The demicellization of the cationic detergents dodecyltrimethylammonium bromide, tetradecyltrimetylammonium bromide, and cetyltrimethylammonium bromide was studied at temperatures between 20 and 60 degrees C in 0.1 M NaCl (pH 6.4) using isothermal titration calorimetry (ITC). We determined the critical micellization concentration (cmc) of the cationic detergents which show a minimum at temperatures between 20 and 34 degrees C. In accordance with the lengthening of the hydrophobic tail of the detergents the cmc decreases with increasing alkyl chain length. The thermodynamic parameters describing the changes of enthalpy (DeltaH(demic)), the changes of entropy (DeltaS(demic)) and the Gibbs free energy change (DeltaG(demic)) for demicellization were first obtained using the pseudophase-separation model. The aggregation number n at the cmc as well as the demicellization enthalpy, entropy and Gibbs free energy change were also calculated using a simulation based on the mass-action model. Furthermore, we investigated the demicellization of CTAB in deionized water in comparison to demicellization in sodium chloride solution to determine the influence of counter ion binding on the demicellization.
High-resolution transmission electron microscopy (HRTEM) and ab initio quantum-mechanical calculations of electronic structure were combined to investigate the structure of the hydroxyapatite (HA) (010) surface, which plays an important role in HA interactions with biological media. HA was synthesized by in vitro precipitation at 37°C. HRTEM images revealed thin elongated rod nanoparticles with preferential growth along the  direction and terminations parallel to the (010) plane. The focal series reconstruction (FSR) technique was applied to develop an atomic-scale structural model of the high-resolution images. The HRTEM simulations identified the coexistence of two structurally distinct terminations for (010) surfaces: a rather flat Ca(II)-terminated surface and a zig-zag structure with open OH channels. Density functional theory (DFT) was applied in a periodic slab plane-wave pseudopotential approach to refine details of atomic coordination and bond lengths of Ca(I) and Ca(II) sites in hydrated HA (010) surfaces, starting from the HRTEM model.
In the last years, researches developed with biosurfactants for application in the medical area have been revealing the promising biological activities of these biomolecules. In this work the antimicrobial and anti-adhesive properties of a biosurfactant Rufisan isolated from the yeast Candida lipolytica UCP 0988, growth in a medium supplemented with ground nut refinery residue was determined against several microorganisms. The biosurfactant was able to reduce the water surface tension from 70 to 25.3 mN/m and showed a critical micelle concentration (CMC) of 0.03%. The biosurfactant was isolated after 72 h of fermentation and was tested in concentrations varying from 0.75 to 12 mg/l. The highest antimicrobial activities were observed against Streptococcus agalactiae, Streptococcus mutans, Streptococcus mutans NS, Streptococcus mutans HG, Streptococcus sanguis 12, Streptococcus oralis J22 at a concentration superior to the biosurfactant critical micelle concentration. Moreover, the biosurfactant showed anti-adhesive activity against most of the microorganisms tested. As far as we know, this is the first compilation of data on antimicrobial and anti-adhesive activities of a biosurfactant obtained from a Candida strain against such a broad group of microorganisms. The results obtained in this work showed that the biosurfactant from C. lipolytica is a potential antimicrobial and/or anti-adhesive agent for several biomedical applications.
The characteristics of the binding reaction of surfactant-cobalt(III) complex, cis-[Co(phen)₂(C₁₄H₂₉NH₂)]Cl₂·3H₂O (phen=1,10-phenanthroline, C₁₄H₂₉NH₂=tetradecylamine) with human serum albumin (HSA) were studied by fluorescence and UV-vis absorption spectroscopy. In addition, the effect of the surfactant-cobalt(III) complex on the conformation of HSA was analysed using synchronous fluorescence spectroscopy. The experimental results showed that surfactant-cobalt(III) complex caused the fluorescence quenching of HSA through a combination of static and dynamic quenching. The number of binding sites (n) and apparent binding constant (K(a)) of surfactant-cobalt(III) complex (above and below the critical micelle concentration (cmc) were determined at various temperatures. According to the thermodynamic parameters, it is likely that hydrophobic interactions are involved in the binding process. The cancer chemotherapeutic potential of surfactant-cobalt(III) complex on ME-180 cervical cancer cell was determined using MTT assay and specific staining techniques. The complex affected the viability of the cells significantly and the cells succumbed through an apoptosis process as seen in the nuclear morphology and cytoplasmic features. In addition, single-cell electrophoresis indicated DNA damage.
The interaction of hem agglutininneuraminidase (HN) and fusion (F) glycoproteins with swollen vesicles of 1,2-dihexadecyl-sn-glycero-3-phosphatidylcholine (DHPC) was investigated under transition from gel to fluid phase. X-ray studies of the structure of lipid/HN-F mixtures in normal and swollen vesicles have shown that the lamellar bilayer structure predominate in the gel and liquid crystalline phases. A swollen lipid phase, in which the mean repeat distance of lipid bilayers is larger than in the other phases was found. The nature of this phase is similar to the anomalous bilayer swelling reported in literature. The presence of HN and F in the vesicles led to the coexistence of structures with low and high lamellar order, showing larger repeat distance in comparison with the pure lipid. This finding was attributed to the increase in the lipid bilayer thickness due to the HN-F included in the free water layer. The thermal behaviour of the system was not affected by the vesicle swelling. The data showed the existence of gel and liquid crystalline lamellar phases and changes in lipid/HN-F specific heats, mainly due to the concentration effect of the HN-F and its location in the free water layer.
Small-angle neutron scattering on extruded unilamellar vesicles in water was used to study bilayer thickness when cholesterol (CHOL) was added at 44.4 mol% to 1,2-dimyristoleoylphosphatidylcholine (diC14:1PC) and 1,2-dierucoylphosphatidylcholine (diC22:1PC) bilayers. Using the (1)H(2)O/(2)H(2)O contrast variation and the small-angle form of Kratky-Porod approximation, the bilayer gyration radii at infinite contrast R(g,infinity) and the bilayer thickness parameters d(g,infinity) = 12(0.5)R(g,infinity) were obtained at 30 degrees C. Addition of cholesterol to diC14:1PC increased the d(g,infinity) from 3.72 +/- 0.02 to 4.26 +/- 0.01 nm, while in the diC22:1PC bilayers the d(g,infinity) change observed was within the experimental error: +0.23 +/- 0.23 nm.
A carbon paste electrode (CPE) modified with 2,2'-[1,2 butanediylbis(nitriloethylidyne)]-bis-hydroquinone (BBNBH) and TiO(2) nanoparticles was used for the sensitive voltammetric determination of epinephrine (EP). The electrochemical response characteristics of the modified electrode toward EP and acetaminophen (AC) were investigated by cyclic and differential pulse voltammetry (CV and DPV). The results showed an efficient catalytic activity of the electrode for the electrooxidation of EP, which leads to a reduction in its overpotential by more than 270mV. The effects of pH and potential sweep rate on the mechanism of the electrode process were investigated. The modified electrode exhibits an efficient electron-mediating behavior together with well-separated oxidation peaks for EP and AC. At the optimum pH of 8.0 in a 0.1M phosphate buffer solution, the DPV anodic peak currents showed a linear relationship versus EP concentrations in the range of 1.0-600.0microM and a detection limit of 0.2microM.
The in vitro transfection activity of a novel series of N,N'-diacyl-1,2-diaminopropyl-3-carbamoyl-(aminoethane) derivatives was evaluated against a mouse melanoma cell line at different +/- charge ratios, in the presence and absence of helper lipids. Only the unsaturated derivative N,N'-dioleoyl-1,2-diaminopropyl-3-carbamoyl-(aminoethane), (1,2lmp) mediated significant increase in the reporter gene level which was significantly boosted in the presence of DOPE peaking at +/- charge ratio of 2. The electrostatic interactions between the cationic liposomes and plasmid DNA were investigated by gel electrophoresis, fluorescence spectroscopy, dynamic light scattering and electrophoretic mobility techniques. In agreement with the transfection results, 1,2lmp/DOPE formulation was most efficient in associating with and retarding DNA migration. The improved association between the dioleoyl derivative and DNA was further confirmed by ethidium bromide displacement assay and particle size distribution analysis of the lipoplexes. Differential scanning calorimetry studies showed that 1,2lmp was the only lipid that exhibited a main phase transition below 37 degrees C. Likewise, 1,2lmp was the only lipid found to form all liquid expanded monolayers at 23 degrees C. In conclusion, the current findings suggest that high in vitro transfection activity is mediated by cationic lipids characterized by increased acyl chain fluidity and high interfacial elasticity.
Chemical modification of spin-cast chitosan films has been performed. This modification involves the attachment of 1,2 Epoxy-3-phenoxy-propane, commonly known as glycidyl phenyl ether (GPE), to the amine group of the chitosan molecule. Optical properties of modified films have been determined in the infrared region of the spectrum using spectroscopic ellipsometry, and are reported in this paper. Special attention is paid to the infrared region where the index of refraction and extinction coefficients from 750 to 4000 cm(-1) were determined. Difference plots of IR optical data before and after chemical modification were generated to confirm that modification had occurred. Optical modeling of infrared spectroscopic ellipsometry (IRSE) data with respect to chemical bond vibrations has also been performed. This modeling involved curve fitting of resonant chemical bond absorptions using Lorentz oscillators. These oscillator models allow for comparison of modified chitosan to unmodified chitosan. The purpose of this research was to determine infrared optical constants of chemically modified chitosan films This work shows that surface chemistry of biomaterials can be studied quite sensitively with spectroscopy ellipsometry, detecting as little as 100 ng/cm(2) of GPE.
The aqueous solubilization of the organoselenium compound viz., 1,2-bis(bis(4-chlorophenyl)methyl)diselane [(ClC(6)H(4))(2)CHSe](2) has been investigated experimentally in micellar solutions of two cationic (hexadecyltrimethylammonium bromide, CTAB, hexadecyltrimethylammonium chloride, CTAC) and one nonionic (polyoxyethylene(20)mono-n-hexadecyl ether, Brij 58) surfactants possessing the same hydrocarbon "tail" length and in their single as well as equimolar binary and ternary mixed states. Solubilization capacity determined with spectrophotometry and tensiometry has been quantified in terms of molar solubilization ratio and micelle-water partition coefficient. FTIR, UV-vis, fluorescence and zeta potential measurements have been utilized to ascertain the interaction of organochalcogen compound with surfactants. Equimolar cationic-nonionic surfactant combinations show better solubilization capacity than pure cationics or nonionics, whereas equimolar cationic-cationic-nonionic ternary surfactant systems exhibit intermediate solubilization efficiency between their single and binary counterparts. Locus of solubilization of [(ClC(6)H(4))(2)CHSe](2) in different micellar solutions was probed by UV-visible spectroscopy. The investigation has presented precious information for the preference of mixed surfactants for solubilizing water-insoluble compounds. Indeed the solubilization aptitude of these surfactants is not merely related to molar capacity. The results furnish adequate support to justify comprehensive exploration of the surfactant properties that influence solubilization.
Crosslinked poly(1-vinyl 1,2,4-triazole) (PVTAz) hydrogels in the form of rod and hollow cylinder (thin and thick wall) have been prepared by (60)Co γ-radiation initiated simultaneous polymerization and crosslinking of 1-vinyl 1,2,4-triazole in the presence of water. In binary aqueous systems, the degree of gelation decreased with the increasing water content. The swelling results obtained by using different form of hydrogels showed that the thin wall hollow cylinder hydrogels perform better than other hydrogels. Structural, morphological and thermal characterizations of the hydrogels were carried out with several techniques, including Fourier transform infrared spectroscopy, scanning electron microscopy, swelling measurements, thermogravimetry, and differential scanning calorimetry. The effects of time, pH, temperature, ionic strength, and salt and solvent type on the swelling behavior of thin wall hollow cylinder hydrogels were also investigated. Swelling equilibrium was attained in 2 days. Thin wall hollow cylinder PVTAz hydrogels originally swelled to 3000% (by volume), and depending on the pH value, no sharp change was observed on the swelling curve. Swelling values of hydrogels showed that the swelling ratio increase with the increasing temperature in the range of 4-20°C. The swelling ratios of the gels decreased with increasing ionic strength. Hydrogels have shown the maximum contraction in the presence of LiCl. 5-Fluorouracil (5-FU) was loaded on PVTAz hydrogels by using incorporation and adsorption methods. Both methods show the similar release profiles. Time-dependent release of 5-FU was achieved within 48 h by the success of 65%. All these results showed that PVTAz hydrogels are suitable for the release of 5-FU in the applications of long-term cancer treatment.
The physicochemical properties of a novel series of symmetric 1,3-dialkylamidopropane-based cationic amphiphiles [M. Sheikh, J. Feig, B. Gee, S. Li, M. Savva, In vitro lipofection with novel series of symmetric 1,3-dialkoylamidopropane-based cationic surfactants containing single primary and tertiary amine polar head groups, Chem. Phys. Lipids 124 (2003) 49-61] were studied by several techniques, in an effort to correlate cationic lipid structure with transfection efficacy. It was found that only the unsubstituted amine and tertiary amine dioleoyl derivatives 1,3lmp5 and 1,3lmt5, respectively, mediated in vitro transfection activity in the absence of helper lipids. This activity pattern was consistent with ethidium bromide fluorescence quenching studies, which indicated that only these two derivatives bound to and efficiently condense plasmid DNA at physiological pH. Dynamic light scattering indicated that lipoplexes made by these two cationic lipids were relatively small particles below 1 microm, in sharp contrast to lipoplexes bigger than 3 microm composed of saturated cationic derivatives. Transmission electron microscopy studies clearly indicated that cationic lipid dispersions made by saturated derivatives form multilamellar tubules at physiological pH. Calorimetric studies showed that cationic amphiphiles with saturated acyl chains longer than 12 carbons exhibit solid-to-liquid crystalline phase transitions above 37 degrees C. In agreement with the microscopy and calorimetry studies, Langmuir film balance experiments indicated that saturated derivatives with hydrophobic chains longer that 12 carbons are not well hydrated and exist at a chain-ordered state at ambient temperature. Calculation of compressibility moduli from monolayer compression isotherms at 23 degrees C suggested that monolayers made by cationic lipids bearing saturated acyl chains are less compressible relative to those of the dioleoyl derivatives 1,3lmp5 and 1,3lmt5. In conclusion, high hydration, increased fluidity and high elasticity of cationic lipid assemblies in isolation, all correlate with high in vitro transfection activity.
In this study, electrochemical polymerized 5-amino-2-mercapto-1,3,4-thiadiazole (poly-AMT)-modified single-use graphite electrodes were fabricated for electrochemical monitoring of bisflavonoid, quercetin. The surfaces of p-AMT modified pencil graphite electrodes (PGEs) were firstly characterized by scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS) analysis. The conditions for electrochemical polymerization by using cyclic voltammetry (CV), such as scan rate and different potential cycles were optimized. Electrochemical behaviors of these electrodes were also investigated using differential pulse voltammetry (DPV) and EIS. A six-fold increase at quercetin signal was obtained by using p-AMT modified PGEs compared to unmodified electrodes.
In this study, single-use graphite electrodes modified with 5-amino-2-mercapto-1,3,4-thidiazole (AMT) were fabricated for electrochemical monitoring of DNA. The surfaces of AMT modified pencil graphite electrodes (PGEs) were characterized by scanning electron microscopy (SEM). Electrochemical behaviors of these electrodes were investigated using differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). The effects of pretreatment of PGE and immobilization time and concentration of AMT were studied to facilitate an effective immobilization on PGE surface. The efficiency of immobilization is verified by measuring the oxidation signals of either AMT itself or adenine base of DNA, immobilized onto AMT modified electrodes.
The size and structure of aggregates formed by interaction of DNA with homologous series of cationic gemini surfactants butane-1,4-diyl-bis(alkyldimethylammonium bromide) (CnGS, n=10-16 is the number of alkyl carbons) were investigated using UV-vis turbidity, dynamic light scattering and small-angle synchrotron X-ray (SAX) diffraction. The detailed analysis of turbidity in the range of lambda=450-600 nm indicates an anomaly in the growth of CnGS+DNA aggregates with increasing concentration of CnGS, possibly involving changes of structure and size of aggregates. Using dynamic light scattering, changes of the effective diameter of CnGS+DNA (n=12 and 16) aggregates formed in the CnGS concentration range 0.002-0.140 mmol/l were observed. SAX diffractograms show the presence of long-range organization of CnGS+DNA (n=12, 13, 14 and 16) aggregates due to DNA interaction with CnGS above the critical micellar concentration. The CnGS+DNA (n=12, 13 and 14) aggregates at 25 degrees C are packed in a lattice of two-dimensional hexagonal symmetry. With increasing C14GS:DNA molar ratio the changes of the lattice parameter in the range of 4.80-5.27 nm are observed at 25 degrees C. The aggregates undergo structural changes induced by temperature in the range 60-95 degrees C, which are accompanied by changes of the diffraction patterns, namely in the region of reciprocal spacing s=0.15-0.30 nm(-1).
Ultrasonic signals propagated through medium were directly applied to unicellular cyanobacterium cell surfaces to investigate the biological effects induced by ultrasound. The gas-vacuolate cyanobacterium Microcystis aeruginosa and the gas-vacuole negative cyanobacterium Synechococcus PCC 7942 responded differently to ultrasound. When M. aeruginosa was irradiated by 1.7 MHz ultrasound at 0.6 W cm(-2) every day, it showed a decrease of nearly 65% in biomass increment, and this group's generation time increased twice as much as the control. While Synechococcus culture irradiated every day still grew as fast as the control, and its final biomass was as much as the control. The value of the electric conductivity change (Deltasigma) sharply increased in Microcystis suspension during the exposure process, which revealed more ultrasonic cavitation yield in liquid related to the gas-vacuolate cyanobacteria. The relative malondialdehyde (MDA) content, a quantitative indicator of lipid peroxidation, increased by 65% in Microcystis cells and 9% in Synechoccus cells after ultrasonic irradiation. Moreover, the membrane permeability, quantified by measuring the relative amount of electrolyte leaking out of cells, increased to more than 60% in the Microcystis cells. The results indicated that Microcystis cells were susceptible to ultrasonic stress. According to Rayleigh-Plesset's bubble activation theory, 1.7 MHz ultrasound approached the eigenfrequency of gas-vacuolate cells. The present investigation suggested the importance of the cavitational effect relative to intracellular gas-vacuoles in the loss of cell viability. In summary, 1.7 MHz ultrasonic irradiation was effective in preventing water-bloom forming cyanobacteria from growing rapidly due to changes in the functioning and integrity of cellular and subcellular structures.
In the present study, the effects of an amphiphilic polymer, d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) on model surfactant monolayers dipalmitoylphosphatidylcholine (DPPC), a binary mixture of DPPC with palmitoyloleoyl phosphatidylglycerol (DPPC-POPG) 9:1 (w/w) and binary mixture of DPPC and oleic acid (DPPC-OA) were evaluated. The ability of TPGS to act as an antioxidant adjuvant for pulmonary surfactants was also evaluated. Compression isotherms of surfactant monolayers at 37 °C in a Langmuir-Blodgett trough showed that DPPC and DPPC:TPGS mixed monolayers (1:0.25-1:1, w/w) exhibited low minimum surface tensions (MST) of 1-2 mN/m. Similarly [DPPC:POPG (9:1, w/w)]:TPGS mixed films of 1:0.25-1:1 weight ratios reached 1-2 mN/m MST. DPPC:POPG:TPGS liposomes adsorbed to surface tensions of 29-31 mN/m within 1s. While monolayers of DPPC:OA (1:1, w/w) reached high MST of ∼11 mN/m, DPPC:OA:TPGS (1:1:0.25, w/w) film reached near zero MST suggesting that low concentrations of TPGS reverses the effect of OA on DPPC monolayer. Capillary surfactometer studies showed DPPC:TPGS and [DPPC:POPG (9:1, w/w)]:TPGS liposomes maintained 84-95% airway patency. Fluorescence spectroscopy of Laurdan loaded DPPC:TPGS and DPPC:POPG:TPGS liposomes revealed no segregation of lipid domains in the lipid bilayer. Addition of TPGS to soybean liposome significantly reduced thiobarbituric acid reactive substance (TBARS) by 29-39% confirming its antioxidant nature. The results suggest a potential use of TPGS as an adjuvant to improve the surfactant activity as well as act as an antioxidant by scavenging free radicals.
Mg-based materials are promising for orthopedic, dental, and cardiovascular applications but their high degradation rate in vivo (release of Mg ions and debris particles) is cause of great concern. Protective treatments involving fluoride conversion coatings have been proposed in order to reduce corrosion rates. The aim of this study was to evaluate Mg debris biodegradation and its possible cytotoxic effects on osteoblastic cells in situ. Neutral Red dying and Acridine Orange staining techniques were used as endpoints to analyse the cytotoxic effects at 100-2000 μg/mL concentration range. Results showed a marked variation of Mg ion concentration in the culture medium after different exposure periods (1, 2, or 24h). Interestingly, the release rate of magnesium ions was dependent on the presence or absence fluoride treatment. Adverse effects induced by ≥1000 μg/mL MP doses and Mg ion concentrations higher than 480 μg/mL were observed on cells. Results showed significant differences between the concentration of Mg ions in the presence and absence of cells. This fact reveals a dynamic equilibrium mediated by Mg ion input and output in the cells that leads to the change in MP corrosion rates. Fluoride release from conversion coatings did not show cytotoxic effects.
Plants are exploited as a potential source for the large-scale production of noble gold nanoparticles in the recent years owing to their various potential applications in nanobiotechnology and nanomedicine. The present work describes green biosynthetic procedures for the production of gold nanoparticles for the first time by using an aqueous extract of the Dysosma pleiantha rhizome. The biosynthesized gold nanoparticles were confirmed and characterized by ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, and scanning electron microscopy equipped with energy dispersive spectroscopy. The results revealed that aqueous extract of D. pleiantha rhizome has potential to reduce chloroauric ions into gold nanoparticles and the synthesized gold nanoparticles were showed spherical in shape with an average of 127nm. Further, we investigated the anti-metastatic activity of biosynthesized gold nanoparticles against human fibrosarcoma cancer cell line HT-1080. The results showed that the biosynthesized gold nanoparticles were non-toxic to cell proliferation and, also it can inhibit the chemo-attractant cell migration of human fibrosarcoma cancer cell line HT-1080 by interfering the actin polymerization pathway. Thus, the usage of gold nanoparticles biosynthesized from D. pleiantha rhizome can be used as a potential candidate in the drug and gene delivery to metastatic cancer.
In this study, a surfactin was extracted from a novel surfactant producing bacterial strain Bacillus amyloliquifaciens KSU-109, isolated from rhizosphere of date palm (Phoenix dactylifera), and characterized based on 16Sr RNA and sfp genes using Blastn, Blastx and phylogenetic analyses. The study was performed to obtain a renewable bioresource for surfactin production, and its application in nanotechnology as a non-hazardous and environmentally compatible nanoparticle (NP) stabilizer. The strain KSU-109 produced the surfactin with an average yield of 160 mg/L with strong surfactant activity, reducing the surface tension of the medium from 72 mN/m to 29.3 mN/m. The surfactin preparation was used for synthesizing the cadmium sulfide nanoparticles (CdS-NPs) by mixing 0.005% surfactin with 1mM Cd(NO(3))(2) in 1:1 ratio (v/v) and 10mM Na(2)S solution at pH 7.2 and ambient temperature, which were stable up to 120 days. The surfactin stabilized CdS-NPs were characterized using XRD, TEM, and spectroscopic techniques. The data revealed a significant role of surfactin as a stabilizer and capping agent, which also causes phase transition to yield the cubic/hexagonal CdS-NPs of average size of 3-4 nm. The results elucidated the significance of biocompatible and biodegradable surfactin as an effective and inexpensive stabilizing agent for developing stable CdS nanoparticles.
An extracellular lipase produced by the strain of Y-11 Trichosporon capitatum was purified to homogeneity by ammonium sulfate precipitation and four chromatographic steps. The purified lipase showed enhanced activity when it was immobilized in gelatin-containing microemulsion-based organogels. Furthermore, the resolution of (R,S)-(+/-)-glycidyl butyrate by this immobilized lipase gave a product of (S)-(-)-glycidol with approximately 98% ee (E=96). By using chiral HPLC separation, (S)-(-)-glycidol was obtained in enantiopure form. Scaled-up reaction in 2l shake flask was also performed and the repeated use of 15 times of immobilized Y-11 T. capitatum lipase resulted in little loss in its activity (4.8%).
In this study the surface composition of 7S and 11S globulin powders from soybean proteins by aqueous buffer and reverse micelle extractions had been examined using X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM). Analysis by XPS revealed that the O and N atomic percentage of 7S and 11S globulin surfaces from bis(2-ethylhexyl) sodium sulfosuccinate (AOT) reverse micelle was higher than from aqueous buffer, but the C atomic percentage was lower. The O/C ratio of the 7S globulin powder from aqueous buffer and reverse micelle was similar while significant differences were obtained in the O/C ratio of the 11S globulin powder, N/C atom ratios of the 7S and 11S globulin powders and high-resolution XPS C 1s, N 1s, O 1s spectra. Powder microstructure after reverse micelle treatment showed the presence of small pores, indicating the effect of reverse micelle on the 7S and 11S globulin structure. The obtained results indicated that the reverse micelle could affect the C, O and N components on the surface of soybean proteins.
In this work, we investigated the interactions of PrP106-126 amide with 1-palmitoyl-2-oleoyl-3-phosphocholine (POPC) and POPC/bovine brain sphingomyelin (BSM) membranes in the presence of calcium ions by in situ time-lapse atomic force microscopy (AFM) and circular dichroism (CD). The CD results show that Ca(2+) has no obvious effects on the random coil conformation of PrP106-126 amide. The tapping mode AFM results demonstrate that electrostatic interaction decreases the measured heights of supported lipid bilayers (SLBs) in HBS-Ca(2+) solution. Electrostatic interaction analysis also can be used to determine the applied force in liquid tapping mode AFM. The interactions of PrP106-126 amide with membranes by AFM demonstrate the following: (i) Ca(2+) inhibits the interaction of PrP106-126 amide with POPC lipid and (ii) the co-interaction between Ca(2+) and BSM increases the poration ability of PrP106-126 amide. These results imply that the main role of Ca(2+) in the interactions of PrP106-126 amide with membranes is changing the surface properties of the membranes.
A functionalized multi-wall nanotube (MWNT) modified glass carbon electrode (GCE) was used to study the effects of aluminum species on glutamate dehydrogenase (GLDH) activity by monitoring amperometric i-t curve for the oxidation of the enzymatically generated NADH. The conformational changes of the coenzyme nicotinamide adenine dinucleotide (NAD(+)) induced by Al(III) and nanometer-sized tridecameric aluminum polycation (nano-Al(13)) were investigated by the fluorescence technique. The results showed that the electrochemical method may be a potential tool to investigate the activity of enzymes in the biological system. It may also be useful in studying the effects of nano-sized aluminum compounds on biomolecules in order to discuss their safety to the environment and human.
During any microbial enhanced oil recovery process, both cells and the metabolic products of bacteria govern the tertiary oil recovery efficiency. However, very accurate examination is needed to find the functionality of these tiny creatures at different reservoir conditions. In this regard, the effect of cell structure on ultimate microbial recovery efficiency which is the most dominant mechanism based on the microorganism types (gram-negative or gram-positive) was systematically investigated. At the first stage, possible different active mechanisms using Bacillus stearothermophilus SUCPM#14 strain were tested using specially designed injection protocol, in situ and ex situ core flooding experiments, interfacial tension, viscosity, pH and Amott wettability index measurements. At the second stage, comparing functionality of B. stearothermophilus SUCPM#14 (a gram-positive type) with the previously examined strain namely Enterobacter cloacae as a gram-negative type, proposed this hypothesis that the cell structure significantly affects the interfacial behaviors. New designed protocols were utilized to check the individual effects of cells, bioproducts and interaction of these together on the oil/water and also fluids/rock interfaces. The final results showed that the cells of B. stearothermophilus SUCPM#14 adhere more into the oil/water interface compared to E. cloacae and change its rheological properties; e.g. its elastic properties which affect the ultimate microbial oil recovery efficiency. Eventually, contradicting results revealed that biosurfactant produced by E. cloacae was able to considerably reduce the interfacial tension and alter the wettability of the rock (to neutral conditions) while biosurfactant produced by B. stearothermophilus SUCPM#14 was not very effective.
Polyacrylamide (PAM) was used as a matrix material for fabricating novel nanocomposite hydrogels reinforced with natural chitosan nanofibers (CNFs) via in situ free-radical polymerization. The nanocomposite's structure, strength, morphology and rheological properties were investigated. The results showed that the CNFs had a strong interaction with PAM through hydrogen and covalent bondings. The CNFs acted as a multifunctional cross-linker and a reinforcing agent in the hydrogel system. The compression strength and storage modulus of the nanocomposite hydrogels were significantly higher than those of the pure PAM hydrogels and the corresponding PAM/chitosan semi-interpenetrating polymer network (PAM-SIPN) hydrogels. The swelling ratio (SR) of the nanocomposite hydrogels was lower than that of the PAM hydrogel, but was similar to that of the PAM-SIPN hydrogel. Among the CNF contents used, the 1.5 wt% CNF loading level showed the best combined swelling and mechanical properties for the hydrogels.
For the first time, chitosan-polyaniline/ZnO hybrids were prepared through a polymerization of aniline hydrochloride in the presence of ZnCl2 and chitosan. The hybrid materials were characterized by FT-IR, BET, SEM, UV-vis spectra and XRD analysis. From the BET and SEM micrographs, the introduction of ZnO nanoparticles into chitosan-polyaniline hybrid could obviously increase the porosity due to good possibility for dye adsorption. Adsorption experiments were carried out as a function of contact time, concentration of dye, adsorbent dosage and pH using reactive orange 16 as a model pollutant. The adsorption equilibrium data were fitted well to the Langmuir isotherm equation, with maximum adsorption capacity value was found to be 476.2mgg(-1). Adsorption kinetics was best described by the pseudo-second-order model agreed well with the experimental data and good correlation (R(2)>0.999). Photocatalytic degradation of dye under UV irradiation at pH 5.8 has also been examined. FT-IR spectrum clearly indicates that before adsorption of hybrid showed the functional groups of chitosan and polyaniline, whereas the dye adsorbed hybrid only present the dye molecules and ZnO. Based on the results of present investigation, the introduction of ZnCl2 into chitosan-polyaniline hybrid will enhance the adsorption of reactive dyes and photocatalytic degradation.
Low water solubility and hepatotoxicity limited the clinical use of 17-allylamino-17-demethoxy geldanamycin (17-AAG), an inhibitor of heat shock protein 90 (HSP90). Folate targeted polylactide-co-glycolide-polyethylene glycol-folic acid (PLGA-PEG-FA) nanoparticles containing 17-AAG were prepared and characterized. Cellular uptake and in vitro cytotoxicity of the prepared nanoparticles were determined in MCF-7 human breast cancer cells. The particle size of 17-AAG loaded folate targeted nanoparticles was 238.67±3.52 nm, drug loading was 8.25±2.49% and about 80% of drug was released from the nanoparticles over 10 days. Cellular uptake studies showed much higher intracellular uptake of folate targeted nanoparticle as compared to nontargeted nanoparticles. Cytotoxicity study showed 2 fold increase (P<0.05, n=3) in the cytotoxicity of folate targeted nanoparticle in comparison to free drug or nontargeted nanoparticles. Due to their targeting ability, nanometer size, high drug loading and controlled release behavior, 17-AAG loaded PLGA-PEG-FA nanoparticles might be developed as a targeted delivery system for breast and other cancer treatment.
Glioblastoma multiforme (GBM) is the most common and aggressive malignant primary brain tumor in human. 17-Allylamino-17-demethoxy geldanamycin (17-AAG) is an inhibitor of heat shock protein 90 (HSP90). The highly lipophilic nature and selective targeting of tumor cells makes 17-AAG a promising candidate for therapy of GBMs but poor water solubility, short biological half-life and hepatotoxicity limited its clinical use. Polymeric mixed micelles composed of Pluronic® P-123 and F-127 (2:1 (w/w)) containing 17-AAG were prepared and characterized. Cellular uptake and in vitro cytotoxicity of the prepared micelles were determined in U87MG human glioblastoma cells. The particle size of 17-AAG loaded Pluronic(®) P-123 and F-127 mixed micelles was 22.2 ± 0.1 nm; drug loading was about 4.0 ± 0.5% (w/w) with 88.2 ± 3.1% (w/w) encapsulation efficiency. About 90% of drug was released from the nanoparticles over 8 days. Cellular uptake studies showed intracellular uptake of mixed micelles. Cytotoxicity study showed 5-fold increase (P < 0.05, n = 3) in the cytotoxicity of 17-AAG-loaded mixed micelles to free 17-AAG. Due to their targeting ability, size, high drug loading and controlled release behavior, 17-AAG loaded Pluronic(®) P-123 and F-127 mixed micelles might be developed as a delivery system for GBM treatment.
ABA triblock copolymer, poly(ethylene glycol)-poly(butylene adipate)-poly(ethylene glycol) (PEG-PBA-PEG) could form micelle-like nanoparticles due to their amphiphilic characteristic. The mechanism of nanoparticle formation is based on the modified oil in water (O/W) emulsion by emulsifying acetone solution of copolymer containing different amount of dissolved 17beta-estradiol valerate (17beta-EV) in olive oil into the external water phase i.e. oil(1) and oil(2) in water (O & O/W). The acetone-evaporated nanodroplets involved an inner oil phase, an outer water phase and a solid shell between two phases, so called O/S/W nanocapsules. In order to control the nanocapsules size, the effect of various emulsion parameters such as the concentration of polymer, drug content, polymer/oil weight ratio and concentration of surfactant on morphology were studied. Particle size, their distribution and release profile of nanocapsules were investigated too. This study gave some important information about the structure, mechanism of formation, release profile and the physical properties (i.e. encapsulation efficiency, drug loading, nanoparticles yield) of nanocapsules. To investigate the influence of PEG chain on the stability of produced nanodroplets, nanocapsules were also prepared with and without Tween 80 as surfactant.
Gossypol, a secondary metabolite stored in the glands of cotton, protecting cottonseed from consumption of human and monogastric animal. This ability is unique to the tribe Gossypieae. Although the relationship between gossypol and pigment gland has been studied for a long time, the development mechanism of pigment gland has not been investigated at molecular level. Here we described a simple and efficient method for constructing a normalized cDNA library from a cotton mutant, Xiangmian-18, during its pigments gland forming stage. It combined switching mechanism at 5'-end of RNA transcript (SMART) technique and duplex-specific nuclease (DSN) normalization methods. In a model experiment, double-stranded cDNAs were synthesized from mRNAs, processed by normalization and Sfi I restriction endonuclease, and finally the cDNAs were ligated to pDNR-LIB vector. The ligation mixture was transformed into E. coli JM109 by electroporation. Counting the number of colonies, the titer of the original library was 5.86x10(5)cfu/ml in this library. Electrophoresis gel results indicated the fragments ranged from 800bp to 2kb, with the average size of 1400bp. Random picking clones showed that the recombination rate was 94%. The results showed that the cDNA library constructed successfully was a full-length library with high quality, and could be used to screen the genes related to development of pigments gland cottons.
Bolas surfactants can be inserted into bi-layers and may operate as permanent holes in such membranes. Significant synthetic work and an exhaustive characterisation of their properties in the bulk was performed. On this purpose, the phase diagram of the system composed by water and 1,16-hexadecanoyl-bis-(2-aminomethyl)-18-crown-6 (termed Bola A16) was investigated in a wide temperature and concentration range. No liquid crystalline phases were observed and a large micellar solution was present, up to about 50 surfactant wt%. Surface tension experiments defined adsorption and micelle formation. The low observed cmc value is important for pharmacological applications, in fact, considering intravenous administration, only micelles with low cmc value can exist in blood. Nuclear magnetic resonance experiments determined both water and surfactant self-diffusion. According to the aforementioned experiments, slight, if any, modifications in the structure of micelles were inferred on increasing Bola A16 content. Dynamic rheological experiments probed the solution micro-structure. The observed rheological behaviour is newtonian. The solution viscosity and the shear relaxation processes were rationalized assuming the presence of spherical aggregates, occurring up to high surfactant content. The viscometric behaviour was rationalised in terms of a former theory of flow as a cooperative phenomenon. The number of micelles coordinated each other during the viscous flow and the interaction strength between them was obtained as a function of Bola A16 concentration. Such value is close to unity and practically independent of surfactant content in the whole concentration range we investigated. This behaviour points out that little, or none, interactions among micellar aggregates occur. The absence of shear induced changes in the aggregate shape implies no change in drug delivery properties under flow, this is useful in the pharmaco-dynamics field, since drug delivery usually operates in mechanically stressed conditions. Thanks to the above properties, the material results particularly suitable for application in pharmaceutical field, may solubilize lipid membranes and selectively transport ions across them. Ancillary effects, such as the uptake of counter-ions in the crown ether, are to be considered.
Abnormal red blood cell (RBC) adhesion to endothelial cells (ECs) has been correlated with vascular complications in diseases such as sickle cell anemia and diabetes. Poloxamer 188 (P188) has been clinically tested to treat vaso-occlusion. However, the underlying mechanism(s) have not been clarified, making a methodical application difficult. In this study, we investigate how and to what extent P188 reduces RBC adhesion to ECs in plasma-like solutions. RBC adhesion to ECs is studied in solutions containing dextran, which is known to induce adhesion via macromolecular depletion interaction. It is demonstrated that P188 itself does not induce adhesion of normal RBCs to ECs but significantly reduces the adhesion in solutions containing high molecular mass-dextran. In addition, it is shown that P188 can reduce the adhesion of RBCs with enhanced exposure of phosphatidylserine (PS). Measurements of the electrophoretic mobility indicate that P188 increases the local viscosity inside the electric double layer of RBCs. Based on these results this study suggests that P188 reduces macromolecular depletion interaction, via penetrating into the depletion layer. Taking into consideration that dextran mimics the effects of pro-adhesive non-adsorbing plasma proteins and macromolecules, our study therefore suggests a mechanism for the adhesion reducing effect of P188 and should thus be of potential value for a detailed understanding of how cell-cell interactions in pathological conditions can be reduced.
In nanoparticulate engineering for drug delivery systems, poloxamers tri block copolymers are employed as adsorbing molecules to modify the aggregation state and impart stability to products. The aim was to prepare nanoparticles using poloxamer188 as stabiliser and investigate the mechanism of stabilisation of the prepared particles. Nanoparticles were prepared by solvent diffusion method with poloxamer 188 as stabiliser. Hydrodynamic thickness and zeta potential of the prepared nanoparticles were determined by photon correlation spectroscopy. To study the extent of adsorption of poloxamer onto the prepared nanoparticles, adsorption isotherms were constructed. The adsorbed amount of poloxamer 188 onto the particles was determined by depletion method. Macrophageal uptake study was performed to assess the uptake of the prepared nanoparticles using RAW 264.7 cell lines. Nanoparticles were prepared with slight increase in particle size and in absolute value of zeta potential compared to uncoated particles suggesting that this effect was due to adsorption of poloxamer 188. TEM studies and surface area analysis supported the results obtained from particle size analysis indicating preparation of particles with a thin layer of adsorbed poloxamer 188. Adsorption kinetics modeling suggested that at low concentrations (0.001-0.010g/L), Langmuir monolayer equation fits quite well and at higher concentrations (above 0.010g/L) multilayer adsorption of poloxamer 188 onto the prepared particles occurred. Thus the nanoparticles had multilayer of poloxamer 188 adsorbed onto the non uniform surface of PLGA. Results of macrophageal uptake and liver cell study exhibits adsorbed concentration dependent bypass of RES uptake of nanoparticles. Hence, results substantiate the application of adsorption isotherms for designing nanoparticles possessing potential to exhibit prolonged circulation when administered in vivo.
Current discovery demonstrates the rapid formation of gold nanoparticles with guavanoic acid a phytochemical of Psidium guajava (Pg). The pharmacological capabilities of the phytochemicals present in the leaves of Pg and their ability to generate gold nanoparticles is presented herein. The new genre of green nanoparticles exhibit remarkable Protein Tyrosine Phosphatase 1B (PTP 1B) inhibitory activity and in vitro stability in various physiological medium including saline, histidine, cysteine, bovine serum albumin (BSA), human serum albumin (HSA) and buffers (pH 5, 7 and 9). It is predicted that this new technology will be felt greatly in several routes of pharmaceuticals.
A low-resolution 1H NMR relaxometry study on the dynamics of an n-decane/water emulsion stabilized by beta-casein is presented. Spin-spin (transverse) relaxation time constants (T2) were used to assess relative mobilities of emulsion components, by a selective deuteration procedure. Data analysis allowed the emulsion investigated to be described by a heterogeneous collection of dynamically distinct populations. A major population of n-decane molecules presented an average mobility that very nearly approached that of pure solvent, which is compatible with its occurrence in the emulsion continuous microphase. beta-Casein molecules displayed a prevalent population with significantly decreased mobility as compared to the free protein in solution, which is in accordance with the protein location at the oil/water interface. Also, a major H2O population with significantly lower average T2 as compared to the pure liquid was detected and has been assigned to interfacial water.
In this work, the photochromic compound 1',3',3'-trimethyl-6-nitrospiro (2H-1-benzopyran-2,2'-indoline) (NOSP) was synthesized by a two step process. The photochromic properties of NOSP were investigated by ultraviolet-visible (UV-Vis) spectrophotometry. The results showed that NOSP was very sensitive to UV irradiation with absorption peaks at about 336 nm and 567 nm. Our hypothesis was that both photochromic nanofibers and photochromic living animal cells could be obtained by combining them with NOSP. To test the hypothesis, photochromic nanofibers were fabricated by electrospinning from various mixed solutions of NOSP and polymers (including a synthetic polymer of poly(methyl methacrylate) and a natural polymer of gelatin); NOSP/ethanol solution was dissolved in culture medium to stain pig iliac endothelial cells (PIEC) and endow them with photochromic capability. Polymer nanofibers from electrospinning were characterized by water contact angle measurements, ultraviolet-visible (UV-Vis) spectrophotometry and fluorescence microscopy. Morphology of photochromic PIEC was observed by fluorescence microscopy after being irradiated. It was shown that nanofibers from electrospun polymers and NOSP-treated PIEC had photochromic properties. The bio-toxicity of the photochromic compound was also evaluated and it was shown that ~50% of PIEC remained viable for at least 20 min. The photochromic compound NOSP could be a potentially powerful tool for development of multi-functional nanofibers and biological applications.
Many proteins form ordered irreversible structural aggregates called amyloid fibrils, which are associated with numerous neurodegenerative diseases. Insulin, a largely α-helical protein associated with type II diabetes, self-assembles to form amyloid fibrils in vitro. Insulin fibrillation goes through a number of intermediate phases that includes a soluble oligomeric phase believed to be the most toxic phase. Small molecules may play a very important role in modulating the fibrillation pathways. It is possible to induce and stabilize helix structures in proteins by a fluorinated alcohol 2,2,2-trifluoro ethanol (TFE). Since fibrillation process of many proteins is associated with conversion of α-helical structures into β-sheet configuration, we thought it would be interesting to study the effect of TFE on the fibrillation of insulin. In absence of TFE, soluble protofibrillar oligomeric intermediates formed directly from the insulin trimer. The protofibrillar aggregates transformed into mature fibrils over time. We demonstrated that although TFE did not prevent the appearance of matured amyloid fibrils, it prevented the appearance of soluble aggregates of insulin. TFE converted the insulin trimer into monomers and fibril formation proceeded from the monomeric state in a cooperative way avoiding the soluble oligomeric phase. At 25% TFE, distinct morphological changes resulting in more discrete fibrils were visible. The effect of the small molecule TFE on the avoidance of the formation soluble oligomeric state during fibrillation may have considerable implications in reducing cellular toxicity.
A water-soluble quaternary ammonium salt of chitosan, chitosan-N-hydroxy-2,3-propyl-N-methyl-N,N-diallylammonium methyl sulfate (MDAACS), was synthesized by reacting chitosan with methyl diallyl ammonium salt (MDAA). The results of water contact angle and swelling ratio showed that the membrane of MDAACS was more hydrophilic than chitosan. The antibacterial activities of MDAACS against Staphylococcus aureus and Klebsiella pneumoniae were evaluated with the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The results showed that the antibacterial activity of MDAACS was higher than that of chitosan. The cytocompatibility was evaluated in vitro with L929 fibroblast proliferation based on MTT colorimetric assay. The results showed that cell growth was much higher on MDAACS than on chitosan.
Biosorption of 2,4,6-trichlorophenol (2,4,6-TCP) from aqueous solution by biomass prepared from Acacia leucocephala bark, an agricultural solid waste has been investigated in the present study. All the experiments are carried out by batch mode technique. The resulting biosorbent was characterized by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) techniques. The effect of experimental parameters such as contact time, effect of pH (2-10), initial concentration of adsorbate (50-200 mg L(-1)) and amount of biosorbent dosage was evaluated. The removal was found to be pH dependent, and maximum removal was found to be at pH 5.0. The equilibrium time was found to be 3h. The biosorbent dose was increased, and the percentage removal of 2,4,6-TCP was increased, while the adsorption capacity at equilibrium q(e) (mg g(-1)) (amount of 2,4,6-TCP loaded per unit weight of adsorbent) decreased. Biosorption kinetic and isotherm studies showed the pseudo-second-order kinetics with a good correlation coefficient (R(2)=0.999), and both Langmuir and Freundlich isotherms were the best choices to describe the adsorption behaviors. The maximum monolayer biosorption capacity of A. leucocephala bark for 2,4,6-TCP was found to be 256.4 mg g(-1), at 30±1°C according to Langmuir model. This study demonstrated for the first time that the A. leucocephala bark could be an alternative for more costly adsorbents used for removal of 2,4,6-TCP from aqueous media.
The direct electrochemistry of hemoglobin (Hb) and application in its interaction with 3,4'-bis-(4-hydro-3-xycoumarin)-2,5-hexanediol (HCH) based on the Hb immobilized in chitosan-room temperature ionic liquid film were investigated by means of cyclic voltammetry, differential pulse voltammetry and amperometry. The binding ratio m and binding constant between HCH and Hb were estimated as 2 and 3.46 M(-1), respectively. The amperometric response showed a linear dependence on the concentration of HCH with the detection limit of 0.06 μM. In addition, the amperometry was a novel method in the field of binding studies and might be taken into account for future binding studies. At last a sensitive and convenient electrochemical method was proposed for the determination of HCH.
Aurein 2.5 (GLFDIVKKVVGAFGSL-NH(2)) is an uncharacterised antimicrobial peptide. At an air/water interface, it exhibited strong surface activity (maximal surface pressure 25mNm(-1)) and molecular areas consistent with the adoption of alpha-helical structure orientated either perpendicular (1.72nm(2)molecule(-1)) or parallel (3.6nm(2)molecule(-1)) to the interface. Aurein 2.5 was strongly antibacterial, exhibiting a minimum inhibitory concentration (MIC) of 30microM against Bacillus subtilis and Escherichia coli. The peptide induced maximal surface pressure changes of 9mNm(-1) and 5mNm(-1), respectively, in monolayers mimicking membranes of these organisms whilst compression isotherm analysis of these monolayers showed DeltaG(Mix)>0, indicating destabilisation by Aurein 2.5. These combined data suggested that toxicity of the peptide to these organisms may involve membrane invasion via the use of oblique orientated alpha-helical structure. The peptide induced strong, comparable maximal surface changes in monolayers of DOPG (7.5mNm(-1)) and DOPE monolayers (6mNm(-1)) suggesting that the membrane interactions of Aurein 2.5 were driven by amphiphilicity rather than electrostatic interaction. Based on these data, it was suggested that the differing ability of Aurein 2.5 to insert into membranes of B. subtilis and E. coli was probably related to membrane-based factors such as differences in lipid packing characteristics. The peptide was active against both sessile E. coli and Staphylococcus aureus with an MIC of 125microM. The broad-spectrum antibacterial activity and non-specific modes of membrane action used by Aurein 2.5 suggested use as an anti-biofilm agent such as in the decontamination of medical devices.
Heat pump systems using treated sewage water as the heat source were used in the Beijing Olympic Village for domestic heating and cooling. However, considerable biofouling occurred in the plate heat exchangers used in the heat pump system, greatly limiting the system efficiency. This study investigates the biofouling characteristics using a plate heat exchanger in parallel with a flow cell system to focus on the effect of calcium ions on the biofilm development. The interactions between the microorganisms and Ca(2+) enhances both the extent and the rate of biofilm development with increasing Ca(2+) concentration, leading to increased heat transfer and flow resistances. Three stages of biofouling development were identified in the presence of Ca(2+) from different biofouling mass growth rates with an initial stage, a rapid growth stage and an extended growth stage. Each growth stage had different biofouling morphologies influenced by the Ca(2+) concentration. The effects of Ca(2+) on the biofouling heat transfer and flow resistances had a synergistic effect related to both the biofouling mass and the morphology. The effect of Ca(2+) on the biofouling development was most prominent during the rapid growth stage.
Microbiologically influenced corrosion (MIC) of stainless steel 304 by a marine aerobic Pseudomonas bacterium in a seawater-based medium was investigated by atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS). AFM was used to observe in situ the proliferation of a sessile Pseudomonas cell by binary fission. The development of a biofilm on the coupon surface and the extent of corrosion damage beneath the biofilm after various exposure times were also characterized by AFM. Results showed that the biofilm formed on the coupon surface increased in thickness and heterogeneity with time, and thus resulting in the occurrence of extensive micro-pitting corrosion; whilst the depth of pits increased linearly with time. The XPS results confirmed that the colonization of Pseudomonas bacteria on the coupon surface induced subtle changes in the alloy elemental composition in the outermost layer of surface films. The most significant feature resulting from microbial colonization on the coupon surface was the depletion of iron (Fe) and the enrichment of chromium (Cr) content as compared to a control coupon exposed to the sterile medium, and the enrichment of Cr increased with time. These compositional changes in the main alloying elements may be correlated with the occurrence of extensive micropitting corrosion on the surface.
In our previous study, the degree of adsorption of 9 representative antimicrobial agents to Toraymyxin(®) PMX-F sheets was quantitatively evaluated . As a result, the adsorption rate was 22.1% for Linezolid in the presence of serum. Therefore, we investigated whether two types of antimicrobial agents (Ciprofroxacin and Linezolid) can be better adsorbed on PMX-F sheets. When the number of PMX-F sheets was increased in a step wise manner, specifically 2, 4, 6, 8 and 12, the adsorption rate increased linearly. In addition, the adsorption to polymyxin-B immobilized fiber (Toraymyxin(®) PMX-20R) cartridges, widely used to remove endotoxins from circulating blood in the treatment of sepsis, was quantitatively evaluated. As a result, in the presence of serum, Linezolid showed adsorption to PMX-20R, and the adsorption rate after 2h was 54.5%, and that after 4h was 65.8%. The results of this study suggest the necessity of monitoring blood antimicrobial concentration during treatment for sepsis with Linezolid, which showed adsorption to PMX-20R in an environment close to a clinical environment.
Drug delivery systems using vesicular carriers such as liposomes or niosomes, have distinct advantages over conventional dosage forms because the vesicles can act as drug containing reservoirs and the modification of the vesicular compositions or surface properties can adjust the drug release rate and/or the affinity for the target site. In recent years, niosomes have been the object of growing scientific attention as an alternative potential drug delivery system to conventional liposomes. The aim of present work was firstly to determine the critical micelle concentration (CMC) and then to analyze the capability of polysorbate 21 (Tween 21) to form niosomal formulations. Non-ionic surfactant vesicles were prepared using Tween 21 and cholesterol (CHOL) at equimolar ratio (15 mM:15 mM) by employing the "film" technique. Cholesterol was used to complete the hydrophobic moiety of single alkyl chain non-ionic surfactant for vesicle formation. Dynamic light scattering was used to determine the size, zeta (ζ)-potential, polydispersity index and colloidal stability of the niosomal formulation. The vesicles were also characterized for their microviscosity and pH-sensitivity using fluorescent probes. The present work led to a simple, but positive result in pharmaceutical technology area. In particular, we have shown that the Tween 21:CHOL vesicles (i) are a homogenous and monodisperse vesicular population; (ii) are characterized by dimension compatible with the transport of drugs across biological barriers especially those whose diameter is about 100nm; (iii) shows a good stability at least 90 days at 4°C and (iv) are pH-sensitive systems. In conclusion, this niosomal formulation could be used as pH-sensitive nanodevices for delivery of drugs to pathological tissues, which exhibit an acidic environment as compared to normal tissues.
The transfer of 22-NBD-cholesterol (22-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-23,24-bisnor-5-cholen-3-ol) between two liposome membranes was quantitatively analyzed by using the fluorescence resonance energy transfer (FRET) method. Liposomes labeled with both 22-NBD-cholesterol and a rhodamine-labeled phosphatidylethanolamine (Rh-DHPE) were used as donor liposomes, and the 22-NBD-cholesterol transfer from these donor liposomes to acceptor liposomes prepared from same type of phosphatidylcholine was monitored. The transfer kinetics was found to be composed of a fast and a slow phase, and all kinetic measurements could be fitted with a bi-exponential model. The results obtained indicate that the 22-NBD-cholesterol transfer kinetics between liposome membranes depends on the fluidity of the liposome used and that the curvature may affect the kinetics. Furthermore, the behavior of 22-NBD-cholesterol in lipid membrane is similar to that of the oxysterol 25-hydroxycholesterol rather than cholesterol. It is proposed that 22-NBD-cholesterol can be a useful fluorescent probe to mimic the intermembrane transfer of oxidized cholesterols like 25-hydroxycholesterol, rather than that of cholesterol itself.
Graphene is a novel carbon-based material widely studied in bio-electrochemical fields because of its high electrical conductivity and excellent electrocatalytic activity. However, its biological applications have been limited due to the lack of understanding of its compatibility with numerous biological entities. In this paper, cytoxicities of MDA-MB-231 breast cancer cells (MDA cells) on carbon paste (CP) and graphene-carbon paste (GCP) substrates are assessed. GCP was prepared by mixing graphene powder into carbon paste with different graphene contents. Cytotoxic effect was evaluated from cell viability, cell adhesion, ROS production and fluorescence staining studies. Cell viability on GCP substrate was found to initially increase as graphene content increases from 0 to 2.5wt% but then decrease as the content increases further. In addition, the viability decreases with time for all substrates. Similarly, graphene concentration affected the number of adherent cells in the same manner as the cell viability. Likewise, reactive oxygen species (ROS) induced by carbon substrate increased with time and decreased with small graphene inclusion, confirming that low graphene content led to lower cytotoxicity. Moreover, confluence of MDA cells on substrate evaluated using Hoechst 33342 fluorescence staining was also found to be enhanced at low graphene concentration. Therefore, low-content graphene incorporation can effectively improve biocompatibility of carbon-based materials with MDA-MB-231 breast cancer cells, enabling potential applications such as electrochemical electrode for cell study.