Determination of 8-Hydroxy-2 Deoxyguanosine in Pseudomonas Fluorescens Freeze-Dried Exposed to the ROS Using HPLC with Electrochemical Detection Method
Abstract— Oxidative DNA damage is involved in the f cell death induced by freeze-dried powder during storage. Cell 8-hydroxy-2’deoxyguanosine (8-oxodG) is widely accepted as a biomarker of the “freeze-dried bacteria” oxidative DNA damage. The aim of this study was to introduce a method for determination 8-oxodG in cell freeze-dried samples using high-performance liquid chromatography with electrochemical detection. In the tested range of 0.5 µmol L-1 to 1.0 nmol L-1, the calibration curve was linear (r2=0.9995) and the limit of detection was 0.05 µmol L-1. The used method did not allow highlighting the presence in the samples of the 8OH within the limits of detection. A more successful method (more sensitive) would be needed to detect possibly the 8OH.
Electrochemical, in-vitro in-vivo study of Co (II)-ofloxacin complexIOSR Journals
Ofloxacin complex has been synthesized and screened for its physicochemical, microbial as well as pharmacological activity have been done in solid and aqueous phase. On the basis of elemental analysis, polarographic studies, amperometric titration and IR spectral studies the probable formula for the complex has been determined at 30±1OC and ionic strength of μ= 1.0[KCl]. Raper’s paper disc method was used for microbial study against various pathogenic bacteria and fungi.Invivo syudy of Swiss mice [25-30gm] were used for antibacterial activity against ofloxacin and its complex on xyline-Alcoholic activity test Kidney, liver and serum of these rats were also studied. On the basis of observed result it could be concluded that Co(II)-Ofloxacin complex were found to be non-toxic and more potent than pure Ofloxacin.(1)
Synthesis, spectroscopic, magnetic properties and superoxide dismutase (SOD) ...IOSR Journals
Three new ternary copper(II) complexes formulated as [Cu(HIda)(bipy)] 1; [Cu(HIda)(phen)] 2; [Cu(HIda)(dmp)] 3; where HIda =N-(2-hydroxyethyl)-2- iminodiacetic acid ; bipy = 2, 2’- bipyridine; phen = 1,10- phenanthroline; dmp = 2,9-dimethyl 1,10-phenanthroline, have been synthesized and characterized by partial elemental analysis, FAB-mass (m/z), EPR, UV-visible and CV measurements. The magnetic and spectroscopic data of all these complexes 1-3 indicate distorted octahedral geometry. The EPR spectra of these complexes in frozen DMSO solutions showed a single at g ca. 2. The trend in g-value (g||>g>2.0023) suggests that the unpaired electron on copper (II) has dx2–y2 character. The SOD activities of the complexes have been investigated. Antibacterial and antifungal activity of these complexes were also measured and discussed.
A STUDY TO EVALUATE THE IN VITRO ANTIMICROBIAL ACTIVITY AND ANTIANDROGENIC E...Dr. Pradeep mitharwal
The present paper deals with synthesis and characterization
of some new chromium (III) Schiff base complexes using microwave irradiation
technique as well as conventional heating. The S∩N donor benzothiazolines, 1-
(2-furanyl) ethanone benzothiazoline (Bzt1N
∩
SH), 1-(2-thienyl) ethanone
benzothiazoline (Bzt2N
∩
SH) and 1-(2-pyridyl) ethanone benzothiazoline
(Bzt3N
∩
SH) were prepared by the condensation of ortho-aminothiophenol with
respective ketones in ethanol.
— In the present work, impact of UV-B radiation (280-
315nm: 0.4 W m-2) on growth, photosynthetic pigments, protein,
ascorbate, proline and lipid peroxidation have been studied in
two cyanobacteria Nostoc muscorum and Synechocystis PCC
6803. UV-B radiation (2 to 6 hrs) leads to 55% inhibition of
growth in Synechocystis PCC 6803 in comparison to control
where as in Nostoc muscorum growth reduces up to 45%. This
UV-B treatment also significantly decreased the contents of
chlorophyll, carotenoids and phycocyanin. Photosynthetic
pigments decreased with increasing doses of UV-B (2 to 6 hrs)
radiation. However, the inhibitory effect in Synechocystis PCC
6803 was more pronounced than in Nostoc muscorum. With
increasing UV-B exposure period, production of ascorbate (19-
45%), proline (12-29%) and lipid peroxidation was significantly
higher in Synechocystis PCC 6803 as compared to control
sample. It was observed that lipid peroxidation enhanced 33 %
than control sample of Synechocystis PCC 6803. Our result shows
that photosynthetic apparatus is the main target of UV-B
radiation causing degradation of photosynthetic pigments. This
study concluded that Synechocystis PCC 6803 was the susceptible
organism for survival in stress condition than Nostoc muscorum.
Electrochemical, in-vitro in-vivo study of Co (II)-ofloxacin complexIOSR Journals
Ofloxacin complex has been synthesized and screened for its physicochemical, microbial as well as pharmacological activity have been done in solid and aqueous phase. On the basis of elemental analysis, polarographic studies, amperometric titration and IR spectral studies the probable formula for the complex has been determined at 30±1OC and ionic strength of μ= 1.0[KCl]. Raper’s paper disc method was used for microbial study against various pathogenic bacteria and fungi.Invivo syudy of Swiss mice [25-30gm] were used for antibacterial activity against ofloxacin and its complex on xyline-Alcoholic activity test Kidney, liver and serum of these rats were also studied. On the basis of observed result it could be concluded that Co(II)-Ofloxacin complex were found to be non-toxic and more potent than pure Ofloxacin.(1)
Synthesis, spectroscopic, magnetic properties and superoxide dismutase (SOD) ...IOSR Journals
Three new ternary copper(II) complexes formulated as [Cu(HIda)(bipy)] 1; [Cu(HIda)(phen)] 2; [Cu(HIda)(dmp)] 3; where HIda =N-(2-hydroxyethyl)-2- iminodiacetic acid ; bipy = 2, 2’- bipyridine; phen = 1,10- phenanthroline; dmp = 2,9-dimethyl 1,10-phenanthroline, have been synthesized and characterized by partial elemental analysis, FAB-mass (m/z), EPR, UV-visible and CV measurements. The magnetic and spectroscopic data of all these complexes 1-3 indicate distorted octahedral geometry. The EPR spectra of these complexes in frozen DMSO solutions showed a single at g ca. 2. The trend in g-value (g||>g>2.0023) suggests that the unpaired electron on copper (II) has dx2–y2 character. The SOD activities of the complexes have been investigated. Antibacterial and antifungal activity of these complexes were also measured and discussed.
A STUDY TO EVALUATE THE IN VITRO ANTIMICROBIAL ACTIVITY AND ANTIANDROGENIC E...Dr. Pradeep mitharwal
The present paper deals with synthesis and characterization
of some new chromium (III) Schiff base complexes using microwave irradiation
technique as well as conventional heating. The S∩N donor benzothiazolines, 1-
(2-furanyl) ethanone benzothiazoline (Bzt1N
∩
SH), 1-(2-thienyl) ethanone
benzothiazoline (Bzt2N
∩
SH) and 1-(2-pyridyl) ethanone benzothiazoline
(Bzt3N
∩
SH) were prepared by the condensation of ortho-aminothiophenol with
respective ketones in ethanol.
— In the present work, impact of UV-B radiation (280-
315nm: 0.4 W m-2) on growth, photosynthetic pigments, protein,
ascorbate, proline and lipid peroxidation have been studied in
two cyanobacteria Nostoc muscorum and Synechocystis PCC
6803. UV-B radiation (2 to 6 hrs) leads to 55% inhibition of
growth in Synechocystis PCC 6803 in comparison to control
where as in Nostoc muscorum growth reduces up to 45%. This
UV-B treatment also significantly decreased the contents of
chlorophyll, carotenoids and phycocyanin. Photosynthetic
pigments decreased with increasing doses of UV-B (2 to 6 hrs)
radiation. However, the inhibitory effect in Synechocystis PCC
6803 was more pronounced than in Nostoc muscorum. With
increasing UV-B exposure period, production of ascorbate (19-
45%), proline (12-29%) and lipid peroxidation was significantly
higher in Synechocystis PCC 6803 as compared to control
sample. It was observed that lipid peroxidation enhanced 33 %
than control sample of Synechocystis PCC 6803. Our result shows
that photosynthetic apparatus is the main target of UV-B
radiation causing degradation of photosynthetic pigments. This
study concluded that Synechocystis PCC 6803 was the susceptible
organism for survival in stress condition than Nostoc muscorum.
Determination of Stability Constants and Gibbs Free Energies of Cefotaxime Zn...ijtsrd
Cefotaxime is a synthetic lactam antibiotic that is active against Gram negative and Gram positive bacteria. Cefotaxime is able to chelate metal ions due to the presence of C=O, NH2, COOH, COOR, NH and NO electron donating groups. The stability constants and Gibbs free energies of cefotaxime Zn II were determined colorimetrically at 25 and 40 oC using continuous variation and mole ratio methods. The formation of Zn II complex with cefotaxime was studied colorimetrically at an absorption maximum of 430 nm at different temperatures. The data showed that Zn II and cefotaxime combine in the molar ratio of 1 2 at pH 7.4 with ionic strength maintained using 0.1M KNO3. Calculated stability constants values were 1.96 x 105 and 1.28 x 105 using continuous variation method and 1.11 x 105 and 1.11 x 105 using mole ratio methods at 25 and 40 oC respectively. Calculated GO for the complex were 3.01 x 104 and 3.06 x 104 J using continuous variation method and 2.88 x 104 J and 3.02 x 104 J using mole ratio method at 25 and 40 oC respectively. The stability constant results suggested that cefotaxime used in the study is a good chelating agent and can be an efficient antidote in the therapy of Zn II overload or poisoning. O. V. Ikpeazu | I. E. Otuokere | K. K. Igwe "Determination of Stability Constants and Gibbs Free Energies of Cefotaxime-Zn(II) Complex at Different Temperatures" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-4 | Issue-5 , August 2020, URL: https://www.ijtsrd.com/papers/ijtsrd31797.pdf Paper Url :https://www.ijtsrd.com/chemistry/other/31797/determination-of-stability-constants-and-gibbs-free-energies-of-cefotaximeznii-complex-at-different-temperatures/o-v-ikpeazu
Dark fermentative hydrogen production is an intermediate microbial process occurring along anaerobic microbial degradation of organic matter. One direct application of this fermentative bioprocess consists in the production of renewable H2 and simultaneous treatment of organic pollutants. Nowadays, high amounts of saline effluents are generated by fish, seafood, petroleum and leather industries. Such saline effluents are rarely treated by biological anaerobic processes that are strongly inhibited by high salt concentrations. Alternative biological processes, such as dark fermentation, still remain to be investigated with the aim of removing organic pollution from such saline effluents. Moreover, more knowledge about the effect of saline conditions on fermentative microbial mixed cultures would provide new insights on the bacterial inhibition resulting from their exposition to saline conditions.
This study deals with the characterization of hydrogen-producing microbial communities after increasing salt concentrations in a range compatible with a marine environment.
A series of batch experiments was performed under anaerobic conditions favorable to hydrogen production, with a NaCl concentration ranging from 9 to 75 gNaCl/L. Marine sediments were used as inoculum. Biogas and bacterial metabolites were monitored over experimental time. The bacterial community structure dynamics were characterized using molecular tools based on the analysis of genomic 16S rDNA (CE-SSCP), and individual bacterial species were further identified by pyrosequencing.
As a result, the significant and highest biohydrogen production yield (0.9±0.04 molH2.molGlucose-1) was observed at the highest NaCl concentration of 75 g.L-1. However, by increasing the NaCl concentration, the bioH2 production rates slowed down gradually, and longer lag phases were observed. A clear and gradual metabolic shift was also observed suggesting a substantial impact of the saline environment on anaerobic bacterial metabolism, as well as a high selection pressure on acidogenic bacteria. As expected, the composition of the bacterial community at 9gNaCl/L (control) was consistent with literature data, with Clostridium sp. and Enterobacter sp as main dominant species. Interestingly, a gradual shift of the bacterial community structure, concomitant to metabolic changes, was observed by increasing NaCl concentration, with Vibrio sp. as new dominant bacteria (87% in abundance) at the highest salinities. This is the first report on the presence of Vibrio sp. as main hydrogen-producing bacteria in such acidogenic mixed-cultures.
Thus, this study provides new insights on anaerobic metabolism occurring in saline conditions with new possibilities of biotechnological applications from such saline effluents.
International Journal of Pharmaceutical Science Invention (IJPSI)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
One Pot Hydrothermal Synthesis Characterizations Of Silver Nanoparticles On R...IOSRJAC
Graphene-based nanocomposite have significant applicability in catalysis, electronics, medicine, and energy. In this report silver nanoparticles (AgNPs) with Reduced Graphene Oxide (RGO) - nanocomposite was prepared by a one-pot hydrothermal process using silver nitrate as a precursor. Under hydrothermal process Graphene oxide (GO) was reduced to reduced graphene oxide (RGO), without using chemical reagents. As synthesized (Ag-RGO) nanocomposite was characterized by XRD, UV Vis-spectroscopy, Scanning electron microscope, and Raman spectroscopy. Antimicrobial activities of the composite were investigated against both Gram-positive and Gram-negative bacteria. The results demonstrate that Ag-RGO nanocomposite was a strong bactericide against Gram-negative bacteria. Antioxidant activity was evaluated for bare GO, Ag and Ag-RGO nanocomposite by DPPH radical scavenging assay. It was observed that Ag/RGO nanocomposite has enhanced antioxidant activity than bare GO and Ag.
Effluents containing heavy metals can be
remediated with the help of dead microorganisms by the process
known as biosorption. In this study the dead biomass 1of fungus
Aspergillus flavus was used for the biosorption of heavy metals
i.e., Zinc and Nickel. The capacity of biosorption by the dead
biomass of Aspergillus flavus was evaluated at room temperature
with different parameters which are; pH, contact time, biomass
concentration and metal ion concentration. The biosorption
capacity for Zn was found to be 47.36% at room temperature, at
pH 6.5, with biomass concentration of 2g/L having contact time of
50 min and solution concentration of 2ppm. Biosorption capacity
for Ni was found to be 61.60% at room temperature, at pH 5,
with biomass concentration of 2g/L having contact time of 60 min
and solution concentration of 2ppm. . In this study, desorption of
the heavy metals by 0.1M HCl was found to be effective. Fungal
biomass was recovered for reuse.
Technique that is used to elucidate mechanism of a reaction or in a metabolic pathway and in a cell. The labeling takes place by exchanging a specific atom with their isotope. The detecting of the isotopes in the product helps to understand the possible mechanism and the stereochemistry in this sequence of the reaction. The detection of the isotopic labels is dependent on the kind of isotope. Radioactive isotopes like 3H 14C are measured radiochemical. Stable isotopes like 2H and 13C are detected for example with NMR- and IR-spectroscopy.
Kristina Melnik & Stephanie Felten (Undergraduate Students)
University of Utah
2014
Is Nano Medicine And Nano Technology The Most Trending Thing Now?science journals
Nano medicine is nothing but application of Nano technologies as medicines. It may include application of non-material as biological devices or nano-electronic biosensors. Molecular nanotechnology as biological machines may have medical applications in future.
In-vitro antioxidant and GC-MS analysis ethanolic extract of poly herbal drugSkyfox Publishing Group
Antioxidants play an important role in inhibiting and scavenging free radicals, thus providing protection to human against
infections and degenerative diseases. Current research is now directed towards natural antioxidants originated from plants due to safe
therapeutics. Poly herbal drugs is used in Indian traditional medicine for a wide range of various ailments. To understand the mechanism
of pharmacological actions, antioxidant properties of the Poly herbal drugs extract were tested using standard in vitro models. The
ethanolic extract of Poly herbal drugs exhibited strong scavenging effect on superoxide, nitric oxide radical and reducing power radical
scavenging assay. The free radical scavenging effect of Poly herbal drugs extract was comparable with that of the reference antioxidants.
The data obtained in the present study suggests that the extract of Poly herbal drugs have potent Invitro antioxidant and Anti Diabetic
activity against free radicals, prevent oxidative damage to major biomolecules and afford significant protection against oxidative damage.
ABSTRACT
The use of Spirulina as a nutraceutical has been popularized owing to its high essential amino acid, vitamin, carotenoid, chlorophyll content, antioxidant and antiinflamatory properties. This organism can also bioaccumulate and biosorb essential and non essential heavy metals. These properties have been exploited in this study using the organism, Spirulina platensis ARM 728. The fortification of the biomass in different concentrations of Selenium (10 ppm, 40 ppm and 100 ppm) and Zinc (1 ppm, 5 ppm and 10 ppm) was carried out and an increased content of proteins, chlorophyll, carotenoids, SOD, CAT and total antioxidant activity was seen. The biosorption and desorption capacity of the organism for antimony at 80 ppm was also seen with fair results.
Keywords: antioxidant properties, bioaccumulation, biosorption, heavy metals, Spirulina fortification.
Determination of Stability Constants and Gibbs Free Energies of Cefotaxime Zn...ijtsrd
Cefotaxime is a synthetic lactam antibiotic that is active against Gram negative and Gram positive bacteria. Cefotaxime is able to chelate metal ions due to the presence of C=O, NH2, COOH, COOR, NH and NO electron donating groups. The stability constants and Gibbs free energies of cefotaxime Zn II were determined colorimetrically at 25 and 40 oC using continuous variation and mole ratio methods. The formation of Zn II complex with cefotaxime was studied colorimetrically at an absorption maximum of 430 nm at different temperatures. The data showed that Zn II and cefotaxime combine in the molar ratio of 1 2 at pH 7.4 with ionic strength maintained using 0.1M KNO3. Calculated stability constants values were 1.96 x 105 and 1.28 x 105 using continuous variation method and 1.11 x 105 and 1.11 x 105 using mole ratio methods at 25 and 40 oC respectively. Calculated GO for the complex were 3.01 x 104 and 3.06 x 104 J using continuous variation method and 2.88 x 104 J and 3.02 x 104 J using mole ratio method at 25 and 40 oC respectively. The stability constant results suggested that cefotaxime used in the study is a good chelating agent and can be an efficient antidote in the therapy of Zn II overload or poisoning. O. V. Ikpeazu | I. E. Otuokere | K. K. Igwe "Determination of Stability Constants and Gibbs Free Energies of Cefotaxime-Zn(II) Complex at Different Temperatures" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-4 | Issue-5 , August 2020, URL: https://www.ijtsrd.com/papers/ijtsrd31797.pdf Paper Url :https://www.ijtsrd.com/chemistry/other/31797/determination-of-stability-constants-and-gibbs-free-energies-of-cefotaximeznii-complex-at-different-temperatures/o-v-ikpeazu
Dark fermentative hydrogen production is an intermediate microbial process occurring along anaerobic microbial degradation of organic matter. One direct application of this fermentative bioprocess consists in the production of renewable H2 and simultaneous treatment of organic pollutants. Nowadays, high amounts of saline effluents are generated by fish, seafood, petroleum and leather industries. Such saline effluents are rarely treated by biological anaerobic processes that are strongly inhibited by high salt concentrations. Alternative biological processes, such as dark fermentation, still remain to be investigated with the aim of removing organic pollution from such saline effluents. Moreover, more knowledge about the effect of saline conditions on fermentative microbial mixed cultures would provide new insights on the bacterial inhibition resulting from their exposition to saline conditions.
This study deals with the characterization of hydrogen-producing microbial communities after increasing salt concentrations in a range compatible with a marine environment.
A series of batch experiments was performed under anaerobic conditions favorable to hydrogen production, with a NaCl concentration ranging from 9 to 75 gNaCl/L. Marine sediments were used as inoculum. Biogas and bacterial metabolites were monitored over experimental time. The bacterial community structure dynamics were characterized using molecular tools based on the analysis of genomic 16S rDNA (CE-SSCP), and individual bacterial species were further identified by pyrosequencing.
As a result, the significant and highest biohydrogen production yield (0.9±0.04 molH2.molGlucose-1) was observed at the highest NaCl concentration of 75 g.L-1. However, by increasing the NaCl concentration, the bioH2 production rates slowed down gradually, and longer lag phases were observed. A clear and gradual metabolic shift was also observed suggesting a substantial impact of the saline environment on anaerobic bacterial metabolism, as well as a high selection pressure on acidogenic bacteria. As expected, the composition of the bacterial community at 9gNaCl/L (control) was consistent with literature data, with Clostridium sp. and Enterobacter sp as main dominant species. Interestingly, a gradual shift of the bacterial community structure, concomitant to metabolic changes, was observed by increasing NaCl concentration, with Vibrio sp. as new dominant bacteria (87% in abundance) at the highest salinities. This is the first report on the presence of Vibrio sp. as main hydrogen-producing bacteria in such acidogenic mixed-cultures.
Thus, this study provides new insights on anaerobic metabolism occurring in saline conditions with new possibilities of biotechnological applications from such saline effluents.
International Journal of Pharmaceutical Science Invention (IJPSI)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
One Pot Hydrothermal Synthesis Characterizations Of Silver Nanoparticles On R...IOSRJAC
Graphene-based nanocomposite have significant applicability in catalysis, electronics, medicine, and energy. In this report silver nanoparticles (AgNPs) with Reduced Graphene Oxide (RGO) - nanocomposite was prepared by a one-pot hydrothermal process using silver nitrate as a precursor. Under hydrothermal process Graphene oxide (GO) was reduced to reduced graphene oxide (RGO), without using chemical reagents. As synthesized (Ag-RGO) nanocomposite was characterized by XRD, UV Vis-spectroscopy, Scanning electron microscope, and Raman spectroscopy. Antimicrobial activities of the composite were investigated against both Gram-positive and Gram-negative bacteria. The results demonstrate that Ag-RGO nanocomposite was a strong bactericide against Gram-negative bacteria. Antioxidant activity was evaluated for bare GO, Ag and Ag-RGO nanocomposite by DPPH radical scavenging assay. It was observed that Ag/RGO nanocomposite has enhanced antioxidant activity than bare GO and Ag.
Effluents containing heavy metals can be
remediated with the help of dead microorganisms by the process
known as biosorption. In this study the dead biomass 1of fungus
Aspergillus flavus was used for the biosorption of heavy metals
i.e., Zinc and Nickel. The capacity of biosorption by the dead
biomass of Aspergillus flavus was evaluated at room temperature
with different parameters which are; pH, contact time, biomass
concentration and metal ion concentration. The biosorption
capacity for Zn was found to be 47.36% at room temperature, at
pH 6.5, with biomass concentration of 2g/L having contact time of
50 min and solution concentration of 2ppm. Biosorption capacity
for Ni was found to be 61.60% at room temperature, at pH 5,
with biomass concentration of 2g/L having contact time of 60 min
and solution concentration of 2ppm. . In this study, desorption of
the heavy metals by 0.1M HCl was found to be effective. Fungal
biomass was recovered for reuse.
Technique that is used to elucidate mechanism of a reaction or in a metabolic pathway and in a cell. The labeling takes place by exchanging a specific atom with their isotope. The detecting of the isotopes in the product helps to understand the possible mechanism and the stereochemistry in this sequence of the reaction. The detection of the isotopic labels is dependent on the kind of isotope. Radioactive isotopes like 3H 14C are measured radiochemical. Stable isotopes like 2H and 13C are detected for example with NMR- and IR-spectroscopy.
Kristina Melnik & Stephanie Felten (Undergraduate Students)
University of Utah
2014
Is Nano Medicine And Nano Technology The Most Trending Thing Now?science journals
Nano medicine is nothing but application of Nano technologies as medicines. It may include application of non-material as biological devices or nano-electronic biosensors. Molecular nanotechnology as biological machines may have medical applications in future.
In-vitro antioxidant and GC-MS analysis ethanolic extract of poly herbal drugSkyfox Publishing Group
Antioxidants play an important role in inhibiting and scavenging free radicals, thus providing protection to human against
infections and degenerative diseases. Current research is now directed towards natural antioxidants originated from plants due to safe
therapeutics. Poly herbal drugs is used in Indian traditional medicine for a wide range of various ailments. To understand the mechanism
of pharmacological actions, antioxidant properties of the Poly herbal drugs extract were tested using standard in vitro models. The
ethanolic extract of Poly herbal drugs exhibited strong scavenging effect on superoxide, nitric oxide radical and reducing power radical
scavenging assay. The free radical scavenging effect of Poly herbal drugs extract was comparable with that of the reference antioxidants.
The data obtained in the present study suggests that the extract of Poly herbal drugs have potent Invitro antioxidant and Anti Diabetic
activity against free radicals, prevent oxidative damage to major biomolecules and afford significant protection against oxidative damage.
ABSTRACT
The use of Spirulina as a nutraceutical has been popularized owing to its high essential amino acid, vitamin, carotenoid, chlorophyll content, antioxidant and antiinflamatory properties. This organism can also bioaccumulate and biosorb essential and non essential heavy metals. These properties have been exploited in this study using the organism, Spirulina platensis ARM 728. The fortification of the biomass in different concentrations of Selenium (10 ppm, 40 ppm and 100 ppm) and Zinc (1 ppm, 5 ppm and 10 ppm) was carried out and an increased content of proteins, chlorophyll, carotenoids, SOD, CAT and total antioxidant activity was seen. The biosorption and desorption capacity of the organism for antimony at 80 ppm was also seen with fair results.
Keywords: antioxidant properties, bioaccumulation, biosorption, heavy metals, Spirulina fortification.
A detailed description about the basic steps involved in the - PCR - Polymerase Chain Reaction, its applications,its limitations and steps to overcome it.
Similar to Determination of 8-Hydroxy-2 Deoxyguanosine in Pseudomonas Fluorescens Freeze-Dried Exposed to the ROS Using HPLC with Electrochemical Detection Method
The Role of Cell Wall-Degrading Enzymes in the Development of Anthracnose Dis...Agriculture Journal IJOEAR
— The ability of Colletotrichumtruncatum CP2 in producing pectinolytic and cellulolytic enzymes was evaluated by shake flask fermentations. The results of enzymatic activity experiment indicated that PG was the first cell wall-degrading enzymes detected and the activities obtained were higher (0.24±0.10 U/mL) than other enzymes, which appeared later and in lower amount. After the cell wall was degraded by the action of PG, further degradation of the cell wall was affected by pectin methylesterases, pectin lyase, pectate lyase and cellulases. The disparity in enzymatic activity at different intervals may suggest their specific role for pathogenesis at proper timings.
Degradation of an organophosphorus insecticide (chlorpyrifos) in simulated wa...Salah Hussein
Induced degradation of chlorpyrifos insecticide in simulated wastewater with advanced oxidation processes (AOPs), using ultraviolet irradiation (UV), ozonation and chemical oxidation using (sodium hypochlorite, calcium hypochlorite, monochloride-isocyanuric acid (MCICA), dichloroiso-cyanuric acid (DCICA), trichloroisocyanuric acid (TCICA) ) was studied. Chlorpyrifos and its degradation products were extracted using solid phase extraction (SPE) method, identified using GC-MS. Results showed that the degradation of chlorpyrifos in simulated wastewater followed the first order reaction, and its half life was 3.34, 5.64, 7.13 and 10.69h under ozonation, UV, 1.5%TCICA and 1.5%DCICA respectively when chlorpyrifos solutions treated for 12 h. The concentrations of chemical oxidative substances, active chlorine content and time of treatments had a significant effect on degradation rate of chlorpyrifos, which increased with increasing of each. The most enhancement of chlorpyrifos degradation was observed in treatment with ozonation, UV, TCICA and DCICA where the dissipations % of the parent compounds were 85.70, 57.71, 43.71 and 35.07 %, respectively. The intermediates products of chlorpyrifos degradation using chemical method were identified as O,O-Diethyl thiophosphate(DEP), 3,5,6-trichloro-2-pyridinol(TCP), 3,5,6-trichloro-2-methoxypyridine(TMP) and 2,3,5,6-tetrachloro-pyridine. UV leads to formation of O,O-Diethyl phosphate, TCP and Chlorpyrifos oxon. Ozonation leads to formation of O,O-Diethyl thiophosphate beside the UV degradation products.
—3-Chloro-1,2-propanediol (3-chloropropanediol) is a well-known food processing contaminant found in a wide range of foods and ingredients and there has been recent concern about the levels of carcinogenic 3-chloropropanediol (3-MCPD) in some soy sauces. This paper reports on the development of an analytical method for the fast determination of 3-MCPD at trace level in commercial soy sauce using novel liquid phase extraction (LPE)/cleanup coupled with microwave-assisted derivatization (MAD) method followed by high performance liquid chromatography-ultraviolet (HPLC-UV) detection. In this method, 3-MCPD was first isolated from soy sauce sample matrix by LPE/cleanup with Extrude NT3 column cartridges and the isolated (eluent) solution was subjected to MAD with acetophenone to form 2-methyl-2-phenyl-4-(chloromethyl)-1,3-dioxolane under microwave irradiation using a specially modified domestic microwave oven, then the derivatizeddioxolane was directly analyzed with a HPLC-UV system. The optimum conditions for MAD such as the ratio of reagents, acidic catalyst, microwave irradiation power and time, as well as the chromatographic conditions were thoroughly investigated. Experimental results indicated that maximum derivatization can be achieved in 10 min under microwave irradiation at 362 watts when compared to 18 hours by conventional refluxing reaction. The proposed method provided a simple and rapid analytical procedure for 3-MCPD analysis in soy sauce with the detection limit of 80 ng mL-1. The relative standard deviations were all below 3.0 % (n = 7). Application was illustrated by the analysis of commercial sauce sample obtained from a local traditional store in central Taiwan.
Micellar Effect On Dephosphorylation Of Bis-4-Chloro-3,5-Dimethylphenylphosph...IOSR Journals
The rate enhancement depends on the hydrophobicity of the nucleophile. The micellar catalyzed reaction between bis-4-chloro-3,5-dimethylphenylphosphate ester and hydroxide or hydroperoxide anions has been examined in buffered medium (pH 8-10). First order rate constant (Kψ) for the reaction of hydroxide ion with bis-4-CDMPP go through maxima with the increasing concentration of cetyltrimethylammoniumbromide (CTABr). Micelles of CTABr very effective catalyst to the reactions of phosphate diesters. Rate constants measured with OH2- ions are approximately twice and thrice than that of OH- ions in presence of CTABr.
Micellar Effect On Dephosphorylation Of Bis-4-Chloro-3,5-Dimethylphenylphosph...IOSR Journals
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Determination of 8-Hydroxy-2 Deoxyguanosine in Pseudomonas Fluorescens Freeze-Dried Exposed to the ROS Using HPLC with Electrochemical Detection Method
1. International Journal of Environmental & Agriculture Research (IJOEAR) ISSN:[2454-1850] [Vol-2, Issue-5, May- 2016]
Page | 225
Determination of 8-Hydroxy-2 Deoxyguanosine in Pseudomonas
Fluorescens Freeze-Dried Exposed to the ROS Using HPLC with
Electrochemical Detection Method
Karl Tshimenga1
, Pascal Nsambu2
, Christina Mputu Tshibadi3
, Eddy Dibwe Finta4
,
Jean-Noël Mputu Kanyinda5*
1
Department of Chemistry and Industry, Faculty of Sciences, University of Kinshasa, BP 190 Kin XI, DR Congo
2
Faculty of Agronomy, Evangelical University in Africa B.P.3323et 266 Bukavu /DR Congo.
3
Clinic of Hematology, Onkology, Immunology and tropical medicine at Klinikum Schwabing. Kölner Platz 1, 80804
München.
4
Division of Natural Drug Discovery, Department of Translational Research, Institute of Natural Medicine, University of
Toyama, 2630 Sugitani, Toyama 930-0194, Japan
Abstract— Oxidative DNA damage is involved in the f cell death induced by freeze-dried powder during storage. Cell 8-
hydroxy-2’deoxyguanosine (8-oxodG) is widely accepted as a biomarker of the “freeze-dried bacteria” oxidative DNA
damage. The aim of this study was to introduce a method for determination 8-oxodG in cell freeze-dried samples using high-
performance liquid chromatography with electrochemical detection. In the tested range of 0.5 µmol L-1
to 1.0 nmol L-1
, the
calibration curve was linear (r2
=0.9995) and the limit of detection was 0.05 µmol L-1
. The used method did not allow
highlighting the presence in the samples of the 8OH within the limits of detection. A more successful method (more sensitive)
would be needed to detect possibly the 8OH.
Keywords— Reactive Oxygen Species (ROS), DNA, 8-Hydroxy-2 deoxyguanosine.
I. INTRODUCTION
The drying by lyophilisation of microorganisms has some consequence on the viability of the latter. The powder of
lyophilized bacteria remains vulnerable on the environmental conditions such as temperature, ultraviolet, reactive species of
oxygens. The latter have mainly four consequences: the loss of the cellular viability caused by an oxidation of the biological
materials like lipids, proteins and DNA [1,2,3]. The drying method cause severe damage in the cellular membranes such as
the lipid peroxidation, denaturation of proteins and DNA leading to a loss of viability [4].
Freeze-drying and the storage of cells lead damage in proteins, fatty acids and DNA [5, 6]. Although the DNA damage
undergone by the cellular membrane during the freeze-drying. It still plays an essential role in the loss of viability. The
damage of the cellular components DNA and ARN affects considerably the viability of the freeze-dried bacteria during
storage. The DNA is very sensitive to the drying, as demonstrated on Echericha coli [7, 8].
There are numerous modifications observed after the oxidation of the DNA, the major product of DNA oxidative damage is
8-hydroxyguanine (8-oxoG) and its nucleoside 8-hydroxy-2’deoxyguanosine (8-oxodG). 8-oxodG has widely been accepted
as a biomarker of DNA damage and cellular oxidative stress [9].
The present study will be focus on the development of novel method for the detection and quantification of the 2-
déoxyguanosine 2dG and the 8-hydroxy-2-déoxyguanosine 8OH in extracts of DNA bacterial of Pseudomonas fluorescens
freeze-dried. Because the 8-hydroxy-2-déoxyguanosine is the main oxidized by-product of the DNA used as a biomarker of
the oxidative stress.
II. MATERIALS AND METHODS
2.1 Materials
2.1.1 Bacteria strain.
The Pseudomonas fluorescens BTP1 of Wallon Center of Industrial Biology laboratory (CWBI) was used as the strain in this
study [2].
2.2 Methods
2. International Journal of Environmental & Agriculture Research (IJOEAR) ISSN:[2454-1850] [Vol-2, Issue-5, May- 2016]
Page | 226
2.2.1 Photosensibilisation
The samples (bacterial powder) were exposed to methylene blue (MB) at 40 µM of in Light presence. The methylene blue
and an incandescent bulb produce singlet oxygen [6,10].
2.2.2 Method of extraction of the genomic DNA of bacteria
Take a sample of cells (1,5 ml of night culture) and spin-dry it 5 minutes at 13000 rpm. Rinsing of the nerve with a PCR,
then the sample were taking back the cap with 480 µL of EDTA (50 mm, pH 8). The addition of 30 µl of a solution of
lysozyme to 20 mg/ml and let incubate 1 hour in 37°C by shaking. Add 30 µl of a solution of proteinase K to 20 mg/mL and
let incubate 30 minutes in 37°C under agitation. Spin-dry 3 minutes at 13000 rpm, taken back by the cap in 600 µL of
solution Nuclei Lysis by pipetting and incubate 10 minutes in-80°C. Incubate then 10 minutes in 80°C (dry bath); let it cool
at room temperature [11].
Add 200 µL of solution protein precipitation shake 20 seconds and incubate 10 minutes in the ice. Spin-dry the solution
during 5 minutes to 13000 rpm. Transfer the floats in a containing sterile tube 600 µL of isopropanol, mix delicately by
inversion and spin-dry 5 minutes at 13000 rpm. Wash the cap with 600 µL of ethanol 70 %, spin-dry it 5 minutes in 13000
rpm and dry it under the basket (10-15 minutes). Rehydrate the cap in 50 µL of rehydration solution, incubate 60 minutes at
50°C and store the DNA during 1 month.
2.2.3 Extracts of bacterial DNA
The DNA was extracted according to the process described by P.Duez (Chromatographic determination of 8-Oxo-7,8-
dihydro-2'deoxyguanosine in cellular DNA.
2.2.4 Chemicals preparation
8-oxodG standard potassium chloride, acetonitrile, and methanol were purchased from Sigma Chemicals (St. Louis, MO,
USA) and potassium dihydrogenphosphate and water were from Merck (Darmstadt, Germany). All chemicals were of pro
analysis grade. Water, acetonitrile, KOH and methanol used for mobile phase were of HPLC grade. Cartridges (Bond Elut
C18, Bond Elut C18 OH and Bond Elut Certify) for solid phase extraction (SPE).
2.2.5 Standard solution preparation
Stock solution of 8-oxodG (8-OH) was prepared dissolving the 8-oxodG standard in purified water to give a final
concentration of 1,5 mmol L-1
and 2 mmol L-1
for 2-déoxyguanosine (2dG). The stock solution was aliquoted and stored at -
18 °C.
2.2.6 Electrochemical conditions
HPLC-EC (High-Performance Liquid Chromatography with an Amperometric Detection).
HPLC (stocking) EC " Pump: Gilson 307, system of injection: Rheodyne 7125 (with two positions: LOAD / INJECTIONS),
column Atlantis C18 (internal diameter: 3,0 µm, size: 4,6 mm x 100 mm) curls, locks up 20 µL; detector amperometric in
thin layer BASi (working electrode in glassy carbon), a reference electrode (electrode Ag / AgCl in a gel of NaCl 3 M), a
stainless steel auxiliary electrode, a potentiostat LC-4B (BASi) and a recorder: PowerChrom 280.
2.2.7 Chromatography’s conditions
Different mobile phases were studied. That used consists of: 50 mM KH2PO4, KOH, pH 5.5, 13% Methanol, Water MilliQ
2L with a potential of 1.1 V. If we need to detect the two molecules simultaneously the potential required is 1.1 V; but for
one molecule such as 8OH, the potential required is 700 mV. A scale of 1 to 5 nA, depending on the concentration with a
flow rate of 0.8 mL / min.
2.3 Statistical analysis
Data from three replications were analyzed by using analysis of variance to determine if significant difference (P≤0.05)
existed between mean values.
3. International Journal of Environmental & Agriculture Research (IJOEAR) ISSN:[2454-1850] [Vol-2, Issue-5, May- 2016]
Page | 227
III. RESULTS AND DISCUSSION
3.1 Results
3.1.1 Curves of calibration
Different curved of calibrations were realized with solutions between 0.5µM and 1nM. The dilutions were prepared in the
mobile phase figure and table 1.
TABLE 1
OXIDATION POTENTIAL OF VARIOUS ELECTRODES GCE AND BDD
FIGURE 1: STUDY BY CYCLIC VOLTAMPEROMETRY OF THE 2-DEOXYGUANOSINE. REFERENCE ELECTRODE:
Ag / AgCl, INDICATOR ELECTRODE: BDD (ELECTRODE OF DIAMOND DOPED IN THE BORON) / GCE
(ELECTRODE OF CARBON GLASSY), AUXILIARY ELECTRODE: PLATINUM, BUFFER: ACETIC ACID, ACETATE OF
SODIUM 0,1 M-pH 4,3, POTENTIAL OF 400 mV in 1350 mV, SPEED OF SCANNING OF 10mV.
According two days experiment, we observed a variation of the sensibility, to the high potential of oxidation and to the state
of the surface of the working electrode according table 2 and figure 2.
TABLE 2
CALIBRATION OF THE METHOD BY THE DILUTION OF STANDARDS
Conc. 2dG (day 1) 2dG (day 2) 8OH (day 1) 8OH (day 2)
0.50 µM 2.50 2 1.50 0.80
0.10 µM 0.56 0.52 0.30 0.22
0.05 µM 0.28 0.22 0.14 0.10
1 nM Nd 0.05 Nd nd
Nd : not detected
FIGURE 2: THE SENSIBILITY VARIATION OF THE METHOD
GCE BDD
Potentiel oxydation : (mV) 2.50 0.56
4. International Journal of Environmental & Agriculture Research (IJOEAR) ISSN:[2454-1850] [Vol-2, Issue-5, May- 2016]
Page | 228
3.1.2 Chromatograph before and after the addition of the sample
We performed analysis of a standard 8OH and 2 dG from Sigma Aldrich, the uniques indicators of the oxidation of DNA
and injected samples. The results are shown in Figure 3.
FIGURE 3: STANDARDS BEFORE AND AFTER INJECTION OF A SAMPLE
3.1.3 Dosage of the 8OH and 2dG
The DNA sample was extracted using the method described above on the lyophilized powder of Pseudomonas fluorescens.
The results are illustrated in the chromatogram as shown in the figures 4.
FIGURE 4: SPECTRE HPLC OF THE 8OH AND 2DG OF THE FREEZE-DRIED PSEUDOMONAS FLUORESCENS
BTP1 STRAIN.
3.2 Discussions
3.2.1 Curves of calibration
The mobile phase was not stamped. We need to notice that other buffers were tested but none of them were well satisfactory
due to the loss of sensibility, the important background noise detected during the experiment. The potential was observed
brought up. It is essential to work with a potential of 1.1V to put in evidence the same time both molecules (the 8OH and
2DG) simultaneously figure 1. Indeed, the 2dG in a high potential of oxidation observed at 900 mV on electrode of glassy
carbon as shown in figure 2. The 8OH has a potential of oxidation of 500mV on glassy carbon.
During the analysis of our sample, we observed a doubling of peaks probably due to degradation of the column and more
particularly C18 groups [Figure 3].
The guard column was placed a in order to avoid damaging the column. In the present experiment, we also precipitate
residual proteins by adding methanol. It is important to notice that it is preferable to work in an acid medium for a better
precipitation, which is not possible in our case because of the non-buffered mobile phase.
Then, the analysis of the samples was preceded by using the appropriate dilution for an efficient detection with the HPLC.
The results are shown in the chromatogram in the figures 4.
5. International Journal of Environmental & Agriculture Research (IJOEAR) ISSN:[2454-1850] [Vol-2, Issue-5, May- 2016]
Page | 229
3.2.2 Dosage of the 8OH and 2dG
The curve of calibration was realized every day for the dosage of the 2dG. In the present study, we proceeded that
determination by using the method of standard additions due to the lower amounts of 80H. The sample preparation was
performed as follow: Collect 75 HL then add 25 µl of methanol (total 100 µL), mixer 30 seconds and centrifuge 5 min at
13,000 rpm. It is difficult to detect at the same time 8OH and 2dG in our conditions. Nevertheless, our results support the
hypothesis that the DNA of the lyophilized the powder freeze-dried of bacteria mainly Pseudomonas fluorescens undergoes a
damage during storage.
IV. CONCLUSION
The main purpose of this study was to confirm the hypothesis which supported that DNA also is subject of the oxidation in
the same manner as lipids and proteins. The sensitivity of the 2dG was better measured in our operating conditions compared
to the 8OH one. The current method was not able to highlight within the limits of detection the presence of the 8OH samples.
An additional method would be needed for such to detection. Some scientists describe the tests on the oxidation of the ARN
instead of the DNA, which represent an interesting way to explore [8].
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Biotechnol. Agron. Soc. Environ. 2015: 18, 1-8.
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subjected to freeze-drying. Journal of Applied Microbiology. 2005: 99, 333-338.
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[6] Santivarangkna C, Kulozik U, Foerst P Alternative Drying Processes for the Industrial Preservation of Lactic Acid Starter Cultures.
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[8] Domijan AM and Peraica M. Determination of 8-hydroxy-2’deoxyguanosine in urine using HPLC with electrochemical detection.
Arh Hig Rada Toksikol, 2008; 59:277-282
[9] Yoon S, Park J, Yang J, and Park J. Oxyregulon controls lipid peroxidation-mediated oxidative stress in escherichia coli," J. Biochem.
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Carcinogenesis. 2006: 27: 405–408.
[11] Rasmusen RS and Morrissey MT. DNA-based methods for the identification of commercial fish and seafood species. Comprehensive
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