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Fasciolosis is a zoonotic parasitic disease that seriously endangers the development of animal husbandry and human health. In order to develop a rapid serological diagnostic method for fasciolosis in ruminants, the CatL1D and CatB4 genes of Fasciola hepatica were amplified by reverse transcription polymerase chain reaction (PCR) and cloned, respectively, and then the CatL-B fusion gene (MeCatL-B) was constructed by gene splicing by overlap extension PCR technique. The recombinant rCatL1D, rCatB4 and rMeCatL-B proteins were then prepared by prokaryotic expression, respectively, and the recombinant protein with high specificity and sensitivity was screened via indirect enzyme-linked immunosorbent assay. Using the selected recombinant protein rCatL1D as a diagnostic antigen, we developed a colloidal gold immunochromatographic assay (CGIA) for detecting F. hepatica-specific antibodies, and 426 serum samples of slaughtered sheep were used to evaluate the sensitivity and specificity of F. hepatica CGIA assay. The results showed that the sensitivity and specificity of rCatL1D protein (100%, 96.67%) were higher than those of rCatB4 (94.29%, 80%) and rMeCatL-B (91.43%, 90%). Compared with the gold standard post-mortem inspection, the specificity and sensitivity of the CGIA method was 100% and 97%, respectively, and the consistency rate between these two methods was 99.3%. These results confirmed that the CGIA method based on rCatL1D protein could be a promising approach for rapid diagnosis of sheep fasciolosis because of its high sensitivity and specificity.
Current available methods to detect cow milk adulteration or accidental contamination of goat milk are both laborious and time consuming. The aim of this technical research communication was to develop a simple, rapid, specific and sensitive method for quantitative detection of cow milk in goat milk. A competitive lateral flow immunoassay (LFIA) strip was developed using a specific monoclonal antibody (mAb) labeled with colloidal gold nanoparticles (GNPs) for specifically binding to cow milk casein. The detection limit of this rapid detection was 0.07% of cow milk in goat milk, providing equal specificity and higher sensitivity when compared with a commercial enzyme-linked immunosorbent assay (ELISA). These result suggest that the established rapid GNPs-LFIA strip could be used for monitoring cow milk adulteration/contamination of goat milk.
This chapter deals with protein aggregation, which is a key issue in biopharmaceutical processes. Several experimental techniques to characterize the aggregate size and content are presented and fundamentals on the kinetic modelling of aggregation mechanisms are provided. The impact of operating conditions on the aggregation rate is reviewed and the steps critical for aggregate formation in biopharmaceutical processes are identified. Finally, methods aiming at reducing the aggregate content are proposed. These methods focus either on improving protein stability or on removing the formed aggregates. The former can be achieved by synthesizing aggregation-resistant proteins, tuning operating conditions, or designing processes with a shorter residence time (e.g. perfusion bioreactors or counter-current chromatography). The latter method is mainly achieved by filtration and chromatography. In particular, the simulated moving bed process is shown to be very advantageous for aggregate removal with size exclusion chromatography: it allows improving productivity, decreasing eluent consumption and increasing the outlet protein concentration as compared to single-column processes.
The occurrence of pesticidal pollution in the environment and the resistance in the mosquito species makes an urge for the safer and an effective pesticide. Permethrin, a poorly water-soluble pyrethroid pesticide, was formulated into a hydrodispersible nanopowder through rapid solvent evaporation of pesticide-loaded oil in water microemulsion. Stability studies confirmed that the nanopermethrin dispersion was stable in paddy field water for 5 days with the mean particle sizes of 175.3 ± 0.75 nm and zeta potential of −30.6 ± 0.62 mV. The instability rate of the nanopermethrin particles was greater in alkaline (pH 10) medium when compared with the neutral (pH 7) and acidic (pH 4) dispersion medium. The colloidal dispersion at 45°C was found to be less stable compared with the dispersions at 25 and 5°C. The 12- and 24-h lethal indices (LC50) for nanopermethrin were found to be 0.057 and 0.014 mg l−1, respectively. These results were corroborative with the severity of damages observed in the mosquito larvae manifested in epithelial cells and the evacuation of the midgut contents. Further, the results were substantiated by the decrease in cellular biomolecules and biomarker enzyme activity in nanopermethrin treated larvae when compared to bulk and control treatment.
A free standing 2D PS colloidal crystal with Au nanoshells/hydrogel composite film (CAuHCF) was fabricated by embedding a 2D PS colloidal crystal with Au nanoshells into a polyacrylic acid (PAA) hydrogel film. This CAuHCF can act as a visualized sensor with high diffraction intensity. The 2D PS colloidal crystal with Au nanoshells was prepared by depositing an Au layer on PS colloidal crystal obtained by interfacial self-assembly. The diffraction intensity of the CAuHCF was increased by about 30-fold than that of traditional 2D PS colloidal crystal/hydrogel composite film on transparent substrate due to large scattering cross section of Au shell. Such sensors based Au nanoshells array with the simple preparation process and the strong diffraction signal are promising ones for practical applications in visual detection. Additionally, with the simple preparation process and high diffraction intensity, other visualized sensors based different hydrogel matrix and the 2D PS colloidal crystal with Au nanoshells could be synthesized for monitoring various analysts.
Caprine and bovine milks have a similar overall gross composition, but vary considerably in the ratios of their casein components. These differences in colloidal casein micelles could affect directly or indirectly the heat stability of caprine and bovine milks at their natural pH. In the present work, the differences in colloidal stability of caprine and bovine milk have been studied by analysing the effect of heat treatment and skimming on precipitation of proteins. Raw and heated milk samples (70 °C/5 min, 80°C/5 min and 90°C/5 min) were centrifuged at 600, 2000, and 4500 g. The amount of precipitate formed after skimming was measured and the protein composition of both precipitates and supernatants analysed using the SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis) and densitometry. In caprine milk, the heat treatment prior to skimming had a statistically significant effect on protein precipitation. Centrifugal force had a statistically significant effect on amount of precipitate for both milks, but the amount was 2 to 4 times higher for caprine milk. When defatting the milk for electrophoresis, a centrifugal force of 600 g appeared to be the most appropriate, in order to avoid protein loss and a possible error in the interpretation of results. Results of this study could also serve as the basis for further investigations on adjusting the skimming conditions for caprine milk in industrial dairy processing environment.
The total calcium content of cheese, along with changes in the equilibrium between soluble and casein (CN)-bound calcium during ripening can have a major impact on its rheological, functional and textural properties; however, little is known about the effect of other alkaline earth metals. NaCl was partially substituted with MgCl2 or SrCl2 (8·7 and 11·4 g/kg curd, respectively) at the salting stage of cheesemaking to study their effects on cheese. Three cheeses were produced: Mg supplemented (+Mg), Sr supplemented (+Sr) and a control Cheddar cheese. Ca, Mg and Sr contents of cheese and expressible serum obtained therefrom were determined by atomic absorption spectroscopy. Addition of Mg2+ or Sr2+ had no effect on % moisture, protein, fat and extent of proteolysis. A proportion of the added Mg2+ and Sr2+ became CN-bound. The level of CN-bound Mg was higher in the +Mg cheese than the control throughout ripening. The level of CN-bound Ca and Mg decreased during ripening in all cheeses, as did % CN-bound Sr in the +Sr cheese. The presence of Sr2+ increased % CN-bound Ca and Mg at a number of ripening times. Adding Mg2+ had no effect on % CN-bound Ca. The +Sr cheese exhibited a higher G′ at 70 °C and a lower LTmax than the control and +Mg cheeses throughout ripening. The +Sr cheese had significantly lower meltability compared with the control and +Mg cheeses after 2 months of ripening. Hardness values of the +Sr cheese were higher at week 2 than the +Mg and control cheeses. Addition of Mg2+ did not influence the physical properties of cheese. Supplementing cheese with Sr appeared to have effects analogous to those previously reported for increasing Ca content. Sr2+ may form and/or modify nanocluster crosslinks causing an increase in the strength of the para-casein matrix.
The mainstream commercialization of colloidal quantum dots (QDs) for light-emitting applications has begun: Sony televisions emitting QD-enhanced colors are now on sale. The bright and uniquely size-tunable colors of solution-processable semiconducting QDs highlight the potential of electroluminescent QD light-emitting devices (QLEDs) for use in energy-efficient, high-color-quality thin-film display and solid-state lighting applications. Indeed, this year’s report of record-efficiency electrically driven QLEDs rivaling the most efficient molecular organic LEDs, together with the emergence of full-color QLED displays, foreshadow QD technologies that will transcend the optically excited QD-enhanced products already available. In this article, we discuss the key advantages of using QDs as luminophores in LEDs and outline the 19-year evolution of four types of QLEDs that have seen efficiencies rise from less than 0.01% to 18%. With an emphasis on the latest advances, we identify the key scientific and technological challenges facing the commercialization of QLEDs. A quantitative analysis, based on published small-scale synthetic procedures, allows us to estimate the material costs of QDs typical in light-emitting applications when produced in large quantities and to assess their commercial viability.
Biodegradable chitosan/bentonite composites with three different compositions were synthesized by the intercalation method using cetyltrimethylammonium bromide as the cationic surfactant. The composites were characterized using conductivity, density, particle size measurements, FTIR, TGA, XRD and SEM methods. Colloidal stabilities of the suspensions prepared in silicone oil (SO) were observed to increase with decreasing density. The effects of dispersed particle concentration, shear rate, electric field strength, electric field frequency and temperature on the electrorheological (ER) activities of the suspensions were investigated. The electric field viscosities of the suspensions showed typical shear thinning non-Newtonian viscoelastic behaviour. Yield stresses of the suspensions were observed to change in proportion to the square of applied electric field (E). Further, according to creep and creep-recovery analysis, reversible viscoelastic deformations were observed in the suspensions under E ≠ 0 kV mm–1.
The effect of bio-conjugation of CdSe/ZnS core-shell quantum dots (QDs) with Interleukin 10 (IL-10) antibodies on the aging of photoluminescence (PL) spectra of the QDs was investigated. The aging occurred upon storage of QDs for about 2 years or thermal annealing at 190 oC for up to 12 hours at atmospheric ambience and consisted in “blue” shifting the PL band position, increasing a PL band half-width and decreasing the PL intensity. The bio-conjugation is found to promote PL aging. The aging upon storage is attributed to the oxidation that decreases the QD core dimension, while the aging upon thermal annealing can be due to both oxidation and alloying of CdSe core and ZnS shell.
A green and mild synthesis of colloidal zinc oxide nanocrystals in ethanol/dimethylformamide mixtures was introduced which allows to produce stable crystalline ZnO particles and tailor their average size in the range of 2.8−4.5 nm by varying temperature and duration of post-synthesis ageing. An increase in dimethylformamide fraction in the mixture results in acceleration of ZnO nanocrystals ripening. Colloidal ZnO nanocrystals emit broadband photoluminescence in the range of 2−3 eV with the quantum yields of up to 12 %.
The paper presents a comparative study of the photoluminescence (PL) and Raman scattering spectra of core-shell CdSe/ZnS quantum dots (QDs) in nonconjugated states and after the conjugation to the anti-human papilloma virus (HPV), HPV 16-E7, antibodies. All optical measurements are performed on the dried droplets of the original solution of nonconjugated and bioconjugated QDs located on the Si substrate. CdSe/ZnS QDs with emission at 655 nm have been used. PL spectra of nonconjugated QDs are characterized by one Gaussian shape PL band related to the exciton emission in the CdSe core. PL spectra of bioconjugated QDs have changed essentially: the core PL band shifts into the high energy spectral range (“blue” sift) and becomes asymmetric. A set of physical reasons has been proposed for the “blue” shift explanation of the core PL band in bioconjugated QDs. The variation of PL spectra versus excitation light intensities has been studied to analyses the exciton emission via excited states in QDs. Finally the PL spectrum transformation for the core emission in bioconjugated QDs has been attributed to the electronic quantum confined effects stimulated by the electric charges of bioconjugated antibodies.
Bio-conjugated CdSe/ZnS core/shell quantum dots (QDs) attract essential scientific interest due to their possible nano-medicine applications, including selective highlighting of affected tissues and targeted drug delivery to the certain type of cells. The paper is focused on the theoretical description of the blue shift observed in the luminescence spectra of CdSe/ZnS QDs upon their bio-conjugation with the anti-interleukin-10 antibodies. We propose a model that describes the ground state of the exciton confined in a quantum dot and explaining the bio-conjugation phenomenon by the change of the effective confinement volume.
Colloidal β-In2S3 quantum dots stabilized in water by a number of polymers or sodium polyphosphate and mercaptoacetate were synthesized. An increase in the stabilizer content was found to result in a decrease in the average dot size from 20–30 to 5–10 nm and formation of a narrow absorption band centered at 290 nm. The position and spectral width of the band were found to be independent on stabilizer concentration, synthesis temperature and molar In:S ratio. The band was assumed to belong to a molecular cluster smaller than 1 nm which is a precursor for formation of larger regular indium sulfide quantum dots.
Cadmium sulfide nanoparticles were synthesized by a microwave-assisted route in aqueous dispersion. The cadmium sulfide nanoparticles showed an average diameter around 5 nm and a cubic phase corresponding to hawleyite. The aqueous dispersions of the nanoparticles were characterized by UV-Vis spectroscopy, luminescence analysis, transmission electron microscopy and X-ray diffraction. The addition of sodium hydroxide solutions at different concentrations causes a red-shift in the wavelength of the first excitonic absorption peak of the cadmium sulfide nanoparticles, indicating a reduction of the band gap energy. Besides, the intensity of the luminescence of the nanoparticle dispersions was increased. However, there is a threshold concentration of the hydroxide ion above which the precipitation of the cadmium sulfide nanoparticles occurs.
Cystic fibrosis (CF) is an inherited childhood-onset life-shortening disease. It is characterized by increased respiratory production, leading to airway obstruction, chronic lung infection and inflammatory reactions. The most common bacteria causing persisting infections in people with CF is Pseudomonas aeruginosa. Superparamagnetic Fe3O4 iron oxide nanoparticles (NPs) conjugated to the antibiotic (tobramycin), guided by a gradient of the magnetic field or subjected to an oscillating magnetic field, show promise in improving the drug delivery across the mucus and P. aeruginosa biofilm to the bacteria. The question remains whether tobramycin needs to be released from the NPs after the penetration of the mucus barrier in order to act upon the pathogenic bacteria. We used a zero-length 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC) crosslinking agent to couple tobramycin, via its amine groups, to the carboxyl groups on Fe3O4 NPs capped with citric acid. The therapeutic efficiency of Fe3O4 NPs attached to the drug versus that of the free drug was investigated in P. aeruginosa culture.
The application of palladium nanoparticles as electron-dense markers for labeling in both transmission and scanning electron microscopy requires their conjugation to a specific protein. The conjugation protocol described here includes the dihydrolipoic acid (DHLA) capping of Pd nanoparticles (8 nm equivalent diameter) and their subsequent covalent attachment to functional protein molecules such as streptavidin, protein A, or avidin. The single-step reaction was mediated using the cross-linking agent ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC). The final Pd conjugates were fully functional, as demonstrated by labeling of ultrathin resin sections of either bovine serum albumin or secretory granules of the salivary gland isolated from the partially fed female Ixodes ricinus tick. The results of bovine serum labeling were quantified, statistically evaluated, and compared with results obtained using commercially available gold particle conjugates (10 nm diameter). The highest values of labeling density were achieved using both streptavidin-Pd (106 ± 7 particles/μm2) and protein A-Au conjugates (130 ± 18 particles/μm2) compared to a commercial streptavidin-Au (66 ± 16 particles/μm2) and protein A-Pd conjugates (70 ± 11 particles/μm2). The concentrations of both DHLA and EDC, pH during conjugation, and finally thorough washing away of unbound proteins crucially influenced conjugation.
Introduction. Many desirable properties make cloudy fruit juice widely produced according to consumer acceptance. The pulp of mango (Mangifera indica L.), a good source of carbohydrates and water, has many uses including the production of juice. Nowadays, the use of food enzymes has become widespread in the juice industry for various reasons. However, the application of enzymes has not yet been tested in the context of the production of cloudy mango juice, except for the stabilization of nectars. Thus, the aim of our study was to examine and optimize some parameters involved in the enzymatic production of cloudy mango juice. Materials and methods. To achieve our objective, we used Response Surface Methodology (RSM) by combining five factors: time (30–50 min), temperature (35–55 °C), pH 4–5.5, Pectinexr Ultra SP-L (20–40 μL·L-1) and Cellubrixr L (15–25 μL· L-1). Results and discussion. The study of juice yield, cloud stability and the browning index showed that the coefficients of determination (R2 values) of these traits with the studied parameters were greater than 0.900. The optimized juice yield, cloud stability and browning index varied with specific values chosen for the five parameters mentioned above. Conclusions. Finally, the use of 30 μ L·L-1 Pectinexr Ultra SP-L and 22 μL·L-1 Cellubrix at 45 °C, for 43 min, at pH 5.5, was found to be the optimum set of conditions for processing cloudy mango juice.
The immunogold technique is a valuable method for labeling cellular macromolecules. However, multiple labeling using colloidal gold (cAu) nanoparticles of different sizes presents certain drawbacks; namely, as particle size increases, there is a decreased labeling efficiency and diminished spatial resolution with respect to the locations of labeled epitopes. Both concerns also limit the utility of heavy metal particles for comparative analysis of labeling densities. To minimize the variables due to differential labeling efficiencies, the best solution would be to conduct multiple labeling with particles of similar size. Consequently, some parameter other than size is necessary to distinguish each label type. In this study, we report the synthesis of colloidal palladium (cPd) nanoparticles of similar size but having two distinct shapes, umbonate and faceted, which are readily distinguishable from spherical colloidal gold particles. Their utility and fidelity as labels using a human platelet whole-mount model is also demonstrated.
In this paper, we develop an efficient procedure to solve for the Stokesian fields around a spherical particle in viscous fluid bounded by a cylindrical confinement. We use our method to comprehensively simulate the general creeping flow involving the particle-conduit system. The calculations are based on the expansion of a vector field in terms of basis functions with separable form. The separable form can be applied to obtain general reflection relations for a vector field at simple surfaces. Such reflection relations enable us to solve the flow equation with specified conditions at different disconnected bodies like the sphere and the cylinder. The main focus of this article is to provide a complete description of the dynamics of a spherical particle in a cylindrical vessel. For this purpose, we consider the motion of a sphere in both quiescent fluid and pressure-driven parabolic flow. Firstly, we determine the force and torque on a translating-rotating particle in quiescent fluid in terms of general friction coefficients. Then we assume an impending parabolic flow, and calculate the force and torque on a fixed sphere as well as the linear and angular velocities of a freely moving particle. The results are presented for different radial positions of the particle and different ratios between the sphere and the cylinder radius. Because of the generality of the procedure, there is no restriction in relative dimensions, particle positions and directions of motion. For the limiting cases of geometric parameters, our results agree with the ones obtained by past researchers using different asymptotic methods.