Abstract
The use of biogenic selenium nanoparticles for various purposes is going to be an issue of considerable importance; thus, appropriate simple methods should be developed and tested for the synthesis and recovery of these nanoparticles. In this study, a fungus was isolated from a soil sample, identified as Aspergillus terreus and used for extracellular synthesis of selenium nanoparticles (Se NPs). UV–Vis spectroscopy and energy dispersive X-ray spectrum studies were carried out to confirm Se NPs formation within 60 min. Dynamic light scattering and scan electron microscopic methods were also used to characterize both size and shapes of the Se NPs. The results show that spherical particles with average size of 47 nm were formed by adding a culture supernatant of A. terreus to selenium ions solution. This approach appears to be an easy and appropriate method for extracellular synthesis of small Se NPs. Extracellular synthesis of small Se NPs has not been reported yet.
Crimson Publishers-Synthesis, Characterization, and Evaluation of the In-vitr...CrimsonpublishersMedical
Cinnamomum zeylanicum have lot-of biological activities including antimicrobial, antioxidant and antifungal properties. Furthermore, cytotoxic and apoptotic activities of several constituents were identified throughout its biological properties. Bark of Cinnamomum zeylanicum (Lauraceae) collected respectively at Nanotechnology laboratory (ANGRAU, Tirupathi, India). In this study, microbiological aspects of scale formation in PVC pipelines bacteria and fungi were isolated. Stable Zn nanoparticles were formed by treating 90ml of 1mm zinc nitrate aqueous solution with 10ml of 10% bark extract. The formation of Cinnamomum zeylanicum bark extract mediated zinc nanoparticles (CZnNPs) was confirmed by UV-visible spectroscopic analysis and recorded the localized surface plasmon resonance (LSPR) at 270nm.
Techniques of DNA Extraction, Purification and QuantificationBHUMI GAMETI
Introduction
The overall process…
Uses of isolated genomic DNA
Extraction of DNA from plant material
Components of DNA extraction solutions
Cell Lysis or Cell disruption :
Purification of DNA
CTAB Method
Phenol–chloroform extraction
PROTEINASE K
Salting out
Silica adsorption method
Magnetic beads
FTA Paper
Nucleic acid quantification
Agarose Gel Electrophoresis
UV spectroscopy
DNA quantification using NanoDrop
The technique of molecular biology like DNA isolation, RNA isolation, PCR, Western blot, RFLP, etc was developed with development in science. This presentation includes the method of DNA and RNA isolation and their Quantification techniques.
Crimson Publishers-Synthesis, Characterization, and Evaluation of the In-vitr...CrimsonpublishersMedical
Cinnamomum zeylanicum have lot-of biological activities including antimicrobial, antioxidant and antifungal properties. Furthermore, cytotoxic and apoptotic activities of several constituents were identified throughout its biological properties. Bark of Cinnamomum zeylanicum (Lauraceae) collected respectively at Nanotechnology laboratory (ANGRAU, Tirupathi, India). In this study, microbiological aspects of scale formation in PVC pipelines bacteria and fungi were isolated. Stable Zn nanoparticles were formed by treating 90ml of 1mm zinc nitrate aqueous solution with 10ml of 10% bark extract. The formation of Cinnamomum zeylanicum bark extract mediated zinc nanoparticles (CZnNPs) was confirmed by UV-visible spectroscopic analysis and recorded the localized surface plasmon resonance (LSPR) at 270nm.
Techniques of DNA Extraction, Purification and QuantificationBHUMI GAMETI
Introduction
The overall process…
Uses of isolated genomic DNA
Extraction of DNA from plant material
Components of DNA extraction solutions
Cell Lysis or Cell disruption :
Purification of DNA
CTAB Method
Phenol–chloroform extraction
PROTEINASE K
Salting out
Silica adsorption method
Magnetic beads
FTA Paper
Nucleic acid quantification
Agarose Gel Electrophoresis
UV spectroscopy
DNA quantification using NanoDrop
The technique of molecular biology like DNA isolation, RNA isolation, PCR, Western blot, RFLP, etc was developed with development in science. This presentation includes the method of DNA and RNA isolation and their Quantification techniques.
DNA extraction is an important step in molecular assays and plays a vital role in obtaining highresolution results in gel-based systems, particularly in the case of cereals with high content of interfering components in the early steps of DNA extraction.This is a rapid miniprep DNA extraction method, optimized for rice, which was achieved via creating some modifications in present DNA extraction methods, especially in first step of breaking down and lyses of cell wall, and the use of cheap and frequent chemicals, found in every lab, in the next steps. The normal quality and quantity was obtained by the method. The PCR based assays also revealed the efficiency of the method.
The advantages of this method are: 1- it is applicable with both dry and fresh samples, 2- no need to large weight samples, 3- no need to liquid nitrogen and 4- easy, rapid and applicable in every laboratory.
Effective disruption of the biological matrix (cell, tissue, environmental or biological sample) to release the nucleic acids. Denaturation of structural proteins associated with the nucleic acids (nucleoproteins) Inactivation of nucleases that will degrade the isolated product (RNase and/or DNase).
Once the genomic DNA is bound to the silica membrane, the nucleic acid is washed with a salt/ethanol solution. These washes remove contaminating proteins, lipopolysaccharides and small RNAs to increase purity while keeping the DNA bound to the silica membrane column.
There are five basic steps of DNA extraction that are consistent across all the possible DNA purification chemistries:
disruption of the cellular structure to create a lysate,
separation of the soluble DNA from cell debris and other insoluble material,
binding the DNA of interest to a purification matrix,
washing proteins and other contaminants away from the matrix and
elution of the DNA.
Isolation and Purification of Chromosomal DNA,Plasmid DNA,Bacteriophage DNA used in Recombinant DNA Technology or Biotechnology to produce Recombinant DNA or Desired DNA
DNA extraction is an important step in molecular assays and plays a vital role in obtaining highresolution results in gel-based systems, particularly in the case of cereals with high content of interfering components in the early steps of DNA extraction.This is a rapid miniprep DNA extraction method, optimized for rice, which was achieved via creating some modifications in present DNA extraction methods, especially in first step of breaking down and lyses of cell wall, and the use of cheap and frequent chemicals, found in every lab, in the next steps. The normal quality and quantity was obtained by the method. The PCR based assays also revealed the efficiency of the method.
The advantages of this method are: 1- it is applicable with both dry and fresh samples, 2- no need to large weight samples, 3- no need to liquid nitrogen and 4- easy, rapid and applicable in every laboratory.
Effective disruption of the biological matrix (cell, tissue, environmental or biological sample) to release the nucleic acids. Denaturation of structural proteins associated with the nucleic acids (nucleoproteins) Inactivation of nucleases that will degrade the isolated product (RNase and/or DNase).
Once the genomic DNA is bound to the silica membrane, the nucleic acid is washed with a salt/ethanol solution. These washes remove contaminating proteins, lipopolysaccharides and small RNAs to increase purity while keeping the DNA bound to the silica membrane column.
There are five basic steps of DNA extraction that are consistent across all the possible DNA purification chemistries:
disruption of the cellular structure to create a lysate,
separation of the soluble DNA from cell debris and other insoluble material,
binding the DNA of interest to a purification matrix,
washing proteins and other contaminants away from the matrix and
elution of the DNA.
Isolation and Purification of Chromosomal DNA,Plasmid DNA,Bacteriophage DNA used in Recombinant DNA Technology or Biotechnology to produce Recombinant DNA or Desired DNA
Abstract
Objective(s):
The development of reliable and ecofriendly process for the synthesis of nano-metals is an important aspect in the field of nanotechnology. Nano-metals are a special group of materials with broad area of applications.
Materials and Methods:
In this study, extracellular synthesis of silver nanoparticles (SNPs) performed by use of the gram positive soil Streptomycetes. Streptomycetes isolated from rice fields of Guilan Province, Iran (5 isolates). Initial characterization of SNPs was performed by visual change color. To determine the bacterium taxonomical identity, its colonies characterized morphologically by use of scanning electron microscope. The PCR molecular analysis of active isolate represented its identity partially. In this regard, 16S rDNA of isolate G was amplified using universal bacterial primers FD1 and RP2. The PCR products were purified and sequenced. Sequence analysis of 16S rDNA was then conducted using NCBI GenBank database using BLAST. Also SNPs were characterized by, transmission electron microscopy (TEM) and X-ray diffraction spectroscopy (XRD).
Results:
From all 5 collected Streptomyces somaliensis isolates, isolate G showed highest extracellular synthesis of SNPs via in vitro. SNPs were formed immediately by the addition of (AgNO3) solution (1 mM). UV-visible spectrophotometry for measuring surface plasmon resonance showed a single absorption peak at 450 nm, which confirmed the presence of SNPs. TEM revealed the extracellular formation of spherical silver nanoparticles in the size range of 5-35 nm.
Conclusions:
The biological approach for the synthesis of metal nanoparticles offers an environmentally benign alternative to the traditional chemical and physical synthesis methods. So, a simple, environmentally friendly and cost-effective method has been developed to synthesize AgNPs using Streptomycetes.
Abstract
Objective(s):
In recent years, the biosynthesis of gold nanoparticles has been the focus of interest because of their emerging application in a number of areas such as biomedicine. In the present study we report the extracellular biosynthesis of gold nanoparticles (AuNPs) by using a positive bacterium named Streptomyces fulvissimus isolate U from rice fields of Guilan Province, Iran.
Materials and Methods:
From over 20 Streptomyces isolates collected, isolate U showed high AuNPs biosynthesis activity. To determine its taxonomical identity, its morphology was characterized by scanning electron microscope and partial molecular analysis performed by PCR. In this regard, 16S rDNA of isolate U was amplified using universal bacterial primers FD1 and RP2. The PCR products were purified and sequenced. Sequence analysis of 16S rDNA was then conducted using NCBI BLAST method. In biosynthesis of AuNPs by this bacterium, the biomass of bacterium exposed to the HAuCl4 solution.
Results:
The nanoparticles obtained were characterized by UV-Visible spectroscopy, transmission electron microscopy (TEM) and Energy dispersive X-ray (EDX) spectroscopy and X-ray diffraction spectroscopy (XRD) analyses. Our results indicated that Streptomyces fulvissimus isolateU bio-synthesizes extracellular AuNPs in the range of 20-50 nm.
Conclusions:
This technique of green synthesis of AuNPs by a microbial source may become a promising method because of its environmental safety. Its optimization may make it a potential procedure for industrial production of gold nanoparticles.
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
Detection of Genetic variation in tissue culture clones of date palm using IS...IJSRD
Date palm is a plant having high nutritional value and long life (yielding up to 100 years). Phoenix dactylifera requires 2-5 males for pollination of 100 females’ plant depending up on genetic and environment factors. Therefore paternity variation expected to very low according to PCR based techniques, Even though we have tried to find out genetic variation among tissue culture cloned plant. Tissue culture technique can be used for genetic improvement of date palm. The main purpose of this study was to evaluate the genetic variation in the tissue culture clones of date palm by using ISSR primers among mother and it’s two clones. The plant DNA was extracted and subjected to detection of genetic variation in two groups of date palm using ISSR primers. In this study ISSR primers produced monomorphic bands within group-1 and group-2. Genetic variation in tissue culture clones of date palm was not detecte by UBC primer series.
Cytotoxic | Primary research | Silver nanoparticlesPubrica
Biosynthesis, Antimicrobial, and Cytotoxic Effects of Silver Nanoparticles Using Acacia Concinna POD Extract and Kigelia Africa Leaf Extract- Secondary metabolites found in plants include alkaloids, flavonoids, phenolic compounds, phytosterols, saponins, tannins, carbohydrates, proteins, lipids, and minerals. These secondary metabolites have a wide range of uses, including the production of nanoparticles.
Biosynthesis, Antimicrobial, and Cytotoxic Effects of Silver Nanoparticles Using Acacia Concinna POD Extract and Kigelia Africa Leaf Extract- Secondary metabolites found in plants include alkaloids, flavonoids, phenolic compounds, phytosterols, saponins, tannins, carbohydrates, proteins, lipids, and minerals. These secondary metabolites have a wide range of uses, including the production of nanoparticles.
Visit us @ https://pubrica.com/insights/sample-work/cytotoxic-effects-of-sliver-nanoparticles-using-leaf-extract/
Assessment of genetic fidelity of in vitro propagated clones of Celastrus pan...iosrjce
Celastrus paniculatus Willd belonging to the family Celastaceae is an endangered Indian medicinal
plant having high pharmaceutical application. The objective of the present investigation was to assess the the
clonal fidelity of in vitro propagated clones of Celastrus paniculatus with the field grown mother plant to
confirm their true to type nature. Micropropagation is an alternative method for the large scale production of
endangered medicinal plants. The genetic stability of in vitro raised clones of celastrus paniculatus were
assessed by using RAPD analysis. Genomic DNA was isolated from healthy and fresh leaves of both mother
plant and in vitro raised plants of Celastrus paniculatus by using CTAB method. Based on the reproducibility of
the primers, 15 RAPD primers were selected for the present investigation. The selected primers gave rise to a
total of 75 scorable bands with an average of 5.1 bands ranging from 300-2700 bp. The number of bands varied
from three (OPQ-07, OPA-13) to seven (OPC-20, OPN-16). Randomly selected 10 micropropagated plants
from each culture period was used. Amplification pattern was electrophoresed in 1.5% TBE, revealing that all
the bands produced by micropropagated plants were monomorphic and similar to that of the field grown plant.
No polymorphism was detected by RAPD analysis.
Hydroxyl capped silver-gold alloy nanoparticles: characterization and their c...Nanomedicine Journal (NMJ)
Objective(s):
Metal nanoparticles (NPs) offer a wide variety of potential applications in pharmaceutical sciences due to the unique advances in nanotechnology research. In this work, bimetal Ag-Au alloy NPs were prepared and their combinations with other antibiotics were tested against Staphylococcus aureus.
Materials and Methods:
Firstly, Ag-Au alloy NPs with Au/Ag molar ratio of 1:1 was fabricated and was purified by agarose gel electrophoresis system. The morphology and size of the purified NPs were confirmed by transmission electron microscopy. Chemical composition and surface chemistry of these NPs were studied with atomic absorption spectophotometry and Fourier transforms infrared spectroscopy, respectively. The size of purified Ag-Au alloy NPs was less than 200 nm. Also the presence of organic compounds with a hydroxyl residue was detected on the surface of these purified NPs. In next step the effect of purified Ag-Au alloy NPs on the antibacterial activity of different antibiotics was evaluated at sub-inhibitory content (5 μg/disk) using disk diffusion method against S. aureus. Ag NPs and Au NPs were also tested at same content (5 μg) using mentioned method.
Results:
The most enhancing effect of Ag-Au alloy NPs was observed for penicillin G and piperacillin. No enhancing effects on the antibacterial activity of different antibiotics were observed at 5 μg/disk for the mono-metal nanoparticles (Ag NPs and Au NPs) against S. aureus.
Conclusion:
These results signify that the Ag-Au alloy NPs potentiates the antimicrobial action of certain antibiotics suggesting a possible utilization of this nano material in combination therapy against resistant S. aureus.
Nanotechnology has become one of the most promising technologies applied in
all areas of science. Metal nanoparticles produced by nanotechnology have
received global attention due to their extensive applications in the biomedical
and physiochemical
fields. Recently, synthesizing metal nanoparticles using
microorganisms and plants has been extensively studied and has been recog-
nized as a green and efficient way for further exploiting microorganisms as
convenient nanofactories. Here, we explore and detail the potential uses of
various biological sources for nanoparticle synthesis and the application of
those nanoparticles. Furthermore, we highlight recent milestones achieved for
the biogenic synthesis of nanoparticles by controlling critical parameters,
including the choice of biological source, incubation period, pH, and
temperature.
To study of the genetic variations among the Azospirillum lipoferu isolates u...ijsrd.com
Among free-living microorganisms, which can be practically used in agriculture, bacteria from the Azospirillum genus as well as other endophytes are nowadays thought of as the most active component of associative dinitrogen fixation. The investigation was carried out to study the characterization of Azospirillum lipoferu found in the soils of the ten agro-climatic zones which Karnataka, is classified. By using RAPD markers, 75 bands were scored out of which 78.6 % were found to be polymorphic. Statistical analysis of RAPD data enabled the classification of 10 Azospirillum isolates into two major groups. . In this, the cluster analysis based on 75 RAPD bands revealed that the ten A. lipoferu isolates examined clustered at a linkage distance of about 40 units on the dendrogram. There was no correlation between RAPD and geographical origin of isolates.
Evaluation of the effect of crocetin on antitumor activity of doxorubicin enc...Nanomedicine Journal (NMJ)
Objective(s): The current study reports investigation of codelivery by PLGA nanoparticles (NPs) loaded with crocetin (Cro), a natural carotenoid dicarboxylicHYPERLINK “http://en.wikipedia.org/wiki/Carboxylic_acid” acid that is found in the crocus flower, and Doxorubicin (DOX).
Materials and Methods: Double emulsion/solvent evaporation method was used for preparation of PLGA nanoparticles containing Dox and Cro. Characterizations of prepared NPs were investigated by atomic force microscopy (AFM) and dynamic light scattering analysis. In vitro Cytotoxicity of DOX and Cro loaded PLGA NPs (PLGA-DOX-Cro) on MCF-7 cell line was evaluated using MTT test. Flow cytometry experiments were implemented to distinguish cells undergoing apoptosis from those undergoing necrosis. Furthermore the expression of caspase 3 was examined by western blot analysis.
Results: The prepared formulations had size of 150- 300 nm. Furthermore, PLGA-DOX-Cro nanoparticles inhibited MCF-7 tumor cells growth more efficiently than either DOX or Cro alone at the same concentrations, as quantified by MTT assay and flow cytometry. Studies on cellular uptake of DOX-Cro-NPs demonstrated that NPs were effectively taken up by MCF-7 tumor cells.
Conclusion: This study suggested that DOX-Cro-NPs may have promising applications in breast cancer therapy.
Effects of combination of magnesium and zinc oxide nanoparticles and heat on ...Nanomedicine Journal (NMJ)
Objective: The objective of this study was to investigate the antibacterial activities of combination of MgO and ZnO nanoparticles in the presence of heat against Escherichia coli and Staphylococcus aureus.
Materials and Methods:Bacteria were grown on either agar or broth media followed by the addition of ZnO and MgO nanoparticles. Then the combined effect of ZnO and MgO nanoparticles was investigated. Furthermore, the media containing nanoparticles were treated with mild heat and their synergistic antibacterial activity was investigated against E. coli and S. aureus in milk.
Results: The data showed that the nanoparticles used in this study had no effect on the bacteria in the agar medium. However, the results showed that ZnO and MgO nanoparticles resulted in a significant decrease in the number of E. coli (P<0.000) and S. aureus (Pd”0.05) in the broth medium. The combination of nanoparticles and mild heat exhibited a significant decrease in the number of E. coli and S. aureus indicating the synergistic effects of nanoparticles and heat.
Conclusion: Using a combination of mild heat, ZnO and MgO nanoparticles, E. coli and S. aureus can be controlled successfully in the milk. Mild heating plus ZnO and MgO nanoparticles has a synergistic effect which would reduce the need for high temperature and also the concentrations of ZnO and MgO nanoparticles required for pathogen control in minimally processed milk during maintaining.
Preparation and evaluation of electrospun nanofibers containing pectin and ti...Nanomedicine Journal (NMJ)
Objective(s):The aim of this study was to prepare electrospun nanofibers of celecoxib using combination of time-dependent polymers with pectin to achieve a colon-specific drug delivery system for celecoxib.
Materials and Methods:Formulations were produced based on two multilevel 22 full factorial designs. The independent variables were the ratio of drug:time-dependent polymer (X1) and the amount of pectin in formulations (X2). Electrospinning process was used for preparation of nanofibers. The spinning solutions were loaded in 5 mL syringes. The feeding rate was fixed by a syringe pump at 2.0 mL/h and a high voltage supply at range 10-18 kV was applied for electrospinning. Electrospun nanofibers were collected and evaluated by scanning electron microscopy and drug release in the acid and buffer with pH 6.8 with and without pectinase.
Results:Electrospun nanofibers of celecoxib with appropriate morphological properties were produced via electrospinning process. Drug release from electrospun nanofibers was very low in the acidic media; while, drug release in the simulated colonic media was the highest from formulations containing pectin.
Conclusion: Formulation F2 (containing drug:ERS with the ratio of 1:2 and 10% pectin) exhibited acceptable morphological characteristics and protection of drug in the upper GI tract and could be a good candidate as a colonic drug delivery system for celecoxib.
The combined effects of Aloe vera gel and silver nanoparticles on wound heali...Nanomedicine Journal (NMJ)
Objective(s): This study was aimed at investigating the synergy effects of Aloe vera gel and silver nanoparticles on the healing rate of the cutting wounds.
Materials and Methods: In order to determine the concentration of silver nanoparticles in Aloe vera gel, the MBC methods were applied on the most common bacteria infecting wounds, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa. The cutting wounds with Full-thickness skin were dorsally created on rats; then the rats were divided into 4 groups. The treatments groups included: mixture of Aloe vera gel and silver nanoparticles, Aloe vera gel alone and silver nanoparticles alone in addition to control groups. The treatment was carried out for 2 weeks and the size of the wound closures were measured by an image software analysis.
Results:There was no significant difference (p<0.05) in healing rate between the control and mixture group. However, there were significant differences between the silver nanoparticles and Aloe vera groups using Tukey’s analysis on the 6th, 8th and 10th days.
Conclusion:The Aloe vera gel increased the rate of wound healing whereas the silver nanoparticles had a delay effect; and when they were mixed, it was similar to the average effect of both Aloe vera gel and silver nanoparticles.
Simultaneous loading of 5-florouracil and SPIONs in HSA nanoparticles: Optimi...Nanomedicine Journal (NMJ)
Objective(s): Over the past two decades, considerable interest has been focused on utilizing biocompatible magnetic nanoparticles (MNPs) for biomedical applications. In this study, production of human serum albumin (HSA) nanoparticles using desolvation technique that were simultaneous loaded with high amounts of superparamagnetic iron oxide nanoparticles (SPIONs) and 5-flourouracil (5-FU) was investigated.
Materials and Methods: 5-FU loading (%) and SPIONs entrapment efficiency (%) were optimized using response surface methodology (RSM). The design expert software used to analyse the interactive effects of pH, 5-FU and SPIONs concentrations.
Results:The optimum conditions found to be pH of 8.2, drug concentration of 1.5 mg/ml and SPIONs concentration of 2.79 mg/ml. Under the mentioned optimum conditions, particles with the size of 111.8 nm, zeta potential of -37.1 mV, 5-FU loading of 15.8% and SPIONs entrapment efficiency of 41.1% were obtained. In vitro cumulative release of 5-FU from the nanoparticles was evaluated in phosphate buffer saline (pH 7.4, 37 °C). Results indicated that 85% of the 5-FU released during 95 h, which revealed a sustained release profile. In addition, Vibrating Sample Magnetometer (VSM) analyses confirmed the superparamagnetic properties of magnetic albumin nanoparticles manufactured under the optimum conditions.
Conclusion: According to the findings,SPIONs and 5-FU loaded HAS nanoparticles arepromising for use as novel targeted delivery system due to proper magnetic and drug release behaviours.
Antimicrobial and cytotoxicity effect of silver nanoparticle synthesized by C...Nanomedicine Journal (NMJ)
Objective(s): For the development of reliable, ecofriendly, less expensive process for the synthesis of silver nanoparticles and to evaluate the bactericidal, and cytotoxicity properties of silver nanoparticles synthesized from root extract of Croton bonplandianum, Baill.
Materials and Methods: The synthesis of silver nanoparticles by plant part of Croton bonplandianum was carried out. The formation of nanoparticles was confirmed by Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM), XRD and UV-Vis spectrophotometric analysis. The biochemical properties were assayed by antibacterial study, cytotoxicity assay using cancer cell line.
Results: The formation of silver nanoparticles was confirmed by UV-VIS spectroscopic analysis which showed absorbance peak at 425 nm. X-ray diffraction photograph indicated the face centered cubic structure of the synthesized AgNPs. TEM has displayed the different dimensional images of biogenic silver nanoparticles with particle size distribution ranging from 15-40 nm with an average size of 32 nm. Silver particles are spherical in shape, clustered. The EDX analysis was used to identify the elemental composition of synthesized AgNPs. Antibacterial activity of the synthesized AgNPs against three Gram positive and Gram negative bacteria strains like Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa carried out showed significant zones of inhibition. The cytotoxicity study by AgNPS also showed cytotoxicity on ovarian cancer cell line PA-1 and lung epithelial cancer cell line A549.
Conclusion: The present study confirms that the AgNPs have great promise as antibacterial, and anticancer agent.
Investigation of the effect of different parameters on the phase inversion te...Nanomedicine Journal (NMJ)
Objective(s): Nanoemulsions are a kind of emulsions that can be transparent, translucent (size range 50-200 nm) or “milky” (up to 500 nm). Nanoemulsions are adequatly effective for transfer of active component through skin which facilitate the entrance of the active component . The transparent nature of the system and lack of the thickener and fluidity are among advantages of nanoemulsion.
Materials and Methods: In this study, a nanoemulsion of lemon oil in water was prepared by the phase inversion temperature (PIT) emulsification method in which the tween 40 was used as surfactant. The effect of concentration of NaCl in aqueous phase, pH and weight percent of surfactant and aqueous on the PIT and droplet size were investigated. Results: The results showed that with increasing of concentration of NaCl from 0.05 M to 1 M, PIT decrease from 72 to 50. The average droplet sizes, for 0.1, 0.5 and 1 M of NaCl in 25 ºC are 497.3, 308.1 and 189.9 nm, respectively and the polydispersity indexes are 0.348, 0.334 and 0.307, respectively.
Conclusion: Considering the characteristics of nanoemulsions such as being transparent, endurance of solution and droplet size can provide suitable reaction environment for polymerization process used in making hygienic and medical materials.
Mechanism of oxidative stress involved in the toxicity of ZnO nanoparticles a...Nanomedicine Journal (NMJ)
ZnO NPs (zinc oxide nanoparticles) has generated significant scientific interest as a novel antibacterial and anticancer agent. Since oxidative stress is a critical determinant of ZnO NPs-induced damage, it is necessary to characterize their underlying mode of action. Different structural and physicochemical properties of ZnO NPs such as particle surface, size, shape, crystal structure, chemical position, and presence of metals can lead to changes in biological activities including ROS (reactive oxygen species) production. However, there are some inconsistencies in the literature on the relation between the physicochemical features of ZnO NPs and their plausible oxidative stress mechanism. Herein, the possible oxidative stress mechanism of ZnO NPs was reviewed. This is worthy of further detailed evaluations in order to improve our understanding of vital NPs characteristics governing their toxicity. Therefore, this study focuses on the different reported oxidative stress paradigms induced by ZnO NPs including ROS generated by NPs, oxidative stress due to the NPs-cell interaction, and role of the particle dissolution in the oxidative damage. Also, this study tries to characterize and understand the multiple pathways involved in oxidative stress induced by ZnO NPs. Knowledge about different cellular signaling cascades stimulated by ZnO NPs lead to the better interpretation of the toxic influences induced by the cellular and acellular parameters. Regarding the potential benefits of toxic effects of ZnO NPs, in-depth evaluation of their toxicity mechanism and various effects of these nanoparticles would facilitate their implementation for biomedical applications.
Combined effects of PEGylation and particle size on uptake of PLGA particles ...Nanomedicine Journal (NMJ)
Abstract
Objective:
At the present study, relationship between phagocytosis of PLGA particles and combined effects of particle size and surface PEGylation was investigated.
Materials and Methods:
Microspheres and nanospheres (3500 nm and 700 nm) were prepared from three types of PLGA polymers (non-PEGylated and PEGylation percents of 9% and 15%). These particles were prepared by solvent evaporation method. All particles were labeled with FITC-Albumin. Interaction of particles with J744.A.1 mouse macrophage cells, was evaluated in the absence or presence of 7% of the serum by flowcytometry method.
Results:
The study revealed more phagocytosis of nanospheres. In the presence of the serum, PEGylated particles were phagocytosed less than non-PEGylated particles. For nanospheres, this difference was significant (P<0/05) and their uptake was affected by PEGylation degree. In the case of microsphere formulation, PEGylation did not affect the cell uptake. In the serum-free medium, the bigger particles had more cell uptake rate than smaller ones but the cell uptake rate was not influenced by PEGylation.
Conclusion:
The results indicated that in nanosized particles both size and PEgylation degree could affect the phagocytosis, but in micron sized particles just size, and not the PEGylation degree, could affect this.
Synthesis of silver nanoparticles and its synergistic effects in combination ...Nanomedicine Journal (NMJ)
Abstract
Objectives:
Biofilms are communities of bacteria attached to surfaces through an external polymeric substances matrix. In the meantime, Acinetobacterbaumannii is the predominant species related to nosocomial infections. In the present study, the effect of silver nanoparticles alone and in combination with biocides and imipenem against planktonic and biofilms of A. baumannii was assessed.
Materials and Methods:
Minimum inhibitory concentrations (MICs) of 75 planktonic isolates of A. baumannii were determined by using the microdilution method as described via clinical and laboratory standards institute (CLSI). Among all strains, 10 isolates which formed strong biofilms were selected and exposed to silver nanoparticles alone and in combination with imipenem, bismuth ethandithiol (BisEDT) and bismuth propanedithiol (BisPDT) to determine minimum biofilm inhibitory concentrations (MBIC). Subsequently, minimum biofilm eradication concentrations (MBECs) of silver nanoparticles alone and in combination with imipenem against mature biofilm of the isolates were evaluated.
Results:
Results showed that 29.3% of isolates were susceptible to silver nanoparticles and could inhibit the growth and eradicate biofilms produced by the isolates. For this reason, ∑FIC, ∑FBIC and ∑FBEC ≤ 0.05 were reported which shows synergism between silver nanoparticles and imipenem against not only planktonic cells but also inhibition and eradication of biofilms. The results of ∑FBIC >2 indicated to antagonistic impacts between silver nanoparticles and BisEDT/BisPDT against biofilms.
Conclusion:
It can be concluded that silver nanoparticles alone can inhibit biofilm formation but in combination with imipenem are more effective against A. baumannii in planktonic and biofilm forms.
Abstract
Objective(s):
Zinc oxide nanoparticles (ZNP) are increasingly used in sunscreens, biosensors, food additives and pigments. In this study the effects of ZNP on liver of rats was investigated.
Materials and Methods:
Experimental groups received 5, 50 and 300 mg/kg ZNP respectively for 14 days. Control group received only distilled water. ALT, AST and ALP were considered as biomarkers to indicate hepatotoxicity. Lipid peroxidation (MDA), SOD and GPx were detected for assessment of oxidative stress in liver tissue. Histological studies and TUNEL assay were also done.
Results:
Plasma concentration of zinc (Zn) was significantly increased in 5 mg/kg ZNP-treated rats. Liver concentration of Zn was significantly increased in the 300 mg/kg ZNP-treated animals. Weight of liver was markedly increased in both 5 and 300 mg/kg doses of ZNP. ZNP at the doses of 5 mg/kg induced a significant increase in oxidative stress through the increase in MDA content and a significant decrease in SOD and GPx enzymes activity in the liver tissue. Administration of ZNP at 5 mg/kg induced a significant elevation in plasma AST, ALT and ALP. Histological studies showed that treatment with 5 mg/kg of ZNP caused hepatocytes swelling, which was accompanied by congestion of RBC and accumulation of inflammatory cells. Apoptotic index was also significantly increased in this group. ZNP at the dose of 300 mg/kg had poor hepatotoxicity effect.
Conclusion:
It is concluded that lower doses of ZNP has more hepatotoxic effects on rats, and recommended to use it with caution if there is a hepatological problem.
Synthesis of graphene oxide-TiO2 nanocomposite as an adsorbent for the enrich...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
In our study, graphene oxide-TiO2 nanocomposite (GO/TiO2) was prepared and used for the enrichment of rutin from real samples for the first time.
Materials and Methods:
The synthesized GO/TiO2 was characterized by X-ray diffraction, scanning electron microscopy, and FT-IR spectra. The enrichment process is fast and highly efficient. The factors including contact time, pH, and amount of GO/TiO2 affecting the adsorption process were studied.
Results:
The maximum adsorption capacity for ciprofloxacin was calculated to be 59.5 mg/g according to the Langmuir adsorption isotherm. The method yielded a linear calibration curve in the concentration ranges from 15 to 200 μg/L for the rutin with regression coefficients (r2) of 0.9990. The limits of detection (LODs, S/N=3) and limits of quantification (LOQs, S/N=10) were found to be 8 μg/Land 28 μg/L, respectively. Both the intra-day and inter-day precisions (RSDs) were < 10% .
Conclusion:
The developed approach offered wide linear range, and good reproducibility. Owing to the diverse structures and unique characteristic, GO/TiO2 possesses great potential in the enrichment and analysis of trace rutin in real aqueous samples.
Preparation and evaluation of vitamin A nanosuspension as a novel ocular drug...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
The aim of this study was to prepare a nanosuspension formulation as a new vehicle for the improvement of the ocular delivery of vitamin A.
Material and Methods:
Formulations were designed based on full factorial design. A high pressure homogenization technique was used to produce nanosuspensions. Fifteen formulations were prepared by the use of different combinations of surfactants Tween 80, benzalkonium chloride and Pluronic and evaluated for pH, particle size, entrapment efficiency, differential scanning calorimetry (DSC), stability and drug release. Also, Draize test was used to evaluate the irritation of rabbit eye by formulations.
Results:
All formulations showed a small mean size that is well suited for ocular application. Also it was observed that the particle size decreased with increase in the amount of surfactant. Drug entrapment increased with increasing amount of surfactant. It was shown that initial and final drug release can be controlled by the ratio and the total amount of surfactants, respectively.
Conclusion:
It was concluded that the use of Tween 80 and Pluronic in the formualtions with a proper ratio does not show eye irritation and could be useful to achieve a suitable nanosuspension of vitamin A as a novel ocular delivery system.
A comparative study about toxicity of CdSe quantum dots on reproductive syste...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
Medicinal benefits of quantum dots have been proved in recent years but there is little known about their toxicity especially in vivo toxicity. In order to use quantum dots in medical applications, studies ontheir in vivo toxicity is important.
Materials and Methods:
CdSe:ZnS quantum dots were injected in 10, 20, and 40 mg/kg doses to male mice10 days later, mice were sacrificed and five micron slides were prepared structural and optical properties of quantum dots were evaluated using XRD.
Results:
Histological studies of testis tissue showed high toxic effect of CdSe:ZnS in 40 mg/kg group. Histological studies of epididymis did not show any effect of quantum dots in terms of morphology and tube structure. Mean concentration of LH and testosterone and testis weight showed considerable changes in mice injected with 40 mg/kg dose of CdSe:ZnS compared to control group. However, FSH and body weight did not show any difference with control group.
Conclusion:
Although it has been reported that CdSe is highly protected from the environment by its shell, but this study showed high toxicity for CdSe:ZnS when it is used in vivo which could be suggested that shell could contribute to increased toxicity of quantum dots. Considering lack of any previous study on this subject, our study could potentially be used as an basis for further extensive studies investigating the effects of quantum dots toxicity on development of male sexual system.
Functionalization of carbon nanotubes and its application in nanomedicine: A ...Nanomedicine Journal (NMJ)
Abstract
This review focuses on the latest developments in applications of carbon nanotubes (CNTs) in medicine. A brief history of CNTs and a general introduction to the field are presented.
Then, surface modification of CNTs that makes them ideal for use in medical applications is highlighted. Examples of common applications, including cell penetration, drug delivery, gene delivery and imaging, are given. At the same time, there are concerns about their possible adverse effects on human health, since there is evidence that exposure to CNTs induces toxic effects in experimental models. However, CNTs are not a single substance but a growing family of different materials possibly eliciting different biological responses. As a consequence, the hazards associated with the exposure of humans to the different forms of CNTs may be different. Understanding the structure–toxicity relationships would help towards the assessment of the risk related to these materials. Finally, toxicity of CNTs, are discussed. This review article overviews the most recent applications of CNTs in Nanomedicine, covering the period from 1991 to early 2015.
The role of surface charge of ISCOMATRIX nanoparticles on the type of immune ...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
ISCOMATRIX vaccines have now been shown to induce strong antigen-specific cellular or humoral immune responses to a broad range of antigens of viral, bacterial, parasite or tumor. In the present study, we investigated the role of ISCOMATRIX charge in induction of a Th1 type of immune response and protection against Leishmania major infection in BALB/c mice.
Materials and Methods:
Positively and negatively charged ISCOMATRIX were prepared. BALB/C mice were immunized subcutaneously, three times with 2-week intervals, with different ISCOMATRIX formulations. Soluble Leishmania antigens (SLA) were mixed with ISCOMATRIX right before injection. The extent of protection and type of immune response were studied in different groups of mice.
Results:
The group of mice immunized with negatively charged ISCOMATRIX showed smaller footpad swelling upon challenge with L. major and the highest IgG2a production compared with positively charged one. The mice immunized with positively charged ISCOMATRIX showed the lowest splenic parasite burden compared to the other groups. Cytokine assay results indicated that the highest level of IFN- γ and IL-4 secretion was observed in the splenocytes of mice immunized with negatively charged ISCOMATRIX as compared to other groups.
Conclusion:
The results indicated that ISCOMATRIX formulations generate an immune response with mixed Th1/Th2 response that was not protective against challenge against L. major.
Abstract
In the last decade, developments in nanotechnology have provided a new field in medicine called “Nanomedicine”. Nanomedicine has provided new tools for photodynamic therapy. Quantum dots (QDs) are approximately spherical nanoparticles that have attracted broad attention and have been used in nanomedicine applications. QDs have high molar extinction coefficients and photoluminescence quantum yield, narrow emission spectra, broad absorption, large effective stokes shifts. QDs are more photostable and resistant to metabolic degradation. These photosensitizing properties can be used as photosensitizers for Photodynamic Therapy (PDT). PDT has been recommended for its unique characteristic, such as low side effect and more efficiency. Therefore, nanomedicine leads a promising future for targeted therapy in cancer tumor. Furthermore, QDs have recently been applied in PDT, which will be addressed in this review letter. Also this review letter evaluates key aspects of nano-particulate design and engineering, including the advantage of the nanometer scale size range, biological behavior, and safety profile.
Preparation of protein-loaded PLGA-PVP blend nanoparticles by nanoprecipitati...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
Despite of wide range applications of polymeric nanoparticles in protein delivery, there are some problems for the field of protein entrapment, initial burst and controlled release profile.
Materials and Methods:
In this study, we investigated the influence of some changes in PLGA nanoparticles formulation to improve the initial and controlled release profile. Selected parameters were: pluronic F127, polysorbate 80 as surfactant, pH of inner aqueous phase, L/G ratio of PLGA polymer, volume of inner aqueous phase and addition of polyvinylpyrrolidone as an excipient. FITC-HSA was used as a model hydrophilic drug. The nanoparticles were prepared by nanoprecipitation.
Results:
Initial release of FITC-HSA from PLGA-tween 80 nanoparticles (opt-4, 61%) was faster than control (PLGA-pluronic) after 2.30 h of incubation. Results showed that decrease in pH of inner aqueous phase to pI of protein can decrease IBR but the release profile of protein is the same as control. Release profile with three phases including a) initial burst b) plateau and c) final release phase was observed when we changed volume of inner aqueous phase and L/G ratio in formulation. Co-entrapment of HSA with PVP and pluronic reduced the IBR and controlled release profile in opt-19. Encapsulation efficiency was more than 97% and nanoparticles size and zeta potentials were mono-modal and -18.99 mV, respectively.
Conclusion:
In this research, we optimized a process for preparation of PLGA-PVP-pluronic nanoparticles of diameter less than 300 nm using nanoprecipitation method. This formulation showed a decreased initial burst and long lasting controlled release profile for FITC-HSA as a model drug for proteins.
How to Make a Field invisible in Odoo 17Celine George
It is possible to hide or invisible some fields in odoo. Commonly using “invisible” attribute in the field definition to invisible the fields. This slide will show how to make a field invisible in odoo 17.
Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
Macroeconomics- Movie Location
This will be used as part of your Personal Professional Portfolio once graded.
Objective:
Prepare a presentation or a paper using research, basic comparative analysis, data organization and application of economic information. You will make an informed assessment of an economic climate outside of the United States to accomplish an entertainment industry objective.
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
For more information, visit-www.vavaclasses.com
Francesca Gottschalk - How can education support child empowerment.pptxEduSkills OECD
Francesca Gottschalk from the OECD’s Centre for Educational Research and Innovation presents at the Ask an Expert Webinar: How can education support child empowerment?
Biological screening of herbal drugs: Introduction and Need for
Phyto-Pharmacological Screening, New Strategies for evaluating
Natural Products, In vitro evaluation techniques for Antioxidants, Antimicrobial and Anticancer drugs. In vivo evaluation techniques
for Anti-inflammatory, Antiulcer, Anticancer, Wound healing, Antidiabetic, Hepatoprotective, Cardio protective, Diuretics and
Antifertility, Toxicity studies as per OECD guidelines
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...
Isolation and characterization of a fungus for extracellular synthesis of small selenium nanoparticles
1. Nanomed J, Vol. 1, No. 1, Autumn 2013 13
Received: Jun. 15, 2013; Accepted: Jul. 12, 2013
Vol. 1, No. 1, Autumn 2013, page 13-19
Online ISSN 2322-5904
http://nmj.mums.ac.ir
Original Research
Isolation and characterization of a fungus for extracellular synthesis of
small selenium nanoparticles
Bijan Zare, Shabnam Babaie, Neda Setayesh, Ahmad Reza Shahverdi*
Department of Pharmaceutical Biotechnology, Faculty of Pharmacy and Biotechnology Research Center, Tehran
University of Medical Sciences, Tehran, Iran
Abstract
Objective(s): The use of biogenic selenium nanoparticles for various purposes is going to be
an issue of considerable importance; thus, appropriate simple methods should be developed
and tested for the synthesis and recovery of these nanoparticles.
Materials and Methods: In this study, a fungus was isolated from a soil sample, identified as
Aspergillus terreus and used for extracellular synthesis of selenium nanoparticles (Se NPs).
UV–Vis spectroscopy and energy dispersive X-ray spectrum studies were carried out to
confirm Se NPs formation within 60 min. Dynamic light scattering and scan electron
microscopic methods were also used to characterize both size and shapes of the Se NPs.
Results: The results show that spherical particles with average size of 47 nm were formed by
adding a culture supernatant of A. terreus to selenium ions solution.
Conclusion: This approach appears to be an easy and appropriate method for extracellular
synthesis of small Se NPs. Extracellular synthesis of small Se NPs has not been reported yet.
Keywords: Aspergillus terreus, Extracellular, Selenium nanoparticle, Soil, Synthesis
*Corresponding author: Ahamdreza Shahverdi, Department of Pharmaceutical Biotechnology, Faculty of
Pharmacy and Biotechnology Research Center, Tehran University of Medical Sciences, Tehran, Iran.
Tel: +98 216461178, Email: shahverd@sina.tums.ac.ir
2. Extracellular synthesis of selenium nanoparticles using fungi
14 Nanomed J, Vol. 1, No. 1, Autumn 2013
Introduction
Metal nanoparticles (MNPs) have found many
applications in medicine, electronics and other
industries (1-3). During past decades, many
reports have been published on the preparation
of different MNPs by biological and chemical
processes (4-8). Among these methods, there
is an increasing interest to use biological
techniques to produce different MNPs (6, 9,
10). Biogenic MNPs show frequently lower
toxicity and exhibit better stability comparing
to those nanoparticles (NPs) which fabricated
by chemical methods (11). Different
microorganisms can reduce toxic metal ions to
elemental NPs (5, 12, 13). These toxic metal
ions are often absorbed into the cells, reduced
and deposited in intracellular space of
microorganisms (4). Isolation of these NPs
needs to release of NPs from the cells and set
up an appropriate extraction method to purify
them for further applications (14). During
extraction method, liquid nitrogen is needed
for disruption of the cells and some organic
solvents (i.e. n-octanol, chloroform, ether)
should be used for purification of generated
MNPs from cell debris (5, 14). These
mentioned processes are time consuming and
may be tedious. Few papers have been recently
published which report the syntheses of MNPs
using cell-free supernatants of many bacterial
or fungal cultures (15, 16). In this strategy the
reduction of metal ions take place in the
presence of the metabolites secreted in culture
broth by living microorganisms. So no
expensive treatments such as cell disruption
method or solvent extraction are required to
isolate the generated NPs (15, 17). In current
investigation, different soil samples were
collected from the around of Tehran, Iran and
screened for finding a fungus to convert
selenium (Se) ions to selenium nanoparticles
(Se NPs). Se NPs have been exhibited
different biological activities in cell culture or
animal models which make this NPs candidate
for different therapeutic purposes (18-21). In
the basis of our literature survey, the
extracellular synthesis of Se NPs by cell-free
culture broth has not been investigated so far
and this is the first report to describe the
synthesis of Se NPs using culture supernatant
of a fungus strain isolated from a soil sample.
Materials and Methods
Screening procedure
Different soil samples were collected from
different parts of the around of Tehran (Iran).
All samples were milled, suspended in sterile
normal saline and subjected to screening
process. Sabouraud dextrose agar (SDA) cul-
ture medium was prepared, sterilized and
supplemented with solution of SeO2 (10 g/l) to
final concentration equal to 100 mg/l SeO2. 1
ml of above suspensions which prepared from
the collected soil samples was 10 times diluted
and spread on the surface of the Se
supplemented SDA plates (100 mg/l). The
plates were incubated aerobically at 30°C, and
after one week, all red-colored colonies which
observed on the Se supplemented SDA plates,
were picked up and transferred to Se free SDB
medium. In next step, all SDB media were
further incubated for 10 days at 30o
C in an
incubator (150 rpm). During incubation
periods several samples (0.5 ml) were
aseptically withdrawn from culture flasks,
centrifuged at 4000×g for 20 min and added to
4.5 ml Se+4
ions solution ( 100 mg/l). After 60
min, the formation of red colloid was checked
in all reaction vessels. Red is the color of
generated elemental Se NPs and thus serves as
a provisional marker to identify a culture
supernatant of an isolate is capable to form Se
NPs or not.
Identification of the isolate
The identification of the isolate was carried
out by 28s ribosomal deoxyribonucleic acid
(rDNA) sequence analysis (22). To earn
genomic DNA, fungal cells harvested from 72
hrs culture mediums, washed three times with
sterile distilled water. The cells were disrupted
by grinding with liquid nitrogen and the slurry
was subjected to phenol-chloroform DNA
extraction procedure. An equal volume of TE-
saturated phenol: chloroform (1:1) added to
the sample contained in a 1.5 ml micro tube
and vortex for 20 seconds. The sample was
centrifuged for 5 minutes at room temperature
3. Zare B, et al.
Nanomed J, Vol. 1, No. 1, Autumn 2013 15
to separate the phases. The upper, aqueous
layer was passed to a new tube. DNA
materials were precipitated by adding cold
absolute ethanol. The sample centrifuged at
12,000 rpm for 15 minutes at 4O
C. The super-
natant decanted, and drained by inverting on a
paper towel. For more purification, ethanol
(80%v/v) added (corresponding to about two
volume of the original sample), incubated at
room temperature for 5-10 minutes and
centrifuged again for 5 minutes, and decanted
and drained the tube, as above. The DNA
materials was re-suspended in diethyl-
pyrocarbonate (DEPC) solution in water bath
(60o
C) and subjected to 28s rDNA polymerase
chain reaction (PCR) amplification.
The PCR amplification included an initial
denaturation at 94°C for 180 sec, 30 cycles of
denaturation 94°C for 60 sec, annealing 52°C
for 45 sec and extension 72°C for 90 sec and a
final extension 72°C for 300 sec. A fragment
of the 28s rDNA gene was amplified using
forward primer 5'-AAGCATATCAATAAGC-
GGAGG-3' AND reverse primer 5'- GGTCC-
GTGTTTCAAGACGG-3'. The amplified
DNA fragment was purified from agarose 1%
gel using the QIAquick Gel Extraction Kit
(Qiagen, USA) according to the supplier's
instructions and was sent for auto-mated
sequencing using the above primers (Cinagen
Co., Iran). Sequence similarity searches were
done with the BLAST database (National
Center for Biotechnology Infor-mation), and
the sequence was submitted to NCBI GenBank
Nucleotide Sequence Data-base (accession
number KC145152).
Preparation of Se NPs and their characterize-
ation
Fungus isolate from the soil sample was used
for extracellular synthesis of the Se NPs.
Sterile Sabouraud dextrose broth (SDB)
medium was prepared, and 100 ml of this
medium was transferred to a sterile 500-mL
Erlenmeyer flask. The medium was inoculated
with 1 ml of the fresh inoculums (OD600, 0.1)
and incubated aerobically at 30°C in a shaker
incubator (150 rpm). After 7 days, the fungal
cells were removed from the culture medium
by centrifugation at 4,000 ×g for 10 min. The
supernatant was collected and filtered through
a 0.22 μm filter (Millipore, USA). Subse-
quently 20 ml of filtered supernatants was
added to 80 ml of Se4+
ions solution (100
mg/ml) and reaction mixture was incubated at
room temperature for 60 min to complete the
Se NPs formation. The isolation of Se NPs
from culture supernatant was carried out by
high speed centrifuging process (20000g, 10
min). In next step, the red sediments was
dispersed in distilled water, centrifuged at
above mentioned condition and washed for
tree times. The collected Se NPs were re-
suspended in distilled water to form a colloid
and characterized using different techniques.
The Se colloid tested for their optical
absorption property by using a UV-Vis
spectrophotometer (UVD-2950, Labomed,
USA) operated at a resolution of 1 nm. The
particle size distribution patterns of generated
NPs were also obtained by the Zetasizer
MS2000 (Malvern Instruments, UK). The
morphology of the prepared Se NPs was
studied using Scanning Electron Microscopy
(SEM). For this purpose, above Se colloid was
further dispersed by ultrasonication method,
and a drop of suspension was located on
carbon-coated SEM grid. Subsequently grid
was dried under an infrared lamp and further
coated by gold. Micrographs were achieved
using a SEM (vega tescan, Czech) operated at
an accelerating voltage at 15 kV. To determine
the elemental composition of the NPs, energy
dispersive X-ray spectrum (EDX) micro-
analysis (vega tescan, Czech) was also per-
formed.
Results and Discussion
Among more than 50 fungus strains which
were isolated from soil samples and cultivated
in SDB media only the culture supernatant of a
fungus isolate could produce exo-metabolites
to convert Se+4
ions to Se NPs. BLAST search
of the amplified 28S rDNA sequence against
the NCBI Nucleotide database was conducted
to identify isolate. Alignment results show
some characters which had identity with
several members of the genus Aspergillus
4. Extracellular synthesis of selenium nanoparticles using fungi
16 Nanomed J, Vol. 1, No. 1, Autumn 2013
terreus. The isolate was designated as A.
terreus BZ1 and its 28S rDNA sequence was
submitted to the NCBI nucleotide database and
registered with accession number of
KC145152.
Figure 1. Left illustration shows a flask contains normal
culture of A. terrus BZ1 in the plain SDB. Right illustration
demonstrates test tubes contain Se4+
ions before (a) and
after (b) addition of culture supernatant of A. terrus BZ1.
As mentioned in materials and methods part,
this isolate was cultivated in liquid media
(SDB) and its filtered supernatant was used for
preparation of Se NPs. Left illustration in
Figure 1 shows normal culture broth of A.
terreus incubated for 1 week. Collected super-
natant of this isolate has been demonstrated in
right illustration of Figure 1 before (tube A)
and after addition to Se ions solution (tube B).
The UV–visible spectrum of the prepared Se
NPs is shown in Figure 2. A band observed in
the spectrum, corresponding to surface
plasmon resonance and occurred at 245 nm,
indicating the formation of Se NPs (23).
Figure 3 shows the corresponding size distri-
bution of Se NPs measured by laser light
scattering method. For separated Se NPs, one
modal peak was clearly revealed in the range
between 18 and 105 nm, and NPs in the size of
47.5 nm had the most frequency. The zeta
potential value for fabricated Se NPs was also
measured by Zetasizer and reported as -22.9
mV. According to SEM image demonstrated
in Figure 4, the prepared Se NPs show
spherical shapes with particle size of ≤ 100
nm.
Figure 2. UV–visible spectrum of the Se NPs synthesized
by culture supernatant of A. terrus BZ1.
Figure 3. Size distribution pattern of Se NPs synthesized
by culture supernatant of A. terrus BZ1.
Furthermore, EDX microanalysis of the separ-
ated NPs exhibited Se absorption peaks
consisting of SeLα, SeKα and SeKβ at 1.37,
11.22 and 12.49 keV, respectively (Figure 5)
and confirms the presence of Se in the
samples.
Elemental composition analysis also showed
the presence of signals from the gold which
are related to the gold which has been used in
grid coating process MNPs such as Se NPs and
silver NPs have found much attention in
different branches of science and technology
(1, 24, 25). Both of these NPs are important
elements in healthcare issue (19, 26). Zhang
and his co-workers showed that Se NPs has a
size-dependent effect in directly scavenging
free radicals in vitro (20).
5. Zare B, et al.
Nanomed J, Vol. 1, No. 1, Autumn 2013 17
Figure 4. SEM image of the Se NPs synthesized by culture
supernatant of A. terrus BZ1.
.
Figure 5. EDX spectrum of the Se NPs synthesized by
culture supernatant of A. terrus BZ1.
Nano-Se has comparable efficacy in up-
regulating selenoenzymes but is less toxic
compared to selenite (20, 27). Nano-elements
such as Se NPs can be produced intracellular
or extracellular by different microorganisms.
Although the intracellular biosynthesis of Se
NPs using different bacteria and their isolation
and purification have been recently reported in
literature and well described [28-30] but there
is no report on the synthesis of Se NPs using
cell-free culture supernatant obtained from a
fungus strain. In contrast, the syntheses of Ag
NP using culture supernatants of some
bacterial or some plant extracts have been
published in literature (16, 31). To obtain a
culture supernatant for extra-cellular
biosynthesis of Se NPs, in this research, we
screened many soil samples and isolated a
fungus which identified as A. terrus. Culture
supernatant of this fungus convert Se4+
ion to
red Se NPs with spherical shape and mean size
diameter of 47 nm. The size of Se NPs which
have been previously prepared in intracellular
spaces of Bacillus Sp. were greater than 100
nm so other alternative microorganisms should
be explored for fabrication of Se NPs with
smaller size (30). This study is the first report
showing the extracellular biosynthesis of small
Se NPs using culture supernatants of A. terrus.
Conclusion
Recently there is an increasing interest to
prepare Se NPs for different medical and
industrial purposes. The synthesis of Se NPs
using a culture supernatant of A. terrus appears
to be simple and an appropriate method for
synthesis of Se NPs with particle size less than
100 nm. Also this approach would be suitable
for developing a biotechnological process for
large-scale production of small Se NPs.
Acknowledgments
This work has been financially supported by
Tehran University of Medical Sciences,
Tehran, Iran.
References
1. Sahoo SK, Parveen S, Panda JJ. The present
and future of nanotechnology in human health
care. Nanomedicine. 2007; 3: 20–31.
2. Doria G, Conde J, Veigas B, Giestas L,
Almeida C, Assuncao M, Rosa J, Baptista PV.
Noble metal nanoparticles for biosensing
applications. Sensors. 2012; 12: 1657-1687.
3. De Gusseme B, Sintubin L, Baert L, Thibo E,
Hennebel T, Vermeulen G, Uyttrendaele M,
Verstraete W, Boon N. Biogenic silver
nanoparticles for disinfection of viral
contaminated water. Commun Agric Appl Biol
Sci. 2011; 76: 73-76.
4. Shaun MB, Thomas DB, James D, Donghui Z,
Seamus C, Farhana SI, Terry JB, Ronald SO.
Formation of Tellurium Nanocrystals during
Anaerobic Growth of Bacteria That Use Te
Oxyanions as Respiratory Electron Acceptors.
Appl Environ Microb. 2007; 73: 2135-2143.
5. Zare B, Faramarzi MA, Sepehrizadeh Z,
Shakibaie M, Rezaie S, Shahverdi AR.
Biosynthesis and recovery of rod-shaped
tellurium nanoparticles and their bactericidal
activities. Mat Res Bull. 2012; 47: 3719–3725.
6. Extracellular synthesis of selenium nanoparticles using fungi
18 Nanomed J, Vol. 1, No. 1, Autumn 2013
6. Duran N, Marcato PD, Duran M, Yadav A,
Gade A, Rai M. Mechanistic aspects in the
biogenic synthesis of extracellular metal
nanoparticles by peptides, bacteria, fungi, and
plants. Appl Microbiol Biotechnol. 2011; 90:
1609-1624.
7. Duran N, Seabra AB. Metallic oxide
nanoparticles: state of the art in biogenic
syntheses and their mechanisms. Appl
Microbiol Biotechnol. 2012; 95: 275-288.
8. Klaus-Joerger T, Joerger R, Olsson E,
Granqvist CG. Bacteria as workers in the living
factory: metal-accumulating bacteria and their
potential for materials science. Trends
Biotechnol. 2001; 19: 15-20.
9. Bahrami K, Nazari P, Sepehrizadeh Z, Zarea
B, Shahverdi AR. Microbial synthesis of
antimony sulfide nanoparticles and their
characterization. Ann Microbiol. 2012; 62:
1419-1425.
10. Mandal D, Bolander M, Mukhopadhyay D,
Sarkar G, Mukherjee P. The use of
microorganisms for the formation of metal
nanoparticles and their application. Appl
Microbiol Biotechnol. 2006; 69: 485-492.
11. Shakibaie M, Shahverdi AR, Faramarzi MA,
Hassanzadeh GR, Rahimi HR, Sabzevari O.
Acute and subacute toxicity of novel biogenic
selenium nanoparticles in mice. Pharm Biol.
2013; 51(1): 58-63
12. Bahrami K, Nazari P, Sepehrizadeh Z, Zarea
B, Shahverdi A. Microbial synthesis of
antimony sulfide nanoparticles and their
characterization. Ann Microbiol. 2012; 62:
1419-1425.
13. Bhattacharya D, G. R. Nanothechnology and
potential of microorganisms. Crit Rev
Biotechnol. 2005; 25: 199-204.
14. Shahverdi A-R, Shakibaie M, Nazari P. Basic
and Practical Procedures for Microbial
Synthesis of Nanoparticles. In: M. Rai, N.
Duran editors. Metal Nanoparticles in
Microbiology. Berlin: Springer;Heidelberg.
2011. 177-195.
15. Kasthuria. j, Veerapandianb. s, N. R.
Biological synthesis of silver and gold
nanoparticles using apiin as reducing agent.
Colloids Surf B. 2009; 68: 55-60.
16. Shahverdi AR, Minaeian S, Shahverdi HR,
Jamalifar H, Nohi AA. Rapid synthesis of
silver nanoparticles using culture supernatants
of Enterobacteria: A novel biological approach.
Process Biochem. 2007; 42: 919-923.
17. Moshfegh M, Forootanfar H, Zare B,
Shahverdi AR, Zarrini G, Faramarzi MA.
Biological synthesis of au, ag and au-ag
bimetallic nanoparticles by α-amylase. Dig J
Nanomater Bios. 2011; 6: 1419-1426.
18. Chen T, Wong Y-S, Zheng W, Bai Y, Huang
L. Selenium nanoparticles fabricated in
Undaria pinnatifida polysaccharide solutions
induce mitochondria-mediated apoptosis in
A375 human melanoma cells. Colloids Surf B.
2008; 67: 26-31.
19. Yazdi MH, Mahdavi M, Varastehmoradi B,
Faramarzi MA, Shahverdi AR. The
Immunostimulatory Effect of Biogenic
Selenium Nanoparticles on the 4T1 Breast
Cancer Model: an In Vivo Study. Biol Trace
Elem Res. 2012; 149(1): 22-28.
20. Zhang JS, Gao XY, Zhang LD, Bao YP.
Biological effects of a nano red elemental
selenium. Biofactors. 2001; 15: 27-38.
21. Prasad KS, Patel H, Patel T, Patel K, Selvaraj
K. Biosynthesis of Se nanoparticles and its
effect on UV-induced DNA damage. Colloids
Surf B. 2013; 103: 261-266.
22. Sandhu GS, Kline BC, Stockman L, Roberts
GD. Molecular probes for diagnosis of fungal
infections. J Clin Microbiol. 1995; 33: 2913-
2919.
23. Yang LB, Shen YH, Xie AJ, Liang JJ, Zhang
BC. Synthesis of Se nanoparticles by using
TSA ion and its photocatalytic application for
decolorization of cango red under UV
irradiation. Mat Res Bull. 2008; 43: 572-582.
24. Thirumurugan D, Dhanaraju MD. Novel
Biogenic Metal Nanoparticles for
Pharmaceutical Applications. Adv Sci Lett.
2011; 4: 339-348.
25. Yang F, Jin C, Subedi S, Lee CL, Wang Q,
Jiang YJ, Li J, Di Y, Fu DL. Emerging
inorganic nanomaterials for pancreatic cancer
diagnosis and treatment. Cancer Treat Rev.
2012; 38: 566-579.
26. Zhang H, Smith JA, Oyanedel-Craver V. The
effect of natural water conditions on the anti-
bacterial performance and stability of silver
nanoparticles capped with different polymers.
Water Res. 2012; 46: 691-699.
27. Wang H, Zhang J, Yu H. Elemental selenium
at nano size possesses lower toxicity without
compromising the fundamental effect on
selenoenzymes: Comparison with
selenomethionine in mice. Free Radical Bio
Med. 2007; 42: 1524–1533.
28. Siddique T, Zhang Y, Okeke BC,
Frankenberger Jr WT. Characterization of
sediment bacteria involved in selenium
reduction. Bioresour Technol. 2006; 97: 1041-
1049.
29. Narayanan KB, Sakthivel N. Biological
synthesis of metal nanoparticles by microbes.
Adv Colloid Interface Sci. 2010; 156: 1-13.
30. Shakibaie M, Khorramizadeh MR, Faramarzi
MA, Sabzevari O, Shahverdi AR. Biosynthesis
7. Zare B, et al.
Nanomed J, Vol. 1, No. 1, Autumn 2013 19
and recovery of selenium nanoparticles and the
effects on matrix metalloproteinas e-2
expression. Biotechnol Appl Biochem. 2010;
56: 7-15.
31. Kumar V, Yadav KS. Plant-mediated synthesis
of silver and gold nanoparticles and their
applications. J Chem Technol Biotechnol.
2009; 84: 151-157.