Purification and Characterization of Recombinant Darbepoetin Alfa from Leishmania tarentolae

Darbepoetin alfa is a biopharmaceutical glycoprotein that stimulates erythropoiesis and is used to treat anemia, which associated with renal failure and cancer chemotherapy. We herein describe the structural characterization of recombinant darbepoetin alfa produced by Leishmania tarentolae T7-TR host. The DNA expression cassette was integrated into the L. tarentolae genome through homologous recombination. Transformed clones were selected by antibiotic resistance, diagnostic PCRs, and protein expression analysis. The structure of recombinant darbepoetin alfa was analyzed by isoelectric focusing, ultraviolet–visible spectrum, and circular dichroism (CD) spectroscopy. Expression analysis showed the presence of a protein band at 40 kDa, and its expression level was 51.2 mg/ml of culture medium. Darbepoetin alfa have 5 isoforms with varying degree of sialylation. The UV absorption and CD spectra were analogous to original drug (Aranesp), which confirmed that the produced protein was darbepoetin alfa. Potency test results revealed that the purified protein was biologically active. In brief, the structural and biological characteristics of expressed darbepoetin alfa were very similar to Aranesp which has been normally expressed in CHO. Our data also suggest that produced protein has potential to be developed for clinical use. © 2016, Springer Science+Business Media New York

Dynamical complexity of short and noisy time series: Compression-Complexity vs. Shannon entropy

Shannon entropy has been extensively used for characteriz- ing complexity of time series arising from chaotic dynamical systems and stochastic processes such as Markov chains. However, for short and noisy time series, Shannon entropy performs poorly. Complexity measures which are based on lossless compression algorithms are a good substitute in such scenarios. We evaluate the performance of two such Compression-Complexity Measures namely Lempel-Ziv complexity(LZ)andEffort-To-Compress( ETC)onshorttimeseriesfrom chaoticdynamicalsystemsinthepresenceofnoise.Both LZ and ETC outperform Shannon entropy (H) in accurately characterizing the dynamical complexity of such systems. For very short binary sequences (which arise in neuroscience applications), ETC has higher number of distinct complexity values than LZ and H, thus enabling a finer resolution. For two-state ergodic Markov chains, we empirically show that ETC converges to a steady state value faster than LZ. Compression-Complexity measures are promising for applications which involve short and noisy time series

Review of research on weed management of chickpea in Iran: challenges, strategies and perspectives

Weeds are one of the most important limiting factors in the production of chickpea (Cicer arietinum) in Iran, especially in autumn sown chickpea. Weed density and biomass in au- tumn chickpea are seven and two and a half times higher than the spring chickpea, respec- tively. The weed damage to chickpea in Tabriz, Kermanshah and West Azerbaijan was esti- mated at 48.3, 57 and 36%, respectively. Sixty-four weed species were identified in chickpea fields. Convolvulus arvensis L. and Galium tricornutum Dandy have the highest presence in chickpea fields. Pyridate and linuron are the only herbicides registered for use in chick- pea fields in Iran. However, research results show that fomesafen and isoxaflutole are the most appropriate herbicides for chickpea fields. Oxyfluorfen, imazethapyr, metribuzin, tri- fluralin, simazine, terbutryn and pendimethalin are the major herbicides studied in weed control research. The combination of herbicides and mechanical control is one of the effec- tive methods to reduce weeds. Hand weeding and cultivation between rows are the most effective mechanical methods of weed control. High nitrogen enhances weed dry weight. Safflower and barley residues reduce weed populations and biomass. Barley-chickpea and wheat-chickpea intercropping systems increase chickpea yield together with proper weed control. In future research, more attention should be paid to surfactants to reduce the use of herbicides, rotation crops and integrated weed management in chickpea

Modified magnetic nanoparticles by PEG-400-immobilized Ag nanoparticles (Fe3O4@PEG–Ag) as a core/shell nanocomposite and evaluation of its antimicrobial activity

Kamiar Zomorodian,1,2 Hamed Veisi,3 Seyed Mahmoud Mousavi,4 Mahmoud Sadeghi Ataabadi,5 Somayeh Yazdanpanah,1,2 Jafar Bagheri,1 Ali Parvizi Mehr,1 Saba Hemmati,3 Hojat Veisi3 1Department of Medical Mycology, Basic Sciences in Infectious Diseases Research Center, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; 2Department of Medical Mycology and Parasitology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; 3Department of Chemistry, Payame Noor University, Tehran, Iran; 4Department of Medical Parasitology, Shiraz University of Medical Sciences, Shiraz, Iran; 5Department of Reproductive Biology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran Background: Noble metal nanoparticles, due to their good physicochemical properties, have been exploited in biological applications. Among these metals, nanosilver has attracted great attention because of its optical properties and broad-spectrum antimicrobial activities with no drug tolerance. Purpose: The present study has attempted to conduct chemical synthesis of Fe3O4@PEG-Ag core/shell nanocomposites in aqueous solutions through co-precipitation of Fe3+ and Fe2+ ions, encapsulating the iron oxide core by poly (ethylene-glycol) (PEG) improve its hydrophilicity and biocompatibility, and immobilizing silver ions by application of NaBH4 as a reducing agent. Patients and methods: The synthesized structures were characterized by Fourier-transform infrared (FT-IR), field emission scanning electron microscopy, energy-dispersive X-ray spectrum, wavelength-dispersive X-ray, vibrating sample magnetometer, inductively coupled plasma-mass spectrometry and transmission electron microscopy methods. Antimicrobial activity of the nanostructures against Staphylococcus aureus, Escherichia coli and Candida albicans was evaluated by broth microdilution based on the methods suggested by Clinical Laboratory Standard Institute. Furthermore, the nanocomposite was tested for possible anti-parasitic effects against Leishmania major promastigotes by MTT assay. Also, its impacts on bacterial cell morphology were defined using atomic force microscopy. Moreover, toxicity of the nanostructure related to animal cell line was determined based on MTT assay. Results: In general, the synthesized core/shell nanostructure can demonstrate noticeable activity against the evaluated representative microorganisms while its toxicity against animal cell line is not considerable. Conclusion: This nanostructure can be applied as a smart drug delivery system with the help of an external magnetic field or it can be used as a powerful antibiotic agent along with other antibiotics that can form a shell on its structure. Keywords: Poly-ethylene-glycol, nanocomposite, AgNPs, antimicrobial propertie

The effect of integrated control methods of broadleaf weeds density on sugar beet yield in kermanshah zone

In order to study the impacts of mechanical (using cultivation), chemical (using herbicides) methods and their integration in controlling broadleaf weeds in sugar beet fields, this experiment was carried out as a randomized complete block design with 4 replications in Mahidasht-Kermanshah in 2006.  In this experiment, 9 different treatments consisting a mixture of post-emergence herbicides phenmedipham + desmedipham +  Ethofumesate and Triflusulfuron Methyl with Sittogate combined with cultivation and 2 check treatments with and without control of the weeds were investigated. The results indicated that cultivation treatment accompanied by Triflusulfuron Methyl reduced dry weight of weeds by 54.91% and showed 100% control of Amaranthus spp. In comparison with control treatment. Cultivation treatment combined with a mixture of phenmedipham+desmedipham+Ethofumesate and Triflusulfuron Methyl with Sittogate increased the sugar beet root dry weight by 159.89% and caused 100% control of the density of Conringia orientalis. Phenmedipham+desmedipham+Ethofumesate plus Triflusulfuron Methyl with sittogate treatment increased root yield by 121.86% compared to the check. In conclusion, the highest root yield was obtained in the cultivation treatment combined with phenmedipham + desmedipham+Ethofumesate, a mixture of phenmedipham + desmedipham + Ethofumesate and Triflusulfuron Methyl with Sittogate and cultivation treatments in sugar beet

Improved high-order bending analysis of double curved sandwich panels subjected to multiple loading conditions

For the first time, the bending analysis of a double curved sandwich panel was presented which was subjected to point load, uniform distributed load on a patch, and harmonic distributed loads and was based on a new improved higher order sandwich panel theory. Since the cross-sectional warping was accurately modeled by this theory, it did not require any shear correction factor. Also, the present analysis incorporated trapezoidal shape factor (the 1+z/R terms) of a curved panel element. Geometry was used for the consideration of different radii curvatures of the face sheets, while the core was unique. Unlike most of other reference works, the core can have non-uniform thickness. The governing equations were derived by the principle of minimum potential energy. The effects of types of boundary conditions, types of applied loads, core to panel, and radii curvatures ratios on the bending response were also studied

Pistacia atlantica leaf extract mediated synthesis of silver nanoparticles and their antioxidant, cytotoxicity, and antibacterial effects under in vitro condition

Recently, researchers have investigated the therapeutical properties of metal nanoparticles especially silver nanoparticles in vitro and in vivo conditions. The aim of the experiment was green synthesis and chemical characterization of silver nanoparticles from aqueous extract of Pistacia atlantica leaf (Ag NPs) and evaluation of their cytotoxicity, antioxidant, and antibacterial effects under in vitro condition. Ag NPs were spherical with a size range of 40-60 nm and characterized using various analysis techniques including UV-Vis absorption spectroscopy to determine the presence of Ag NP in the solution. We studied functional groups of Pistacia atlantica extract in the reduction and capping process of Ag NP by FT-IR, crystallinity and FCC planes by XRD pattern, elemental analysis of the sample by EDS, and surface morphology, shapes, and size of Ag NPs by SEM, AFM, and TEM. Destroy initiation and termination temperatures of the Ag NPs were determined by TGA. DPPH free radical scavenging test was done to evaluate the antioxidant potentials, which indicated similar antioxidant potentials for Ag NPs and butylated hydroxytoluene. The synthesized Ag NPs had great cell viability dose-dependently and indicated this method was nontoxic. Agar diffusion tests were done to determine the antibacterial characteristic. Ag NPs revealed similar antibacterial property to the standard antibiotic. Also, Ag NPs prevented the growth of all bacteria at 1-7 mu g/ml concentrations and removed them at 3-15 mu g/ml concentrations. Finally, synthesized Ag NPs revealed non-cytotoxicity, antioxidant and antibacterial activities in a dose-depended manner

Production and Characterization of Single Chain Nimotuzumab: An In Vitro Study

Recombinant expression and EGFR-binding activity assay of single chain nimotuzumab (nimotuzumab scFv) is reported in this study. The scFv was produced in VH-linker-VL format in Origami™ 2(DE3)pLysS bacterial cells and purified using Ni-TNA resin column. 3-D structure prediction using I-TASSER (Iterative Threading Assembly Refinement) server and analyzing the predicted models using YASARA (Yet Another Scientific Artificial Reality Application) viewer revealed that VH and VL domains assemble into a correctly-folded single chain antibody that is able to bind EGFR. The scFv was evaluated in ELISA and western blot tests and proven to be able to bind EGFR-overexpressing cancer cells (A-431 cells in an efficient manner but unable to recognize cancer cells expressing low levels of EGFR (MCF-7 breast cancer cells). © 2017, Springer Science+Business Media New York

Green synthesis of silver nanoparticles using aqueous extract of Stachys lavandulifolia flower, and their cytotoxicity, antioxidant, antibacterial and cutaneous wound-healing properties

In a biological process where the herbal tea (Stachys lavandulifolia) aqueous extract was applied as a capping and reducing agent, nanoparticles (NPs) of silver (Ag) were synthesized. These AgNPs were characterized using Fourier transform-infrared spectroscopy, field emission-scanning electron microscopy, energy-dispersive X-ray spectroscopy, transmission electron microscopy and ultraviolet-visible spectroscopy. The synthesized AgNPs had great cell viability dose-dependently investigating the effect of the plant on human umbilical vein endothelial cell line and indicated this method was non-toxic. In this study, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging test was carried out to examine antioxidant properties, which revealed similar antioxidant properties for AgNPs and butylated hydroxytoluene. Agar diffusion tests were applied to determine the antibacterial characteristics. The macro-broth tube test was run to determine minimum inhibitory concentration. All data of antibacterial and cutaneous wound-healing examinations were analyzed by SPSS 21 software (Duncan post hoc test). AgNPs showed higher antibacterial property than all standard antibiotics (p <= 0.01). Also, AgNPs prevented the growth of all bacteria at 2-8 mg/ml concentrations and destroyed them at 2-16 mg/ml concentrations (p <= 0.01). For the in vivo experiment, after creating the cutaneous wound, the rats were randomly divided into six groups: untreated control; treatment with Eucerin basal ointment; treatment with 3% tetracycline ointment; treatment with 0.2% AgNO3 ointment; treatment with 0.2% S. lavandulifolia ointment; and treatment with 0.2% AgNPs ointment. These groups were treated for 10 days. For histopathological and biochemical analysis of the healing trend, a 3 x 3-cm section was prepared from all dermal thicknesses at day 10. Use of AgNPs ointment in the treatment groups substantially reduced (p <= 0.01) the wound area, total cells, neutrophil, macrophage and lymphocyte, and remarkably raised (p <= 0.01) the wound contracture, hydroxyl proline, hexosamine, hexuronic acid, fibrocyte and fibrocytes/fibroblast rate compared with other groups. Seemingly, AgNPs can be used as a medical supplement owing to their non-cytotoxic, antioxidant, antibacterial and cutaneous wound-healing properties