Search results for: two-photon fluorescence microscopy

Authors: E. Kilicay, Z. Karahaliloglu, B. Hazer, E. B. Denkbas

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In this study, we have prepared concanavaline A (ConA) functionalized curcumin (CUR) loaded PHBHHx (poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)) nanoparticles as a novel and efficient drug delivery system. CUR is a promising anticancer agent for various cancer types. The aim of this study was to evaluate therapeutic potential of curcumin loaded PHBHHx nanoparticles (CUR-NPs) and concanavaline A conjugated curcumin loaded NPs (ConA-CUR NPs) for ovarian cancer treatment. ConA was covalently connected to the carboxylic group of nanoparticles by EDC/NHS activation method. In the ligand attachment experiment, the binding capacity of ConA on the surface of NPs was found about 90%. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) analysis showed that the prepared nanoparticles were smooth and spherical in shape. The size and zeta potential of prepared NPs were about 228±5 nm and −21.3 mV respectively. ConA-CUR NPs were characterized by FT-IR spectroscopy which confirmed the existence of CUR and ConA in the nanoparticles. The entrapment and loading efficiencies of different polymer/drug weight ratios, 1/0.125 PHBHHx/CUR= 1.25CUR-NPs; 1/0.25 PHBHHx/CUR= 2.5CUR-NPs; 1/0.5 PHBHHx/CUR= 5CUR-NPs, ConA-1.25CUR NPs, ConA-2.5CUR NPs and ConA-5CUR NPs were found to be ≈ 68%-16.8%; 55%-17.7 %; 45%-33.6%; 70%-15.7%; 60%-17%; 51%-30.2% respectively. In vitro drug release showed that the sustained release of curcumin was observed from CUR-NPs and ConA-CUR NPs over a period of 19 days. After binding of ConA, the release rate was slightly increased due to the migration of curcumin to the surface of the nanoparticles and the matrix integrities was decreased because of the conjugation reaction. This functionalized nanoparticles demonstrated high drug loading capacity, sustained drug release profile, and high and long term anticancer efficacy in human cancer cell lines. Anticancer activity of ConA-CUR NPs was proved by MTT assay and reconfirmed by apoptosis and necrosis assay. The anticancer activity of ConA-CUR NPs was measured in ovarian cancer cells (SKOV-3) and the results revealed that the ConA-CUR NPs had better tumor cells decline activity than free curcumin. The nacked nanoparticles have no cytotoxicity against human ovarian carcinoma cells. Thus the developed functionalized nanoformulation could be a promising candidate in cancer therapy.

Keywords: curcumin, curcumin-PHBHHx nanoparticles, concanavalin A, concanavalin A-curcumin PHBHHx nanoparticles, PHBHHx nanoparticles, ovarian cancer cell

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Authors: Satoshi Nishimura

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Although much information has been garnered from the genomes of humans and mice, it remains difficult to extend that information to explain physiological and pathological phenomena. This is because the processes underlying life are by nature stochastic and fluctuate with time. Thus, we developed novel "in vivo molecular imaging" method based on single and two-photon microscopy. We visualized and analyzed many life phenomena, including common adult diseases. We integrated the knowledge obtained, and established new models that will serve as the basis for new minimally invasive therapeutic approaches.

Keywords: two photon microscope, intravital visualization, thrombus, artery

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Authors: Guna Raj Dhungana, Roshan Nepal, Apshara Parajuli, , Archana Maharjan, Shyam K. Mishra, Pramod Aryal, Rajani Malla

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Introduction: Klebsiella pneumoniae is a well-known opportunistic human pathogen, primarily causing healthcare-associated infections. The global emergence of carbapenemase-producing K. pneumoniaeis a major public health burden, which is often extensively multidrug resistant.Thus, because of the difficulty to treat these ‘superbug’ and menace and some term as ‘apocalypse’ of post antibiotics era, an alternative approach to controlling this pathogen is prudent and one of the approaches is phage mediated control and/or treatment. Objective: In this study, we aimed to isolate novel bacteriophage against carbapenemase-producing K. pneumoniaeand characterize for potential use inphage therapy. Material and Methods: Twenty lytic phages were isolated from river water using double layer agar assay and purified. Biological features, physiochemical characters, burst size, host specificity and activity spectrum of phages were determined. One most potent phage: Phage TU_Kle10O was selected and characterized by electron microscopy. Whole genome sequences of the phage were analyzed for presence/absence of virulent factors, and other lysin genes. Results: Novel phage TU_Kle10O showed multiple host range within own genus and did not induce any BIM up to 5th generation of host’s life cycle. Electron microscopy confirmed that the phage was tailed and belonged to Caudovirales family. Next generation sequencing revealed its genome to be 166.2 Kb. bioinformatical analysis further confirmed that the phage genome ‘did not’ contain any ‘bacterial genes’ within phage genome, which ruled out the concern for transfer of virulent genes. Specific 'lysin’ enzyme was identified phages which could be used as 'antibiotics'. Conclusion: Extensively multidrug resistant bacteria like carbapenemase-producing K. pneumoniaecould be treated efficiently by phages.Absence of ‘virulent’ genes of bacterial origin and presence of lysin proteins within phage genome makes phages an excellent candidate for therapeutics.

Keywords: bacteriophage, Klebsiella pneumoniae, MDR, phage therapy, carbapenemase,

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Authors: Afzal Hussain, Mohammad A. Altamimi, Syed Sarim Imam, Mudassar Shahid, Osamah Abdulrahman Alnemer

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Exhaustive application of sulfamethoxazole (SUX) became as a global threat for human health due to water contamination through diverse sources. The addressed combined application of Hansen solubility (HSPiP software) parameters and Quality by Design tool for developing various green nanoemulsions. HSPiP program assisted to screen suitable excipients based on Hansen solubility parameters and experimental solubility data. Various green nanoemulsions were prepared and characterized for globular size, size distribution, zeta potential, and removal efficiency. Design Expert (DoE) software further helped to identify critical factors responsible to have direct impact on percent removal efficiency, size, and viscosity. Morphological investigation was visualized under transmission electron microscopy (TEM). Finally, the treated was studied to negate the presence of the tested drug employing ICP-OES (inductively coupled plasma optical emission microscopy) technique and HPLC (high performance liquid chromatography). Results showed that HSPiP predicted biocompatible lipid, safe surfactant (lecithin), and propylene glycol (PG). Experimental solubility of the drug in the predicted excipients were quite convincing and vindicated. Various green nanoemulsions were fabricated, and these were evaluated for in vitro findings. Globular size (100-300 nm), PDI (0.1-0.5), zeta potential (~ 25 mV), and removal efficiency (%RE = 70-98%) were found to be in acceptable range for deciding input factors with level in DoE. Experimental design tool assisted to identify the most critical variables controlling %RE and optimized content of nanoemulsion under set constraints. Dispersion time was varied from 5-30 min. Finally, ICP-OES and HPLC techniques corroborated the absence of SUX in the treated water. Thus, the strategy is simple, economic, selective, and efficient.

Keywords: quality by design, sulfamethoxazole, green nanoemulsion, water treatment, icp-oes, hansen program (hspip software

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Authors: Amanda C. Q. M. Vieira, Cybelly M. Melo, Camila B. M. Figueirêdo, Giovanna C. R. M. Schver, Salvana P. M. Costa, Magaly A. M. de Lyra, Ping I. Lee, José L. Soares-Sobrinho, Pedro J. Rolim-Neto, Mônica F. R. Soares

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Epiisopiloturine (EPI) is a drug candidate that is extracted from Pilocarpus microphyllus and isolated from the waste of Pilocarpine. EPI has demonstrated promising schistosomicidal, leishmanicide, anti-inflammatory and antinociceptive activities, according to in vitro studies that have been carried out since 2009. However, this molecule shows poor aqueous solubility, which represents a problem for the release of the drug candidate and its absorption by the organism. The purpose of the present study is to investigate the extent of enhancement of kinetic solubility of a solid solution (SS) of EPI in hipromellose phthalate HP-55 (HPMCP), an enteric polymer carrier. SS was obtained by the solvent evaporation methodology, using acetone/methanol (60:40) as solvent system. Both EPI and polymer (drug loading 10%) were dissolved in this solvent until a clear solution was obtained, and then dried in oven at 60ºC during 12 hours, followed by drying in a vacuum oven for 4 h. The results show a considerable modification in the crystalline structure of the drug candidate. For instance, X-ray diffraction (XRD) shows a crystalline behavior for the EPI, which becomes amorphous for the SS. Polarized light microscopy, a more sensitive technique than XRD, also shows completely absence of crystals in SS sample. Differential Scanning Calorimetric (DSC) curves show no signal of EPI melting point in SS curve, indicating, once more, no presence of crystal in this system. Interaction between the drug candidate and the polymer were found in Infrared microscopy, which shows a carbonyl 43.3 cm-1 band shift, indicating a moderate-strong interaction between them, probably one of the reasons to the SS formation. Under sink conditions (pH 6.8), EPI SS had its dissolution performance increased in 2.8 times when compared with the isolated drug candidate. EPI SS sample provided a release of more than 95% of the drug candidate in 15 min, whereas only 45% of EPI (alone) could be dissolved in 15 min and 70% in 90 min. Thus, HPMCP demonstrates to have a good potential to enhance the kinetic solubility profile of EPI. Future studies to evaluate the stability of SS are required to conclude the benefits of this system.

Keywords: epiisopiloturine, hipromellose phthalate HP-55, pharmaceuticaltechnology, solubility

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Authors: M. Areekijseree, W. Pongsawat, M. Pumipaiboon, C. Thepsithar, S. Sengsai, T. Chuen-Im

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Morphological interaction of porcine cumulus-oocyte complexes (pCOCs) was investigated on in vitro condition using electron microscope (SEM and TEM). The totals of 1,923 oocytes were round in shape, surrounded by zona pellucida with layer of cumulus cells ranging between 59.29-202.14 µm in size. They were classified into intact-, multi-, partial cumulus cell layer oocyte, and completely denuded oocyte, at the percentage composition of 22.80% 32.70%, 18.60%, and 25.90 % respectively. The pCOCs classified as intact- and multi cumulus cell layer oocytes were further culturing at 37°C with 5% CO2, 95% air atmosphere and high humidity for 44 h in M199 with Earle’s salts supplemented with 10% HTFCS, 2.2 mg/mL NaHCO3, 1 M Hepes, 0.25 mM pyruvate, 15 µg/mL porcine follicle-stimulating hormone, 1 µg/mL LH, 1µg/mL estradiol with ethanol, and 50 µg/mL gentamycin sulfate. On electron microscope study, cumulus cells were found to stick their processes to secrete substance from the sac-shape end into zona pellucida of the oocyte and also communicated with the neighboring cells through their microvilli on the beginning of incubation period. It is believed that the cumulus cells communicate with the oocyte by inserting the microvilli through this gap and embedded in the oocyte cytoplasm before secreting substance, through the sac-shape end of the microvilli, to inhibit primary oocyte development at the prophase I. Morphological changes of the complexes were observed after culturing for 24-44 h. One hundred percentages of the cumulus layers were expanded and cumulus cells were peeling off from the oocyte surface. In addition, the round-shape cumulus cells transformed themselves into either an elongate shape or a columnar shape, and no communication between cumulus neighboring cells. After 44 h of incubation time, diameter of oocytes surrounded by cumulus cells was larger than 0 h incubation. The effect of hormones in culture medium is exerted by their receptors present in porcine oocyte. It is likely that all morphological changes of the complexes after hormone treatment were to allow maturation of the oocyte. This study demonstrated that the association of hormones in M199 could promote porcine follicle activation in 44 h in vitro condition. This culture system should be useful for studying the regulation of early follicular growth and development, especially because these follicles represent a large source of oocytes that could be used in vitro for cell technology.

Keywords: cumulus cells, electron microscopy, in vitro, porcine oocyte

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Authors: A. Ojha, Y. K. Gupta

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Organophosphate (OP) pesticides are among the most widely used synthetic chemicals for controlling a wide variety of pests throughout the world. Chlorpyrifos (CPF), methyl parathion (MPT), and malathion (MLT) are among the most extensively used OP pesticides in India. DNA strand breaks and DNA-protein crosslinks (DPC) are toxic lesions associated with the mechanisms of toxicity of genotoxic compounds. In the present study, we have examined the potential of CPF, MPT, and MLT individually and in combination, to cause DNA strand breakage and DPC formation. Peripheral blood lymphocytes of rat were exposed to 1/4 and 1/10 LC50 dose of CPF, MPT, and MLT for 2, 4, 8, and 12h. The DNA strand break was measured by the comet assay and expressed as DNA damage index while DPC estimation was done by fluorescence emission. There was significantly marked increase in DNA damage and DNA-protein crosslink formation in time and dose dependent manner. It was also observed that MPT caused the highest level of DNA damage as compared to other studied OP compounds. Thus, from present study, we can conclude that studied pesticides have genotoxic potential. The pesticides mixture does not potentiate the toxicity of each other. Nonetheless, additional in vivo data are required before a definitive conclusion can be drawn regarding hazard prediction to humans.

Keywords: organophosphate, pesticides, DNA damage, DNA protein crosslink, genotoxic

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Authors: P. Slepicka, N. Slepickova Kasalkova, I. Michaljanicova, O. Nedela, Z. Kolska, V. Svorcik

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Polymers exposed to laser or plasma treatment or modified with different wet methods which enable the introduction of nanoparticles or biologically active species, such as amino-acids, may find many applications both as biocompatible or anti-bacterial materials or on the contrary, can be applied for a decrease in the number of cells on the treated surface which opens application in single cell units. For the experiments, two types of materials were chosen, a representative of non-biodegradable polymers, polyethersulphone (PES) and polyhydroxybutyrate (PHB) as biodegradable material. Exposure of solid substrate to laser well below the ablation threshold can lead to formation of various surface structures. The ripples have a period roughly comparable to the wavelength of the incident laser radiation, and their dimensions depend on many factors, such as chemical composition of the polymer substrate, laser wavelength and the angle of incidence. On the contrary, biopolymers may significantly change their surface roughness and thus influence cell compatibility. The focus was on the surface treatment of PES and PHB by pulse excimer KrF laser with wavelength of 248 nm. The changes of physicochemical properties, surface morphology, surface chemistry and ablation of exposed polymers were studied both for PES and PHB. Several analytical methods involving atomic force microscopy, gravimetry, scanning electron microscopy and others were used for the analysis of the treated surface. It was found that the combination of certain input parameters leads not only to the formation of optimal narrow pattern, but to the combination of a ripple and a wrinkle-like structure, which could be an optimal candidate for cell attachment. The interaction of different types of cells and their interactions with the laser exposed surface were studied. It was found that laser treatment contributes as a major factor for wettability/contact angle change. The combination of optimal laser energy and pulse number was used for the construction of a surface with an anti-cellular response. Due to the simple laser treatment, we were able to prepare a biopolymer surface with higher roughness and thus significantly influence the area of growth of different types of cells (U-2 OS cells).

Keywords: cell response, excimer laser, polymer treatment, periodic pattern, surface morphology

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Authors: Evdokia K. Oikonomou, Nikolay Christov, Galder Cristobal, Graziana Messina, Giovani Marletta, Laurent Heux, Jean-Francois Berret

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Fabric softeners are aqueous formulations that contain ~10 wt. % double tailed cationic surfactants. Here, a formulation in which 50% surfactant was replaced with low quantities of natural guar polymers was developed. Thanks to the reduced surfactant quantity this product has less environmental impact while the guars presence was found to maintain the product’s performance. The objective of this work is to elucidate the effect of the guar polymers on the softener deposition and the adsorption mechanism on the cotton surface. The surfactants in these formulations are assembled into large distributed (0.1 – 1 µm) vesicles that are stable in the presence of guars and upon dilution. The effect of guars on the vesicles adsorption on cotton was first estimated by using cellulose nanocrystals (CNC) as a stand-in for cotton. The dispersion of CNC in water permits to follow the interaction between the vesicles, guars, and CNC in the bulk. It was found that guars enhance the deposition on CNC and that the vesicles are deposited intactly on the fibers driven by electrostatics. The mechanism of the vesicles/guars adsorption on cellulose fibers was identified by quartz crystal microbalance with dissipation monitoring. It was found that the guars increase the surfactant deposited quantity, in agreement with the results in the bulk. Also, the structure of the adsorbed surfactant on the fibers' surfaces (vesicle or bilayer) was influenced by the guars presence. Deposition studies on cotton fabrics were also conducted. Attenuated total reflection and scanning electron microscopy were used to study the effect of the polymers on this deposition. Finally, fluorescent microscopy was used to follow the adsorption of surfactant vesicles, labeled with a fluorescent dye, on cotton fabrics in water. It was found that, in the presence or not of polymers, the surfactant vesicles are adsorbed on fiber maintaining their vesicular structure in water (supported vesicular bilayer structure). The guars influence this process. However, upon drying the vesicles are transformed into bilayers and eventually wrap the fibers (supported lipid bilayer structure). This mechanism is proposed for the adsorption of vesicular conditioner on cotton fiber and can be affected by the presence of polymers.

Keywords: cellulose nanocrystals, cotton fibers, fabric softeners, guar polymers, surfactant vesicles

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Authors: Zhaojun Zheng, Yuanfa Liu, Jiaxin Li, Jinwei Li, Yong-jiang Xu, Chen Cao

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Black bean is an excellent protein source for preparing hydrolysates, which attract much attention due to their biological activity. The objective of this study was to characterize the physicochemical and antioxidant properties of black bean protein, hydrolyzed by ficin, bromelain or alcalase until 300 min of hydrolysis. Results showed that bromelain and alcalase hydrolysates possessed a higher degree of hydrolysis (DH) than that of ficin, thereby presenting different ultraviolet absorption, fluorescence intensity, and circular dichroism. Moreover, all hydrolysates possessed the capacity to scavenge DPPH radical with the lowest IC₅₀ of 21.11 µg/mL, as well as to chelate ferrous ion (Fe²⁺) with the IC₅₀ values ranging from 6.82 to 30.68 µg/mL. Intriguingly, the oxidation of linoleic acid, sunflower oil, and sunflower oil-in-water emulsion was remarkedly retarded by the three selected protein hydrolysates, especially by bromelain-treated protein hydrolysate, which might attribute to their high hydrophobicity and emulsifying properties. These findings can provide strong support for black bean protein hydrolysates to be employed in food products acting as natural antioxidant alternatives.

Keywords: antioxidant activity, black bean protein hydrolysate, emulsion physicochemical properties, sunflower oil

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Authors: Eve S. A. Loiseau, Marion Frey, Yves Blickenstorfer, Fabian Niedermair, André R. Studart

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Porous microcapsules have a broad range of applications that require a robust shell. We propose a new method to produce macroporous polymer capsules with controlled size, shell thickness, porosity and mechanical properties using co-flow flow-focusing glass capillary devices. The porous structure was investigated through SEM and the permeability through confocal microscopy. Compression tests on single capsules were performed. We obtained microcapsules with tailored permeability from open to close pores structures and able to withstand loads up to 150 g.

Keywords: microcapsules, micromechanics, porosity, polymer shells

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Authors: Geeta Sharma, Suma Chandra

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Sugarcane is one of the major commercial crop playing roles in agriculture and industrial economy of India. Globally sugarcane is affected by approximately 240 diseases caused by various plant pathogenic organisms. Among them, red rot disease caused by the fungus Colletotrichum falcatum, is one of the most important diseases. In the present investigation, one fungal bioagent of Trichoderma harzianum, Pant Bioagent 1 and one bacterial bioagent Pseudomonas fluorescence, Pant Bioagent 2 (PBAT 1 and PBAT 2, respectively) were tested by dual culture method against the pathogen under laboratory conditions. The effectiveness of biocontrol agents was observed against four isolates of C. falcatum. In the case of PBAT1 maximum percent inhibition of pathogen was recorded in isolated Cf 0238 (61.05%), followed by Cf 09 (60.62%) whereas, minimum percent inhibition was recorded in Cf 3220 (48.55%) and in case of PBAT2 maximum mycelial growth inhibition percent was recorded in Cf 767 (50.50%) followed by Cf 088230(48.83%), whereas minimum percent inhibition was recorded in Cf 08 (40.16%) followed by Cf 0238 (41.83%). The present study showed that these biocontrol agents have the potential of controlling the pathogen and can further be used for the management of red rot disease in field.

Keywords: biocontrol agents, Colletotrichum falcatum, isolates, sugarcane

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Authors: Samiul Islam Hazarika, Domngam Boje, Ananta Kumar Atta

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It is well familiar that solvents play a significant role in modern chemistry. Solvents can change the reactivity and physicochemical properties of molecules in a solution. Keeping this in mind, we have designed and synthesized a mono-triazolyl-linked pyrenyl-appended xylofuranose derivative for the detection of metal ions with changing solvent systems. The incorporation of a sugar backbone in the sensor increases the water solubility and biocompatibility. The experimental study revealed that the xylofuranose-based fluorescence probe did not exhibit any specific selectivity towards metal ions in acetonitrile (CH₃CN) solvent. Whereas, we revealed that triazole-linked pyrenyl-appended xylofuranose-based fluorescent sensor would exhibit high selectivity and sensitivity towards Au³⁺ ions in CH₃CN-H₂O (1/1, v/v) system. This observation might be explained by the viscosity and polarity differences of CH₃CN and CH₃CN-H₂O solvent systems. The formation of the sensor-Au³⁺ complex was also established by high-resolution mass spectrometry (HRMS) data of the complex.

Keywords: triazole, furanose, fluorometric, solvent dependent

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Authors: Rahim Sabbaghizadeh, Mansor Hashim, Nooshin Shourcheh

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Nanocrystalline Nd8Pr2Fe79-xCo5B6Alx (x=0, 1, 2, 3) magnets were prepared by mechanical alloying and respective heat treatment, and the effects of the addition of Al on the microstructure and magnetic properties of Nd-Fe-Co-B alloy were studied. The changes in the nanostructure and magnetic properties were examined by X-Ray diffraction, combined with Field Emission Scanning electron microscopy (FeSEM) and vibrating sample magnetometer (VSM). Addition of Al was found to be effective for improving the coercivity and the hysteresis squareness in Nd–Fe–Co–B magnets without decreasing much the remanent magnetization.

Keywords: mechanical alloying, nanocrystalline, Nd-Fe-B, vibrating sample magnetomete

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Authors: S. K. M. Nzaba, H. H. Nyoni, B. Ntsendwana, B. B. Mamba, A. T. Kuvarega

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Azo dye effluents, released into water bodies are not only toxic to the ecosystem but also pose a serious impact on human health due to the carcinogenic and mutagenic effects of the compounds present in the dye discharge. Conventional water treatment methods such as adsorption, flocculation/coagulation and biological processes are not effective in completely removing most of the dyes and their natural degradation by-products. Advanced oxidation processes (AOPs) have proven to be effective technologies for complete mineralization of these recalcitrant pollutants. Therefore, there is a need for new technology that can solve the problem. Thus, this study examined the photocatalytic degradation of an azo dye brilliant black (BB) using non-metal/metal codoped TiO₂. N, Pt co-doped TiO₂ photocatalysts were prepared by a modified sol-gel method using amine-terminated polyamidoamine dendrimer generation 0 (PAMAM G0), amine-terminated polyamidoamine dendrimer generation 1 ( PAMAM G1) and hyperbranched polyethyleneimine (HPEI) as templates and source of nitrogen. Structural, morphological, and textural properties were evaluated using scanning electron microscopy coupled to energy dispersive X-ray spectroscopy (SEM/EDX), high-resolution transmission electron microscopy (HRTEM), X-ray diffraction spectroscopy (XRD), X-ray photoelectron spectroscopy (XPS), thermal gravimetric analysis (TGA), Fourier- transform infrared (FTIR), Raman spectroscopy (RS), photoluminescence (PL) and ultra-violet /visible spectroscopy (UV-Vis). The synthesized photocatalysts exhibited lower band gap energies as compared to the Degussa P-25 revealing a red shift in band gap towards the visible light absorption region. Photocatalytic activity of N, Pt co-doped TiO₂ was measured by the reaction of photocatalytic degradation of brilliant black (BB) dye. The N, metal codoped TiO₂ containing 0.5 wt. % of the metal consisted mainly of the anatase phase as confirmed by XRD results of all three samples, with a particle size range of 13–30 nm. The particles were largely spherical and shifted the absorption edge well into the visible region. Band gap reduction was more pronounced for the N, Pt HPEI (Pt 0.5 wt. %) codoped TiO₂ compared to PAMAM G0 and PAMAM G1. Consequently, codoping led to an enhancement in the photocatalytic activity of the materials for the degradation of brilliant black (BB).

Keywords: codoped TiO₂, dendrimer, photodegradation, wastewater

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Authors: Luan Trong Nguyen, M. Azizur Rahman, Yusuke Tamenori, Toshihiro Yoshimura, Nozomu Iwasaki, Hiroshi Hasegawa

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This study investigated the distribution of magnesium (Mg), phosphorus (P), sulfur (S) and strontium (Sr) using micro X-ray fluorescence (µ-XRF) along the annual growth rings in the skeleton of Japanese red coral Paracorallium japonicum. The Mg, P and S distribution in µ-XRF mapping images correspond to the dark and light bands along the annual growth rings observed in microscopic images of the coral skeleton. The µ-XRF mapping data showed a positive correlation (r = 0.6) between P and S distribution in the coral skeleton. A contrasting distribution pattern of S and Mg along the axial skeleton of P. japonicum indicates a weak negative correlation (r = -0.2) between these two trace elements. The distribution pattern of S, P and Mg reveals linkage between their distributions and the formation of dark/light bands along the annual growth rings in the axial skeleton of P. japonicum. Sulfur and P were distributed in the organic matrix rich dark bands, while Mg was distributed in the light bands of the annual growth rings.

Keywords: µ-XRF, trace element, precious coral, Paracorallium japonicum

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Authors: Parvathy Anitha, Nilesh J. Vasa, M. S. Ramachandra Rao

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ZnO is a semiconducting material with a direct wide band gap of 3.37 eV and a large exciton binding energy of 60 meV at room temperature. Microspheres with high sphericity and symmetry exhibit unique functionalities which makes them excellent omnidirectional optical resonators. Hence there is an advent interest in fabrication of single crystalline semiconductor microspheres especially magnetic ZnO microspheres, as ZnO is a promising material for semiconductor device applications. Also, ZnO is non-toxic and biocompatible, implying it is a potential material for biomedical applications. Room temperature Photoluminescence (PL) spectra of the fabricated ZnO microspheres were measured, at an excitation wavelength of 325 nm. The ultraviolet (UV) luminescence observed is attributed to the room-temperature free exciton related near-band-edge (NBE) emission in ZnO. Besides the NBE luminescence, weak and broad visible luminescence (~560nm) was also observed. This broad emission band in the visible range is associated with oxygen vacancies related to structural defects. In transition metal (TM) ion-doped ZnO, 3d levels emissions of TM ions will modify the inherent characteristic emissions of ZnO. A micron-sized ZnO crystal has generally a wurtzite structure with a natural hexagonal cross section, which will serve as a WGM (whispering gallery mode) lasing micro cavity due to its high refractive index (~2.2). But hexagonal cavities suffers more optical loss at their corners in comparison to spherical structures; hence spheres may be a better candidate to achieve effective light confinement. In our study, highly smooth spherical shaped micro particles with different diameters ranging from ~4 to 6 μm were grown on different substrates. SEM (Scanning Electron Microscopy) and AFM (Atomic Force Microscopy) images show the presence of uniform smooth surfaced spheres. Raman scattering measurements from the fabricated samples at 488 nm light excitation provide convincing supports for the wurtzite structure of the prepared ZnO microspheres. WGM lasing studies from TM-doped ZnO microparticles are in progress.

Keywords: laser ablation, microcavity, photoluminescence, ZnO microsphere

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Authors: Varun Kumar, Chandra Shakher

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Micro-optical components such as microlenses and microlens array have numerous engineering and industrial applications for collimation of laser diodes, imaging devices for sensor system (CCD/CMOS, document copier machines etc.), for making beam homogeneous for high power lasers, a critical component in Shack-Hartmann sensor, fiber optic coupling and optical switching in communication technology. Also micro-optical components have become an alternative for applications where miniaturization, reduction of alignment and packaging cost are necessary. The compliance with high-quality standards in the manufacturing of micro-optical components is a precondition to be compatible on worldwide markets. Therefore, high demands are put on quality assurance. For quality assurance of these lenses, an economical measurement technique is needed. For cost and time reason, technique should be fast, simple (for production reason), and robust with high resolution. The technique should provide non contact, non-invasive and full field information about the shape of micro- optical component under test. The interferometric techniques are noncontact type and non invasive and provide full field information about the shape of the optical components. The conventional interferometric technique such as holographic interferometry or Mach-Zehnder interferometry is available for characterization of micro-lenses. However, these techniques need more experimental efforts and are also time consuming. Digital holography (DH) overcomes the above described problems. Digital holographic microscopy (DHM) allows one to extract both the amplitude and phase information of a wavefront transmitted through the transparent object (microlens or microlens array) from a single recorded digital hologram by using numerical methods. Also one can reconstruct the complex object wavefront at different depths due to numerical reconstruction. Digital holography provides axial resolution in nanometer range while lateral resolution is limited by diffraction and the size of the sensor. In this paper, Mach-Zehnder based digital holographic interferometric microscope (DHIM) system is used for the testing of transparent microlenses. The advantage of using the DHIM is that the distortions due to aberrations in the optical system are avoided by the interferometric comparison of reconstructed phase with and without the object (microlens array). In the experiment, first a digital hologram is recorded in the absence of sample (microlens array) as a reference hologram. Second hologram is recorded in the presence of microlens array. The presence of transparent microlens array will induce a phase change in the transmitted laser light. Complex amplitude of object wavefront in presence and absence of microlens array is reconstructed by using Fresnel reconstruction method. From the reconstructed complex amplitude, one can evaluate the phase of object wave in presence and absence of microlens array. Phase difference between the two states of object wave will provide the information about the optical path length change due to the shape of the microlens. By the knowledge of the value of the refractive index of microlens array material and air, the surface profile of microlens array is evaluated. The Sag of microlens and radius of curvature of microlens are evaluated and reported. The sag of microlens agrees well within the experimental limit as provided in the specification by the manufacturer.

Keywords: micro-optics, microlens array, phase map, digital holographic interferometric microscopy

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Authors: Abdelghani Boucheham, Ahcene Karaali, Amar Manseri

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Nanostructured Titania layers formed on the surface of titanium and titanium alloys by anodic oxidation play an important role in the enhancement of their biocompatibility and osseointegration in the human body. In the current work, highly ordered titania nanotube array films were elaborated on Ti₆Al₄V medical grade alloys in organic electrolyte containing ethylene glycol, 0.2 wt. % NH₄F and 4 vol. % H₂O at an applied potential of 60 V for different durations. The diameters, lengths and wall thicknesses of the obtained nanotubes were characterized by scanning electronic microscopy (SEM).

Keywords: anodization, dental implants, titania nanotubes, titanium alloys, SEM

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Authors: Mechri hanane, Azzaz Mohammed

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Polycrystalline CuFe thin film was prepared by thermal evaporation process (Physical vapor deposition), using the nanocrystalline CuFe powder obtained by mechanical alloying After 24 h of milling elemental powders. The microscopic study of nanocrystalline powder and the thin film of Cu70Fe30 binary alloy were examined using transmission electron microscopy (TEM) and scanning electron microscope (SEM). The cross-sectional TEM images showed that the obtained CuFe layer was polycrystalline film of about 20 nm thick and composed of grains of different size ranging from 4 nm to 18 nm.

Keywords: nanomaterials, thin films, TEM, SEM

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Authors: Lu-Chen Yeh‚ Jui-Ming Yeh

Abstract:

In this manuscript, we produced neat electrospun poly(o-methoxyaniline) (POMA) fibers and utilized it for applying the growth of neural stem cells. The transparency and morphology of as-prepared POMA fibers were characterized by UV-visible spectroscopy and scanning electron microscopy, respectively. It was found to have no adverse effects on the long-term proliferation of the neural stem cells (NSCs), retained the ability to self-renew, and exhibit multi-potentiality. Results of immunofluorescence staining studies confirmed that POMA electrospun fibers could provide a great environment for NSCs and enhance its differentiation.

Keywords: electrospun, polyaniline, neural stem cell, differentiation

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Authors: Zeynep Busra Velioglu, Deniz Pulat, Beril Demirbakan, Burak Ozcan, Ece Bayrak, Cevat Erisken

Abstract:

Bone tissue has the ability to perform a wide array of functions including providing posture, load-bearing capacity, protection for the internal organs, initiating hematopoiesis, and maintaining the homeostasis of key electrolytes via calcium/phosphate ion storage. The most common cause for bone defects is extensive trauma and subsequent infection. Bone tissue has the self-healing capability without a scar tissue formation for the majority of the injuries. However, some may result with delayed union or fracture non-union. Such cases include reconstruction of large bone defects or cases of compromised regenerative process as a result of avascular necrosis and osteoporosis. Several surgical methods exist to treat bone defects, including Ilizarov method, Masquelete technique, growth factor stimulation, and bone replacement. Unfortunately, these are technically demanding and come with noteworthy disadvantages such as lengthy treatment duration, adverse effects on the patient’s psychology, repeated surgical procedures, and often long hospitalization times. These limitations associated with surgical techniques make bone substitutes an attractive alternative. Here, it was hypothesized that a 3D printed scaffold will mimic trabecular bone in terms of biomechanical properties and that such scaffolds will support cell attachment and survival. To test this hypothesis, this study aimed at fabricating poly(lactic acid), PLA, structures using 3D printing technology for trabecular bone defects, characterizing the scaffolds and comparing with bovine trabecular bone. Capacity of scaffolds on human bone marrow stem cell (hBMSC) attachment and survival was also evaluated. Cubes with a volume of 1 cm³ having pore sizes of 0.50, 1.00 and 1.25 mm were printed. The scaffolds/grafts were characterized in terms of porosity, contact angle, compressive mechanical properties as well cell response. Porosities of the 3D printed scaffolds were calculated based on apparent densities. For contact angles, 50 µl distilled water was dropped over the surface of scaffolds, and contact angles were measured using ‘Image J’ software. Mechanical characterization under compression was performed on scaffolds and native trabecular bone (bovine, 15 months) specimens using a universal testing machine at a rate of 0.5mm/min. hBMSCs were seeded onto the 3D printed scaffolds. After 3 days of incubation with fully supplemented Dulbecco’s modified Eagle’s medium, the cells were fixed using 2% formaldehyde and glutaraldehyde mixture. The specimens were then imaged under scanning electron microscopy. Cell proliferation was determined by using EZQuant dsDNA Quantitation kit. Fluorescence was measured using microplate reader Spectramax M2 at the excitation and emission wavelengths of 485nm and 535nm, respectively. Findings suggested that porosity of scaffolds with pore dimensions of 0.5mm, 1.0mm and 1.25mm were not affected by pore size, while contact angle and compressive modulus decreased with increasing pore size. Biomechanical characterization of trabecular bone yielded higher modulus values as compared to scaffolds with all pore sizes studied. Cells attached and survived in all surfaces, demonstrating higher proliferation on scaffolds with 1.25mm pores as compared with those of 1mm. Collectively, given lower mechanical properties of scaffolds as compared to native bone, and biocompatibility of the scaffolds, the 3D printed PLA scaffolds of this study appear as candidate substitutes for bone repair and regeneration.

Keywords: 3D printing, biomechanics, bone repair, stem cell

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Authors: Felora Rafiei, Shahram Shoaei

Abstract:

The aim of this studies was to evaluate effect of nitroxin, super nitro plus and biophosphorus on yield and yield components of maize (Zea mays) under different levels of chemical fertilizers in the condition of normal and difficiet irrigation. Experiment laid out as split plot factorial based on randomized complete block design with three replications. Main plots includes two irrigation treatments of 70 (I1), 120(I2) mm evaporation from class A pan. Sub plots were biological fertilizer and chemical fertilizer as factorial biological fertilizer consisting of nitroxin: Azospirillium lipoferum, Azospirillium brasilens, Azotobacter chroococcum Azotobacter agilis (108 CFU ml-1) (B1), super nitro plus (Azospirillium spp, + Pseudomonas fluorescence + Bacillus subtilis (108 CFU ml-1) + biological fungicide) (B2), biophosphorus (Pseudomonas spp + Bacillus spp (107 CFU ml-1) (B3), and chemical fertilizer consisting of NPK (C1), N5oP5oK5o (C2) and NoPoKo (C3).The results showed that usage of biological fertilizer have positive effects on chemical fertilizers use efficiency and tolerance to drought stress in maize. Also with use of biological fertilizer can decrease usage of chemical fertilizers.

Keywords: biological fertilizer, chemical fertilizer, yield component, yield, corn

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Authors: Hasan Ashkanani

Abstract:

Pottery assemblages from the site of Al-Khidr on Failaka Island, Kuwait, were analysed in order to reconstruct the chemical composition of Bronze Age wares and to build a mineralogical database of Bronze Age pottery dated from Failaka Periods 1–3B (2000–1650 BCE). A total of 145 ceramic sherds from Al-Khidr, as well as reference groups, were analysed by non-destructive portable X-ray fluorescence (pXRF) spectrometry. Preliminarily petrographic thin-section analysis was applied to four samples to reconstruct possible clay paste recipes and to identify raw materials. The results indicate that geochemical analyses can successfully distinguish subgroups within a typological category of ceramic assemblages. The results identified two subgroups within the Al-Khidr typological category: the Dilmun Barbar tradition and the Mesopotamian tradition. Future comparative compositional studies can be conducted to explore other aspects of craft specialisation, such as ceramic technological choices and possibly the influence of sociopolitical units

Keywords: Kuwait archaeology, pottery, pXRF, Dilmun

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Authors: I. Yalcin Enis, J. Vojtech, T. Gok Sadikoglu

Abstract:

In this study, polycaprolactone (PCL) was dissolved in chloroform: ethanol solvent system at a concentration of 18 w/v %. 1, 2, 4, and 6 droplets of formic acid were added to the prepared 10ml PCL-chloroform:ethanol solutions separately. Fibrous webs were produced by electrospinning technique. Morphology of the webs was investigated by using scanning electron microscopy (SEM) whereas fiber diameters were measured by Image J Software System. The effect of formic acid addition to the mostly used chloroform solvent on fiber morphology was examined.

Keywords: chloroform, electrospinning, formic acid polycaprolactone, fiber

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Authors: Gülşah Çelik Gül, Figen Kurtuluş

Abstract:

Cesium molybdates with general formula CsMIII(MoO4)2, where MIII = Bi, Dy, Pr, Er, exhibit rich polymorphism, and crystallize in a layered structure. These properties cause intensive studies on cesium molybdates. CsBi(MoO4)2 was synthesized by microwave method by using cerium sulphate, bismuth oxide and molybdenum (VI) oxide in an appropriate molar ratio. Characterizations were done by x-ray diffraction (XRD), fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy/energy dispersive analyze (SEM/EDS), thermo gravimetric/differantial thermal analysis (TG/DTA).

Keywords: cesium bismuth dimolybdate, microwave synthesis, powder x-ray diffraction, rare earth dimolybdates

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Authors: K. C. Gayithri, S. K. Naveen Kumar

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ZnO is one of the most important semiconductor materials, non toxic, biocompatible, antibacterial properties for research and it is used in many biomedical applications. MnxZn1-xO nano thin films were prepared by a spin coating sol-gel method on silicon substrate. The structural, optical, electrical properties of Mn Doped ZnO are studied by using X-rd, FESEM, UV-Visible spectrophotometer. The X-rd reveals that the sample shows hexagonal wurtzits structure. Surface morphology and thickness of the sample are characterized by field emission scanning electron microscopy. Absorption and transmission spectra are studied by UV-Visible spectrophotometer. The electrical properties are measured by TCR meter.

Keywords: transition metals, Mn doped ZnO, Sol-gel, x-ray diffraction

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Authors: Mahdi Rahaie

Abstract:

MicroRNAs are a novel class of small RNAs which regulate gene expression by translational repression or degradation of messenger RNAs. To produce sensitive, simple and cost-effective assays for microRNAs, detection is in urgent demand due to important role of these biomolecules in progression of human disease such as Alzheimer’s, Multiple sclerosis, and some other neurodegenerative diseases. Herein, we report several novel, sensitive and specific microRNA nanobiosensors which were designed based on colorimetric and fluorescence detection of nanoparticles and hybridization chain reaction amplification as an enzyme-free amplification. These new strategies eliminate the need for enzymatic reactions, chemical changes, separation processes and sophisticated equipment whereas less limit of detection with most specify are acceptable. The important features of these methods are high sensitivity and specificity to differentiate between perfectly matched, mismatched and non-complementary target microRNAs and also decent response in the real sample analysis with blood plasma. These nanobiosensors can clinically be used not only for the early detection of neuro diseases but also for every sickness related to miRNAs by direct detection of the plasma microRNAs in real clinical samples, without a need for sample preparation, RNA extraction and/or amplification.

Keywords: hybridization chain reaction, microRNA, nanobiosensor, neurodegenerative diseases

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Authors: Stepan Papacek, Jiri Jablonsky, Radek Kana, Ctirad Matonoha, Stefan Kindermann

Abstract:

FRAP (Fluorescence Recovery After Photobleaching) is a widely used measurement technique to determine the mobility of fluorescent molecules within living cells. While the experimental setup and protocol for FRAP experiments are usually fixed, data processing part is still under development. In this paper, we formulate and solve the problem of data selection which enhances the processing of FRAP images. We introduce the concept of the irrelevant data set, i.e., the data which are almost not reducing the confidence interval of the estimated parameters and thus could be neglected. Based on sensitivity analysis, we both solve the problem of the optimal data space selection and we find specific conditions for optimizing an important experimental design factor, e.g., the radius of bleach spot. Finally, a theorem announcing less precision of the integrated data approach compared to the full data case is proven; i.e., we claim that the data set represented by the FRAP recovery curve lead to a larger confidence interval compared to the spatio-temporal (full) data.

Keywords: FRAP, inverse problem, parameter identification, sensitivity analysis, optimal experimental design

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Authors: Babak Sadeghi, Rony Snyders, Marie-Paule.Delplancke-Ogletree

Abstract:

Nitrogen fixation (NF) is one of the crucial industrial processes. Many attempts have been made in order to artificially fix nitrogen, and among them, the Haber-Bosch’s (H-B) process is widely used. However, it presents two major drawbacks: huge fossil feedstock consumption and noticeable greenhouse gases emission. It is, therefore, necessary to develop alternatives. Plasma technology, as an inherent “green” technology, is considered to have a great potential for reducing the environmental impacts and improving the energy efficiency of the NF process. In this work, we have studied the catalyst assisted microwave plasma for NF application. Heterogeneous catalysts of MoO₃, with various loads 0, 5, 10, 20, and 30 wt%, supported on γ-alumina were prepared by conventional wet impregnation. Crystallinity, surface area, pore size, and microstructure were obtained by X-ray diffraction (XRD), Brunauer–Emmett–Teller (BET) adsorption isotherm, Scanning electron microscopy (SEM), and Transmission electron microscopy (TEM). The XRD patterns of calcined alumina confirm the γ- phase. Characteristic picks of MoO₃ could not be observed for low loads (< 20 wt%), likely indicating a high dispersion of metal oxide over the support. The specific surface area along with pores size are decreasing with increasing calcination temperature and MoO₃ loading. The MoO₃ loading does not modify the microstructure. TEM and SEM results for loading inferior to 20 wt% are coherent with a monolayer of MoO₃ on the support as proposed elsewhere. For loading of 20 wt% and more, TEM and Electron diffraction (ED) show nanocrystalline ₃-D MoO₃ particles. The catalytic performances of these catalysts were investigated in the post-discharge of a microwave plasma for NOx formation from N₂/O₂ mixtures. The plasma is sustained by a surface wave launched in a quartz tube via a surfaguide supplied by a 2.45 GHz microwave generator in pulse mode. In-situ identification and quantification of the products were carried out by Fourier-transform infrared spectroscopy (FTIR) in the post-discharge region. FTIR analysis of the exhausted gas reveal NO and NO₂ bands in presence of catalyst while only NO band were assigned without catalyst. On the other hand, in presence of catalyst, a 10% increase of NOₓ formation and of 20% increase in energy efficiency are observed.

Keywords: γ-Al2O₃-MoO₃, µ-waveplasma, N2 fixation, Plasma-catalysis, Plasma diagnostic

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