cinnamomum zeylanicum (CZ), belonging to family Lauraceae and commonly known as cinnamon is one such spice known to have diverse medicinal properties since time immemorial.

Aim Of The Study: In the present study, apoptotic and anti-microbial activity of ethanolic extract of CZ was evaluated against human breast cancer cell line MDA-MB-231 and compared for its effect on normal kidney epithelial cell line Vero.

Materials And Methods: Ethanolic extract of tree bark of CZ was used to determine the cytotoxic effect on MDA-MB-231 using Trypan blue dye exclusion method and cytometry. The tested dose of the extract was 10-100 µg/mL. Antibacterial activity was determined using disc diffusion method against Staphylococcus aureus and Escherichia coli in the range 2-10 mg/mL. Apoptotic activity was determined using DNA fragmentation assay.

Results: Ethanolic extract of CZ was found to have an IC value of 25 µg/mL against MDA cell line. On the other hand, CZ extract did not have any significant effect on Vero cells even at 100 µg/mL (IC > 100 µg/mL). The ethanolic extract of CZ bark showed significant antibacterial activity against S. aureus at 10 mg/mL while no appreciable activity was detected against E. coli. DNA isolated from extract treated cancer cells showed a fragmentation pattern characteristic of apoptosis. However, no DNA fragmentation was observed in DNA isolated from extract treated Vero cells.

Conclusion: Ethanolic bark extract of CZ could be potentially beneficial in treating breast cancer and may be of interest for future studies in developing integrative cancer therapy against proliferation, metastasis, and migration of breast cancer cells.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jep.2018.02.001DOI Listing
June 2018

Evaluation of apoptotic activity of Withania coagulans methanolic extract against human breast cancer and Vero cell lines.

J Ayurveda Integr Med 2017 Jul - Sep;8(3):177-183. Epub 2017 Jul 6.

Chairman Research, Era's Lucknow Medical College & Hospital, Sarfarazganj, Hardoi Road, Lucknow, 226003, India.

Background: The genus Withania (Family: Solanaceae) holds an important position in Ayurveda, the Indian traditional system of medicine. Withania somnifera Dunal and Withania coagulans Dunal have been documented in folklore as panaceas for various ailments since time immemorial. W. coagulans (WC), commonly called as Indian cheese maker is used for fermenting milk for cheese production in various parts of India.

Objectives: In the study, in vitro cytotoxicity of methanolic extract of dried fruits (berries) of WC was evaluated in a dose dependent manner using trypan blue dye exclusion method against human breast cancer cell line MDA-MB-231 and normal kidney epithelial cell line Vero in the range 20-200 μg/ml.

Material And Methods: The percentage viability of the cell lines was determined by using MTT assay and cytometry.

Results: Methanolic extract of WC showed significant anticancer activity against MDA-MB-231 cell line. Cell viability was reduced to about 50% at 40 μg/ml of methanolic extract in 50% DMSO. Cytotoxicity of the extract was lower in 10% and 1% DMSO. On the other hand, methanolic extract of WC did not exhibit any significant in vitro activity against Vero cells at 170 and 200 μg/ml. AGE of isolated DNA from treated cancer cells revealed characteristic ladder like fragmentation, a hallmark of apoptosis. HPLC profiling was carried out for identification of the active components, which demonstrated the presence of Withaferin A in the methanolic extract.

Conclusion: Methanolic extract of WC possesses apoptotic activity against human breast cancer cells in vitro albeit lower in comparison to W. somnifera and warrants further investigation.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jaim.2017.01.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5607390PMC
July 2017

Molecular docking, PASS analysis, bioactivity score prediction, synthesis, characterization and biological activity evaluation of a functionalized 2-butanone thiosemicarbazone ligand and its complexes.

J Chem Biol 2017 Jul 4;10(3):91-104. Epub 2017 Apr 4.

Department of Chemistry, Integral University, Dasauli, P.O. Bas-ha, Kursi Road, Lucknow, UP 226026 India.

2-Butanone thiosemicarbazone ligand was prepared by condensation reaction between thiosemicarbazide and butanone. The ligand was characterized by H NMR, C NMR, FT-IR, mass spectrometry and UV spectroscopic studies. Docking studies were performed to study inhibitory action against topoisomerase II (Topo II) and ribonucleoside diphosphate reductase (RR) enzymes. Inhibition constants ( ) of the ligand were 437.87 and 327.4 μM for the two enzymes, respectively. The ligand was tested for its potential anticancer activity against two cancer cell lines MDA-MB-231 and A549 using MTT assay and was found to exhibit good activity at higher doses with an IC = 80 μM against human breast cancer cell line MDA-MB-231. On the other hand, no significant activity was obtained against the lung carcinoma cell line A549. Antibacterial activity of the ligand was tested against and using the disc diffusion method. Ligand did not exhibit any significant antibacterial activity. Four complexes of Co(III), Fe(II), Cu(II), and Zn(II) were prepared with the ligand and characterized by various spectroscopic studies. Low molar conductance values were obtained for all complexes displaying non-electrolyte nature except in Co(III) complex. As expected, complexation with metal ions significantly increased the cytotoxicity of the ligand against the tested cell lines viz. IC values of <20 μM for Co, Fe, and Zn complexes and approx. 80 μM against MDA cells versus IC value of <20 μM for Co and Cu complexes and that of 30 and 50 μM for Fe and Zn complexes, respectively, against A549 cells. The Cu complex was found to be active against and with MIC values in the range of 6-10 mg/mL. Other than Cu, only Co complex was found to possess antibacterial activity with MIC values of 5-10 mg/mL when tested against . Bioactivity score and Prediction of Activity Spectra for Substances (PASS) analysis also depicted the drug-like nature of ligand and complexes.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s12154-017-0167-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5480261PMC
July 2017

Effect of ethyl acetate aroma on viability of human breast cancer and normal kidney epithelial cells .

Integr Med Res 2017 Mar 12;6(1):47-59. Epub 2016 Dec 12.

Department of Pathology, Era's Lucknow Medical College and Hospital, Lucknow, India.

Background: Aromatherapy is used in clinical settings for patients suffering from several chronic and critical diseases such as cancer. Ethyl acetate (EA) is a colorless liquid with a characteristic fruity smell and is naturally present in fruits and wines.

Methods: In the present study, the effect of the aroma of EA was evaluated on human breast cancer cell line MDA-MB-231 and normal cell line, Vero. Cell line viability and mechanism of EA cytotoxicity were determined by Trypan blue dye exclusion assay and phase contrast microscopy.

Results: It was found that EA at a concentration of 0.026 M was effective in causing considerable cytotoxicity in breast cancer cells (without even coming in contact with the culture medium and cells), while showing no effect on normal cells. Mechanism of action of EA on cancer and Vero cells was investigated by DNA fragmentation and dye binding assays using agarose gel electrophoresis (AGE) and fluorescence microscopy/cytometry, respectively. It was found that EA aroma induced predominantly necrosis in the cancer cells exposed to it.

Conclusion: A study such as this has not been attempted before and results need further investigation before EA aroma can be used as a complementary therapy.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.imr.2016.11.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5395688PMC
March 2017

An efficient method for native protein purification in the selected range from prostate cancer tissue digests.

Chin Clin Oncol 2016 Dec;5(6):78

Department of Urology, University of Washington, Seattle, WA 98195, USA; Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98195, USA.

Background: Prostate cancer (CP) cells differ from their normal counterpart in gene expression. Genes encoding secreted or extracellular proteins with increased expression in CP may serve as potential biomarkers. For their detection and quantification, assays based on monoclonal antibodies are best suited for development in the clinical setting. One approach to obtain antibodies is to use recombinant proteins as immunogen. However, the synthesis of recombinant protein for each identified candidate is time-consuming and expensive. It is also not practical to generate high quality antibodies to all identified candidates individually. Furthermore, non-native forms (e.g., recombinant) of proteins may not always lead to useful antibodies. Our approach was to purify a subset of proteins from CP tissue specimens for use as immunogen.

Methods: In the present investigation, ten cancer specimens obtained from cases scored Gleason 3+3, 3+4 and 4+3 were digested by collagenase to single cells in serum-free tissue culture media. Cells were pelleted after collagenase digestion, and the cell-free supernatant from each specimen were pooled and used for isolation of proteins in the 10-30 kDa molecular weight range using a combination of sonication, dialysis and Amicon ultrafiltration. Western blotting and mass spectrometry (MS) proteomics were performed to identify the proteins in the selected size fraction.

Results: The presence of cancer-specific anterior gradient 2 (AGR2) and absence of prostate-specific antigen (PSA)/KLK3 were confirmed by Western blotting. Proteomics also detected AGR2 among many other proteins, some outside the selected molecular weight range, as well.

Conclusions: Using this approach, the potentially harmful (to the mouse host) exogenously added collagenase was removed as well as other abundant prostatic proteins like ACPP/PAP and AZGP1 to preclude the generation of antibodies against these species. The paper presents an optimized scheme for convenient and rapid isolation of native proteins in any desired size range with minor modifications.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.21037/cco.2016.12.03DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5627993PMC
December 2016

Neurocysticercosis: a review on status in India, management, and current therapeutic interventions.

Parasitol Res 2017 Jan 24;116(1):21-33. Epub 2016 Oct 24.

Division of Biochemistry, CSIR-Central Drug Research Institute, Lucknow, 226031, UP, India.

Tapeworms (cestodes) are segmented flatworms responsible for causing diseases that may prove fatal and difficult to treat in the absence of proper treatment and efficient drugs. Neurocysticercosis (NCC) is a common parasitic infection of the central nervous system and a major contributor to epilepsy caused by the metacestode (larva) of the human tapeworm Taenia solium, characterized by a range of pathological symptoms including epileptic seizures, headaches, and hydrocephalus. Cysticercosis is considered as a "biological imprint" of the socioeconomic development of a community in general and a country in particular. It is the single most common cause of epilepsy in the resource-poor endemic regions of the world, including most of South and Central America, India, Southeast Asia, China, and sub-Saharan Africa. A vast degree of variation in the neuropathology and clinical symptoms of NCC often makes it difficult to diagnose and manage. To add to it, emerging drug resistance to known anti-parasitic agents, together with the inability of these agents to prevent re-infection and relapse, further complicates the disease scenario. The aim of the current review was to provide the latest update on NCC with special emphasis on the Indian scenario, along with current and novel methods of diagnosis as well as scope of development for novel detection techniques, novel targets for drug development, and therapeutic interventions, as well as future challenges.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00436-016-5278-9DOI Listing
January 2017

Evaluation and Comparison of the In Vitro Cytotoxic Activity of Withania somnifera Methanolic and Ethanolic Extracts against MDA-MB-231 and Vero Cell Lines.

Sci Pharm 2016 Jan-Mar;84(1):41-59. Epub 2015 Sep 26.

Chairman Research, Era's Lucknow Medical College & Hospital, Sarfarazganj, Hardoi Road, Lucknow-226003, India.

Withania somnifera Dunal (WS), commonly known as Ashwagandha in India, belongs to the family Solanaceae. It is extensively used in most of the Indian herbal pharmaceuticals and nutraceuticals. In the current study, the in vitro cytotoxic activity of methanolic, ethanolic, and aqueous extracts of WS stems was evaluated using cytometry and the MTT assay against the MDA-MB-231 human breast cancer cell line. Methanolic and ethanolic extracts of WS showed potent anticancer activity on the MDA-MB-231 human breast cancer cell line, whereas the aqueous extract did not exhibit any significant activity at 100 µg/ml. The percentage viability of the cell lines was determined by using the Trypan blue dye exclusion method. Cell viability was reduced to 21% and 0% at 50 and 100 µg/ml of the methanolic extract, respectively, as compared to 19% and 0% at 50 and 100 µg/ml for the ethanolic extract and 37% at 100 µg/ml in sterile Milli-Q water after 48 hours of treatment. Methanolic and ethanolic extracts of WS were shown to possess IC50 values of 30 and 37 µg/ml, respectively, by the MTT assay and cytometer-based analysis, with the methanolic extract being more active than the other two. On the other hand, methanolic and ethanolic extracts of WS did not exhibit any significant in vitro activity against the normal epithelial cell line Vero at 50 µg/ml. HPLC was carried out for the analysis of its phytochemical profile and demonstrated the presence of the active component Withaferin A in both extracts. The methanolic and ethanolic extracts of Withania should be studied further for the isolation and characterization of the active components to lead optimization studies.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3797/scipharm.1507-13DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4839554PMC
April 2016

Effect of methyl butyrate aroma on the survival and viability of human breast cancer cells in vitro.

J Egypt Natl Canc Inst 2016 Jun 19;28(2):81-8. Epub 2016 Apr 19.

Dept. of Pathology, Era's Lucknow Medical College & Hospital, Sarfarazganj, Hardoi Road, Lucknow-226003, India.

Background: Aroma can have far reaching effects on mind, body and soul. Pleasant aromas are known to have a soothing effect on the mind and are known to relieve stress and enhance concentration. Recently, it has been demonstrated that aroma may also have some curative effects as well as benefits and can be used both for prophylaxis and therapy of diseases. Our aim was to test our hypothesis whether aroma can cure or prevent cancer. Methyl butyrate (MB) is the methyl ester of butyric acid having a characteristic sweet and fruity odor like that of apples and pineapples. It occurs in many plant products in minute quantities and in pineapple oil.

Methods: In the present study, the effect of aroma of MB has been evaluated on human breast cancer cell line MDA-MB-231 in vitro. The percentage viability of the cell line was determined by using Trypan blue dye exclusion assay.

Results: It was found that MB at a concentration of 0.01M was effective in causing considerable cytotoxicity (40%) in breast cancer cells (without even coming in contact with cells) while at 0.02M, % cytotoxicity was found to be 50%. Mechanism of action of MB on cancer cells was investigated by acridine orange-ethidium bromide assay using fluorescence microscopy and DNA fragmentation assay. MB aroma appeared to induce necrosis in cancer cells exposed to it.

Conclusion: No study involving the effect of aroma/smell on cancer cells has ever been reported before and warrants further investigation on other cancer and normal cell lines.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jnci.2016.02.005DOI Listing
June 2016

Purification and characterization of glutathione reductase (E.C. 1.8.1.7) from bovine filarial worms Setaria cervi.

J Parasit Dis 2013 Apr 18;37(1):94-104. Epub 2012 Jul 18.

Division of Biochemistry, Central Drug Research Institute, P.O. Box No. 173, Chattar Manzil Palace, Lucknow, 226 001 UP India.

Antioxidant enzymes are the parasite's premier resource to defend themselves against reactive oxygen species generated by macrophages, neutrophils and eosinophils of the host. These enzymes may be particularly important for parasites involved in chronic infections, such as parasitic helminths. Glutathione (GSH) and glutathione reductase (GR) are parts of the GSH redox cycle, which protects cells against damage by oxidants. Both GSH and GR are present in significant amounts in Setaria cervi female worms. GR has a central role in glutathione metabolism and as such is a potential target for chemotherapy. The aim of the work was to purify and characterize GR from S. cervi and to compare the properties of the helminth enzyme with its mammalian counterpart. GR was purified from filarial parasites S. cervi and preliminary steady state kinetics was performed. The purified protein was observed to be a dimer of 55 kDa subunit as evident from SDS-PAGE analysis. Kinetic studies revealed significant differences in the properties of S. cervi GR from its mammalian counterpart which may be exploited in chemotherapy of filariasis. Filarial GR is thus proposed as a potential drug target.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s12639-012-0138-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3590390PMC
April 2013

An economic and ecological perspective of ethanol production from renewable agro waste: a review.

AMB Express 2012 Dec 7;2(1):65. Epub 2012 Dec 7.

Department of Life Sciences, Institute of Technology & Management University, Gwalior, Madhya Pradesh, 475001, India.

Agro-industrial wastes are generated during the industrial processing of agricultural products. These wastes are generated in large amounts throughout the year, and are the most abundant renewable resources on earth. Due to the large availability and composition rich in compounds that could be used in other processes, there is a great interest on the reuse of these wastes, both from economical and environmental view points. The economic aspect is based on the fact that such wastes may be used as low-cost raw materials for the production of other value-added compounds, with the expectancy of reducing the production costs. The environmental concern is because most of the agro-industrial wastes contain phenolic compounds and/or other compounds of toxic potential; which may cause deterioration of the environment when the waste is discharged to the nature. Although the production of bioethanol offers many benefits, more research is needed in the aspects like feedstock preparation, fermentation technology modification, etc., to make bioethanol more economically viable.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/2191-0855-2-65DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3547755PMC
December 2012

Development of an ELISA to detect the secreted prostate cancer biomarker AGR2 in voided urine.

Prostate 2012 Jun 9;72(9):1023-34. Epub 2011 Nov 9.

Antibody Resource, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA.

Background: Comparative transcriptomics between sorted cells identified AGR2 as one of the highest up-regulated genes in cancer. Overexpression in primary tumors was verified by tissue microarray analysis. AGR2 encodes a 19-kDa secreted protein that might be found in urine.

Methods: Monoclonal antibodies were generated against AGR2. One antibody pair, P1G4 (IgG1) to capture and P3A5 (IgG2a) to detect, showed good performance characteristics in a sandwich ELISA. This assay could detect AGR2 at sub ng/ml quantities.

Results: AGR2 was detected in tissue digestion media of tumor specimens and culture media of AGR2-secreting prostate cancer cell lines. Additional testings involved frozen section immunohistochemistry, immunoprecipitation, and Western blot analysis. Voided urine samples were collected from pre-operative cancer patients, and urinary protein was desalted and concentrated by filtration. The amount of AGR2 detected was scored as pg/100 µg total protein, and then converted to pg/ml urine. The developed ELISA detected AGR2 protein, ranging from 3.6 to 181 pg/ml, in an initial cohort of samples. AGR2 was not detected in the urine of non-cancer and a bladder cancer patient.

Conclusions: For prostate cancer, an AGR2 urine test could be used for diagnosis. The data, although derived from a small number of samples assayed, showed that developing such a test for clinical application is viable because AGR2 is specific to cancer cells, and apparently secreted into urine.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/pros.21508DOI Listing
June 2012

Purification and biochemical characterization of cytosolic glutathione-S-transferase from filarial worms Setaria cervi.

Comp Biochem Physiol B Biochem Mol Biol 2008 Nov 8;151(3):237-45. Epub 2008 Apr 8.

Division of Biochemistry, P.O. Box No. 173, Central Drug Research Institute, Chattar Manzil Palace, Lucknow-226001, India.

The present study reports the purification and characterization of GST from cytosolic fraction of Setaria cervi. GST activity was determined in various subcellular fractions of bovine filarial worms S. cervi (Bubalus bubalis Linn.) and was found to be localized mainly in the cytosolic and microsomal fractions. The soluble enzyme from S. cervi was purified to homogeneity using a combination of salt precipitation, centrifugation, cation exchange and GSH-Sepharose affinity chromatography followed by ultrafiltration. SDS-PAGE analysis revealed a single band and activity staining was also detected on PAGE gels. Gel filtration and MALDI-TOF studies revealed that the native enzyme is a homodimer with a subunit molecular mass of 24.6 kDa. Comparison of kinetic properties of the parasitic and mammalian enzymes revealed significant differences between them. The substrate specificity and inhibitor profile of cytosolic GST from S. cervi appeared to be different from GST from mammalian sources.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.cbpb.2008.03.019DOI Listing
November 2008

Cytosolic and microsomal glutathione-S-transferases from bovine filarial worms Setaria cervi.

J Parasitol 2007 Dec;93(6):1285-90

Division of Biochemistry, P.O. Box No. 173, Central Drug Research Institute, Chattar Manzil Palace, Lucknow 226001, India.

The work presented here deals with the status of glutathione-S-transferase (GST; E.C. 2.5.1.18), the major enzyme of the phase II detoxification pathway, in bovine filarial worms Setaria cervi. GST activity was determined in various subcellular fractions of bovine filarial worms S. cervi (Bubalus bubalis Linn.) and was found to be mainly associated with cytosolic and microsomal fractions. The respective specific activities of the enzyme from cytosolic and microsomal fractions of S. cervi females were determined to be 0.122 +/- 0.024 and 0.010 +/- 0.0052 micromol/min/mg protein, respectively. Cytosolic enzyme was found to possess optimal activity between pH 6.5 and 7.5, whereas the microsomal enzyme showed a broad pH optima, centered at pH 6.0. Kinetic studies on the cytosolic and microsomal forms of the enzyme revealed significant differences between them, thereby indicating that microsomal GST from S. cervi is quite distinct to the cytosolic protein catalyzing the same reaction.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1645/GE-1119.1DOI Listing
December 2007

Inhibition of glutathione-S-transferase from Plasmodium yoelii by protoporphyrin IX, cibacron blue and menadione: implications and therapeutic benefits.

Parasitol Res 2008 Mar 5;102(4):805-7. Epub 2008 Jan 5.

Division of Biochemistry, Central Drug Research Institute, Chattar Manzil Palace, P.O. Box No. 173, Lucknow 226001, India.

The rapidly developing resistance to drugs used for prophylaxis and treatment of malaria makes the identification of novel drug targets necessary. Glutathione-S-transferase (GST, E.C. 2.5.1.18), an important enzyme of the glutathione (GSH) cycle, is considered to be an essential detoxification enzyme in malarial parasites. Selective inhibition of this enzyme from malarial parasites by various classes of inhibitors may be viewed as a potential chemotherapeutic strategy to combat malaria. Purified GST from Plasmodium yoelii was inhibited by compounds like protoporphyrin IX, cibacron blue, as well as by the GSH depletor menadione. Cytosolic GST was inhibited to varying degrees by each compound. A characteristic inhibitor constant (Ki) was obtained for each inhibitor. The possible consequences of selective inhibition of parasitic GST to that of the host are discussed in relation to the chemotherapy of malaria.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00436-007-0836-9DOI Listing
March 2008

Biochemical composition and metabolic pathways of filarial worms Setaria cervi: search for new antifilarial agents.

J Helminthol 2007 Sep;81(3):261-80

Division of Biochemistry, Po Box 173, Central Drug Research Institute, Chattar Manzil Palace, Lucknow-226001, India.

The main problem regarding the chemotherapy of filariasis is that no safe and effective drug is available yet to combat the adult human filarial worms. Setaria cervi, the causal organism of setariasis and lumbar paralysis in cattle, is routinely employed as a model organism for conducting biochemical and enzymatic studies on filarial parasites. In view of the practical difficulties in procuring human strains of Wuchereria bancrofti and Brugia malayi for drug screening, the bovine filarial parasite S. cervi, resembling the human species in having microfilarial periodicity and chemotherapeutic response to known antifilarial agents, is widely used as a model in such studies. For a rational approach to antifilarial chemotherapy, knowledge of the biochemical composition and metabolic pathways of this helminth parasite may be of paramount importance, so that more potent antifilarial agents based on specific drug targets can be identified in drug discovery programmes. The present review provides an update on the biochemistry of the important metabolic pathways functioning within this potentially important bovine parasite, that have so far been studied, and on those that need to be investigated further so as to identify novel drug targets that can be exploited for designing new antifilarial drugs.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1017/S0022149X07799133DOI Listing
September 2007

Synthesis and biological evaluation of potential modulators of malarial glutathione-S-transferase(s).

J Enzyme Inhib Med Chem 2007 Jun;22(3):327-42

Division of Biochemistry, Central Drug Research Institute, Lucknow, India.

Glutathione-S-transferase(s) (E.C.2.5.1.18, GSTs) have been investigated in parasitic protozoans with respect to their biochemistry and they have been identified as potential vaccine candidates in protozoan parasites and as a target in the synthesis of new antiparasitic agents. In a search towards the identification of novel biochemical targets for antimalarial drug design, the area of Plasmodium glutathione metabolism provides a number of promising chemotherapeutic targets. GST activity was determined in various subcellular fractions of malarial parasites Plasmodium yoelii and was found to be localized mainly in the cytosolic fraction (specific activity, c. 0.058 +/- 0.016 micromol/min/mg protein). Hemin, a known inhibitor of mammalian GST(s), maximally inhibited this enzyme from P. yoelii to nearly 86%. In a search towards synthetic modulators of malarial GST(s), 575 compounds belonging to various chemical classes were screened for their effect on crude GST from P. yoelii and 92 compounds belonging to various chemical classes were studied on recombinant GST from P. falciparum. Among all the compounds screened, 83 compounds inhibited/stimulated the enzyme from P. yoelii/P. falciparum to the extent of 40% or more.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/14756360601072676DOI Listing
June 2007

Purification and biochemical characterization of cytosolic glutathione-S-transferase from malarial parasites Plasmodium yoelii.

Parasitol Res 2007 Feb 6;100(3):581-8. Epub 2006 Oct 6.

Division of Biochemistry, Central Drug Research Institute, Lucknow, 226001, India.

Glutathione (GSH) metabolism represents a potential target for antiparasitic drug design. Glutathione-S-transferase (GST), an important enzyme of the GSH cycle, is considered to be an essential detoxification enzyme in parasitic species. Soluble GST from rodent malarial parasites Plasmodium yoelii was purified to homogeneity using a combination of salt precipitation, affinity chromatography on GSH-sepharose 6B and ultrafiltration. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed a single band and activity staining was also detected on PAGE gels. Kinetic studies on the purified enzyme revealed significant differences between the parasitic and mammalian enzymes. The purified enzyme exhibited an optimum pH of 8.2 and K (m) values of 0.2+/-0.213 and 3.3+/-0.056 mM with respect to co-substrate GSH and substrate 1-chloro-2, 4-dinitrobenzene (CDNB), respectively. Hemin, the known mammalian GST inhibitor was found to be a potent inhibitor of P. yoelii GST, with a K (i) of 4.0 microM.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00436-006-0295-8DOI Listing
February 2007