Layered Media Research Articles

Directional enhancement of underwater impact and bubble loads in neighbor two-phase fluid domains charge

The loading characteristics of underwater explosions and the dynamic behavior of bubbles are directly related to the charge structure. This study proposes a unique charge structure in a gas–liquid two-phase fluid domain. Numerical methods are used to investigate the effects of fluid layer thickness and gas–liquid ratio on underwater explosion shock wave load, bubble dynamics, and bubble pulsation load. The results show that the two-phase fluid layer significantly enhances the directional release of shock wave energy and bubble pulsation load. During the shock wave phase, a lagging wave effect appears in the liquid layer direction, causing a secondary high-energy shock, significantly increasing the specific impulse. The gas layer direction may form a pressure relief channel effect, enhancing the shock wave peak pressure. For the bubble motion phase, differences in the physical properties of the fluid layer medium lead to irregular bubble boundary movements, promoting bubble tearing and rupture. The gaseous medium converts the accumulated shock wave energy into the internal energy of the bubble, increasing its volume potential. Although this characteristic reduces the pulsation frequency, it significantly increases the specific impulse. Altering the fluid layer medium can control explosion loads and bubble movement, offering new insights for ocean engineering applications.

A high efficiency wideband co‐polarised metalens antenna

AbstractThe traditional co‐polarised metasurface achieves 360° phase coverage only at the resonant frequency but not full phase coverage across a wide bandwidth. This paper proposes a multilayer structure metasurface, which composes of five layers of metal patch and four layers of medium, forming the Fabry–Perry resonator, using the principle of the cancellation of two polarisation conversion, which can achieve a wideband efficient co‐polarisation transmission. Based on the metasurface transmission phase principle, 360° phase coverage has been achieved by changing the geometric parameters of the intermediate metal patches. Through phase compensation arrangement, a metalens is designed using the proposed metasurface unit cell. At the same time, horn antenna is used as a feed to feed the metalens, which can significantly improve the gain of the antenna within a wideband. Simulated and measured results show that the proposed metalens antenna operates at 22–26 GHz, and the maximum measured gain of the metalens antenna reaches 24.16 dBi at 25 GHz, which is 10.85 dB higher than the horn antenna without metasurface compensation, showing good focusing performance. The proposed multilayer metasurface breaks the narrow band limit of the traditional stacked metasurface in principle, opening a way in the broadband metasurface design.

Prediction of Broadband and Highly-Efficient Electromagnetic Wave-Absorbing SiC@SiO2 Nanowire Aerogel by Genetic Algorithm.

Searching for lightweight and high-temperature stable electromagnetic wave-absorbing materials with broad absorbing bandwidth and high efficiency is of significance for applications in daily life and industry. Optimizing the dielectric properties of SiC nanowire aerogel by both compositional and structural designs is an efficient way to obtain simultaneous efficient wave-dissipation ability and good impendence matching and thus the desired properties. However, due to the complex effects of dielectric parameters on the wave-absorbing properties, rational design of high-performance electromagnetic wave-absorbing materials remains challenging. Herein, we propose a genetic algorithm-based approach to predict broadband and highly efficient electromagnetic wave-absorbing materials in a SiC@SiO2 nanowire aerogel-based system. The obtained SiC@SiO2 nanowire aerogels exhibit a gradient multilayered structure with a low dielectric outer layer, a medium layer with alternatively distributed electromagnetic wave transparent and attenuation layers, and an inner high attenuation layer, giving it a broadband electromagnetic wave-absorbing performance covering almost all the 2-18 GHz bandwidth and simultaneous high efficiency. The results show that the genetic algorithm-based approach is efficient in predicting high-performance electromagnetic wave-absorbing ceramic aerogels.

Long-term-survival phase cells of Salmonella enteritidis ATCC 13076 exhibit significantly greater tolerance to atmospheric cold plasma treatment of shell eggs

The main objective of this study was to evaluate the tolerance of stationary phase (STAT) and long-term survival phase (LTS) Salmonella Enteritidis ATCC 13076 to atmospheric cold plasma (ACP) in phosphate buffered saline (PBS, pH 7.0) and on shell eggs. Salmonella Enteritidis was cultured in tryptic soy broth supplemented with 0.6% (w/v) yeast extract (35°C) for 20 h (STAT) and 21 days (LTS). Cell morphology was determined by light microscopy. The PBS and shell eggs were inoculated with STAT or LTS cells to obtain ∼7.0 log10 CFU/mL or egg. The ACP was applied at 45 kV (PBS) and 60 kV (shell eggs) for 1–4 min and 1–5 min, respectively. Pathogen survivors were enumerated on thin agar layer (TAL) medium and on xylose lysine tergitol-4 (XLT-4) agar after 48 h of incubation (35°C). For survivors on shell eggs, R2 and mean square error values were obtained using Log-linear with Tail and Weibull models. The STAT cells were predominantly rod-shaped whereas LTS cells were coccoid. In PBS, reductions (log10 CFU/mL) of STAT cells were 1.0, 0.95, 1.45, and 1.44 after exposure to ACP for 1, 2, 3, and 4 min, respectively. In contrast, reductions in LTS cells were significantly lower (p< 0.05) at 0.04 (1 min), 0.06 (2 min) 0.01 (3 min), and 0.11 (4 min). A similar pattern was observed for shell eggs whereby LTS cells exhibited much higher tolerance to ACP than STAT cells (p < 0.05). The Log-linear with Tail model produced a better fit of the survival data for STAT cells; times to achieve a 4- and 5- log reduction were 5.29 and 5.78 min, respectively. Sub-lethal injury occurred in both STAT and LTS survivors; however, differences were not significant (P > 0.05). Additionally, there were no observed differences in shell strength and yolk color between ACP-treated and control eggs. Based on these results, LTS cells of S. Enteritidis are more tolerant to ACP than STAT cells and should be considered when developing process validation protocols involving application of ACP to inactivate Salmonella on shell eggs.

Control resonant excitation of surface plasmon polariton in quantum-dot molecule system via tunneling induced transparency and Autler–Townes splitting

Electromagnetic induced transparency (EIT) and Autler–Townes splitting (ATS) both produce a transparency window in an absorption profile, yet EIT is distinct in that it can create a transparency window for a weak control field. To elucidate the distinct physical mechanisms underlying EIT and ATS effects, we explore the controlled resonant excitation of surface plasmon polaritons (SPPs) within these regimes. Our setup is composed of three layers: a top vacuum or air layer, a central metal film, and a bottom layer of dielectric medium based on the quantum-dot molecules (QDMs) system. Unlike typical systems, that employ prism or grating coupling scheme, our setup takes advantage of the unique features of the QDMs medium to investigate the direct resonance excitation of SPPs. A weak probe and a strong control field are carried through air or vacuum to the top layer. Our results demonstrate that the permittivity of the bottom medium displays tunneling induced transparency (TIT) if the tunneling strength (Te) and the decay rate of the direct exciton (Γ1) follow the condition Te/Γ1⩽0.5; otherwise, it represents the ATS effect. In both instances, the permittivity of the QDMs medium satisfy the low-loss (high transmission) criteria, i.e., Re [ɛd]<1 and Im [ɛd]≪1, providing a framework for exploring the control resonant excitation of SPPs. We simulate the reflection and transmission spectra using the Fresnel’s formula and noticed sharp dips in reflection and peaks in the transmission spectra, which are the signature of coupler free resonance excitation of SPPs. We noticed that the narrow peak in TIT allows for strong light transmission and thus a high peak in SPP resonance excitation, resulting in higher sensitivity in plasmonic sensors. Conversely, the large spacing between the two peaks in the ATS effect broadens the range of effective excitation frequencies for SPPs, increasing the system’s adaptability and efficiency in applications requiring broad spectrum responses.

Transmission characteristics of high-order vortex beams in layered media

Transmission characteristics of high-order vortex beams in layered media

Successful Pedal Bypass in a Patient With Pseudoxanthoma Elasticum.

Pseudoxanthoma elasticum (PXE) is a rare metabolic disease, causing calcification in the arterial media layer and further peripheral artery disease (PAD). A high rate of failure has been reported after endovascular and open surgical management of PAD among patients with PXE. Critical limb ischemia (CLI) rarely develops in PXE, and there are only few reports of its treatment. We present a case report of a 57year-old female diagnosed with pseudoxanthoma elasticum (PXE). She presented with critical limb ischemia (CLI) and was successfully treated with pedal bypass using the great saphenous vein. Despite obtaining suboptimal outcomes through the initial approach of percutaneous transluminal angioplasty to treat critical limb ischemia, the subsequent bypass operation proved to be a success. At the first follow-up appointment at 1month, the patient was asymptomatic and the ulceration had almost healed. The patient underwent an ultrasound examination at 3, 6, 12, and 24months after discharge, and the surveillance was uncomplicated. With a clear indication for surgery, limb-threatening ischemia can be successfully treated with distal bypass, if necessary, in patients with PXE similarly to atherosclerotic PADs. Appropriate diagnostic and surveillance imaging and the utilization of a multidisciplinary team are key components for effective management of PAD in patients with PXE.

Effects of Restoration Strategies on Soil Health after Lycium chinense Removal in the Qaidam Basin

Ecological restoration of arid land plays a pivotal role in maintaining ecological sustainability and enhancing the resilience of local communities. As an ecologically significant arid land, the Qinghai Qaidam Basin has been severely impacted by human activities such as the widespread planting of Lycium chinense, leading to considerable degradation of vegetation and soil carbon and nutrients. Currently, this vital area is undergoing extensive ecological restoration through the employment of a variety of strategies, but the impact remains inadequately understood. This study seeks to compare the effects of different restoration strategies in the Qinghai Qaidam Basin, focusing on soil properties across five scenarios: a controlled desert area, natural restoration after the removal of L. chinense, continued planting of L. chinense, restoration through the planting of Haloxylon ammodendron, and mixed reseeding after four years of restoration. Our findings indicate that mixed reseeding significantly improved soil water storage to 4.26%, especially in the deep soil layer. The planting of H. ammodendron strategy efficiently reduced soil pH in such an alkaline environment. Soil nutrients, including total nitrogen (TN), total phosphorus (TP), and total potassium (TK), were predominantly concentrated in the top soil layer, with reduced concentrations observed in the medium and deep soil layers. Although soil organic matter remained relatively stable across all restoration strategies, its content was notably lower in the deeper layers. Overall, mixed reseeding proved to be the most efficient strategy for enhancing soil water retention and nutrient levels. In contrast, despite achieving high vegetative coverage to 62.6%, planting of L. chinense was less ecologically beneficial due to its extensive irrigation requirements and adverse effects on soil structure. These findings suggest that restoring degraded areas to an ideal ecological state cannot be achieved within a few years, underscoring the importance of sustained restoration efforts. This study offers valuable insights and practical guidance for the ecological restoration of arid lands, contributing to the development of sustainable land management practices in similar regions.

Nesfatin-1 enhances vascular smooth muscle calcification through facilitating BMP-2 osteogenic signaling

Vascular calcification (VC) arises from the accumulation of calcium salts in the intimal or tunica media layer of the aorta, contributing to higher risk of cardiovascular events and mortality. Despite this, the mechanisms driving VC remain incompletely understood. We previously described that nesfatin-1 functioned as a switch for vascular smooth muscle cells (VSMCs) plasticity in hypertension and neointimal hyperplasia. In this study, we sought to investigate the role and mechanism of nesfatin-1 in VC. The expression of nesfatin-1 was measured in calcified VSMCs and aortas, as well as in patients. Loss- and gain-of-function experiments were evaluated the roles of nesfatin-1 in VC pathogenesis. The transcription activation of nesfatin-1 was detected using a mass spectrometry. We found higher levels of nesfatin-1 in both calcified VSMCs and aortas, as well as in patients with coronary calcification. Loss-of-function and gain-of-function experiments revealed that nesfatin-1 was a key regulator of VC by facilitating the osteogenic transformation of VSMCs. Mechanistically, nesfatin-1 promoted the de-ubiquitination and stability of BMP-2 via inhibiting the E3 ligase SYTL4, and the interaction of nesfatin-1 with BMP-2 potentiated BMP-2 signaling and induced phosphorylation of Smad, followed by HDAC4 phosphorylation and nuclear exclusion. The dissociation of HDAC4 from RUNX2 elicited RUNX2 acetylation and subsequent nuclear translocation, leading to the transcription upregulation of OPN, a critical player in VC. From a small library of natural compounds, we identified that Curculigoside and Chebulagic acid reduced VC development via binding to and inhibiting nesfatin-1. Eventually, we designed a mass spectrometry-based DNA-protein interaction screening to identify that STAT3 mediated the transcription activation of nesfatin-1 in the context of VC. Overall, our study demonstrates that nesfatin-1 enhances BMP-2 signaling by inhibiting the E3 ligase SYTL4, thereby stabilizing BMP-2 and facilitating the downstream phosphorylation of SMAD1/5/9 and HDAC4. This signaling cascade leads to RUNX2 activation and the transcriptional upregulation of MSX2, driving VC. These insights position nesfatin-1 as a potential therapeutic target for preventing or treating VC, advancing our understanding of the molecular mechanisms underlying this critical cardiovascular condition.

A Novel YOLOv5_ES based on lightweight small object detection head for PCB surface defect detection

In the manufacturing process of printed circuit boards (PCBs), surface defects have a significant negative impact on product quality. Considering that traditional object detection algorithms have low accuracy in handling PCB images with complex backgrounds, various types, and small-sized defects, this paper proposes a PCB defect detection algorithm based on a novel YOLOv5 multi-scale attention mechanism(EMA) spatial pyramid dilated Convolution (SPD-Conv) (YOLOv5_ES) network improved YOLOv5s framework. Firstly, the detection head is optimized by removing medium and large detection layers, fully leveraging the capability of the small detection head to identify minor target defects. This approach not only improves model accuracy but also achieves lightweighting. Secondly, in order to further reduce the number of parameters and computational costs, the SPD-Conv is introduced to improve the feature extraction capability by reducing information loss. Thirdly, a EMA module is introduced to fuse context information of different scales, enhancing the model’s generalization ability. Compared to the YOLOv5s model, there is a 3.1% improvement in mean average precision (mAP0.5), a 55.8% reduction in model parameters, and a 4.8% reduction in giga floating-point operations per second (GFLOPs). These results demonstrate a significant improvement in both accuracy and model parameter efficiency.

Novel brittleness index construction and pre‐stack seismic prediction for gas hydrate reservoirs

AbstractReservoir transformation is essential for developing gas hydrate reservoirs. Predicting sediment brittleness is key to optimizing drilling design and evaluating engineering sweet spots. Constructing a brittleness index reflecting the brittle mineral content of a rock based on elastic parameters and predicting it using seismic data is a feasible solution for assessing reservoir brittleness. In addition, the elastic brittleness index can characterize the effect of complex pore types, fractures and pore fillings on rock brittleness. With the shallow hydrate reservoir in the sea as the research target. First, a novel brittleness index characterized by multiplying the Lamé parameter () by Poisson's ratio () is proposed. Its superiority in indicating brittle mineral content is verified by a rock‐physics model. Second, a reflection coefficient approximation equation including the novel brittleness index is derived, enabling direct estimation of reservoir brittleness from seismic data. The new brittleness index has proven to better reflect brittle mineral content and effectively indicate the high brittleness characteristics of hydrate reservoirs. The accuracy of the proposed approximate equation is verified by a layered medium model, and the viability of predicting the new brittleness index using seismic data is also theoretically supported by the model test. Finally, the proposed method has obtained favourable results in the application of hydrate work area data collected at the South China Sea, confirming its availability and practicality.

Adipocytokine levels in patients with atherosclerosis and high triglyceride – glucose index

Aim. To study the levels of adipocytokines and their associations with stable and unstable atherosclerotic plaques in patients with a high triglyceride – glucose (TyG) index.Materials and methods. The study included 109 men aged 38–79 years (mean age 62.28 ± 8.19 years) with atherosclerosis hospitalized for coronary artery bypass grafting (CABG). After microscopy of the intima – media layer, the type of atherosclerotic plaque was determined: stable / unstable. The TyG index ≥ 4.49 was considered as high. Fifty-eight (60%) men had stable plaques in the CA (28 (56%) of them had TyG ≥ 4.49); 39 (40%) men had unstable plaques in the CA (15 (39%) had TyG ≥ 4.49). Blood adipocytokine level was studied using the multiplex assay and the Human Metabolic Hormone Panel V3.Results. The final analysis included 97 patients. The level of glucose-dependent insulinotropic polypeptide (GIP) was 1.53 times greater in patients with TyG ≥ 4.49 (34.16 [18.71; 54.98] vs. 22.34 [15.02; 34.77], p = 0.004). In patients with TyG &lt; 4.49, the adipsin level was 1.2 times higher in patients with unstable plaques than in patients with stable ones. In patients with stable plaques and TyG ≥ 4.49, the GIP level was 1.88 times higher than in patients with TyG &lt; 4.49 (42.13 [25.34; 68.95] vs. 22.39 [17.00; 28.60], p = 0.003). In patients with unstable plaques and TyG ≥ 4.49, the level of peptide tyrosine – tyrosine (PYY) was 1.46 times greater than in patients with TyG &lt; 4.49 (46.14 [30.49; 70.66] vs. 31.53 [24.71; 43.01], p = 0.048).Conclusion. Men with atherosclerosis and TyG ≥ 4.49 had higher blood levels of GIP and PYY. Blood adipsin levels were higher in patients with unstable plaques without insulin resistance.

Near-infrared multi-band small-angle TE-polarization nonreciprocal thermal emitter

Multi-band small-angle irreversible radiation is significantly demanded in applications of high-resolution infrared sensing and infrared filtering. However, current nonreciprocal thermal radiation is either multichannel radiating with large incident angles or radiating at small angles within one single band. To address this issue, a multi-band nonreciprocal thermal emitter operated by exciting multiple resonances within the near-infrared band is proposed. The adopted microstructure arrays consist of periodic silicon square-ring arrays that are supported by magneto-optical medium and metal layer. With the physical mechanisms study through electromagnetic field energy distribution, the TE-polarized multi-band nonreciprocity of thermal radiation is over 70 % at small incident angle of 4°. Equally important, this strategy is effective and generalizable for dual-polarized small-angle multi-band nonreciprocal radiation with adopted microstructure arrays. It is believed that this scheme can promote the development of nonreciprocal thermal photonics.

A novel pretreatment approach on titanium alloys for electroless nickel

To augment the wear resistance of the titanium alloy, electroless nickel (EN) is an effective method to strengthen the surface properties. However, the oxide layer on the surface of titanium alloy will inhibit the adhesion of EN. Meanwhile, the traditional pretreatment methods are cumbersome steps and time-consuming. To overcome these limitations, this study systematically explores a one-step pretreatment approach through amino tris(methylenephosphonic acid) (ATMPA) and HF, which is designed to improve the binding strength of EN. The results exhibit that the compact composite membrane with Ti-ATMPA and TiF3 are successfully prepared under pH 3.0, which has provided a pretreatment layer to replace the oxidation of titanium alloy during EN process, and the adhesion of EN coating (EN-3.0) based on the pretreatment layer reaches 22.46 MPa. The wear resistance of titanium alloy and EN plating system studied via a friction measurement, and the lowest wear rate exhibit in EN-3.0 plating (880.9 × 10-10mm3/Nm) that is lower than EN-traditional pretreatment with a decrease of 32 %. Moreover, the formation and catalytic model of medial layer are also discussed.

Efficient temporal compression of 10-µJ pulses in periodic layered Kerr media.

Pulse compression based on periodic layered Kerr media has been demonstrated as an effective technique for femtosecond pulses with energies around 100 µJ or more. We report such a compressor designed for pulses in the 10-µJ range, which is valuable for many applications. Pulses from a fiber chirped-pulse amplifier are compressed from 300 fs to as short as 60 fs, with good pulse and beam quality. The compressor is a simple and efficient way to extend the performance of common Yb-based sources of pulses with microjoule energies.

Comprehensive analysis of wave interactions and resulting spall damage in layered heterogeneous medium under impact

The effect of dynamic spall damage on the response of layered medium subjected to one-dimensional impact involving an impactor and a multi-layered target is investigated. The study examines the damage caused by elastic waves below the Hugoniot Elastic Limit (HEL) during impact events. Analytical expressions of stress and particle velocity derived from mass and momentum conservation principles are utilized to solve the wave interactions within the impactor-target system, considering reflections, transmissions, and wave interactions between layers. These analytical expressions, integrated into a computational framework, facilitate the monitoring of material responses following each wave interaction, thereby analysing the impact response of the layered medium. Wave interactions, resulting in tensile stresses, that can cause damage to the target have been identified. A strain-based damage initiation and evolution law is incorporated into the developed computer program to predict damage in each layer of a medium. Damage alters the medium’s impact behaviour by decreasing stress magnitude and inducing temporal delays in the response due to attenuated wave propagation speed. The occurrences of tensile stress are explored, analyzing spatial and temporal variations of multiple damage incidents within the layered medium. The tension at the interface between two layers can induce damage in one or both adjacent layers, leading to debonding between layers. The effect of damage on the impact response of the medium is analysed by comparing the results for the damaged and undamaged target scenarios. The impact behaviour of a single-layer and a multi-layer target obtained from the present model, in terms of stress and particle velocity, is verified through Finite Element simulations of the identical impact problems, where the damage evolution criterion is incorporated using a VUMAT subroutine. The outcomes derived from this study align closely with Finite Element (FE) results.

Full-Wave Analysis of Thermal Noise in Antenna Arrays on Top of Layered Medium

Full-Wave Analysis of Thermal Noise in Antenna Arrays on Top of Layered Medium

An ion soft-landing apparatus for ion transport study with surface potential measurement.

An apparatus for explorations of ion transport in a medium and across an interface has been constructed. The ion soft-landing technique is used to deposit low-energy ions onto a pre-adsorbed medium layer on a metal substrate. The designed low-energy ion source can produce a mass-filtered ion beam with tens of nanoampere from solid sources such as bulk metals and salts. The kinetic energy of the ion beam can be lower than 1.0eV, enabling the ions to be soft-landed onto the medium at the surface. A Kelvin probe with a resolution of less than 32 mV is incorporated to measure the surface potential (SP) variation of the ion-landed sample to monitor the ion transport process in the medium. Temperature-programmed SP measurements on an Ag+-adsorbed ice film prepared on Pt(111) reveal that the temperature threshold for the Ag+-induced SP change of the ice film is about 110K. The apparatus performance demonstrates its potential in studies of ion transport and related phenomena at both macroscopic and microscopic levels.

Drying model and drying characteristic analysis of multiphase porous medium for flake materials

The main purpose of this paper is to optimize the drying process and gain a fundamental understanding of the movement of different phases during the drying process of cigarette flakes. Therefore, the drying model of a multiphase porous medium for cigarette flakes has been established in this paper. Binary diffusion of water vapor and gas pressure transport, as well as capillary diffusion of liquid water and gas pressure transport, are considered in this model. The non-equilibrium equations are used to calculate the evaporation rate, and the fluxes of water vapor and liquid water are also obtained. The results indicated that the stochastic stacking model is suitable for studying cigarette flake drying. Because water evaporation absorbs more heat than hot air, the temperature of the cigarette flakes decreases during the drying process. The water activity, vapor density, and vapor mole fraction of the medium layer of cigarette flakes were significantly higher than those of the upper and lower layers. The highest water vapor and liquid water fluxes occur in the medium of the drying process. Additionally, it was observed that the saturated vapor pressure was higher than the equilibrium vapor pressure and steam pressure, non-equilibrium evaporation occurs in the medium of drying. The correlation between the evaporation rate and the drying temperature and humidity of the cigarette flakes was found to be positive.

Initiation of Medial Calcification: Revisiting Calcium Ion Binding to Elastin.

Pathological calcification of elastin, a key connective tissue protein in the medial layers of blood vessels, starts with the binding of calcium ions. This Mini-Review focuses on understanding how calcium ions interact with elastin to initiate calcification at a molecular level, and emphasizes water's critical role in mediating this interaction. In the past decade, great strides have been made in understanding and modeling ion-specific hydration and its effects on biomolecule interactions. However, these advances have been largely absent from our understanding of elastin calcification. Historically, charge-neutral backbone carbonyls and negatively charged carboxyl groups have been proposed as elastin's calcium binding sites. Recently, tropoelastin's only four carboxyl groups have been identified as binding sites from classical molecular dynamics (MD). While carboxyl groups have a much higher affinity for binding calcium ions than backbone carbonyls, conflicting evidence persists for both functional group's importance in elastin calcification. This can be attributed to the fact that divalent ions strongly polarize water, leading to a hydration shell that shields electrostatic forces. The hydration shell surrounding both a calcium ion and either of the proposed binding sites must be displaced to enable binding. Providing our own extended X-ray absorption fine structure (EXAFS) data and complementary simulations, we discuss the potential structures of calcium binding in elastin and review prior knowledge regarding the relative importance of the two proposed binding sites.