The document describes a study that isolated and characterized anti-cancer compounds from marine bacteria. Bacteria were isolated from soil samples and their extracts were screened on cancer cell lines. The ethyl acetate extract of one isolate inhibited cancer cell growth by 87.34% and was non-toxic. 16S rRNA sequencing identified the bacterium as Micrococcus luteus. GC-MS analysis identified the active compound as pyrrolo(1,2-alpha)pyrazine-1,4-dione hexahydro-3-(2-methylpropyl). This compound from M. luteus shows potential as a natural anti-cancer agent.
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is an open access international journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Onion (Allium Cepa) Genotoxicity Test
Laboratory of Ecotoxicology and LCA
Department of Environmental Chemistry, ICT Prague
References:
1. FERETTI, D., ZERBINI, I., ZANI, C., CERETTI, E., MORETTI,M.,MONARCA, S. (2007): Allium cepa chromosome
abberation and micronucleus tests applied to study genotoxicity of extracts from pesticide-treated vegetables and
grapes. Food Addit. Contam. 24 (6): 561-572.
2. RANK, J., NIELSEN, M.H. (1997): Allium anaphase-telophase genotoxicity assay. Department of Environment,
Technology and Social Studies, Roskilde University, Denmark.
Anti neoplastic effect of Eclipta prostrata L. (HepG2) cell lines. Hepatocellular carcinoma (HCC) is a tumor of the liver. HCC is responsible for over 12,000 deaths per year in the United States. It is one of the serious health problems in most developing countries. The present probe proved that ethanol extract of Eclipta prostrata L. significantly suppressed the growth and induced the apoptosis in the liver cancer (HepG2) cell lines. IC50 dose was measured with methyl thiazolyl tetrazolium. 100 μg of extract showed 50% reduction of in HepG2 cell line growth at 48 h of incubation. The whole plant of E. prostrata L. extract-induced apoptotic features of cell death was stained with acridine orange. The intracellular enzymatic antioxidants such as superoxide dismutase, glutathione peroxidase, and catalase were slightly decreased in their activities when compared to control. Thus, the study resolves that E. prostrata L. extract is an effective to prevent or retard the spread of malignant cells and antineoplastic effect.
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is an open access international journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Onion (Allium Cepa) Genotoxicity Test
Laboratory of Ecotoxicology and LCA
Department of Environmental Chemistry, ICT Prague
References:
1. FERETTI, D., ZERBINI, I., ZANI, C., CERETTI, E., MORETTI,M.,MONARCA, S. (2007): Allium cepa chromosome
abberation and micronucleus tests applied to study genotoxicity of extracts from pesticide-treated vegetables and
grapes. Food Addit. Contam. 24 (6): 561-572.
2. RANK, J., NIELSEN, M.H. (1997): Allium anaphase-telophase genotoxicity assay. Department of Environment,
Technology and Social Studies, Roskilde University, Denmark.
Anti neoplastic effect of Eclipta prostrata L. (HepG2) cell lines. Hepatocellular carcinoma (HCC) is a tumor of the liver. HCC is responsible for over 12,000 deaths per year in the United States. It is one of the serious health problems in most developing countries. The present probe proved that ethanol extract of Eclipta prostrata L. significantly suppressed the growth and induced the apoptosis in the liver cancer (HepG2) cell lines. IC50 dose was measured with methyl thiazolyl tetrazolium. 100 μg of extract showed 50% reduction of in HepG2 cell line growth at 48 h of incubation. The whole plant of E. prostrata L. extract-induced apoptotic features of cell death was stained with acridine orange. The intracellular enzymatic antioxidants such as superoxide dismutase, glutathione peroxidase, and catalase were slightly decreased in their activities when compared to control. Thus, the study resolves that E. prostrata L. extract is an effective to prevent or retard the spread of malignant cells and antineoplastic effect.
In Vitro Antibacterial Activities of Cochlospermum planchonii Roots Crude Ext...iosrjce
The antibacterial activities of the methanolic, hot water, chloroform and petroleum ether of
Cochlospermum planchonii root extracts on some clinical bacterial isolates and reference organisms were
investigated using conventional microbiological and microdilution indicator technique. Phytochemical
screenings were also carried on the extracts. The root extracts of the plant exhibited antibacterial activities
against reference strains and clinical isolates of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus
aureus, Shigella flexneri, and Salmonella typhii. However, the susceptibility pattern of the bacteria did not
differ significantly from each other (p>0.05). The methanolic root extracts exhibited the highest antibacterial
activity, its minimum inhibitory concentration (MIC) ranging between 1.25 mg/ml and 5.00mg/ml; and its zones
of inhibition diameter on the various test microorganisms ranging between 8mm and 12mm. The petroleum
ether extracts had the weakest antibacterial activity, with minimum inhibitory concentration of 5.00mg/ml and
its zones of inhibition diameter ranging between 4mm and 7mm. The bioactive constituents in the plant were
alkaloids, tannins, saponins, cardiac glycosides, and sterols. The methanolic extracts of root appeared to be
more biologically active than other extracts and may be more useful in treating human infections caused by
these pathogens.
Preliminary evaluation of the larvicidal efficacy of coelomic fluid of Eudril...inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
In vitro studies on Efflux pump Inhibition of Catharanthus roseus and piperin...inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
DOI:10.21276/ijlssr.2016.2.4.2
neoplastic progression through the action of viral oncoproteins, mainly E6 and E7.Cervical cancer remains the second
most common cancer in women worldwide with India as a major contributor to global burden with an annual incidence of
132,000 new cases and mortality rate of 74,000 deaths annually. In this study turmeric, neem, tulasi and ginger were
selected as natural anticancer drugs. The objective of the study was to analyze the anticancer property of turmeric
(Curcuma longa), neem (Azadirachta indica), tulasi (Occimum sanctum) and ginger (Zingiber officinale) on HeLa cells.
Turmeric, neem, tulasi and ginger capsules (Himalaya’s Company) were used and aqueous and methanolic extracts of the
turmeric, neem, tulasi and ginger were obtained using a soxhlet extraction. To check the efficacy of these drug MTT assay
was performed, that determines % viability and/or cytotoxicity. IC50 of aqueous turmeric, neem, tulasi and ginger extracts
in case of HeLa cells were 17.8, 22, 79.4, 27.86 respectively and in case of methanolic turmeric, neem, tulasi and ginger
extracts 17, 7.35, 75.24 and 16.1 respectively. To confirm apoptosis as the sole reason behind cell death
immunofluorescence based apoptosis assay was performed using TALI image based cytometer. The study has led to
postulate hypothesis that natural drugs e.g. turmeric, neem, tulasi and ginger are potent anti-cancer compound that are
capable of inhibiting the growth of immortal cells by apoptosis. Key-words- Cervical cancer, Human papillomavirus (HPV), Oncoproteins E6 and E7, Natural compounds, HeLa cell
line (adherent), Cell viability and MTT assay, Apoptosis assay
Phytochemical screening and antibacterial properties from extract of Alchorne...Uploadworld
This study involved a survey on the use of extract of Alchornea cordifolia a medicinal plant used locally in Cameroon as traditional medicine for the treatment of infectious diseases.
In vitro experiments of prokaryotic and eukaryotic antimicrobial peptide cyto...AI Publications
These proteinaceous molecules, called antimicrobial peptides (AMPs), are a varied collection of antimicrobial peptides. The ability of AMPs to combat gut infections necessitates further study of the AMP-GI tract interaction. These peptides need to be tested in vitro for cytotoxicity before they may be considered for use in clinical infections. Using the MTT conversion assay, neutral red dye absorption assay, and a comparison to vancomycin, researchers examined the cytotoxicity of gallidermin, nisin A, natural magainin peptides, and melittin in two gastrointestinal cell types (HT29 and Caco-2). Sheep erythrocyte hemolytic activity was also studied, and the influence of AMPs on paracellular permeability was assessed using transepithelial resistance (TEER) and TEM. Gallidermin, nisin A, magainin I, magainin II, and melittin were the least cytotoxic AMPs. To our knowledge, only Melittin and NIS caused considerable hemolysis. There are two distinct ways that melittin and nisin differ in their ability to kill bacteria. It was the only AMP that had an effect on the permeability of the paracellular space. Intestinal tight junctions and cell–cell adhesion were destroyed by long-term melittin therapy, as were microvilli, cell debris, and cell–cell adhesion. Antimicrobial activity and low cytotoxicity make Gallidermin a promising therapeutic drug. The antibacterial properties of Melittin are limited, but its ability to transport poorly bioavailable medicines may be useful.
Design and Analysis of Bolted Joint in Composite LaminatedIJMER
In this work plate was designed for single and four bolted joint with two different materials such as mild steel and E-glass fiber. The aim of this work is to examine the distribution of tensile and crushing stress among the different bolts by changing material of plates and bolt. The bolted joints for mild steel plate and composite laminate were analyzed by using FEA. The result shows that tensile stress and crushing stress is less for composite laminate compare to mild steel .It is concluded that Weight reduction of structure is also achieved for e-glass fiber structure. The stress concentration was reduced in composite laminate bolted joints compare to mild steel so this will improve strength of structure.
Bolts are widely used as critical load transferring components. Despite the importance to integrity and safety, little attention has been paid to the correct use of bolts and bolting materials.
In Vitro Antibacterial Activities of Cochlospermum planchonii Roots Crude Ext...iosrjce
The antibacterial activities of the methanolic, hot water, chloroform and petroleum ether of
Cochlospermum planchonii root extracts on some clinical bacterial isolates and reference organisms were
investigated using conventional microbiological and microdilution indicator technique. Phytochemical
screenings were also carried on the extracts. The root extracts of the plant exhibited antibacterial activities
against reference strains and clinical isolates of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus
aureus, Shigella flexneri, and Salmonella typhii. However, the susceptibility pattern of the bacteria did not
differ significantly from each other (p>0.05). The methanolic root extracts exhibited the highest antibacterial
activity, its minimum inhibitory concentration (MIC) ranging between 1.25 mg/ml and 5.00mg/ml; and its zones
of inhibition diameter on the various test microorganisms ranging between 8mm and 12mm. The petroleum
ether extracts had the weakest antibacterial activity, with minimum inhibitory concentration of 5.00mg/ml and
its zones of inhibition diameter ranging between 4mm and 7mm. The bioactive constituents in the plant were
alkaloids, tannins, saponins, cardiac glycosides, and sterols. The methanolic extracts of root appeared to be
more biologically active than other extracts and may be more useful in treating human infections caused by
these pathogens.
Preliminary evaluation of the larvicidal efficacy of coelomic fluid of Eudril...inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
In vitro studies on Efflux pump Inhibition of Catharanthus roseus and piperin...inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
DOI:10.21276/ijlssr.2016.2.4.2
neoplastic progression through the action of viral oncoproteins, mainly E6 and E7.Cervical cancer remains the second
most common cancer in women worldwide with India as a major contributor to global burden with an annual incidence of
132,000 new cases and mortality rate of 74,000 deaths annually. In this study turmeric, neem, tulasi and ginger were
selected as natural anticancer drugs. The objective of the study was to analyze the anticancer property of turmeric
(Curcuma longa), neem (Azadirachta indica), tulasi (Occimum sanctum) and ginger (Zingiber officinale) on HeLa cells.
Turmeric, neem, tulasi and ginger capsules (Himalaya’s Company) were used and aqueous and methanolic extracts of the
turmeric, neem, tulasi and ginger were obtained using a soxhlet extraction. To check the efficacy of these drug MTT assay
was performed, that determines % viability and/or cytotoxicity. IC50 of aqueous turmeric, neem, tulasi and ginger extracts
in case of HeLa cells were 17.8, 22, 79.4, 27.86 respectively and in case of methanolic turmeric, neem, tulasi and ginger
extracts 17, 7.35, 75.24 and 16.1 respectively. To confirm apoptosis as the sole reason behind cell death
immunofluorescence based apoptosis assay was performed using TALI image based cytometer. The study has led to
postulate hypothesis that natural drugs e.g. turmeric, neem, tulasi and ginger are potent anti-cancer compound that are
capable of inhibiting the growth of immortal cells by apoptosis. Key-words- Cervical cancer, Human papillomavirus (HPV), Oncoproteins E6 and E7, Natural compounds, HeLa cell
line (adherent), Cell viability and MTT assay, Apoptosis assay
Phytochemical screening and antibacterial properties from extract of Alchorne...Uploadworld
This study involved a survey on the use of extract of Alchornea cordifolia a medicinal plant used locally in Cameroon as traditional medicine for the treatment of infectious diseases.
In vitro experiments of prokaryotic and eukaryotic antimicrobial peptide cyto...AI Publications
These proteinaceous molecules, called antimicrobial peptides (AMPs), are a varied collection of antimicrobial peptides. The ability of AMPs to combat gut infections necessitates further study of the AMP-GI tract interaction. These peptides need to be tested in vitro for cytotoxicity before they may be considered for use in clinical infections. Using the MTT conversion assay, neutral red dye absorption assay, and a comparison to vancomycin, researchers examined the cytotoxicity of gallidermin, nisin A, natural magainin peptides, and melittin in two gastrointestinal cell types (HT29 and Caco-2). Sheep erythrocyte hemolytic activity was also studied, and the influence of AMPs on paracellular permeability was assessed using transepithelial resistance (TEER) and TEM. Gallidermin, nisin A, magainin I, magainin II, and melittin were the least cytotoxic AMPs. To our knowledge, only Melittin and NIS caused considerable hemolysis. There are two distinct ways that melittin and nisin differ in their ability to kill bacteria. It was the only AMP that had an effect on the permeability of the paracellular space. Intestinal tight junctions and cell–cell adhesion were destroyed by long-term melittin therapy, as were microvilli, cell debris, and cell–cell adhesion. Antimicrobial activity and low cytotoxicity make Gallidermin a promising therapeutic drug. The antibacterial properties of Melittin are limited, but its ability to transport poorly bioavailable medicines may be useful.
Design and Analysis of Bolted Joint in Composite LaminatedIJMER
In this work plate was designed for single and four bolted joint with two different materials such as mild steel and E-glass fiber. The aim of this work is to examine the distribution of tensile and crushing stress among the different bolts by changing material of plates and bolt. The bolted joints for mild steel plate and composite laminate were analyzed by using FEA. The result shows that tensile stress and crushing stress is less for composite laminate compare to mild steel .It is concluded that Weight reduction of structure is also achieved for e-glass fiber structure. The stress concentration was reduced in composite laminate bolted joints compare to mild steel so this will improve strength of structure.
Bolts are widely used as critical load transferring components. Despite the importance to integrity and safety, little attention has been paid to the correct use of bolts and bolting materials.
Influence of Mineralogy and Fabric on the Engineering Properties of the Miang...IJMER
Abstract: The major problem associated with the use of rock aggregates in engineering construction, is the difficulty of
predicting their probable field performance. This is mainly due to the inadequate understanding of the decisive factors that
control their engineering behavior. The mineralogy and fabric of the Miango granite porphyry was studied to assess their
influence on engineering properties. The uniaxial compressive strength and aggregate crushing values show that the rocks are
weak while other tests such as aggregate impact strength, water absorption, and absorption under saturation, soundness and
specific gravity values are fairly good. However, thin section studies revealed three distinctive features which greatly influence
the physico-mechanical properties: (a) abundant fractures of varying sizes (b) Sericitization of the orthoclase and or plagioclase
feldspars (c) intergrowth of quartz and or orthoclase feldspars. The quartz grains shows extensive cracking which further reduces
their mechanical strength. The strength loss of the granite porphyry could be attributed to the presence of the fractures on the
quartz grains and the sericitization of the orthoclase and plagioclase feldspars. Geotechnical characterization of the rocks shows
that they can be utilized as roadstone or could be polished and used as facing stones because of their non disintegration to
sulphate attack and the large feldspar phenocrysts in the rock.
Key Words: Aggregate Impact Value Tests; Aggregate Crushing Value; Water Absorption; Crushing Load
Study of Performance of Different Blends of Biodiesel Prepared From Waste Co...IJMER
The use of biodiesel is rapidly expanding around the world, making it imperative to fully
understand the impacts of biodiesel on the diesel engine combustion process and pollutant formation.
Biodiesel was made by the well-known transesterification process. Waste cottonseed oil was selected for
biodiesel production. Three different blends of biodiesel were prepared i.e. B10, B20 and B30. These three
blends were fuelled in a compression ignition (C.I.) engine. A maximum of 77% biodiesel was produced
with 20% methanol in presence of 0.5% sodium hydroxide. Different parameters for the optimization of
biodiesel production were investigated in the first phase of this study, while in the next phase of the study
performance test of a diesel engine with neat diesel fuel and biodiesel mixtures are to be carried out. The
performance characteristics like brake power (B.P.), brake specific fuel consumption (BSFC) and brake
thermal efficiency. This performance was then compared with that of petro diesel.
Regression analysis of shot peening process for performance characteristics o...IJMER
International Journal of Modern Engineering Research (IJMER) is Peer reviewed, online Journal. It serves as an international archival forum of scholarly research related to engineering and science education.
An Enhance PSO Based Approach for Solving Economical Ordered Quantity (EOQ) P...IJMER
The Meta-heuristic approaches can provide a sufficiently good solution to an
optimization problem, especially with incomplete or imperfect information and with lower
computational complexity especially for numerical solutions. This paper presents an enhanced PSO
(Particle Swarm Optimization) technique for solving the same problem. Through the PSO performs well
but it may require some more iteration to converge or sometimes many repetitions for the complex
problems. To overcome these problems an enhanced PSO is presented which utilizes the PSO with
double chaotic maps to perform irregular velocity updates which forces the particles to search greater
space for best global solution. Finally the comparison between both algorithms is performed for the
EOQ problem considering deteriorating items, shortages and partially backlogging. The simulation
results shows that the proposed enhanced PSO converges quickly and found much closer solution then
PSO.
Modified Procedure for Construction and Selection of Sampling Plans for Vari...IJMER
Linear Trend is Technique to generate the values for observerd frequency distribution and it
will give the accuracy of the smoothing obtained depends on the number of available data sets. In this
article ,an attempt was made to estimate the modified liner trend value generator for construction and
selection of sampling plans for variable inspection scheme indexed through the MAAOQ over the Liner
Trend .We compare the constructed sampling plans indexed through MAAOQ over Linear Trend with
the basic sampling plans indexed with AOQL. And also obtain the performance of the operating
characteristic curves.
Content Cytotoxicity Studies of Colorectal Carcinoma Cells Using Printed Impe...journalBEEI
Monitoring the effectiveness of drugs on cancer cells is crucial for chemotherapeutics studies. In-vitro cell-based biosensors can be used as an alternative for characteristic studies of cells’ response to drugs. Cell-based sensors provide real-time measurements and require smaller sample volumes compared to conventional T-flask measurement methods. This paper presents a biosensor that detects in real-time, impedance variations of human colon cancer, HCT-116 cells when treated with anti-cancer agent, 5-Fluorouracil (5-FU). Two different extracellular matrix (ECM); polyaniline and gelatin were tested and evaluated in terms of attachment quality. Polyaniline was found to provide the best attachment for HCT-116 cells and was used for cytotoxicity studies. Cytokinetic behavior indicated that 5-FU inhibited HCT-116 cells at IC50 of 6.8 µg/mL. Trypan blue exclusion method for testing cell viability was used to validate the impedance measurements, where the cancer cell concentrations were reduced to ~35% when treated with 2.5 µg/mL, and 50% when treated with 6.8 µg/mL. The results generated by the microfabricated impedance biosensor are comparable to the Trypan blue method since both gave similar cell growth trend. It can be concluded that the impedance biosensor has potential to be used as an alternative method in drug testing applications.
Phytochemical screening and antimicrobial studies of uapaca togoensis (pax) s...theijes
The International Journal of Engineering & Science is aimed at providing a platform for researchers, engineers, scientists, or educators to publish their original research results, to exchange new ideas, to disseminate information in innovative designs, engineering experiences and technological skills. It is also the Journal's objective to promote engineering and technology education. All papers submitted to the Journal will be blind peer-reviewed. Only original articles will be published.
Isolation, Screening and Selection of Fungal Strains for Potential Cellulase ...inventionjournals
The present study was aimed to isolate, screen and identify the potential cellulase and xylanase producing fungi from the soil samples collected from different areas of Haryana. Total one hundred fifty one fungal isolates were isolated from these soil samples were then screened by using selective media (i.e. CMC and Xylan agar) in order to determine the potency of microbes in producing cellulase and xylanase which were indicated by clear zones formation around the cultures. This qualitative screening which showing greater cellulase and xylanase indexes were subjected to enzyme activity tests by Dinitrosalicylic acid (DNS) method. Maximum enzyme production was achieved at 30°C, pH of 6.0 by Trichoderma atroviride on 5th day of incubation.
Isolation, Screening and Selection of Fungal Strains for Potential Cellulase ...inventionjournals
The present study was aimed to isolate, screen and identify the potential cellulase and xylanase producing fungi from the soil samples collected from different areas of Haryana. Total one hundred fifty one fungal isolates were isolated from these soil samples were then screened by using selective media (i.e. CMC and Xylan agar) in order to determine the potency of microbes in producing cellulase and xylanase which were indicated by clear zones formation around the cultures. This qualitative screening which showing greater cellulase and xylanase indexes were subjected to enzyme activity tests by Dinitrosalicylic acid (DNS) method. Maximum enzyme production was achieved at 30°C, pH of 6.0 by Trichoderma atroviride on 5th day of incubation.
Bacterial pigments have many applications in current day to day life. The pigments produced by chromobacteria can be used for various applications like dairy, pharmaceutical, and food etc. In this study, three types of pigments were isolated i.e. yellow from Xanthomonas sp., pinkish Red from Rhodotorula sp., and orange from Sarcina sp. Pigmented bacterial isolates were obtained from the soil samples and used for the pigment extraction study. We studied that the pigment producing bacteria and identified the color producing pigments. Soil samples from Pondicherry, Cuddalore, Chennai, and Andhra sea coast were collected and used for isolation of microbes producing pigments. Purification of extracted pigments were done by column chromatography, whereas identification and characterization of purified pigment done by UV-Visible spectrophotometry and GC/MS analysis etc. The pigment isolated from bacterial sp. were used for the antimicrobial activity, antioxidant, and anticancer & transformation studies. The bacterial extracts of carotenoid pigment extracted and used as natural colorants for food products and dying of cloth.
Key-words: - Soil samples, GC/MS analysis, UV-Visible spectrophotometry, Carotenoid, Pigment extraction
Phytochemical Screening and Evaluation of Cytotoxicity and Thrombolytic Prope...IOSR Journals
The rural and marginal people of Bangladesh are deprived of modern treatment facilities and hence greatly depend on medicinal plants. Besides, the higher cost and toxicity of synthetic drugs drives scientists towards search for natural source of medication for a number of diseases. Cost-effectiveness, easy availability and fewer side effects are making the herbal medicine more popular both among rural and city people. Plants with Cytotoxic and clot lysis potential are good candidate as source of novel anti-tumor agents and thrombolytic drugs. This study aimed at screening out of phytochemical constituents and evaluation of cytotoxicity and thrombolytic potential of an important medicinal plant Achyranthes aspera methanolic leaf extract. In vitro phytochemical screening of A. aspera leaf extract carried out by qualitative tests revealed the presence of alkaloids, glycosides, cardiac glycosides, flavonoids, tannins, terpenoids, steroids and saponins while phlobatannins were absent. Cytotoxicity test of A. aspera leaf extract carried out by Brine shrimp Lethality (BSL) Bioassay showed the highest percentage of mortality (90%) in 1250 μg/ml and LC50 value was 50.12 μg/ml. Thrombolytic test showed 32.87 ± 9.42% clot lytic activity for A. aspera while positive control (streptokinase) and negative control (water) showed 81.19 ± 3.78% and 6.67 ± 2.58% clot lysis, respectively. Synergistic effect of streptokinase and A. aspera extract also produced better result (56.30 ± 6.95%) than A. aspera alone.
Antibacterial Effect of Endophytic Actinomycetes from Marine Algae against Multi Drug Resistant Gram Negative Bacteria by Manoharan N in Examines in Marine Biology & Oceanography
In vitro controlling of selected human diarrhea causing bacteria by clove ext...Open Access Research Paper
Antibacterial activity of clove extracts (Syzygium aromaticum L.) was proven against five diarrhea causing bacteria. This was further confirmed when compared with commonly used three commercial antibiotics (ciprofloxacin, tetracycline and erythromycin) as a positive control. Significant differences (P<0.0001) were observed in the effect of the antimicrobial agents (clove extracts and antibiotics), and in the sensitivities of the bacterial species (P<0.0001) to the antimicrobial agents. Clove extracts had significant (P<0.001) activity with the acetone extract demonstrating highest activity followed by antibiotics and other extracts against tested bacteria. The zone of inhibition of clove extracts was ranged from 7.33 to 12.00 mm whereas in antibiotics, it was 0.00 to 11.67 mm. Of all the bacteria, Salmonella typhimurium was the most susceptible against all of the extracts as well as concentrations of clove, while low MIC (180 mgml-1) and MBC (680 mgml-1) of the extracts were observed against Shigella dysenteriae. Consequently, clove has a significant antidiarrheal activity and it could be used as an effective antibacterial agent, alternative to the use of antibiotics.
Degradation of Nevirapine and Trimethoprim from Aqueous Solutions using Selec...Agriculture Journal IJOEAR
Together with pharmaceutical residues, personal care products encompassing prescription drugs, fragrances, and cosmetics have been detected in groundwater and other aquatic environments, hence compromising the quality of water. Their classification as micropollutants is due to their antibacterial resistance potential, persistence, and ecotoxicity. Biodegradation has been identified as a potential mechanism in their removal. The focus of this study focus was bioaugmentation; (Bacillus subtilis, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeroginosa) to enhance the degradation of Nevirapine and Trimethoprim in model aqueous solutions. A liquid chromatography-tandem mass spectrometer (LC-MS/MS) was used to determine the pharmaceuticals. The efficacy of the bacterial strains to degrade selected drugs was evaluated by making the two drugs the sole source of energy and carbon. From the experimental data, the highest percentage biodegradation was recorded; Pseudomonas aeroginosa (86 %) and Staphylococcus aureus (79 %) for TMP and NVP respectively.
A Study on Translucent Concrete Product and Its Properties by Using Optical F...IJMER
- Translucent concrete is a concrete based material with light-transferring properties,
obtained due to embedded light optical elements like Optical fibers used in concrete. Light is conducted
through the concrete from one end to the other. This results into a certain light pattern on the other
surface, depending on the fiber structure. Optical fibers transmit light so effectively that there is
virtually no loss of light conducted through the fibers. This paper deals with the modeling of such
translucent or transparent concrete blocks and panel and their usage and also the advantages it brings
in the field. The main purpose is to use sunlight as a light source to reduce the power consumption of
illumination and to use the optical fiber to sense the stress of structures and also use this concrete as an
architectural purpose of the building
Developing Cost Effective Automation for Cotton Seed DelintingIJMER
A low cost automation system for removal of lint from cottonseed is to be designed and
developed. The setup consists of stainless steel drum with stirrer in which cottonseeds having lint is mixed
with concentrated sulphuric acid. So lint will get burn. This lint free cottonseed treated with lime water to
neutralize acidic nature. After water washing this cottonseeds are used for agriculter purpose
Study & Testing Of Bio-Composite Material Based On Munja FibreIJMER
The incorporation of natural fibres such as munja fiber composites has gained
increasing applications both in many areas of Engineering and Technology. The aim of this study is to
evaluate mechanical properties such as flexural and tensile properties of reinforced epoxy composites.
This is mainly due to their applicable benefits as they are light weight and offer low cost compared to
synthetic fibre composites. Munja fibres recently have been a substitute material in many weight-critical
applications in areas such as aerospace, automotive and other high demanding industrial sectors. In
this study, natural munja fibre composites and munja/fibreglass hybrid composites were fabricated by a
combination of hand lay-up and cold-press methods. A new variety in munja fibre is the present work
the main aim of the work is to extract the neat fibre and is characterized for its flexural characteristics.
The composites are fabricated by reinforcing untreated and treated fibre and are tested for their
mechanical, properties strictly as per ASTM procedures.
Hybrid Engine (Stirling Engine + IC Engine + Electric Motor)IJMER
Hybrid engine is a combination of Stirling engine, IC engine and Electric motor. All these 3 are
connected together to a single shaft. The power source of the Stirling engine will be a Solar Panel. The aim of
this is to run the automobile using a Hybrid engine
Fabrication & Characterization of Bio Composite Materials Based On Sunnhemp F...IJMER
The present day technology demands eco-friendly developments. In this era the
composite material are playing a vital roal in different field of Engineering .The composite materials
are using as a principle materials. Nowaday the composite materials are utilizing as a important
component of engineering field .Where as the importance of the applications of composites is well
known, but thrust on the use of natural fibres in it for reinforcement has been given priority for some
times. But changing from synthetic fibres to natural fibres provides only half green-composites. A
partial green composite will be achieved if the matrix component is also eco-friendly. Keeping this in
view, a detailed literature surveyed has been carried out through various issues of the Journals
related to this field. The material systems used are sunnhemp fibres. Some epoxy and hardener has
been also added for stability and drying of the bio-composites. Various graphs and bar-charts are
super-imposed on each other for comparison among themselves and Graphs is plotted on MAT LAB
and ORIGIN 6.0 software. To determining tensile strengths, Various properties for different biocomposites
have been compared among themselves. Comparison of the behaviour of bio-composites of
this work has been also compare with other works. The bio-composites developed in this work are
likely to get applications in fall ceilings, partitions, bio-degradable packagings, automotive interiors,
sports things (e.g. rackets, nets, etc.), toys etc.
Geochemistry and Genesis of Kammatturu Iron Ores of Devagiri Formation, Sandu...IJMER
The Greenstone belts of Karnataka are enriched in BIFs in Dharwar craton, where Iron
formations are confined to the basin shelf, clearly separated from the deeper-water iron formation that
accumulated at the basin margin and flanking the marine basin. Geochemical data procured in terms of
major, trace and REE are plotted in various diagrams to interpret the genesis of BIFs. Al2O3, Fe2O3 (T),
TiO2, CaO, and SiO2 abundances and ratios show a wide variation. Ni, Co, Zr, Sc, V, Rb, Sr, U, Th,
ΣREE, La, Ce and Eu anomalies and their binary relationships indicate that wherever the terrigenous
component has increased, the concentration of elements of felsic such as Zr and Hf has gone up. Elevated
concentrations of Ni, Co and Sc are contributed by chlorite and other components characteristic of basic
volcanic debris. The data suggest that these formations were generated by chemical and clastic
sedimentary processes on a shallow shelf. During transgression, chemical precipitation took place at the
sediment-water interface, whereas at the time of regression. Iron ore formed with sedimentary structures
and textures in Kammatturu area, in a setting where the water column was oxygenated.
Experimental Investigation on Characteristic Study of the Carbon Steel C45 in...IJMER
In this paper, the mechanical characteristics of C45 medium carbon steel are investigated
under various working conditions. The main characteristic to be studied on this paper is impact toughness
of the material with different configurations and the experiment were carried out on charpy impact testing
equipment. This study reveals the ability of the material to absorb energy up to failure for various
specimen configurations under different heat treated conditions and the corresponding results were
compared with the analysis outcome
Non linear analysis of Robot Gun Support Structure using Equivalent Dynamic A...IJMER
Robot guns are being increasingly employed in automotive manufacturing to replace
risky jobs and also to increase productivity. Using a single robot for a single operation proves to be
expensive. Hence for cost optimization, multiple guns are mounted on a single robot and multiple
operations are performed. Robot Gun structure is an efficient way in which multiple welds can be done
simultaneously. However mounting several weld guns on a single structure induces a variety of
dynamic loads, especially during movement of the robot arm as it maneuvers to reach the weld
locations. The primary idea employed in this paper, is to model those dynamic loads as equivalent G
force loads in FEA. This approach will be on the conservative side, and will be saving time and
subsequently cost efficient. The approach of the paper is towards creating a standard operating
procedure when it comes to analysis of such structures, with emphasis on deploying various technical
aspects of FEA such as Non Linear Geometry, Multipoint Constraint Contact Algorithm, Multizone
meshing .
Static Analysis of Go-Kart Chassis by Analytical and Solid Works SimulationIJMER
This paper aims to do modelling, simulation and performing the static analysis of a go
kart chassis consisting of Circular beams. Modelling, simulations and analysis are performed using 3-D
modelling software i.e. Solid Works and ANSYS according to the rulebook provided by Indian Society of
New Era Engineers (ISNEE) for National Go Kart Championship (NGKC-14).The maximum deflection is
determined by performing static analysis. Computed results are then compared to analytical calculation,
where it is found that the location of maximum deflection agrees well with theoretical approximation but
varies on magnitude aspect.
In récent year various vehicle introduced in market but due to limitation in
carbon émission and BS Séries limitd speed availability vehicle in the market and causing of
environnent pollution over few year There is need to decrease dependancy on fuel vehicle.
bicycle is to be modified for optional in the future To implement new technique using change in
pedal assembly and variable speed gearbox such as planetary gear optimise speed of vehicle
with variable speed ratio.To increase the efficiency of bicycle for confortable drive and to
reduce torque appli éd on bicycle. we introduced epicyclic gear box in which transmission done
throgh Chain Drive (i.e. Sprocket )to rear wheel with help of Epicyclical gear Box to give
number of différent Speed during driving.To reduce torque requirent in the cycle with change in
the pedal mechanism
Integration of Struts & Spring & Hibernate for Enterprise ApplicationsIJMER
The proposal of this paper is to present Spring Framework which is widely used in
developing enterprise applications. Considering the current state where applications are developed using
the EJB model, Spring Framework assert that ordinary java beans(POJO) can be utilize with minimal
modifications. This modular framework can be used to develop the application faster and can reduce
complexity. This paper will highlight the design overview of Spring Framework along with its features that
have made the framework useful. The integration of multiple frameworks for an E-commerce system has
also been addressed in this paper. This paper also proposes structure for a website based on integration of
Spring, Hibernate and Struts Framework.
Microcontroller Based Automatic Sprinkler Irrigation SystemIJMER
Microcontroller based Automatic Sprinkler System is a new concept of using
intelligence power of embedded technology in the sprinkler irrigation work. Designed system replaces
the conventional manual work involved in sprinkler irrigation to automatic process. Using this system a
farmer is protected against adverse inhuman weather conditions, tedious work of changing over of
sprinkler water pipe lines & risk of accident due to high pressure in the water pipe line. Overall
sprinkler irrigation work is transformed in to a comfortableautomatic work. This system provides
flexibility & accuracy in respect of time set for the operation of a sprinkler water pipe lines. In present
work the author has designed and developed an automatic sprinkler irrigation system which is
controlled and monitored by a microcontroller interfaced with solenoid valves.
On some locally closed sets and spaces in Ideal Topological SpacesIJMER
In this paper we introduce and characterize some new generalized locally closed sets
known as
δ
ˆ
s-locally closed sets and spaces are known as
δ
ˆ
s-normal space and
δ
ˆ
s-connected space and
discussed some of their properties
Intrusion Detection and Forensics based on decision tree and Association rule...IJMER
This paper present an approach based on the combination of, two techniques using
decision tree and Association rule mining for Probe attack detection. This approach proves to be
better than the traditional approach of generating rules for fuzzy expert system by clustering methods.
Association rule mining for selecting the best attributes together and decision tree for identifying the
best parameters together to create the rules for fuzzy expert system. After that rules for fuzzy expert
system are generated using association rule mining and decision trees. Decision trees is generated for
dataset and to find the basic parameters for creating the membership functions of fuzzy inference
system. Membership functions are generated for the probe attack. Based on these rules we have
created the fuzzy inference system that is used as an input to neuro-fuzzy system. Fuzzy inference
system is loaded to neuro-fuzzy toolbox as an input and the final ANFIS structure is generated for
outcome of neuro-fuzzy approach. The experiments and evaluations of the proposed method were
done with NSL-KDD intrusion detection dataset. As the experimental results, the proposed approach
based on the combination of, two techniques using decision tree and Association rule mining
efficiently detected probe attacks. Experimental results shows better results for detecting intrusions as
compared to others existing methods
Natural Language Ambiguity and its Effect on Machine LearningIJMER
"Natural language processing" here refers to the use and ability of systems to process
sentences in a natural language such as English, rather than in a specialized artificial computer
language such as C++. The systems of real interest here are digital computers of the type we think of as
personal computers and mainframes. Of course humans can process natural languages, but for us the
question is whether digital computers can or ever will process natural languages. We have tried to
explore in depth and break down the types of ambiguities persistent throughout the natural languages
and provide an answer to the question “How it affects the machine translation process and thereby
machine learning as whole?” .
Today in era of software industry there is no perfect software framework available for
analysis and software development. Currently there are enormous number of software development
process exists which can be implemented to stabilize the process of developing a software system. But no
perfect system is recognized till yet which can help software developers for opting of best software
development process. This paper present the framework of skillful system combined with Likert scale. With
the help of Likert scale we define a rule based model and delegate some mass score to every process and
develop one tool name as MuxSet which will help the software developers to select an appropriate
development process that may enhance the probability of system success.
Material Parameter and Effect of Thermal Load on Functionally Graded CylindersIJMER
The present study investigates the creep in a thick-walled composite cylinders made
up of aluminum/aluminum alloy matrix and reinforced with silicon carbide particles. The distribution
of SiCp is assumed to be either uniform or decreasing linearly from the inner to the outer radius of
the cylinder. The creep behavior of the cylinder has been described by threshold stress based creep
law with a stress exponent of 5. The composite cylinders are subjected to internal pressure which is
applied gradually and steady state condition of stress is assumed. The creep parameters required to
be used in creep law, are extracted by conducting regression analysis on the available experimental
results. The mathematical models have been developed to describe steady state creep in the composite
cylinder by using von-Mises criterion. Regression analysis is used to obtain the creep parameters
required in the study. The basic equilibrium equation of the cylinder and other constitutive equations
have been solved to obtain creep stresses in the cylinder. The effect of varying particle size, particle
content and temperature on the stresses in the composite cylinder has been analyzed. The study
revealed that the stress distributions in the cylinder do not vary significantly for various combinations
of particle size, particle content and operating temperature except for slight variation observed for
varying particle content. Functionally Graded Materials (FGMs) emerged and led to the development
of superior heat resistant materials.
Energy Audit is the systematic process for finding out the energy conservation
opportunities in industrial processes. The project carried out studies on various energy conservation
measures application in areas like lighting, motors, compressors, transformer, ventilation system etc.
In this investigation, studied the technical aspects of the various measures along with its cost benefit
analysis.
Investigation found that major areas of energy conservation are-
1. Energy efficient lighting schemes.
2. Use of electronic ballast instead of copper ballast.
3. Use of wind ventilators for ventilation.
4. Use of VFD for compressor.
5. Transparent roofing sheets to reduce energy consumption.
So Energy Audit is the only perfect & analyzed way of meeting the Industrial Energy Conservation.
An Implementation of I2C Slave Interface using Verilog HDLIJMER
The focus of this paper is on implementation of Inter Integrated Circuit (I2C) protocol
following slave module for no data loss. In this paper, the principle and the operation of I2C bus protocol
will be introduced. It follows the I2C specification to provide device addressing, read/write operation and
an acknowledgement. The programmable nature of device provide users with the flexibility of configuring
the I2C slave device to any legal slave address to avoid the slave address collision on an I2C bus with
multiple slave devices. This paper demonstrates how I2C Master controller transmits and receives data to
and from the Slave with proper synchronization.
The module is designed in Verilog and simulated in ModelSim. The design is also synthesized in Xilinx
XST 14.1. This module acts as a slave for the microprocessor which can be customized for no data loss.
Discrete Model of Two Predators competing for One PreyIJMER
This paper investigates the dynamical behavior of a discrete model of one prey two
predator systems. The equilibrium points and their stability are analyzed. Time series plots are obtained
for different sets of parameter values. Also bifurcation diagrams are plotted to show dynamical behavior
of the system in selected range of growth parameter
Discrete Model of Two Predators competing for One Prey
Ds2645104515
1. International Journal of Modern Engineering Research (IJMER)
www.ijmer.com Vol.2, Issue.6, Nov-Dec. 2012 pp-4510-4515 ISSN: 2249-6645
Isolation and Molecular Characterization of Anti-Cancerous Compound
Producing Marine Bacteria by Using 16S rRNA Sequencing and GC-MS
Techniques
Sai Lakshman Mithun.V1, C.S.V.Ramachandra Rao2
Department of Biotechnology, DVR & Dr.HS MIC College of Technology, Kanchikacherla, Andhra Pradesh, India.
Abstract: Extracts from microorganisms have served as a valuable source of diverse molecules in many drug discoveries.
Identification of microbial strains having promising biological activities and purifying the bio-molecules which are
responsible for the biological activities, have led to the discovery of many bioactive molecules. Extracts of bacteria in vitro
tested on various cancer cell lines. The lyophilized bacterial extract powder was dissolved in various chemical solvents like
Methanol, Chloroform and Ethyl acetate. From them cytotoxic assays was performed the extracts were screened on HCT 15
and MES- SA cancer cell lines to study the cytotoxic potential. Where the Ethyl acetate showed the inhibition 87.34% when
compared to other solvents. The Ethyl acetate extract of isolate showed promising results by MTT assay and Trypan blue
staining. Identification of the microorganism, the selected isolates was characterized by using the 16s rRNA sequencing
based on microbial characterization, the compound produced by bacteria was analyzed by GC-MS technique. The organism
was identified as Micrococcus luteus and biochemical tests of the extracts were also carried out.
Key words: Anticancer, Bacterial extracts, MTT assay, HCT 15 cell line, MES-SA cell line.
I. INTRODUCTION
Cancer is a disease in which a cell, or a group of cells represents uncontrolled growth invasion (intrusion on and
distortion of adjacent tissues), and metastasis (spread from one part to another part in the body through lymph or blood).
These three malignant properties differentiate cancer from benign tumors, which are self-limited and do not invade or
metastasize while the malignant tumors are not self limited and metastasize. Most of cancers occur form a tumor; the
oncology is deals with the study, diagnosis, treatment, and prevention of cancer and it’s a branch of medicine. Cancer is a
human tragedy that affects people at all ages with the risk in most types increasing with age. Cancers are primarily an
environmental disease with 90-95% of cases due to modification in lifestyle and environmental factors and 5-10% due to
genetics Cancer is caused.
By both external factors (tobacco, chemicals, radiation and infectious organisms) and internal factors (inherited
mutations, hormones, immune conditions and mutation that occurs from metabolism). Common environmental factors
leading to cancer death include: tobacco 25-30%, diet and obesity 30-35%, infections 15-20%, radiation, stress, lack of
physical activity and environmental pollutants. [B Dhorajiya et al., 2011].
Early discovery of carcinogens by John Hill, an English physician, 1761. In it he made the first causal link between
substances in the environment and cancer when he described a relationship between tobacco snuff and nasal cancer. This
brought about the awareness of carcinogens (chemical agents that have been demonstrated to cause cancer). That associated
particular occupations with an increased risk of developing specific forms of cancer the forerunner to the field of public
health and cancer.[C. A. Almeida and S. A. Barry, 2010].
Natural products have played an important role in treating and preventing human diseases. Natural products with
medicinal value have come from various sources viz., terrestrial plants, terrestrial microorganisms, marine organisms, and
terrestrial vertebrates and invertebrates the microbial extracts have served as a valuable source of diverse molecules in many
drug discovery efforts and led to the isolation of several important drugs. [Newman et al., 2002]. The chemical composition
of bioactive compounds of microbial origin is often highly complex. Organic compounds from aqua to terrestrial
microorganisms have extensive use in the treatment of many diseases and serve as compounds of interest both in their
natural form and as templates for synthetic modification. These compounds provided important contributions to the
discovery of antibacterial agents like penicillins, cephalosporins, tetracyclines, and polypeptides, immunosuppressive agents
like cyclosporine, ant diabetic agent like acarbose, cholesterol-lowering agents like lovastatin and mevastatin and anti cancer
agents like peplomycin, pentostatin, and epirubicin [Butler, 2005; Sneader, 2005]
II. MATERIALS AND METHODS
Screening and isolation of bacterial strains:
Collection of soil sample:
Marine soil samples were collected from Bay of Bengal coast of Machilipatnam, Krishna district, AP, India. About
15 grams of soil sediments were collected in a sterile polythene bags and stored at 4 oC until further use. The soil samples
were stored under sterile conditions for preventing the bacterial cross contamination. Then the sample was serially diluted to
till 10-6 dilution by adding 1 gram of soil to 10ml of distilled water.
www.ijmer.com 4510 | Page
2. International Journal of Modern Engineering Research (IJMER)
www.ijmer.com Vol.2, Issue.6, Nov-Dec. 2012 pp-4510-4515 ISSN: 2249-6645
Screening for bacterial single isolates:
From the serially diluted samples, 100µl of sample was poured on Nutrient Agar plates (Himedia). From the 10 -5
-6
and 10 dilutions the single isolated bacterial colonies were obtained on agar plates by the procedure according to [Shirai et
al. (1989).], The bacterial pure cultures were isolated.
Sub culturing of bacterial isolates:
The inoculum was prepared by transferring a single colony of the isolates into 5 ml of the corresponding medium.
This inoculum was transferred to 95 ml of the corresponding broth and bacterial isolates were incubated at c for 2 -3 days
.this broth was used for the extraction of bioactive metabolites.
Preparation of bacterial extracts:
The culture broth was centrifuged at 3000 rpm for 15 min to obtain a clear supernatant. Components were extracted
successively dissolved with chemical solvents such as chloroform (C), Methanol (M) and Ethyl acetate (E) and followed by
vacuum evaporation of these extracts to obtain the dry extracts by vacuum rotary evaporator. [Angel TreasaThomas etal.,
2011]
Sample preparation for biological studies:
The bacterial crude extracts were dissolved in 1 ml DMSO and transferred to sterile vials of 2 ml capacity; these
samples were stored at room temperature for further use, protected from light.
Maintenance of cancer cell lines:
Human colorectal adenocarcinoma Cancer cells (HCT 15) and Human uterine cancer cells (MES-SA) were
maintained in Dulbecco’s modified essential medium (DMEM) supplemented with 4.5 g/l glucose and 2mm 1-glutamine and
5% fetal bovine serum (FBS) (growth medium) at 370c in 5% Co2 incubator.
Trypan blue dye exclusion technique:
A cell suspension was made with a fixed volume of cells (e.g. 1ml). Although an aseptic technique is not essential
in all stages of this procedure, 50uL of cell suspension was taken and mixed it with an equal volume of trypan blue. Solution
was well mixed using a pipette. It transfers to a hemocytometer and then counted live cell as clear form and dead cell as blue
cells. After staining with trypan blue solution counting should commence <5minutes as after that time the cells will begin to
take up the dye. The hemocytometer was placed on the stage of an inverted microscope.
Focus was adjusted and power until a single counting square fills the field. The number of cells per ml, and the total number
of cells were counted using the following formula:
Calculate percent viability by using formula:
% viability = (live cell count/total cell count) ×100
Anticancer assay:
Micro culture tetrazolium (MTT) assay:
The monolayer cell culture was trypsinized and the cell count was adjusted to 1ml using medium. To each well of
96 well microtitre plates, 0.1ml of diluted cell suspension was added. After 72 hour, the sample solution in wells was flicked
off and 50μl of MTT dye was added to each well. The plates were gently shaken and incubated for 4 hours at 37ₒ c in 5%
CO2 incubator. The supernatant was removed, 50 μl of Propanol was added, and the plates were gently shaken to solubilize
the formed formazan. The absorbance was measured using a micro plate reader at a wavelength of 490 nm. The percentage
growth inhibition was calculated using the Formula below:
The percentage growth inhibition was calculated using following formula:
%cell inhibition= 100-{(At-Ab)/ (Ac- Ab)} x100
Where,
At= Absorbance value of test compound
Ab= Absorbance value of blank
Ac=Absorbance value of control [T. Mosmann, J. Immunol. Methods]
Identification of microorganism:
Biomass culture for anticancer activity assays was carried out in 500 mL conical flasks at 37°C temperature. Total
DNA was extracted and PCR amplification was performed according to the method described by [Fawley and Fawley
(2004)] using following primers:
Forward primer- 51 GGCGAACGGGTGAGTAA 31
Reverse primer- 51 ACTGCTGCCTCCCGTAG 31
The Genomic DNA was isolated from the overnight grown bacterial culture was amplified with universal bacterial
primers. A 25μl of reaction mixture contains 15μl of master mix (10 x assay buffer, dntp’s, Taq polymerease and MgCl 2),
1μl of forward primer, 1μl of reverse primer, 2μl of template DNA and 6μl of distilled water. PCR was carried out by the
thermal cycler under the following conditions- An intialization step at 94ºC for 4min followed by 30 cycles of 94ºC for
1min, 52ºC for 1min, 72ºC for 1.15min followed by final extension at 72ºC for 1min and holding temperature at 10ºC for
www.ijmer.com 4511 | Page
3. International Journal of Modern Engineering Research (IJMER)
www.ijmer.com Vol.2, Issue.6, Nov-Dec. 2012 pp-4510-4515 ISSN: 2249-6645
1min. The amplified DNA fragments were observed by agarose gel electrophoresis in 2% agarose gel and sequenced. The
unknown bacterium was identified using
GenBank database. The amplified PCR products were directly sequenced in an ABM Prism 3100-Avant
Sequencer. The obtained sequences were analyzed using BLAST tool to get the relative identification of each
bacterial species. PCR samples were purified, after that the bacterial extracts were subjected to thin layer
chromatography for detecting the activity of extract and their components
Screening of Bacterial metabolite with GC-MS:
The active bacterial extract which was showing the maximum inhibition on cancer cell lines growth was
analyzed by Gas Chromatography and Mass Spectrophotometer, using this technique the main compound
having the anticancerous activity can be detected
III. RESULTS AND DISCUSSION
Cancer cells were tested with Chloroform, Methanol and Ethyl acetate bacterial extracts cell viability was
evaluated by MTT assay. A gradual decrease in the viability of HCT 15 and MES-SA cancer cells were observed in for all
the bacterial extracts used in the study. The bacteria was identified as Micrococcus luteus by 16s r RNA sequencing.
MTT cell growth inhibition assay was taken as in vitro measure of anticancer activity of bacterial extracts by using
different cancer cell lines. Ethyl acetate extracts of bacteria are effective in inhibiting cancer cell growth of the two cancer
cell lines. Therefore present invitro studies on ethyl acetate bacterial extracts demonstrate the remarkable anticancer
potentials due to the presence of active principles may responsible for this anti cancer activity. Hence Micrococcus luteus
Ethyl acetate extracts can be used as a potent natural source of anticancer agent. The GC-MS analyses the bacterial extract
and compound identified as pyrrolo (1, 2-alpha) pyrazine1,4 dione hexahydro 3-(2-methylpropyl)
Isolation of pure cultures:
The bacterial pure cultures were isolated by serial dilution and streak plating on nutrient agar medium and selected
bacterial colonies were sub cultured in to nutrient broth, and then the bacterial cultures were lyophilized and dissolved in
chemical solvents. These bacterial extracts were dissolved in DMSO for long period further use.
FIGURE 1: Isolation of pure culture
Trypan blue staining Report:
The trypan blue staining was performed and the cancer cell line viability cell proliferation as shown below
Table 1: Trypan blue staining % of Viability of cancer cells
Cell line % viability Live cell Total cell PH
count count
MES-SA 81.1% 1.72×105 2.12×105 7.5
HCT 15 72% 1.728×105 2.40×105 6.9
GC-MS Report:
Gas chromatography and mass spectrophotometer analyses the bacterial extract and the metabolite produced by
bacteria is identified as Pyrrolo [1, 2-a] pyrazine-1, 4-dione, hexahydro-3-(2-methylpropyl)
www.ijmer.com 4512 | Page
4. International Journal of Modern Engineering Research (IJMER)
www.ijmer.com Vol.2, Issue.6, Nov-Dec. 2012 pp-4510-4515 ISSN: 2249-6645
FIGURE 1: GC-MS analysis report with bacterial extract
16s rRNA sequencing report:
The selected bacterial extract sample was analyzed by using 16s rRNA sequencing, the isolate was identified as Micrococcus
luteus
>mt_16srRNA
CGCATGGTGGGTGTTGGAAAGATTTATCGGTTTTGGATGGACTCGCGGCCTATCAGCTTGTTGGTGAGGT
AATGGCTCACCAAGGCGACGACGGGTAGCCGGCCTGAGAGGGTGACCGGCCACACTGGGACTGAGACACG
GCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCGACGCC
GCGTGAGGGATGACGGCCTTCGGGTTGTAAACCTCTTTCAGTAGGGAAGAAGCGAAAGTGACGGTACCTG
CAGAAGAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCGAGCGTTATCCGGAAT
TATTGGGCGTAAAGAGCTCGTAGGCGGTTTGTCGCGTCTGTCGTGAAAGTCCGGGGCTTAACCCCGGATC
TGCGGTGGGTACGGGCAGACTAGAGTGCAGTAGGGGAGACTGGAATTCCTGGTGTAGCGGTGGAATGCGC
AGATATCAGGAGGAACACCGATGGCGAAGGCAGGTCTCTGGGCTGTAACTGACGCTGAGGAGCGAAAGCA
TGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGTTGGGCACTAGGTGTGGGGACCA
TTCCACGGTTTCCGCGCCGCAGCTAACGCATTAAGTGCCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAA
CTCAAAGGAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGCGGATTAATTCGATGCAACGCGAAGAAC
CTTACCAAGGCTTGACATGTTCTCGATCGCCGTAGAGATACGGTTTCCCCTTTGGGGCGGGTTCACAGGT
GGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGTT
CCATGTTGCCAGCACGTCGTGGTGGGGACTCATGGGAGACTGCCGGGGTCAACTCGGAGGAAGGTGAGGA
CGACGTCAAATCATCATGCCCCTTATGTCTTGGGCTTCACGCATGCTACAATGGCCGGTACAATGGGTTG
CGATACTGTGAGGTGGAGCTAATCCCAAAAAGCCGGTCTCAGTTCGGATTGGGGTCTGCAACTCGACCCC
MTT assay Report:
From the Table 2 and graph 1: we can say that as the concentration of Chloroform, Methanol and Ethyl
acetate bacterial extracts increased from 20 to 120 µl/ml, the % HCT 15 cancer cell growth inhibition was
increased from 26.19 % to 69.76 % in chloroform bacterial extract, 29.45% to 73.56 % in Methanol bacterial
extract and 33.24 % to 87.34% in Ethyl acetate bacterial extract, that means they induces a cell arrest to inhibit
the growth of the HCT 15cancer cells. The Ethyl acetate bacterial extract showing the promising results
www.ijmer.com 4513 | Page
5. International Journal of Modern Engineering Research (IJMER)
www.ijmer.com Vol.2, Issue.6, Nov-Dec. 2012 pp-4510-4515 ISSN: 2249-6645
Table 2: Percentage of HCT 15 cell inhibition at various concentrations
Concentration Percentage of HCT 15 cell inhibition
µg/ml
Chloroform Methanol Bacterial Ethyl acetate
Bacterial extract extract Bacterial extract
20 26.19 29.45 33.24
40 32.12 38.62 41.77
60 38.47 47.56 57.33
80 45.28 58.08 64.73
100 54.11 62.9 70.65
120 69.76 73.56 87.34
Graph 1: Effect of Chloroform, Methanol and Ethyl acetate bacterial extracts on HCT 15 cancer cell growth
inhibition
IV. CONCLUSION
Bacterial extracts of the metabolites were in vitro tested for their cytotoxic potential on various cancer cell lines.
The extracts were screened on HCT 15 and MES-SA cell lines. The Ethyl acetate extract of bacterial isolate showed
promising results by MTT assay and Trypan blue staining. The isolate was identified as Micrococcus luteus sps by using 16s
rRNA typing, GC-MS analyses the bacterial extract and the metabolite is identified as Pyrrolo [1, 2-a] pyrazine-1,4-dione,
hexahydro-3-(2-methylpropyl) showing promising results were obtained with anti cancerous activity
V. ACKNOWLEDGEMENTS
SAI LAKSHMAN MITHUN.V would like to thank Department of Biotechnology, DVR& Dr.HS
MIC College of Technology, Kanchikacherla, and Andhra Pradesh for their cooperation to complete this work
For Masters degree
REFERENCES
[1] Angel TreasaThomas , Josyula Venkata Rao, Volety Mallikarjuna Subramanian , Hariharapura Raghu Chandrasekhar,
Naseer Maliyakkal1, Tukaram Kedar Kisan, Alex Joseph, Nayanabhirama Udupa, In vitro anticancer activity of
microbial isolates from diverse habitats, Brazilian Journal of Pharmaceutical Sciences, vol. 47, n. 2, apr./jun., 2011
[2] B Dhorajiya, M Malani, B Dholakiya, Extraction and Preservation Protocol of Anti-Cancer Agents from Marine
World, Chemical Sciences Journal, CSJ-38, accepted version, Oct 22, 2011
[3] Angel.T.T, Venkata rao.J, Jesilm.A, Subrahmanyam.V.M, Venkatesh.K.B, Udupa.N. Antimicrobial profile of
extremophiles from aqua to terrestrial habitats. Pharmacology online, v.1, p.111-126, 2009.
[4] T. Mossman, J. Immunol. Methods; 1983, 65, 55.
www.ijmer.com 4514 | Page
6. International Journal of Modern Engineering Research (IJMER)
www.ijmer.com Vol.2, Issue.6, Nov-Dec. 2012 pp-4510-4515 ISSN: 2249-6645
[5] Sanjay patel. Nirav gheewala, Ashok suthar, Anand shah, In-vitro cytotoxicity activity of solanum nigrum extract
against Hela cell line and vero cell line, International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 1, Suppl
1, Nov.-Dec. 2009
[6] CLARDY. J. Discovery of new compounds in nature. Proc. Amer. Phil. Soc., v.151, p.201-210, 2007.
[7] BUTLER, M.S. Natural products to drugs: natural products derived compounds in clinical trials. Nat. Prod. Rep., v.25,
p.475-516, 2008.
[8] HARLEY, J.P.; PRESCOTT, L.M. Laboratory exercises in microbiology. 5. ed. Columbus: The MC Graw Hill
Companies, 2002. P.13-16, 125-207.
[9] ATTA–UR-RAHMAN.; IQBAL, C.E.; WILLIAM, J.T. Bioassay techniques for drug development. Amsterdam:
Harwood Academic Publishers, 2001. P.1-5
[10] Phillips HJ and Terryberry JE. Counting actively metabolizing tissue cultured cells. Cell Research 1957; 13: 341-347.
[11] Masters RW. Animal cell culture, Trypan Blue Assay sop. 3rd ed. 2000, p. 1 – 3
[12] Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D et al. Evaluation of Colorimetric Protein and
Biomass Stains for Assaying Drug Effects upon Human Tumor Cell Lines. Proceedings of the American Association
for Cancer Research 1989; 30: 612.
[13] Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D et al. New Colorimetric Cytotoxicity Assay for
Anticancer-Drug Screening. Journal National Cancer Institute 1990; 82(13), 1107-1112.
[14] Masters RW. Animal cell culture, Cytotoxicity and viability assays. 3rd ed. 2000, p. 202-203.
[15] JEFFREY, M.E.; LINDA, S.A.; ANDREW, O.M. A rapid and simple MTT based spectrophotometric assay for
determining drug sensitivity in monolayer culture. Tissue Cult. Meth, v.11, p.15-17.
[16] McCloud TG, 2010. High Throughput Extraction of Plant, Marine and Fungal Specimens for preservation of
Biologically Active Molecules. Marine Drugs, 15: 4526-4563
[17] MARIANNA, J.; JAMES, P.K.; RONALD, R.R. Macquarimicins, microbial metabolites from Micromonospora I.
Discovery, taxonomy, fermentation and biological properties. J. Antiriot., v.48, p.462-466, 1995
[18] Newman DJ, Cragg GM, 2007. Natural products as sources of new drugs over the last 25 years. Natural Products, 70:
461–477.
[19] Rajeev Kumar Jha, and Xu Zi-rong, 2004 Biomedical Compounds from Marine organisms, Marine drugs 2,123-124
[20] Alejandro M.S. MAYER and Kirk R. GUSTAFSON, Marine pharmacology in 2000: Anti tumor and cytotoxic
compounds, International journal of Cancer: 105, 291–299 (2003)
www.ijmer.com 4515 | Page