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K. Kemparaju - One of the best experts on this subject based on the ideXlab platform.
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anti coagulant activity of a metalloprotease further characterization from the Indian Cobra naja naja venom
Journal of Thrombosis and Thrombolysis, 2010Co-Authors: M S Kumar, B S Vishwanath, V R Devaraj, K. KemparajuAbstract:A high molecular mass, non toxic metalloprotease the NN-PF3 with the bound Ca2+ and Zn2+ from the Naja naja venom has been studied further for its anticoagulant property. The molecular mass by MALDI-TOF mass spectrometry was 67.81 kDa. The NN-PF3 exhibited fibrin(ogen)olytic activity. In addition to fibrinogen, NN-PF3 hydrolyzed blood and plasma clot with the later hydrolyzed about one fold higher. The α polymer of fibrin was preferentially hydrolyzed over the α chain but the β chain and γ–γ dimer remained untouched. It was devoid of plasminogen activation property. It prolonged the activated partial thromboplastin time, prothrombin time and the thrombin clotting time of citrated human plasma. It did not affect the thrombin activity. In mice, defibrinogentaion, prolonged bleeding time (P < 0.01) and reduced fibrinogen level were observed following intravenous injection. Human plasma or α2-macroglobulin did not, but the polyvalent anti-venom inhibited the NN-PF3 activity. In contrast to most snake venom metalloproteases, it did not degrade extra cellular matrix proteins.
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region specific neutralization of Indian Cobra naja naja venom by polyclonal antibody raised against the eastern regional venom a comparative study of the venoms from three different geographical distributions
International Immunopharmacology, 2007Co-Authors: R. Shashidharamurthy, K. KemparajuAbstract:Indian Cobra (Naja naja) venoms from different geographical locations vary in their composition, biochemical, and pharmacological properties. Venom samples from eastern, western and southern India are compared in this study. The venom from eastern region was found to be the most lethal of the three regional venoms. Monovalent antivenom (NNEV-IgG) prepared against the eastern venom was found to cross-react with the other two regional venoms. NNEV-IgG at an Ag:Ab ratio of 1:25 completely neutralized the lethality of eastern venom. At this ratio, it did not neutralize the other two venoms, but the survival time of experimental mice was extended significantly. Commercially available polyvalent antivenom neutralized the lethality of western venom at an Ag:Ab ratio of 1:60 and increased the survival time of experimental mice injected with eastern and southern venoms marginally. Further, NNEV-IgG neutralized the tested pharmacological and enzymatic activities of all the three venom samples dose dependently, with neutralization potency varying with the geographic origin of the tested venoms. Thus, the present study demonstrates the diversity in the immunological properties of venom from different geographical regions and underscores the importance of developing region-specific antivenoms for therapeutic purpose.
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a neurotoxic phospholipase a2 variant isolation and characterization from eastern regional Indian Cobra naja naja venom
Toxicon, 2006Co-Authors: R. Shashidharamurthy, K. KemparajuAbstract:CM-Sephadex C-25 column chromatography profile of Indian Cobra (Naja naja) venom from eastern region showed a distinct and a dominant phospholipase peak, peak-10, while it was not seen in either southern or western venom samples. Peak-10 was subjected to CM-Sephadex C-25 and Sephadex G-50 column chromatography to isolate NN-X-PLA(2). NN-X-PLA(2) is a single chain protein with the relative molecular weight of 10kDa by SDS-PAGE. It was toxic to mice with an LD(50) value 0.098 mg/kg body weight (i.p.) and the mice exhibited acute neurotoxic symptoms. Upon indirect stimulation, it inhibited the twitching of frog's gastrocnemius muscle in a dose dependent manner. NN-X-PLA(2) was weakly anticoagulant and devoid of cytotoxicity, myotoxicity, hemorrhage, edema inducing, and directlytic activities and effects on platelet aggregation process. Upon chemical modification independently with p-bromophenacyl bromide and acetic anhydride, NN-X-PLA(2) lost both enzymatic and toxic properties.
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inhibition of naja naja venom hyaluronidase role in the management of poisonous bite
Life Sciences, 2006Co-Authors: K.s. Girish, K. KemparajuAbstract:Hyaluronidase is present virtually in all snake venoms and has been known as a “spreading factor.” The enzyme damages the extracellular matrix at the site of the bite, leading to severe morbidity. In this study, the benefits of inhibiting the hyaluronidase activity of Indian Cobra (Naja naja) venom have been investigated. Anti-NNH1 and aristolochic acid both inhibited the in vitro activity of the purified hyaluronidase, (NNH1) and the hyaluronidase activity of whole venom in a dose-dependent manner. Both anti-NNH1 and aristolochic acid abolished the degradation of hyaluronan in human skin tissue sections by NNH1 and by whole venom. Aristolochic acid quenched the fluorescent emission of NNH1. A non-competitive mechanism of NNH1 inhibition was observed with aristolochic acid. NNH1 potentiates the toxicity of Daboia russellii VRV-PL-VIII myotoxin and hemorrhagic complex-I. However, the potentiation of toxicity was inhibited dose-dependently by anti-NNH1 and aristolochic acid. Further, mice injected with whole venom which had been preincubated with anti-NNH1/aristolochic acid, showed more than a two-fold increase in survival time, compared to mice injected with venom alone. A more moderate increase in survival time was observed when mice were injected with anti-NNH1/aristolochic acid 10 min after whole venom injection. This study illustrates the significance of venom hyaluronidase in the pathophysiology of snake venom poisoning and the therapeutic value of its inhibition.
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a low molecular weight isoform of hyaluronidase purification from Indian Cobra naja naja venom and partial characterization
Biochemistry, 2005Co-Authors: K.s. Girish, K. KemparajuAbstract:A low molecular weight isoform of hyaluronidase (NNH2) has been isolated from Indian Cobra (Naja naja) venom by successive chromatography on Sephadex G-75 and CM-Sephadex C-25 columns. The apparent molecular weight determined by SDS-PAGE is 52 kD, and the pI value is 9.7. NNH2 is an endoglycosidase and exhibits in vitro absolute specificity for hyaluronan; it also hydrolyzed hyaluronan in human skin sections. NNH2 is nontoxic, but it indirectly potentiates the hemorrhagic activity of hemorrhagic complex-I. Curcumin, indomethacin, and tannic acid inhibited dose dependently the degradation of hyaluronan by NNH2.
Antony Gomes - One of the best experts on this subject based on the ideXlab platform.
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nano gold conjugation anti arthritic potential and toxicity studies of snake naja kaouthia lesson 1831 venom protein toxin nkct1 in male albino rats and mice
Indian Journal of Experimental Biology, 2014Co-Authors: Partha Pratim Saha, Tanmoy Bhowmik, Anjan Kumar Dasgupta, Antony GomesAbstract:Nanoscience and Nanotechnology have found their way in the fields of pharmacology and medicine. The conjugation of drug to nanoparticles combines the properties of both. In this study, gold nanoparticle (GNP) was conjugated with NKCT1, a cytotoxic protein toxin from Indian Cobra venom for evaluation of anti-arthritic activity and toxicity in experimental animal models. GNP conjugated NKCT1 (GNP-NKCT1) synthesized by NaBH4 reduction method was stable at room temperature (25 +/- 2 degrees C), pH 7.2. Hydrodynamic size of GNP-NKCT1 was 68-122 nm. Arthritis was developed by Freund's complete adjuvant induction in male albino rats and treatment was done with NKCT1/GNP-NKCT1/standard drug. The paw/ankle swelling, urinary markers, serum markers and cytokines were changed significantly in arthritic control rats which were restored after GNP-NKCT1 treatment. Acute toxicity study revealed that GNP conjugation increased the minimum lethal dose value of NKCT1 and partially reduced the NKCT1 induced increase of the serum biochemical tissue injury markers. Histopathological study showed partial restoration of toxic effect in kidney tissue after GNP conjugation. Normal lymphocyte count in culture was in the order of GNP-NKCT1 > NKCT1 > Indomethacine treatment. The present study confirmed that GNP conjugation increased the antiarthritic activity and decreased toxicity profile of NKCT1.
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Antileukemic potential of PEGylated gold nanoparticle conjugated with protein toxin (NKCT1) isolated from Indian Cobra (Naja kaouthia) venom
Cancer Nanotechnology, 2013Co-Authors: Tanmoy Bhowmik, Partha Pratim Saha, Anjan Dasgupta, Antony GomesAbstract:Limited efficacy of current first-line treatment for leukemia calls attention for further development of efficient strategies. Recently, much attention has been given to nanoparticle-based drug delivery systems loaded with dual drugs to improve current disease therapies by overcoming toxicity. In the present study, we document to explore an approach to conjugate gold nanoparticles (GNPs) with protein toxin (NKCT1), a protein toxin from the Indian Cobra ( Naja kaouthia ) venom, and to establish its antileukemic activity. GNP was prepared by NaBH_4 reduction method. UV–vis spectroscopy of GNP showed the absorbance at 530 nm for plasma resonance. Dynamic light scattering (DLS) size of GNPs was 2–8 nm and the GNP-NKCT1 was 68–122 nm. CD spectra of GNP-NKCT1 showed change in percentage of β-turn as compared with NKCT1. GNP-NKCT1 significantly inhibited leukemic cell growth in dose- and time-dependent manner by two- to threefold more than NKCT1. For human leukemic lymphoma cell line and human myelogenous leukemic cell line, the IC50 dose was found to be 1.2 and 0.75 μg/ml, respectively, observed by trypan blue exclusion method and tetrazolium bromide reduction assay. Flow cytometric analysis showed appreciable number of both cell lines in early and late apoptotic stages and arrested cell cycle in the G1 phase by GNP-NKCT1. Resilient power of leukemic cell line after wound healing and migration or invasive power of the cell line was significantly low in GNP-NKCT1-treated plate than the control plate. These analyses reveal that GNP-NKCT1 possesses significant and selective anticancer activity, likely by inducing programmed cell death through mitochondrial and/or lysosomal pathway.
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inhibition of leukemic u937 cell growth by induction of apoptosis cell cycle arrest and suppression of vegf mmp 2 and mmp 9 activities by cytotoxin protein nn 32 purified from Indian spectacled Cobra naja naja venom
Toxicon, 2013Co-Authors: Tanaya Das, Antony Gomes, Shamik Bhattacharya, Archita Biswas, Shubho Das Gupta, Aparna GomesAbstract:A cytotoxin NN-32 (6.7 kDa) from Indian Cobra (Naja naja) venom inhibited human leukemic U937 cell growth as observed by Trypan blue dye exclusion method and cytotoxicity was confirmed by MTT assay. NN-32 induced apoptosis of U937 cell and cell cycle arrest of sub-G1 phase were revealed by FACS analysis. Increased Bax/Bcl-2 ratio, increased caspase 3 and 9 activities, cleaved PARP, decreased VEGF, MMP-2 and MMP-9 activities were observed after NN-32 treatment of U937 cell. Antileukemic activity of NN-32 on U937 cell may be due to activation of apoptosis, arresting cell cycle and antiangiogenesis activities.
Tv Gowda - One of the best experts on this subject based on the ideXlab platform.
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PRIMARY STRUCTURE OF A CYTOTOXIN-LIKE BASIC PROTEIN FROM NAJA NAJA NAJA (Indian Cobra) VENOM
International journal of peptide and protein research, 2009Co-Authors: A.s. Babu, Kamal D. Puri, Tv GowdaAbstract:The complete amino acid sequence of a cytotoxin-like basic protein (CLBP) from the venom of Naja naja naja (Indian Cobra) was determined by manual degradation using a 4-dimethylaminoazobenzene-4'-isothiocyanate double-coupling method. Peptide fragments obtained by chemical cleavage with cyanogen bromide and enzymic cleavages with trypsin and Staphylococcus aureus proteases for sequence analysis were purified by reversed-phase chromatography. The total number of amino acid residues was 61, with leucine as the C-terminal residue. (C) Munksgaard 1995.
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isolation and characterization of hyaluronidase a spreading factor from Indian Cobra naja naja venom
Biochimie, 2004Co-Authors: K.s. Girish, R. Shashidharamurthy, Tv Gowda, S Nagaraju, K. KemparajuAbstract:Hyaluronidase, ubiquitous enzyme in snake venoms, known originally as "spreading factor", has not been well studied. The present study describes the purification and characterization of hyaluronidase from Indian Cobra (Naja naja) venom and provides systematic evaluation of the spreading property of the enzyme. Hyaluronidase (NNH1) has been purified through gel permeation and ion exchange chromatography. The molecular mass was found to be 70.406 kDa by MALDI-TOF mass spectrometry and with the (p)i pI of 9.2. The amino acid sequence of the N-terminus was found to be NEQSTHGAYV. The enzyme shows absolute specificity for hyaluronan and belongs to the group of neutral active enzymes. Tetrasaccharides are the final product of hyaluronan digestion. The enzyme cleaves beta 1,4-glycosidic linkage and belongs to a group of endo-beta-N-acetyl hexosaminidases. Hyaluronidase indirectly potentiates the myotoxicity of VRV-PL-VIII, a phospholipolytic myotoxin, and also the hemorrhagic potency of a hemorrhagic complex-I. Localization of hyaluronan in human skin section and selective degradation by venom hyaluronidase (NNH1) corroborate the plausible in vivo degradation of hyaluronan in the extracellular matrix (ECM) resulting in easy dissemination of VRV-PL-VIII myotoxin and hemorrhagic complex-I.
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isolation and characterization of hyaluronidase a spreading factor from Indian Cobra naja naja venom
Biochimie, 2004Co-Authors: K.s. Girish, R. Shashidharamurthy, Tv Gowda, S Nagaraju, K. KemparajuAbstract:Hyaluronidase, ubiquitous enzyme in snake venoms, known originally as ``spreading factor'', has not been well studied. The present study describes the purification and characterization of hyaluronidase from Indian Cobra (Naja naja) venom and provides systematic evaluation of the spreading property of the enzyme. Hyaluronidase (NNH1) has been purified through gel permeation and ion exchange chromatography. The molecular mass was found to be 70.406 kDa by MALDI-TOF mass spectrometry and with the Pi pI of 9.2. The amino acid sequence of the N-terminus was found to be NEQSTHGAYV. The enzyme shows absolute specificity for hyaluronan and belongs to the group of neutral active enzymes. Tetrasaccharides are the final product of hyaluronan digestion. The enzyme cleaves beta 1,4-glycosidic linkage and belongs to a group of endo-beta-N-acetyl hexosaminidases. Hyaluronidase indirectly potentiates the myotoxicity of VRV-PL-VIII, a phospholipolytic myotoxin, and also the hemorrhagic potency of a hemorrhagic complex-I. Localization of hyaluronan in human skin section and selective degradation by venom hyaluronidase (NNH1) corroborate the plausible in vivo degradation of hyaluronan in the extracellular matrix (ECM) resulting in easy dissemination of VRV-PL-VIII myotoxin and hemorrhagic complex-I. (C) 2004 Elsevier SAS. All rights reserved.
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Purification and characterization of three acidic, cytotoxic phospholipases A(2) from Indian Cobra (Naja naja naja) venom
1998Co-Authors: L.m.s Rudrammaji, Tv GowdaAbstract:Three acidic phospholipases A(2) (NN-I-2c-PLA(2), NN-I-2d-PLA(2) and NN-I-2e-PLA(2)) were purified by successive chromatography of Indian Cobra (Naja naja naja) venom on CM-Sephadex C-25, Sephadex G-50 and QAE-Sephadex A-25 columns. They have molecular weights of 13 000-14 500 by sodium dodecyl sulphatepolyacrylamide gel electrophoresis. They showed tryptophan specific fluorescence emission spectra (similar to 345 nm). All the three phospholipases A(2) were enzymatically highly active with specific activities 9-17 mu mol min(-1) mg(-1) They were non-lethal to mice when injected intraperitoneally in doses up to 10 mg kg(-1) body weight. They induced edema in mouse foot pads and were cytotoxic to Ehrlich ascites tumour cells. They did not exhibit direct haemolytic and anticoagulant activities. (C) 1998 Elsevier Science Ltd. All rights reserved.
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Purification and characterization of three acidic, cytotoxic phospholipases A2 from Indian Cobra (Naja naja naja) venom
Toxicon : official journal of the International Society on Toxinology, 1998Co-Authors: L.m.s Rudrammaji, Tv GowdaAbstract:Abstract Three acidic phospholipases A 2 (NN-I 2c -PLA 2 , NN-I 2d -PLA 2 and NN-I 2e -PLA 2 ) were purified by successive chromatography of Indian Cobra ( Naja naja naja ) venom on CM–Sephadex C-25, Sephadex G-50 and QAE–Sephadex A-25 columns. They have molecular weights of 13 000–14 500 by sodium dodecyl sulphate–polyacrylamide gel electrophoresis. They showed tryptophan specific fluorescence emission spectra (∼345 nm). All the three phospholipases A 2 were enzymatically highly active with specific activities 9–17 μ mol min −1 mg −1 . They were non-lethal to mice when injected intraperitoneally in doses up to 10 mg kg −1 body weight. They induced edema in mouse foot pads and were cytotoxic to Ehrlich ascites tumour cells. They did not exhibit direct haemolytic and anticoagulant activities.
K.s. Girish - One of the best experts on this subject based on the ideXlab platform.
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inhibition of naja naja venom hyaluronidase role in the management of poisonous bite
Life Sciences, 2006Co-Authors: K.s. Girish, K. KemparajuAbstract:Hyaluronidase is present virtually in all snake venoms and has been known as a “spreading factor.” The enzyme damages the extracellular matrix at the site of the bite, leading to severe morbidity. In this study, the benefits of inhibiting the hyaluronidase activity of Indian Cobra (Naja naja) venom have been investigated. Anti-NNH1 and aristolochic acid both inhibited the in vitro activity of the purified hyaluronidase, (NNH1) and the hyaluronidase activity of whole venom in a dose-dependent manner. Both anti-NNH1 and aristolochic acid abolished the degradation of hyaluronan in human skin tissue sections by NNH1 and by whole venom. Aristolochic acid quenched the fluorescent emission of NNH1. A non-competitive mechanism of NNH1 inhibition was observed with aristolochic acid. NNH1 potentiates the toxicity of Daboia russellii VRV-PL-VIII myotoxin and hemorrhagic complex-I. However, the potentiation of toxicity was inhibited dose-dependently by anti-NNH1 and aristolochic acid. Further, mice injected with whole venom which had been preincubated with anti-NNH1/aristolochic acid, showed more than a two-fold increase in survival time, compared to mice injected with venom alone. A more moderate increase in survival time was observed when mice were injected with anti-NNH1/aristolochic acid 10 min after whole venom injection. This study illustrates the significance of venom hyaluronidase in the pathophysiology of snake venom poisoning and the therapeutic value of its inhibition.
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a low molecular weight isoform of hyaluronidase purification from Indian Cobra naja naja venom and partial characterization
Biochemistry, 2005Co-Authors: K.s. Girish, K. KemparajuAbstract:A low molecular weight isoform of hyaluronidase (NNH2) has been isolated from Indian Cobra (Naja naja) venom by successive chromatography on Sephadex G-75 and CM-Sephadex C-25 columns. The apparent molecular weight determined by SDS-PAGE is 52 kD, and the pI value is 9.7. NNH2 is an endoglycosidase and exhibits in vitro absolute specificity for hyaluronan; it also hydrolyzed hyaluronan in human skin sections. NNH2 is nontoxic, but it indirectly potentiates the hemorrhagic activity of hemorrhagic complex-I. Curcumin, indomethacin, and tannic acid inhibited dose dependently the degradation of hyaluronan by NNH2.
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isolation and characterization of hyaluronidase a spreading factor from Indian Cobra naja naja venom
Biochimie, 2004Co-Authors: K.s. Girish, R. Shashidharamurthy, Tv Gowda, S Nagaraju, K. KemparajuAbstract:Hyaluronidase, ubiquitous enzyme in snake venoms, known originally as "spreading factor", has not been well studied. The present study describes the purification and characterization of hyaluronidase from Indian Cobra (Naja naja) venom and provides systematic evaluation of the spreading property of the enzyme. Hyaluronidase (NNH1) has been purified through gel permeation and ion exchange chromatography. The molecular mass was found to be 70.406 kDa by MALDI-TOF mass spectrometry and with the (p)i pI of 9.2. The amino acid sequence of the N-terminus was found to be NEQSTHGAYV. The enzyme shows absolute specificity for hyaluronan and belongs to the group of neutral active enzymes. Tetrasaccharides are the final product of hyaluronan digestion. The enzyme cleaves beta 1,4-glycosidic linkage and belongs to a group of endo-beta-N-acetyl hexosaminidases. Hyaluronidase indirectly potentiates the myotoxicity of VRV-PL-VIII, a phospholipolytic myotoxin, and also the hemorrhagic potency of a hemorrhagic complex-I. Localization of hyaluronan in human skin section and selective degradation by venom hyaluronidase (NNH1) corroborate the plausible in vivo degradation of hyaluronan in the extracellular matrix (ECM) resulting in easy dissemination of VRV-PL-VIII myotoxin and hemorrhagic complex-I.
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isolation and characterization of hyaluronidase a spreading factor from Indian Cobra naja naja venom
Biochimie, 2004Co-Authors: K.s. Girish, R. Shashidharamurthy, Tv Gowda, S Nagaraju, K. KemparajuAbstract:Hyaluronidase, ubiquitous enzyme in snake venoms, known originally as ``spreading factor'', has not been well studied. The present study describes the purification and characterization of hyaluronidase from Indian Cobra (Naja naja) venom and provides systematic evaluation of the spreading property of the enzyme. Hyaluronidase (NNH1) has been purified through gel permeation and ion exchange chromatography. The molecular mass was found to be 70.406 kDa by MALDI-TOF mass spectrometry and with the Pi pI of 9.2. The amino acid sequence of the N-terminus was found to be NEQSTHGAYV. The enzyme shows absolute specificity for hyaluronan and belongs to the group of neutral active enzymes. Tetrasaccharides are the final product of hyaluronan digestion. The enzyme cleaves beta 1,4-glycosidic linkage and belongs to a group of endo-beta-N-acetyl hexosaminidases. Hyaluronidase indirectly potentiates the myotoxicity of VRV-PL-VIII, a phospholipolytic myotoxin, and also the hemorrhagic potency of a hemorrhagic complex-I. Localization of hyaluronan in human skin section and selective degradation by venom hyaluronidase (NNH1) corroborate the plausible in vivo degradation of hyaluronan in the extracellular matrix (ECM) resulting in easy dissemination of VRV-PL-VIII myotoxin and hemorrhagic complex-I. (C) 2004 Elsevier SAS. All rights reserved.
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Variation in biochemical and pharmacological properties of Indian Cobra (Naja naja naja) venom due to geographical distribution
Molecular and Cellular Biochemistry, 2002Co-Authors: R. Shashidharamurthy, D.k. Jagadeesha, K.s. Girish, K. KemparajuAbstract:Indian Cobra ( Naja naja naja ) venom obtained from three different geographical regions was studied in terms of electrophoretic pattern, biochemical and pharmacological activities. SDS-PAGE banding pattern revealed significant variation in the protein constituents of the three regional venoms. The eastern venom showed highest indirect hemolysis and hyaluronidase activity. In contrast, western and southern venoms were rich in proteolytic activity. All the three regional venoms were devoid of p-tosyl-L-arginine methyl ester hydrolysing activity. The eastern venom was found to be most lethal among the three regional venoms. The lethal potency varied as eastern > western > southern regional venoms. In addition, all the three regional venoms showed marked variations in their ability to induce symptoms/signs of neurotoxicity, myotoxicity, edema and effect on plasma coagulation process. Polyclonal antiserum prepared against the venom of eastern region cross-reacted with both southern and western regional venoms, but varied in the extent of cross-reactivity by ouchterlony immunodiffusion and ELISA.
Tanmoy Bhowmik - One of the best experts on this subject based on the ideXlab platform.
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nano gold conjugation anti arthritic potential and toxicity studies of snake naja kaouthia lesson 1831 venom protein toxin nkct1 in male albino rats and mice
Indian Journal of Experimental Biology, 2014Co-Authors: Partha Pratim Saha, Tanmoy Bhowmik, Anjan Kumar Dasgupta, Antony GomesAbstract:Nanoscience and Nanotechnology have found their way in the fields of pharmacology and medicine. The conjugation of drug to nanoparticles combines the properties of both. In this study, gold nanoparticle (GNP) was conjugated with NKCT1, a cytotoxic protein toxin from Indian Cobra venom for evaluation of anti-arthritic activity and toxicity in experimental animal models. GNP conjugated NKCT1 (GNP-NKCT1) synthesized by NaBH4 reduction method was stable at room temperature (25 +/- 2 degrees C), pH 7.2. Hydrodynamic size of GNP-NKCT1 was 68-122 nm. Arthritis was developed by Freund's complete adjuvant induction in male albino rats and treatment was done with NKCT1/GNP-NKCT1/standard drug. The paw/ankle swelling, urinary markers, serum markers and cytokines were changed significantly in arthritic control rats which were restored after GNP-NKCT1 treatment. Acute toxicity study revealed that GNP conjugation increased the minimum lethal dose value of NKCT1 and partially reduced the NKCT1 induced increase of the serum biochemical tissue injury markers. Histopathological study showed partial restoration of toxic effect in kidney tissue after GNP conjugation. Normal lymphocyte count in culture was in the order of GNP-NKCT1 > NKCT1 > Indomethacine treatment. The present study confirmed that GNP conjugation increased the antiarthritic activity and decreased toxicity profile of NKCT1.
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Antileukemic potential of PEGylated gold nanoparticle conjugated with protein toxin (NKCT1) isolated from Indian Cobra (Naja kaouthia) venom
Cancer Nanotechnology, 2013Co-Authors: Tanmoy Bhowmik, Partha Pratim Saha, Anjan Dasgupta, Antony GomesAbstract:Limited efficacy of current first-line treatment for leukemia calls attention for further development of efficient strategies. Recently, much attention has been given to nanoparticle-based drug delivery systems loaded with dual drugs to improve current disease therapies by overcoming toxicity. In the present study, we document to explore an approach to conjugate gold nanoparticles (GNPs) with protein toxin (NKCT1), a protein toxin from the Indian Cobra ( Naja kaouthia ) venom, and to establish its antileukemic activity. GNP was prepared by NaBH_4 reduction method. UV–vis spectroscopy of GNP showed the absorbance at 530 nm for plasma resonance. Dynamic light scattering (DLS) size of GNPs was 2–8 nm and the GNP-NKCT1 was 68–122 nm. CD spectra of GNP-NKCT1 showed change in percentage of β-turn as compared with NKCT1. GNP-NKCT1 significantly inhibited leukemic cell growth in dose- and time-dependent manner by two- to threefold more than NKCT1. For human leukemic lymphoma cell line and human myelogenous leukemic cell line, the IC50 dose was found to be 1.2 and 0.75 μg/ml, respectively, observed by trypan blue exclusion method and tetrazolium bromide reduction assay. Flow cytometric analysis showed appreciable number of both cell lines in early and late apoptotic stages and arrested cell cycle in the G1 phase by GNP-NKCT1. Resilient power of leukemic cell line after wound healing and migration or invasive power of the cell line was significantly low in GNP-NKCT1-treated plate than the control plate. These analyses reveal that GNP-NKCT1 possesses significant and selective anticancer activity, likely by inducing programmed cell death through mitochondrial and/or lysosomal pathway.