Do hydration dynamics follow the structural perturbation during thermal denaturation of a protein: a terahertz absorption study

We investigate the thermal denaturation of human serum albumin and the associated solvation using terahertz (THz) spectroscopy in aqueous buffer solution. Far- and near-ultraviolet circular dichroism spectroscopy reveal that the protein undergoes a native (N) to extended (E) state transition at temperature ≤55°C with a marginal change in the secondary and tertiary structure. At 70°C, the protein transforms into an unfolded (U) state with significant irreversible disruption of its structures. We measure the concentration- and temperature-dependent THz absorption coefficient (α) of the protein solution using a p-Ge THz difference spectrometer (2.1–2.8 THz frequency range), thereby probing the collective protein-water network dynamics. When the solvated protein is heated up to 55°C and cooled down again, a reversible change in THz absorption is observed. When increasing the temperature up to 70°C, we find a dramatic irreversible change of THz absorption. The increase in THz absorption compared to bulk water is attributed to a blue shift in the spectrum of the solvated protein compared to bulk water. This is supported by measurements of THz absorption coefficients using THz time-domain spectroscopy (0.1–1.2 THz frequency range). We also use picosecond-resolved fluorescence spectroscopy of the tryptophan 214 moiety of human serum albumin. All experimental observations can be explained by a change in the hydration dynamics of the solvated protein due to the additional exposure of hydrophobic residues upon unfolding.     

Label-free amplified bioaffinity detection using terahertz wave technology

A new affinity biosensor based on pulsed terahertz (THz) wave technology has been used to monitor binding between biotin and avidin molecules. Amplified detection of avidin-biotin binding is obtained on supported membranes composed of biotin layers on quartz surface, which is modified with octadecanol. Agarose particles are conjugated with avidin and then applied to biotin, which is already bound to the octadecanol quartz surface, the biotin binds to the conjugate rapidly and causes an enhancement of the THz difference signal between biotin and biotin-avidin complexes by a factor greater than eight fold when compared to the same sample without agarose beads. The technique was able to detect less than 10.3 ng/cm2 avidin, thus, giving the THz system a detection capability of sub-thin solid films better than ellipsometry and reflectometry techniques. Further improvement is underway using highly refractive beads together with appropriate surface chemistry. This newly developed method is being saliently optimized for future application, including the detection of DNA hybridization and ligand-analyte affinity binding.     

Tunable Salisbury Screen Absorber Using Square Lattice of Plasmonic Nanodisk

We propose a novel polarization independent Salisbury screen absorber to provide tunable resonant absorption at terahertz (THz) frequencies. The Salisbury screen absorber is designed by using a planar array of thin gold nanodisks arranged in a square lattice. Certain configurations of Salisbury screen have multiple distinctive absorption bands that support near-unity/FWHM absorption bandwidth reaching 36 THz/169 THz, respectively. Moreover, the absorption bandwidth depends upon the optical thickness of the dielectric spacer between the metasurface and the metallic ground plane. The proposed tunable Salisbury screen absorber can find practical applications in photonic detection, imaging, sensing, and solar cells at optical frequencies.     

Terahertz Plasmonic Quantum Cascade Lasers: a Comprehensive Physics Study

Generation of terahertz (THz) radiation has been a hot research topic in recent years. Plasmonic quantum cascade lasers (QCLs) are among the most compact and efficient sources to generate THz radiation. In this paper, we comprehensively study plasmonic QCLs designed based on the antenna-feedback structure to generate efficient radiation about the center frequency of 3 THz. By changing the geometric structure of the plasmonic cavity and using two-dimensional simulation, a minimum loss less than 5.9 cm^?1 is achieved at the lasing frequency. It is also possible to control the orientation of the output beam either vertically or tilted by changing the geometry of the antenna design via chirped or non-chirped grating scheme. Moreover, the output characteristics of the QCL are simulated based on the three-level rate equations through which the dynamics of the laser, as well as the P-I curve, are investigated. Also, the gain spectra for two laser designs (with chirped and non-chirped gratings) are simulated and compared to each other. The results of this paper may provide deep insight into designing efficient laser sources in the THz region.

     

New dinuclear copper(II) and zinc(II) complexes for the investigation of sugar–metal ion interactions

We have studied the binding interactions of biologically important carbohydrates (d-glucose, d-xylose and d-mannose) with the newly synthesized five-coordinate dinuclear copper(II) complex, [Cu₂(hpnbpda)(μ-OAc)] (1) and zinc(II) complex, [Zn₂(hpnbpda)(μ-OAc)] (2) [H₃hpnbpda = N,N′-bis(2-pyridylmethyl)-2-hydroxy-1,3-propanediamine-N,N′-diacetic acid] in aqueous alkaline solution. The complexes 1 and 2 are fully characterized both in solid and solution using different analytical techniques. A geometrical optimization was made of the ligand H₃hpnbpda and the complexes 1 and 2 by molecular mechanics (MM+) method in order to establish the stable conformations. All carbohydrates bind to the metal complexes in a 1:1 molar ratio. The binding events have been investigated by a combined approach of FTIR, UV–vis and ¹³C NMR spectroscopic techniques. UV–vis spectra indicate a significant blue shift of the absorption maximum of complex 1 during carbohydrate coordination highlighting the sugar binding ability of complex 1. The apparent binding constants of the substrate-bound copper(II) complexes have been determined from the UV–vis titration experiments. The binding ability and mode of binding of these sugar substrates with complex 2 are indicated by their characteristic coordination induced shift (CIS) values in ¹³C NMR spectra for carbon atoms C1, C2, and C3 of sugar substrates.     

Biotin and Zn2+ Increase Xylitol Production by Candida tropicalis

In this study, the medium requirements to increase the production of xylitol by using Candida tropicalis (CT) have been investigated. The technique of single addition or omission of medium components was applied to determine the nutritional requirements. The addition of amino acids such as Asp, Glu, Gln, Asn, Thr, and Gly had no significant effect on CT growth. However, in the absence of other metal ions, there was a higher concentration of cell growth and xylitol production when only Zn²⁺ was present in the medium. The analysis of various vitamins unveiled that biotin and thiamine were the only vitamins required for the growth of CT. Surprisingly, when only biotin was present in the medium as a vitamin, there was less growth of CT than when the medium was complete, but the amount of xylitol released was significantly higher. Overall, this study will increase the xylitol production using the single omission or addtion technique.     

The biotin requirement of HeLa cells

We have examined the effect of alterations in the biotin content of the medium on the growth, viability, biotin content, and the activities of biotin-dependent and biotin-independent enzymes of the HeLa cells. The inclusion in the growth medium of avidin, which almost irreversibly binds with biotin (K_d, 10^?15 M), results in an increase in cellular biotin content and biotin enzyme activity over that seen when the cells are grown in a biotin-depleted medium. The addition of avidin-bound biotin to the growth medium led to a forty-fold increase in cellular biotin when compared to the inclusion of an equivalent amount of free biotin in the medium. HeLa cells are able to internalize avidin-bound biotin. Biotin is released from this complex to function as the prosthetic group of biotin enzymes. HeLa cells do have a nutritional requirement for biotin.     

Multiband Terahertz Metamaterial Absorber Based on Multipolar Plasmonic Resonances

Terahertz metamaterial absorbers (MMA) have found wide scope of research prospective, remarkably in the development of multiband absorbers. Considerable applications are established using these multiband absorbers in THz imaging, wireless communication and bolometric detectors. The MMA was built on a GaAs substrate of 30 µm thickness and the hexagonal metallic pattern was etched out on a gold layer of 0.4 µm thickness on the top surface. The underlying ground layer is metallic backed. This design realizes the multiband (9-bands) of absorption in the spectral region from 0.56 to 0.92 THz. The multiband absorption mechanism of the absorber was examined by electric field dispersion analysis and impedance matching concept. From the established results, the absorber exhibits nine bands within a narrow frequency range and secures promising applications in hyperspectral imaging, clinical sensing and detection.

     

Active Focal Length Control of Terahertz Slitted Plane Lenses by Magnetoplasmons

Active plasmonic devices are mostly designed at visible frequencies. Here, we propose an active terahertz (THz) plasmonic lens tuned by an external magnetic field. Unlike other tunable devices where the tuning is achieved by changing the plasma frequency of materials, the proposed active lens is tuned by changing the cyclotron frequency through manipulating magnetoplasmons (MPs). We have theoretically investigated the dispersion relation of MPs of a semiconductor?Cinsulator?Csemiconductor structure in the Voigt configuration and systematically designed several lenses realized with a doped semiconductor slab perforated with sub-wavelength slits. It is shown through finite?Cdifference time?Cdomain simulations that THz wave propagating through the designed structure can be focused to a small size spot via the control of MPs. The tuning range of the focal length under the applied magnetic field (up to 1?T) is ??3??, about 50% of the original focal length. Various lenses, including one with two focal spots and a tunable lens for dipole source imaging, are realized for the proposed structure, demonstrating the flexibility of the design approach. The proposed tunable THz plasmonic lenses may find applications in THz science and technology such as THz imaging.     

Peptide nucleic acid probe for protein affinity purification based on biotin–streptavidin interaction and peptide nucleic acid strand hybridization

We describe a new method for protein affinity purification that capitalizes on the high affinity of streptavidin for biotin but does not require dissociation of the biotin–streptavidin complex for protein retrieval. Conventional reagents place both the selectively reacting group (the “warhead”) and the biotin on the same molecule. We place the warhead and the biotin on separate molecules, each linked to a short strand of peptide nucleic acid (PNA), synthetic polymers that use the same bases as DNA but attached to a backbone that is resistant to attack by proteases and nucleases. As in DNA, PNA strands with complementary base sequences hybridize. In conditions that favor PNA duplex formation, the warhead strand (carrying the tagged protein) and the biotin strand form a complex that is held onto immobilized streptavidin. As in DNA, the PNA duplex dissociates at moderately elevated temperature; therefore, retrieval of the tagged protein is accomplished by a brief exposure to heat. Using iodoacetate as the warhead, 8-base PNA strands, biotin, and streptavidin-coated magnetic beads, we demonstrate retrieval of the cysteine protease papain. We were also able to use our iodoacetyl–PNA:PNA–biotin probe for retrieval and identification of a thiol reductase and a glutathione transferase from soybean seedling cotyledons.     

Highly Sensitive Plasmonic Sensor Based on Fano Resonance from Silver Nanoparticle Heterodimer Array on a Thin Silver Film

We investigate the optical spectrum of a multilayer metallic slab using multiple-scattering formalism. A thin silver film is attached to a periodic array of heterodimers consisting of two vertically spaced silver nanoparticles of different radii. Depending on the radius of nanoparticles, heterodimer array presents a simple nanoscale geometry which gives rise to remarkable plasmonic properties of multipolar resonances. Due to the coherent interference of the localized nanoparticle plasmons (discrete mode) and surface plasmon polaritons of metallic film (continuous mode), the reflection spectrum represents a sharp asymmetric Fano resonance dip, which is strongly sensitive to the refractive index of the surrounding embedded dielectric host. The physical features contribute to a highly efficient plasmonic sensor for refractive index sensing with sensitivity of ～1.5?×?10^?3 RIU/nm.     

A Simple Design of a Multi-Band Terahertz Metamaterial Absorber Based on Periodic Square Metallic Layer with T-Shaped Gap

We present a multi-band terahertz absorber formed by periodic square metallic ribbon with T-shaped gap and a metallic ground plane separated by a dielectric layer. It is demonstrated that absorption spectra of the proposed structure consist of four absorption peaks located at 1.12, 2.49, 3.45, and 3.91 THz with high absorption coefficients of 98.0, 98.9, 98.7, and 99.6%, respectively. It is demonstrated that the proposed absorber has the tunability from single-band to broadband by changing the length of square metallic ribbon and we can also select or tune the frequencies which we want to use by changing polarization angles. Importantly, the quality factor Q at 3.91 THz is 30.1, which is 5.6 times higher than that of 1.12 THz. These results indicate that the proposed absorber has a promising potential for devices, such as detection, sensing, and imaging.

     

Demonstration of protein-protein interaction specificity by NMR chemical shift mapping.

Chemical shift mapping is becoming a popular method for studying protein-protein interactions in solution. The technique is used to identify putative sites of interaction on a protein surface by detecting chemical shift perturbations in simple (1H, 15N)-HSQC NMR spectra of a uniformly labeled protein as a function of added (unlabeled) target protein. The high concentrations required for these experiments raise questions concerning the possibility for non-specific interactions being detected, thereby compromising the information obtained. We demonstrate here that the simple chemical shift mapping approach faithfully reproduces the known functional specificities among pairs of closely related proteins from the phosphoenolpyruvate:sugar phosphotransferase systems of Escherichia coli and Bacillus subtilis.     

Integrating the QCM detection with magnetic separation for on-line analysis

We investigate the feasibility of coupling the quartz crystal microbalance (QCM) with magnetic separation for on-line analysis. A flow cell was integrated with QCM and magnetic force for the analysis of magnetic and nonmagnetic samples. The resonant frequency change (Deltaf) of QCM was related to the amount of deposited magnetic nanoparticles. This experiment demonstrates that QCM can be used as an on-line detector for magnetic separation. The QCM also gives a characteristic response of the binding between the streptavidin and biotin labeled on the magnetic nanoparticles. Biotin-labeled magnetic nanoparticles were flowed through a gold electrode of QCM to deposit as a matrix for selective capturing streptavidin. The resonant frequency change of QCM was proportional to the amounts of streptavidin captured by biotin. This technique can provide a simple, economic, and automatic method for on-line detection of biomarkers.     

Simulation,Fabrication and Characterization of THz Metamaterial Absorbers

Metamaterials (MM), artificial materials engineered to have properties that may not be found in nature, have been widely explored since the first theoretical¹ and experimental demonstration² of their unique properties. MMs can provide a highly controllable electromagnetic response, and to date have been demonstrated in every technologically relevant spectral range including the optical³, near IR⁴, mid IR⁵ , THz⁶ , mm-wave⁷ , microwave⁸ and radio⁹ bands. Applications include perfect lenses¹⁰, sensors¹¹, telecommunications¹², invisibility cloaks¹³ and filters^14,15. We have recently developed single band¹⁶, dual band¹⁷ and broadband¹⁸ THz metamaterial absorber devices capable of greater than 80% absorption at the resonance peak. The concept of a MM absorber is especially important at THz frequencies where it is difficult to find strong frequency selective THz absorbers¹⁹. In our MM absorber the THz radiation is absorbed in a thickness of ~ λ/20, overcoming the thickness limitation of traditional quarter wavelength absorbers. MM absorbers naturally lend themselves to THz detection applications, such as thermal sensors, and if integrated with suitable THz sources (e.g. QCLs), could lead to compact, highly sensitive, low cost, real time THz imaging systems.     

Mesh considerations for finite element blast modelling in biomechanics

Finite element (FE) modelling is a popular tool for studying human body response to blast exposure. However, blast modelling is a complex problem owing to more numerous fluid–structure interactions (FSIs) and the high–frequency loading that accompanies blast exposures. This study investigates FE mesh design for blast modelling using a sphere in a closed-ended shock tube meshed with varying element sizes using both tetrahedral and hexahedral elements. FSI was consistent for sphere-to-fluid element ratios between 0.25 and 4, and acceleration response was similar for both element types (R² = 0.997). Tetrahedral elements were found to become increasingly volatile following shock loading, causing higher pressures and stresses than predicted with the hexahedral elements. Deviatoric stress response was dependent on the sphere mesh size (p < 0.001), while the pressure response was dependent on the shock tube mesh size (p < 0.001). The results of this study highlight the necessity for mesh sensitivity analysis in blast models.     

Characterizing Far-infrared Laser Emissions and the Measurement of Their Frequencies

The generation and subsequent measurement of far-infrared radiation has found numerous applications in high-resolution spectroscopy, radio astronomy, and Terahertz imaging. For about 45 years, the generation of coherent, far-infrared radiation has been accomplished using the optically pumped molecular laser. Once far-infrared laser radiation is detected, the frequencies of these laser emissions are measured using a three-laser heterodyne technique. With this technique, the unknown frequency from the optically pumped molecular laser is mixed with the difference frequency between two stabilized, infrared reference frequencies. These reference frequencies are generated by independent carbon dioxide lasers, each stabilized using the fluorescence signal from an external, low pressure reference cell. The resulting beat between the known and unknown laser frequencies is monitored by a metal-insulator-metal point contact diode detector whose output is observed on a spectrum analyzer. The beat frequency between these laser emissions is subsequently measured and combined with the known reference frequencies to extrapolate the unknown far-infrared laser frequency. The resulting one-sigma fractional uncertainty for laser frequencies measured with this technique is ± 5 parts in 10⁷. Accurately determining the frequency of far-infrared laser emissions is critical as they are often used as a reference for other measurements, as in the high-resolution spectroscopic investigations of free radicals using laser magnetic resonance. As part of this investigation, difluoromethane, CH₂F₂, was used as the far-infrared laser medium. In all, eight far-infrared laser frequencies were measured for the first time with frequencies ranging from 0.359 to 1.273 THz. Three of these laser emissions were discovered during this investigation and are reported with their optimal operating pressure, polarization with respect to the CO₂ pump laser, and strength.     

Effect of Boundary Condition and Periodical Extension on Transmission Characteristics of Terahertz Filters with Periodical Hole Array Structure Fabricated on Aluminum Slab

Terahertz (THz) filters based on extraordinary optical transmission from periodical hole array structures fabricated on aluminum slab have been experimentally investigated by using THz time-domain spectroscopy. The incident THz pulses with frequency from 0.1 to 2.7 THz could be partly filtered, and the central peak was at ～0.26. The high frequency signal could be observed to decrease, especially for the frequency above ～1 THz. Moreover, the transmission peak from small-size sample with less hole arrays shifts to high frequency at ～0.53 THz due to both the effects of boundary condition and insufficient periodical extension. Furthermore, finite element method with surface plasmon polariton theory is employed to analyze this extraordinary optical transmission and filter phenomena.     

Properties of the Biotin Transport System in Bifidobacterium breve N4

Binding of biotin to resting cells of Bifidobacterium breve N4, which grew in a biotin-deficient medium, was independent of pH from 1 to 9 and of temperature below 50 C. It was not inhibited by metabolic inhibitors including sulfhydryl reagents, but it was inhibited by treatment with 80% ethanol or 5% trichloroacetic acid. It was also competitively inhibited by biotin-sulfone, but not by tetrahydrothiophene nor dethiobiotin. The binding constant was calculated to be 3.3 × 10⁸m^?1. The amount of biotin unextractable with hot water, representing part of the transported biotin, increased gradually for 20 min, this increase was inhibited by NaF, hydroxylamine and low temperature. ¹⁴C-biotin on the cells was displaced by cold biotin and biotin-sulfone; the displacement was not inhibited by metabolic inhibitors, but it was dependent on temperature. A few minutes after binding, the biotin was released to the medium. The release was dependent on pH and temperature, was affected by energy sources and was inhibited by metabolic inhibitors, e.g. NaF, p-chloromercuribenzoic acid and hydroxylamine. It could be stopped at any time by cooling to 0 C or by adding NaF, and the amount of accumulated biotin did not increase under those conditions. These results suggest that the binding sites on the cell surface decreased in number or in their binding affinity for biotin through an energy-dependent process.