M. Narasu | Jawaharlal Nehru Technological University Hyderabad (original) (raw)
Papers by M. Narasu
Tetrahedron Letters, 2017
We present a convenient route for the synthesis of C6-amino-C5’-N-cyclopropyl carboxamido-C2-alky... more We present a convenient route for the synthesis of C6-amino-C5’-N-cyclopropyl carboxamido-C2-alkynylated purine nucleoside analogues 11a-g via Sonogashira coupling reaction. The nine step synthesis is easy to perform, employing commercially available reagents. Compound 9 is used as key intermediate for the synthesis of analogues 11a-g. Synthetic intermediates and final products are appropriately characterized by IR, 1H NMR, 13C NMR and Mass. The modified nucleoside analogues 11a-g is evaluated for in vitro anticancer activity against MDA-MB-231 and Caco-2 cell lines. Screening data reveals that compounds 11b and 11e displayed potent IC50 value of 7.9, 6.8 µg/mL respectively against MDA-MB-231 and of 7.5, 8.3 µg/mL respectively against Caco-2 than the standard drug doxorubicin, thus establishing the potential anti-cancer properties of these newer derivatives.
Tetrahedron Letters
We present a convenient route for the synthesis of C6-amino-C5’-N-cyclopropyl carboxamido-C2-alky... more We present a convenient route for the synthesis of C6-amino-C5’-N-cyclopropyl carboxamido-C2-alkynylated purine nucleoside analogues 11a-g via Sonogashira coupling reaction. The nine step synthesis is easy to perform, employing commercially available reagents. Compound 9 is used as key intermediate for the synthesis of analogues 11a-g. Synthetic intermediates and final products are appropriately characterized by IR, 1H NMR, 13C NMR and Mass. The modified nucleoside analogues 11a-g is evaluated for in vitro anticancer activity against MDA-MB-231 and Caco-2 cell lines. Screening data reveals that compounds 11b and 11e displayed potent IC50 value of 7.9, 6.8 µg/mL respectively against MDA-MB-231 and of 7.5, 8.3 µg/mL respectively against Caco-2 than the standard drug doxorubicin, thus establishing the potential anti-cancer properties of these newer derivatives.
Indian journal of applied research, 2016
Kaempferol is a flavonoid found abundantly in many plants. Many investigations revealed its anti-... more Kaempferol is a flavonoid found abundantly in many plants. Many investigations revealed its anti-microbial, anti-cancer, cardio protective, anti-oxidant activities. In the present study, we screened the effect of kaempferol on human colon cancer cell line (HCT-15) and human normal lymphocytes at different concentrations. The morphological alterations were observed using microscopy and the cytotoxicity was evaluated by MTT assay. Kaempferol showed dose dependent antiproliferative activity on HCT-15 after 24hrs of incubation. The study was compared with chemotherapeutic drugs Doxorubicin and Cisplatin. The IC 50 concentrations were found to be 120 μg/ml , 50 ug/mland 25 ug/ml for Kaempferol, Doxorubucin and Cisplatin respectively. However, normal human lymphocytes were not affected with Kaempferol.
Cancer is the main leading cause of cancer deaths w orld ide and chemotherapy is mainly used to t... more Cancer is the main leading cause of cancer deaths w orld ide and chemotherapy is mainly used to treat c ancer. However, the severe side effects of the drugs led t he researchers to search for an alternative. Gallic c d, being a polyphenols, has been reported for its antiprolifer ative activity against many cancer cell lines. More ov r, its cytoprotective activity made gallic acid a potential compound in c an er therapy. Since colon cancer and breast cancer s ar among the most prevalent, the present study examines the anti cancer property of gallic acid against two these tw o cancers, HCT15, human colon cancer cell line and MDA MB 231, human breast cancer cell line. The finding estimat ed the IC50 of the compound against the two cell lines. The pre sent study also predicted the possible mechanism of the activity to be apoptosis, yet a detailed study is needed to find o ut the molecular targets.
Journal of Biological Chemistry
Indian Journal of Veterinary Pathology, 2008
Physiology and Molecular Biology of Plants, 2016
Journal of Biological Chemistry
ABSTRACT
Abstract The alcohol extract of Thespesia populnea Soland ex. Correa, a traditional medicinal pla... more Abstract The alcohol extract of Thespesia populnea Soland ex. Correa, a traditional medicinal plant, was shown to have anti-hepatotoxicity against carbon tetrachloride (CCl4) toxicity in rats. Follow up studies were carried out to isolate the active principle or fraction and evaluate its efficacy against various toxic chemicals-induced liver damages and hepatitis B virus. An active ethyl acetate fraction was isolated from the alcohol extract of T. Populnea by activity guided solvent fractionation. The fraction was further separated into two major components using column chromatography. The fast moving component on thin layer chromatogram (TLC) with an Rf value of 0.8 was found to be the active component which showed positive reaction to phenolic acids. HPLC analysis showed 3 closely related compounds in the phenolic acid fraction (PF). The hepatoprotective activity in rats of the PF against CCl4, acetaminophen and thioacetamide was evaluated by measuring levels of serum marker enzym...
BioMed research international, 2014
Alpha-galactosidase production in submerged fermentation by Acinetobacter sp. was optimized using... more Alpha-galactosidase production in submerged fermentation by Acinetobacter sp. was optimized using feed forward neural networks and genetic algorithm (FFNN-GA). Six different parameters, pH, temperature, agitation speed, carbon source (raffinose), nitrogen source (tryptone), and K2HPO4, were chosen and used to construct 6-10-1 topology of feed forward neural network to study interactions between fermentation parameters and enzyme yield. The predicted values were further optimized by genetic algorithm (GA). The predictability of neural networks was further analysed by using mean squared error (MSE), root mean squared error (RMSE), mean absolute error (MAE), mean absolute percentage error (MAPE), and R2-value for training and testing data. Using hybrid neural networks and genetic algorithm, alpha-galactosidase production was improved from 7.5 U/mL to 10.2 U/mL.
Indian journal of biochemistry & biophysics, 1988
Biochemical Education, 1999
Malaria continues to be a serious health problem in the tropical world. The fight against malaria... more Malaria continues to be a serious health problem in the tropical world. The fight against malaria is ridden with a number of technical, logistic and economic problems. It is becoming increasingly difficult to manage due to failure of mosquito control methods and development of resistance of parasite to the existing antimalarials. The search for newer antimalarial has led to the development of artemisinin, which is a promising drug. This article discusses the present status in the production and development of artemisinin and its potential as a potent antimalarial drug.
E-Journal of Chemistry, 2008
Solubility and dissolution rate of three antiretroviral drugs such as lamivudine, zidovudine and ... more Solubility and dissolution rate of three antiretroviral drugs such as lamivudine, zidovudine and stavudine was studied in four media having different pH. The samples were analyzed by usingUVVisible spectrophotometer. lamivudine shows more solubility that is 276.08 mg/mL in 0.01 N HCl. Stavudine showing highest solubility that is 101.23 mg/mL in pH 4.5 acetate buffer. Zidovudine showing highest solubility that is 28.90 mg/mL in both water and 0.01 N HCl. All three drugs showing lower solubility in pH 6.8 phosphate buffer. Lamivudine and stavudine showing good dissolution rate in all media and showing similar release profiles and good correlation, whereas in zidovudine it was clearly observed a slower release at initial time points and then faster release profiles. The solubility and dissolution data in various media is helpful in predicting the bioavailability and also in dissolution method development.
Chem-Bio Informatics Journal, 2010
CDK2 (Cyclin Dependent Kinase 2) acts as a potential therapeutic target in cancer and several eff... more CDK2 (Cyclin Dependent Kinase 2) acts as a potential therapeutic target in cancer and several efforts have been made to find more specific, potent and selective ATP competitive CDK2 inhibitors. In this paper, we report a virtual screening approach that resulted in 54,558 Lipinski compliant hits from ZINC database based on the features exhibited by four compounds from our previous study. Docking and scoring of all compounds using GOLD (Genetic Optimisation for Ligand Docking) software, to evaluate the affinity of binding towards CDK2 enzyme 2UZO resulted in dock scores between 41.71-82.33 kcal/mol. The resultant dataset of 392 hits were filtered based on the specificity between CDK2 and GSK-3 (Glycogen Synthase Kinase-3) to obtain 17 compounds that are more specific towards CDK2. Further, re-scoring of 17 best docked poses followed by a consensus scoring approach tested with five different scoring functions such as GOLD score, CHEM score implemented in GOLD 3.1, eHiTS_score (electronic High Throughput Screening), MolDock score of Molegro software and X-Score retrieved top hits. Finally, the top ten compounds were examined for anti-proliferative effects against human lung adenocarcinoma epithelial cell line, A549 using MTT assay.
Tetrahedron Letters, 2014
E-Journal of Chemistry, 2008
New simple, sensitive, rapid, reproducible and economical spectrophotometric method have been dev... more New simple, sensitive, rapid, reproducible and economical spectrophotometric method have been developed for the determination of amoxicillin trihydrate in pharmaceutical bulk and tablet dosage form using citro phosphate buffer pH 7.2. The system obeys Lambert-Beer's law at 231 nm in the concentration range 2.5-50 μg/m. Molar absorptivity, correlation coefficient and Sandell's sensitivity values were found to be 1.0020 x 104mol-1cm,-10.9996 and 0.03906 μg cm-2respectively. The proposed methods have been successfully applied to the analysis of the bulk drug and its tablet dosage form. The methods have been statistically evaluated and were found to be precise and accurate
Volume 3 (2) CONTENTS April-2009 Reviews Molecular farming of Plant Derived Edible Vaccines 113-D... more Volume 3 (2) CONTENTS April-2009 Reviews Molecular farming of Plant Derived Edible Vaccines 113-D.K. Das Regulation of CIP/KIP cell cycle inhibitors and their biological implications 128-Jinhwa Lee Information to Authors The Current Trends in Biotechnology and Pharmacy is an official international journal of Association of Biotechnology and Pharmacy. It is a peer reviewed quarterly journal dedicated to publish high quality original research articles in biotechnology and pharmacy. The journal will accept contributions from all areas of biotechnology and pharmacy including plant, animal, industrial, microbial, medical, pharmaceutical and analytical biotechnologies, immunology, proteomics, genomics, metabolomics, bioinformatics and different areas in pharmacy such as, pharmaceutics, pharmacology, pharmaceutical chemistry, pharma analysis and pharmacognosy. In addition to the original research papers, review articles in the above mentioned fields will also be considered.
World Journal of Gastroenterology, 2008
AIM: To study the hepatoprotective capacity of Sapindus mukorossi (S. mukorossi) and Rheum emodi ... more AIM: To study the hepatoprotective capacity of Sapindus mukorossi (S. mukorossi) and Rheum emodi (R. emodi) extracts in CCl4 treated male rats. METHODS: The dried powder of S. mukorossi and R. emodi was extracted successively with petroleum ether, benzene, chloroform, and ethanol and concentrated in vacuum. Primary rat hepatocyte monolayer cultures were used for in vitro studies. In vivo , the hepatoprotective capacity of the extract of the fruit pericarp of S. mukorossi and the rhizomes of R. emodi was analyzed in liver injured CCl4-treated male rats. RESULTS: In vitro : primary hepatocytes monolayer c u l t u re s w e re t re a t e d w i t h C C l 4 a n d e x t ra c t s o f S. mukorossi & R. emodi. A protective activity could be demonstrated in the CCl4 damaged primary monolayer culture. In vivo : extracts of the fruit pericarp of S. mukorossi (2.5 mg/mL) and rhizomes of R. emodi (3.0 mg/mL) were found to have protective properties in rats with CCl4 induced liver damage as judged from serum marker enzyme activities. CONCLUSION: The extracts of S. mukorossi and R. emodi do have a protective capacity both in vitro
Research in Veterinary Science, 2008
Infectious bovine rhinotracheitis (IBR) virus causes vulvovaginitis, abortion and respiratory dis... more Infectious bovine rhinotracheitis (IBR) virus causes vulvovaginitis, abortion and respiratory disease in cows and heifers. Betapropiolactone (BPL) is a disinfectant, effective against bacteria, fungi and viruses. It is also used to prepare inactivated vaccines because it destroys the nucleic acid core of viruses but does not damage the capsid. For the validation of BPL when used as an inactivant, it is more important to assure the quality of inactivating agent and the validity of the inactivation process. In the present study, the inactivation kinetics of IBR virus was determined with different concentration of BPL (1:250, 1:500, 1:1000, 1:1500, 1:2000 and 1:2500) at 4 and 37 degrees C. The result indicated that the BPL at 4 degrees C was able to inactivate the IBR virus within 4, 5 and 12h with the concentration of 1:250, 1:500 and 1:1000, respectively. BPL at 37 degrees C was able to inactivate virus within 30 min with the concentration of 1:250. BPL with the concentration of 1:500 and 1:1000 were able to inactivate the virus within 120 min at 37 degrees C. Based on the kinetic study seven formulations were prepared and a sero conversion study of IBR inactivated vaccine was carried out. Serological response in animals to different formulations did not differ significantly (P>0.05).
Protein Expression and Purification, 2008
Among the four human malarial species, Plasmodium falciparum causes most of the mortality associa... more Among the four human malarial species, Plasmodium falciparum causes most of the mortality associated with malaria. Several approaches are being pursued to develop a suitable malaria vaccine since it may be the most effective weapon to fight against malaria. A highly immunogenic, synthetic protein consisting of 21 epitopes from pre-erythrocytic and blood stages of P. falciparum (FALVAC-1A) was constructed and expressed in Escherichia coli. This vaccine candidate was highly immunogenic and induced protective antibodies in rabbits when produced through lab-scale processes in milligram quantities. In order to take this vaccine candidate for further clinical trial, we optimized the process for industrial scale production and purification. Here we describe various methods used in pilot scale production and characterization of FALVAC-1A. A fed-batch cultivation process in a bioreactor at 10-L scale was optimized to express the protein in high yields as inclusion bodies in E. coli cells with the recombinant plasmids. Methods to solubilize, capture and purify the target protein from the inclusion bodies were optimized and the resultant protein was >95% pure based on SDS-PAGE and RP-HPLC. This protein was then refolded and nativity was confirmed by Far-UV CD spectroscopy. Final purified protein was characterized to estimate yield, purity, mass and confirmed to be free of host cell proteins, nucleic acids and bacterial endotoxins. This study confirms that industrial scale clinical grade FALVAC-1A can be produced in a cost-effective manner for clinical trials.
Tetrahedron Letters, 2017
We present a convenient route for the synthesis of C6-amino-C5’-N-cyclopropyl carboxamido-C2-alky... more We present a convenient route for the synthesis of C6-amino-C5’-N-cyclopropyl carboxamido-C2-alkynylated purine nucleoside analogues 11a-g via Sonogashira coupling reaction. The nine step synthesis is easy to perform, employing commercially available reagents. Compound 9 is used as key intermediate for the synthesis of analogues 11a-g. Synthetic intermediates and final products are appropriately characterized by IR, 1H NMR, 13C NMR and Mass. The modified nucleoside analogues 11a-g is evaluated for in vitro anticancer activity against MDA-MB-231 and Caco-2 cell lines. Screening data reveals that compounds 11b and 11e displayed potent IC50 value of 7.9, 6.8 µg/mL respectively against MDA-MB-231 and of 7.5, 8.3 µg/mL respectively against Caco-2 than the standard drug doxorubicin, thus establishing the potential anti-cancer properties of these newer derivatives.
Tetrahedron Letters
We present a convenient route for the synthesis of C6-amino-C5’-N-cyclopropyl carboxamido-C2-alky... more We present a convenient route for the synthesis of C6-amino-C5’-N-cyclopropyl carboxamido-C2-alkynylated purine nucleoside analogues 11a-g via Sonogashira coupling reaction. The nine step synthesis is easy to perform, employing commercially available reagents. Compound 9 is used as key intermediate for the synthesis of analogues 11a-g. Synthetic intermediates and final products are appropriately characterized by IR, 1H NMR, 13C NMR and Mass. The modified nucleoside analogues 11a-g is evaluated for in vitro anticancer activity against MDA-MB-231 and Caco-2 cell lines. Screening data reveals that compounds 11b and 11e displayed potent IC50 value of 7.9, 6.8 µg/mL respectively against MDA-MB-231 and of 7.5, 8.3 µg/mL respectively against Caco-2 than the standard drug doxorubicin, thus establishing the potential anti-cancer properties of these newer derivatives.
Indian journal of applied research, 2016
Kaempferol is a flavonoid found abundantly in many plants. Many investigations revealed its anti-... more Kaempferol is a flavonoid found abundantly in many plants. Many investigations revealed its anti-microbial, anti-cancer, cardio protective, anti-oxidant activities. In the present study, we screened the effect of kaempferol on human colon cancer cell line (HCT-15) and human normal lymphocytes at different concentrations. The morphological alterations were observed using microscopy and the cytotoxicity was evaluated by MTT assay. Kaempferol showed dose dependent antiproliferative activity on HCT-15 after 24hrs of incubation. The study was compared with chemotherapeutic drugs Doxorubicin and Cisplatin. The IC 50 concentrations were found to be 120 μg/ml , 50 ug/mland 25 ug/ml for Kaempferol, Doxorubucin and Cisplatin respectively. However, normal human lymphocytes were not affected with Kaempferol.
Cancer is the main leading cause of cancer deaths w orld ide and chemotherapy is mainly used to t... more Cancer is the main leading cause of cancer deaths w orld ide and chemotherapy is mainly used to treat c ancer. However, the severe side effects of the drugs led t he researchers to search for an alternative. Gallic c d, being a polyphenols, has been reported for its antiprolifer ative activity against many cancer cell lines. More ov r, its cytoprotective activity made gallic acid a potential compound in c an er therapy. Since colon cancer and breast cancer s ar among the most prevalent, the present study examines the anti cancer property of gallic acid against two these tw o cancers, HCT15, human colon cancer cell line and MDA MB 231, human breast cancer cell line. The finding estimat ed the IC50 of the compound against the two cell lines. The pre sent study also predicted the possible mechanism of the activity to be apoptosis, yet a detailed study is needed to find o ut the molecular targets.
Journal of Biological Chemistry
Indian Journal of Veterinary Pathology, 2008
Physiology and Molecular Biology of Plants, 2016
Journal of Biological Chemistry
ABSTRACT
Abstract The alcohol extract of Thespesia populnea Soland ex. Correa, a traditional medicinal pla... more Abstract The alcohol extract of Thespesia populnea Soland ex. Correa, a traditional medicinal plant, was shown to have anti-hepatotoxicity against carbon tetrachloride (CCl4) toxicity in rats. Follow up studies were carried out to isolate the active principle or fraction and evaluate its efficacy against various toxic chemicals-induced liver damages and hepatitis B virus. An active ethyl acetate fraction was isolated from the alcohol extract of T. Populnea by activity guided solvent fractionation. The fraction was further separated into two major components using column chromatography. The fast moving component on thin layer chromatogram (TLC) with an Rf value of 0.8 was found to be the active component which showed positive reaction to phenolic acids. HPLC analysis showed 3 closely related compounds in the phenolic acid fraction (PF). The hepatoprotective activity in rats of the PF against CCl4, acetaminophen and thioacetamide was evaluated by measuring levels of serum marker enzym...
BioMed research international, 2014
Alpha-galactosidase production in submerged fermentation by Acinetobacter sp. was optimized using... more Alpha-galactosidase production in submerged fermentation by Acinetobacter sp. was optimized using feed forward neural networks and genetic algorithm (FFNN-GA). Six different parameters, pH, temperature, agitation speed, carbon source (raffinose), nitrogen source (tryptone), and K2HPO4, were chosen and used to construct 6-10-1 topology of feed forward neural network to study interactions between fermentation parameters and enzyme yield. The predicted values were further optimized by genetic algorithm (GA). The predictability of neural networks was further analysed by using mean squared error (MSE), root mean squared error (RMSE), mean absolute error (MAE), mean absolute percentage error (MAPE), and R2-value for training and testing data. Using hybrid neural networks and genetic algorithm, alpha-galactosidase production was improved from 7.5 U/mL to 10.2 U/mL.
Indian journal of biochemistry & biophysics, 1988
Biochemical Education, 1999
Malaria continues to be a serious health problem in the tropical world. The fight against malaria... more Malaria continues to be a serious health problem in the tropical world. The fight against malaria is ridden with a number of technical, logistic and economic problems. It is becoming increasingly difficult to manage due to failure of mosquito control methods and development of resistance of parasite to the existing antimalarials. The search for newer antimalarial has led to the development of artemisinin, which is a promising drug. This article discusses the present status in the production and development of artemisinin and its potential as a potent antimalarial drug.
E-Journal of Chemistry, 2008
Solubility and dissolution rate of three antiretroviral drugs such as lamivudine, zidovudine and ... more Solubility and dissolution rate of three antiretroviral drugs such as lamivudine, zidovudine and stavudine was studied in four media having different pH. The samples were analyzed by usingUVVisible spectrophotometer. lamivudine shows more solubility that is 276.08 mg/mL in 0.01 N HCl. Stavudine showing highest solubility that is 101.23 mg/mL in pH 4.5 acetate buffer. Zidovudine showing highest solubility that is 28.90 mg/mL in both water and 0.01 N HCl. All three drugs showing lower solubility in pH 6.8 phosphate buffer. Lamivudine and stavudine showing good dissolution rate in all media and showing similar release profiles and good correlation, whereas in zidovudine it was clearly observed a slower release at initial time points and then faster release profiles. The solubility and dissolution data in various media is helpful in predicting the bioavailability and also in dissolution method development.
Chem-Bio Informatics Journal, 2010
CDK2 (Cyclin Dependent Kinase 2) acts as a potential therapeutic target in cancer and several eff... more CDK2 (Cyclin Dependent Kinase 2) acts as a potential therapeutic target in cancer and several efforts have been made to find more specific, potent and selective ATP competitive CDK2 inhibitors. In this paper, we report a virtual screening approach that resulted in 54,558 Lipinski compliant hits from ZINC database based on the features exhibited by four compounds from our previous study. Docking and scoring of all compounds using GOLD (Genetic Optimisation for Ligand Docking) software, to evaluate the affinity of binding towards CDK2 enzyme 2UZO resulted in dock scores between 41.71-82.33 kcal/mol. The resultant dataset of 392 hits were filtered based on the specificity between CDK2 and GSK-3 (Glycogen Synthase Kinase-3) to obtain 17 compounds that are more specific towards CDK2. Further, re-scoring of 17 best docked poses followed by a consensus scoring approach tested with five different scoring functions such as GOLD score, CHEM score implemented in GOLD 3.1, eHiTS_score (electronic High Throughput Screening), MolDock score of Molegro software and X-Score retrieved top hits. Finally, the top ten compounds were examined for anti-proliferative effects against human lung adenocarcinoma epithelial cell line, A549 using MTT assay.
Tetrahedron Letters, 2014
E-Journal of Chemistry, 2008
New simple, sensitive, rapid, reproducible and economical spectrophotometric method have been dev... more New simple, sensitive, rapid, reproducible and economical spectrophotometric method have been developed for the determination of amoxicillin trihydrate in pharmaceutical bulk and tablet dosage form using citro phosphate buffer pH 7.2. The system obeys Lambert-Beer's law at 231 nm in the concentration range 2.5-50 μg/m. Molar absorptivity, correlation coefficient and Sandell's sensitivity values were found to be 1.0020 x 104mol-1cm,-10.9996 and 0.03906 μg cm-2respectively. The proposed methods have been successfully applied to the analysis of the bulk drug and its tablet dosage form. The methods have been statistically evaluated and were found to be precise and accurate
Volume 3 (2) CONTENTS April-2009 Reviews Molecular farming of Plant Derived Edible Vaccines 113-D... more Volume 3 (2) CONTENTS April-2009 Reviews Molecular farming of Plant Derived Edible Vaccines 113-D.K. Das Regulation of CIP/KIP cell cycle inhibitors and their biological implications 128-Jinhwa Lee Information to Authors The Current Trends in Biotechnology and Pharmacy is an official international journal of Association of Biotechnology and Pharmacy. It is a peer reviewed quarterly journal dedicated to publish high quality original research articles in biotechnology and pharmacy. The journal will accept contributions from all areas of biotechnology and pharmacy including plant, animal, industrial, microbial, medical, pharmaceutical and analytical biotechnologies, immunology, proteomics, genomics, metabolomics, bioinformatics and different areas in pharmacy such as, pharmaceutics, pharmacology, pharmaceutical chemistry, pharma analysis and pharmacognosy. In addition to the original research papers, review articles in the above mentioned fields will also be considered.
World Journal of Gastroenterology, 2008
AIM: To study the hepatoprotective capacity of Sapindus mukorossi (S. mukorossi) and Rheum emodi ... more AIM: To study the hepatoprotective capacity of Sapindus mukorossi (S. mukorossi) and Rheum emodi (R. emodi) extracts in CCl4 treated male rats. METHODS: The dried powder of S. mukorossi and R. emodi was extracted successively with petroleum ether, benzene, chloroform, and ethanol and concentrated in vacuum. Primary rat hepatocyte monolayer cultures were used for in vitro studies. In vivo , the hepatoprotective capacity of the extract of the fruit pericarp of S. mukorossi and the rhizomes of R. emodi was analyzed in liver injured CCl4-treated male rats. RESULTS: In vitro : primary hepatocytes monolayer c u l t u re s w e re t re a t e d w i t h C C l 4 a n d e x t ra c t s o f S. mukorossi & R. emodi. A protective activity could be demonstrated in the CCl4 damaged primary monolayer culture. In vivo : extracts of the fruit pericarp of S. mukorossi (2.5 mg/mL) and rhizomes of R. emodi (3.0 mg/mL) were found to have protective properties in rats with CCl4 induced liver damage as judged from serum marker enzyme activities. CONCLUSION: The extracts of S. mukorossi and R. emodi do have a protective capacity both in vitro
Research in Veterinary Science, 2008
Infectious bovine rhinotracheitis (IBR) virus causes vulvovaginitis, abortion and respiratory dis... more Infectious bovine rhinotracheitis (IBR) virus causes vulvovaginitis, abortion and respiratory disease in cows and heifers. Betapropiolactone (BPL) is a disinfectant, effective against bacteria, fungi and viruses. It is also used to prepare inactivated vaccines because it destroys the nucleic acid core of viruses but does not damage the capsid. For the validation of BPL when used as an inactivant, it is more important to assure the quality of inactivating agent and the validity of the inactivation process. In the present study, the inactivation kinetics of IBR virus was determined with different concentration of BPL (1:250, 1:500, 1:1000, 1:1500, 1:2000 and 1:2500) at 4 and 37 degrees C. The result indicated that the BPL at 4 degrees C was able to inactivate the IBR virus within 4, 5 and 12h with the concentration of 1:250, 1:500 and 1:1000, respectively. BPL at 37 degrees C was able to inactivate virus within 30 min with the concentration of 1:250. BPL with the concentration of 1:500 and 1:1000 were able to inactivate the virus within 120 min at 37 degrees C. Based on the kinetic study seven formulations were prepared and a sero conversion study of IBR inactivated vaccine was carried out. Serological response in animals to different formulations did not differ significantly (P>0.05).
Protein Expression and Purification, 2008
Among the four human malarial species, Plasmodium falciparum causes most of the mortality associa... more Among the four human malarial species, Plasmodium falciparum causes most of the mortality associated with malaria. Several approaches are being pursued to develop a suitable malaria vaccine since it may be the most effective weapon to fight against malaria. A highly immunogenic, synthetic protein consisting of 21 epitopes from pre-erythrocytic and blood stages of P. falciparum (FALVAC-1A) was constructed and expressed in Escherichia coli. This vaccine candidate was highly immunogenic and induced protective antibodies in rabbits when produced through lab-scale processes in milligram quantities. In order to take this vaccine candidate for further clinical trial, we optimized the process for industrial scale production and purification. Here we describe various methods used in pilot scale production and characterization of FALVAC-1A. A fed-batch cultivation process in a bioreactor at 10-L scale was optimized to express the protein in high yields as inclusion bodies in E. coli cells with the recombinant plasmids. Methods to solubilize, capture and purify the target protein from the inclusion bodies were optimized and the resultant protein was >95% pure based on SDS-PAGE and RP-HPLC. This protein was then refolded and nativity was confirmed by Far-UV CD spectroscopy. Final purified protein was characterized to estimate yield, purity, mass and confirmed to be free of host cell proteins, nucleic acids and bacterial endotoxins. This study confirms that industrial scale clinical grade FALVAC-1A can be produced in a cost-effective manner for clinical trials.