Acousto-dielectric tweezers enable independent manipulation of multiple particles.

Acoustic tweezers have gained substantial interest in biology, engineering, and materials science for their label-free, precise, contactless, and programmable manipulation of small objects. However, acoustic tweezers cannot independently manipulate multiple microparticles simultaneously. This study introduces acousto-dielectric tweezers capable of independently manipulating multiple microparticles and precise control over intercellular distances and cyclical cell pairing and separation for detailed cell-cell interaction analysis. Our acousto-dielectric tweezers leverage the competition between acoustic radiation forces, generated by standing surface acoustic waves (SAWs), and dielectrophoretic (DEP) forces, induced by gradient electric fields. Modulating these fields allows for the precise positioning of individual microparticles at points where acoustic radiation and DEP forces are in equilibrium. This mechanism enables the simultaneous movement of multiple microparticles along specified paths as well as cyclical cell pairing and separation. We anticipate our acousto-dielectric tweezers to have enormous potential in colloidal assembly, cell-cell interaction studies, disease diagnostics, and tissue engineering.

Inhibiting Shuttle Effect and Dendrite Growth in Sodium-Sulfur Batteries Enabled by Applying External Acoustic Field.

The room-temperature sodium-sulfur (RT Na-S) battery is a promising alternative to traditional lithium-ion batteries owing to its abundant material availability and high specific energy density. However, the sodium polysulfide shuttle effect and dendritic growth pose significant challenges to their practical applications. In this study, we apply diverse disciplinary backgrounds to introduce a novel method to stimulate polarized BaTiO3 (BTO) nanoparticles on the separator. This approach generates more charges due to the piezoelectric effect under stronger driving forces produced by applying a controllable acoustic field at the outer edge of the cell. The acoustically stimulated BTO attracts more polysulfides, thus reducing the shuttling effect from the cathode to the anode and ultimately enhancing the battery performance. Meanwhile, the acoustic waves create additional streaming flows, improving the uniformity of the sodium ion dispersion, enhancing the sodium ion transport and reducing the possibility of sodium dendrite development. We believe that this work offers a new strategy for the development of high-performance Na-S batteries.

Zirconium ion mediated collagen nanofibrous hydrogels with high mechanical strength.

Low mechanical strength is still the key question for collagen hydrogel consisting of nanofibrils as hard tissue repair scaffolds with no loss of biological function. In this work, novel collagen nanofibrous hydrogels with high mechanical strength were fabricated based on the pre-protection of trisodium citrate masked Zr(SO4)2 solution for collagen self-assembling nanofibrils and then further coordination with Zr(SO4)2 solution. The mature collagen nanofibrils with d-period were observed in Zr(IV) mediated collagen hydrogels by AFM when the Zr(IV) concentration was ≥ 10 mmol/L, and the distribution of zirconium element was uniform. Due to the coordination of Zr(IV) with ─COOH, ─NH2 and ─OH within collagen and the tighter entanglement of collagen nanofibrils, the elastic modulus and compressive strength of Zr(IV) mediated collagen nanofibrous hydrogel were 208.3 and 1103.0 kPa, which were approximate 77 and 12 times larger than those of pure collagen hydrogel, respectively. Moreover, the environmental stability such as thermostability, swelling ability and biodegradability got outstanding improvements and could be regulated by Zr(IV) concentration. Most importantly, the resultant hydrogel showed excellent biocompatibility and even accelerated cell proliferation.

Rapid identification of chemical components and screening of acetylcholinesterase inhibitors from Dalbergia odorifera based on mass defect and diagnostic ion filtering strategy, affinity ultrafiltration, and liquid chromatography-tandem mass spectrometry.

Dalbergia odorifera is a natural product rich in pharmacological ingredients, but the comprehensive characterization and rapid profiling of active components remain a challenge. Thus, an integrated data mining and identification strategy was exploited to efficiently identify the chemical constituents and screen acetylcholinesterase inhibitors (AChEIs) through affinity ultrafiltration and ultra-high-performance liquid chromatography-mass spectrometry (AUF-UHPLC-MS). As a result, polygonal mass defect filtering, diagnostic product ions, and neutral loss rules were created for rapid structural classification and component identification. A total of 140 flavonoids were tentatively characterized, including 41 isoflavonoids, 23 flavanones, 21 isoflavans, 19 flavones and flavonols, 13 neoflavonoids, 11 isoflavanones, seven flavone glycosides, and five chalcones. Subsequently, six natural AChEIs including tectorigenin, fisetin, dalbergin, pterostilbene, isoliquiritigenin, and biochanin A were screened out using AUF-UHPLC-MS and molecular docking. Meanwhile, the AChE inhibitory activities of the six compounds were assessed in vitro, tectorigenin, fisetinand, and dalbergin have moderate inhibitory activity. In conclusion, a novel strategy for systematic characterization and further screening of active compounds in natural products was established, which provides a material basis for quality control of Dalbergia odorifera.

Investigation of Viscoelastic Guided Wave Properties in Anisotropic Laminated Composites Using a Legendre Orthogonal Polynomials Expansion-Assisted Viscoelastodynamic Model.

This study investigates viscoelastic guided wave properties (e.g., complex-wavenumber-, phase-velocity-, and attenuation-frequency relations) for multiple modes, including different orders of antisymmetric, symmetric, and shear horizontal modes in viscoelastic anisotropic laminated composites. To obtain those frequency-dependent relations, a guided wave characteristic equation is formulated based on a Legendre orthogonal polynomials expansion (LOPE)-assisted viscoelastodynamic model, which fuses the hysteretic viscoelastic model-based wave dynamics and the LOPE-based mode shape approximation. Then, the complex-wavenumber-frequency solutions are obtained by solving the characteristic equation using an improved root-finding algorithm, which leverages coefficient matrix determinant ratios and our proposed local tracking windows. To trace the solutions on the dispersion curves of different wave modes and avoid curve-tracing misalignment in regions with phase-velocity curve crossing, we presented a curve-tracing strategy considering wave attenuation. With the LOPE-assisted viscoelastodynamic model, the effects of material viscosity and fiber orientation on different guided wave modes are investigated for unidirectional carbon-fiber-reinforced composites. The results show that the viscosity in the hysteresis model mainly affects the frequency-dependent attenuation of viscoelastic guided waves, while the fiber orientation influences both the phase-velocity and attenuation curves. We expect the theoretical work in this study to facilitate the development of guided wave-based techniques for the NDT and SHM of viscoelastic anisotropic laminated composites.

Molecular and structural basis of an ATPase-nuclease dual-enzyme anti-phage defense complex.

Coupling distinct enzymatic effectors emerges as an efficient strategy for defense against phage infection in bacterial immune responses, such as the widely studied nuclease and cyclase activities in the type III CRISPR-Cas system. However, concerted enzymatic activities in other bacterial defense systems are poorly understood. Here, we biochemically and structurally characterize a two-component defense system DUF4297-HerA, demonstrating that DUF4297-HerA confers resistance against phage infection by cooperatively cleaving dsDNA and hydrolyzing ATP. DUF4297 alone forms a dimer, and HerA alone exists as a nonplanar split spiral hexamer, both of which exhibit extremely low enzymatic activity. Interestingly, DUF4297 and HerA assemble into an approximately 1 MDa supramolecular complex, where two layers of DUF4297 (6 DUF4297 molecules per layer) linked via inter-layer dimerization of neighboring DUF4297 molecules are stacked on top of the HerA hexamer. Importantly, the complex assembly promotes dimerization of DUF4297 molecules in the upper layer and enables a transition of HerA from a nonplanar hexamer to a planar hexamer, thus activating their respective enzymatic activities to abrogate phage infection. Together, our findings not only characterize a novel dual-enzyme anti-phage defense system, but also reveal a unique activation mechanism by cooperative complex assembly in bacterial immunity.

Study on the hypotensive effect and mechanism of hawthorn (Crataegus pinnatifida) fruits and hyperoside in spontaneously hypertensive rats.

Hawthorn fruits have a sweet and sour taste, besides having beneficial therapeutic effects on hyperlipidemia, hypertension, and coronary heart disease, making them widely used in food and clinical medicine. However, their hypotensive effects and potential mechanisms of anti-hypertension still need to be elucidated. This study aims to explore the antihypertensive effect of hawthorn and its monomer hyperoside on spontaneously hypertensive rats through pharmacodynamics, serum metabolomics, and in vivo mechanism studies. After 7 weeks of intervention with hawthorn extract and hyperoside, the blood pressure was significantly reduced. Aortic vascular staining results showed that the injury was significantly improved after intervention with hawthorn extract and hyperoside. According to the serum metabolomics study, the main metabolic pathway regulating blood pressure in hawthorn extract and hyperoside groups was the primary bile acid biosynthesis pathway. Quantitative experiments confirmed that the level of bile acid in the model group was significantly different from that in the normal group, while that in the hawthorn group and the hyperoside group was close to that in the normal group. Based on the prediction of bile acid-hypertension related targets and the literature, nine genes involved in bile acid metabolism and inflammatory pathways were selected for further study. The FXR, TGR5, ET-1, NOS3, Akt1, TNF-α, Ptgs2, ACE2 and Kdr mRNA expression levels in the hawthorn extract and hyperoside groups were significantly different from those in the model groups. In summary, hawthorn extract and hyperoside have a hypotensive effect on spontaneously hypertensive rats through bile acid and inflammation related targets. Hence, hawthorn extract has the potential to become a functional food or an alternative therapy for hypertension.

Development of a piezo stack - laser Doppler vibrometer sensing approach for characterizing shear wave dispersion and local viscoelastic property distributions.

Laser Doppler vibrometry and wavefield analysis have recently shown great potential for nondestructive evaluation, structural health monitoring, and studying wave physics. However, there are limited studies on these approaches for viscoelastic soft materials, especially, very few studies on the laser Doppler vibrometer (LDV)-based acquisition of time-space wavefields of dispersive shear waves in viscoelastic materials and the analysis of these wavefields for characterizing shear wave dispersion and evaluating local viscoelastic property distributions. Therefore, this research focuses on developing a piezo stack-LDV system and shear wave time-space wavefield analysis methods for enabling the functions of characterizing the shear wave dispersion and the distributions of local viscoelastic material properties. Our system leverages a piezo stack to generate shear waves in viscoelastic materials and an LDV to acquire time-space wavefields. We introduced space-frequency-wavenumber analysis and least square regression-based dispersion comparison to analyze shear wave time-space wavefields and offer functions including extracting shear wave dispersion relations from wavefields and characterizing the spatial distributions of local wavenumbers and viscoelastic properties (e.g., shear elasticity and viscosity). Proof-of-concept experiments were performed using a synthetic gelatin phantom. The results show that our system can successfully generate shear waves and acquire time-space wavefields. They also prove that our wavefield analysis methods can reveal the shear wave dispersion relation and show the spatial distributions of local wavenumbers and viscoelastic properties. We expect this research to benefit engineering and biomedical research communities and inspire researchers interested in developing shear wave-based technologies for characterizing viscoelastic materials.

Peroxynitrite imaging in ferroptosis-mediated drug-induced liver injury with a near-infrared fluorescence probe.

BACKGROUND: Over-consumption of drugs can result in drug-induced liver damage (DILI), which can worsen liver failure. Numerous studies have shown the significant role ferroptosis plays in the pathophysiology of DILI, which is typified by a marked imbalance between the generation and breakdown of lipid reactive oxygen species (ROS). The content of peroxynitrite (ONOO-) rapidly increased during this process and was thought to be a significant marker of early liver injury. Therefore, the construction of fluorescence probe for the detection and imaging of ONOO- holds immense importance in the early diagnosis and treatment of ferroptosis-mediated DILI. RESULTS: We designed a probe DILI-ONOO based on the ICT mechanism for the purpose of measuring and visualizing ONOO- in ferroptosis-mediated DILI processes and associated studies. This probe exhibited significant fluorescence changes with good sensitivity, selectivity, and can image exogenous and endogenous ONOO- in cells with low cytotoxicity. Using this probe, we were able to show changes in ONOO- content in ferroptosis-mediated DILI cells and mice models induced by the intervention of acetaminophen (APAP) and isoniazid (INH). By measuring the concentration of ferroptosis-related indicators in mice liver tissue, we were able to validate the role of ferroptosis in DILI. It is worth mentioning that compared to existing alanine transaminase (ALT) and aspartate aminotransferase (AST) detection methods, this probe can achieve early identification of DILI prior to serious liver injury. SIGNIFICANCE: This work has significant reference value in researching the relationship between ferroptosis and DILI and visualizing research. The results indicate a strong correlation between the progression of DILI and ferroptosis. Additionally, the use of DILI-ONOO shows promise in investigating the DILI process and assessing the effectiveness of medications in treating DILI.

Robot-assisted chirality-tunable acoustic vortex tweezers for contactless, multifunctional, 4-DOF object manipulation.

Robotic manipulation of small objects has shown great potential for engineering, biology, and chemistry research. However, existing robotic platforms have difficulty in achieving contactless, high-resolution, 4-degrees-of-freedom (4-DOF) manipulation of small objects, and noninvasive maneuvering of objects in regions shielded by tissue and bone barriers. Here, we present chirality-tunable acoustic vortex tweezers that can tune acoustic vortex chirality, transmit through biological barriers, trap single micro- to millimeter-sized objects, and control object rotation. Assisted by programmable robots, our acoustic systems further enable contactless, high-resolution translation of single objects. Our systems were demonstrated by tuning acoustic vortex chirality, controlling object rotation, and translating objects along arbitrary-shaped paths. Moreover, we used our systems to trap single objects in regions with tissue and skull barriers and translate an object inside a Y-shaped channel of a thick biomimetic phantom. In addition, we showed the function of ultrasound imaging-assisted acoustic manipulation by monitoring acoustic object manipulation via live ultrasound imaging.

Capillary-based, multifunctional manipulation of particles and fluids via focused surface acoustic waves.

Surface acoustic wave (SAW)-enabled acoustofluidic technologies have recently atttracted increasing attention for applications in biology, chemistry, biophysics, and medicine. Most SAW acoustofluidic devices generate acoustic energy which is then transmitted into custom microfabricated polymer-based channels. There are limited studies on delivering this acoustic energy into convenient commercially-available glass tubes for manipulating particles and fluids. Herein, we have constructed a capillary-based SAW acoustofluidic device for multifunctional fluidic and particle manipulation. This device integrates a converging interdigitated transducer to generate focused SAWs on a piezoelectric chip, as well as a glass capillary that transports particles and fluids. To understand the actuation mechanisms underlying this device, we performed finite element simulations by considering piezoelectric, solid mechanic, and pressure acoustic physics. This experimental study shows that the capillary-based SAW acoustofluidic device can perform multiple functions including enriching particles, patterning particles, transporting particles and fluids, as well as generating droplets with controlled sizes. Given the usefulness of these functions, we expect that this acoustofluidic device can be useful in applications such as pharmaceutical manufacturing, biofabrication, and bioanalysis.

The genus Oxytropis DC: application, phytochemistry, pharmacology, and toxicity.

OBJECTIVES: Oxytropis DC is a perennial plant of Fabaceae family, which is widely distributed in the northern temperate zone. It is known as "locoweed" because of its toxic component swainsonine. However, it is widely used in Tibetan medicine and Mongolian medicine, mainly for the treatment of heat-clearing and detoxifying, pain-relieving, anti-inflammatory, hemostasis, and other diseases. To provide a basis for the further development and utilization of Oxytropis DC, the pieces of literature about the application, phytochemistry, pharmacological action, and toxicity of Oxytropis DC were reviewed and analyzed. KEY FINDINGS: A total of 373 chemical constituents were found from Oxytropis DC, including flavonoids, alkaloids, steroids, terpenoids, and others. Pharmacological actions mainly include antitumor, antioxidation, anti-inflammatory, analgesic, antibacterial, antifibrosis, and other pharmacological actions, among them, the antitumor effect is particularly prominent. SUMMARY: At present, studies on its pharmacological effects are mainly concentrated on the extracts, some flavonoids, and alkaloids. In the follow-up studies, research on the pharmacological activities of the other chemical constituents in Oxytropis should be strengthened. It has the potential to pave the way for research and development of novel Oxytropis medicines.

Isolation, characterization and SARS-CoV-2 3CL protease inhibitory activity of a new methylsulfinyl-butanyl derivative from Raphani Semen.

A new compound named raphanised A (1), along with two known methylsulfinyl -butanyl derivatives (2-3) and seven known indole derivatives (4-10), were isolated from the Raphani Semen. Among the indole derivatives, 5 was identified as a new natural product, and 4, 6, 7, 8, 9, 10 were isolated from the genus of Raphanus for the first time. Their structures were elucidated based on the NMR and HR-EI-MS analysis. Additionally, the inhibitory activity of methylsulfinyl-butanyl derivatives 1-3 on SARS CoV-2 3CL protease was evaluated. The results showed that 1-3 exhibited inhibition of SARS-CoV-2 3CL protease activity at concentrations ranging from 3.3 to 30 µM.

Acoustothermal transfection for cell therapy.

Transfected stem cells and T cells are promising in personalized cell therapy and immunotherapy against various diseases. However, existing transfection techniques face a fundamental trade-off between transfection efficiency and cell viability; achieving both simultaneously remains a substantial challenge. This study presents an acoustothermal transfection method that leverages acoustic and thermal effects on cells to enhance the permeability of both the cell membrane and nuclear envelope to achieve safe, efficient, and high-throughput transfection of primary T cells and stem cells. With this method, two types of plasmids were simultaneously delivered into the nuclei of mesenchymal stem cells (MSCs) with efficiencies of 89.6 ± 1.2%. CXCR4-transfected MSCs could efficiently target cerebral ischemia sites in vivo and reduce the infarct volume in mice. Our acoustothermal transfection method addresses a key bottleneck in balancing the transfection efficiency and cell viability, which can become a powerful tool in the future for cellular and gene therapies.

Hydrophobic association: A facile approach to prepare physical cross-linked gelatin hydrogel with desirable thermal stability, flexibility and self-healing ability.

Methacrylic anhydride was added to 20 % gelatin solution to prepare gelatin methacryloyl (GelMA) but an unexpected gelation process was observed within several minutes. The experimental data revealed that the methacryloyl substitution can increase the hydrophobicity of gelatin and the micellar diameter in solution. Therefore, we speculated that the methacryloyl substitution caused the formation of micellar cross-links based on the hydrophobic residues of gelatin and the methacryloyl groups, thus obtaining the hydrophobic association hydrogels. The thixotropic and tensile experiments confirmed that GelMA hydrogel possessed the features of hydrophobic association hydrogels like self-healing and stretchable abilities. The rheological experiments revealed that the gelation rate and the mechanical strength of the GelMA hydrogels were in direct proportion to the concentration of GelMA and the degree of methacryloyl substitution. GelMA hydrogels possessed desirable thermal stability that it didn't melt after being heated to 90 °C. Furthermore, the MTT assays and calcein AM/PI staining revealed that GelMA hydrogel was biocompatible. These results collectively confirm that the hydrophobic association is a prospective and facile approach to prepare gelatin hydrogel with desirable properties for further application.

Aerosol jet printing of surface acoustic wave microfluidic devices.

The addition of surface acoustic wave (SAW) technologies to microfluidics has greatly advanced lab-on-a-chip applications due to their unique and powerful attributes, including high-precision manipulation, versatility, integrability, biocompatibility, contactless nature, and rapid actuation. However, the development of SAW microfluidic devices is limited by complex and time-consuming micro/nanofabrication techniques and access to cleanroom facilities for multistep photolithography and vacuum-based processing. To simplify the fabrication of SAW microfluidic devices with customizable dimensions and functions, we utilized the additive manufacturing technique of aerosol jet printing. We successfully fabricated customized SAW microfluidic devices of varying materials, including silver nanowires, graphene, and poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS). To characterize and compare the acoustic actuation performance of these aerosol jet printed SAW microfluidic devices with their cleanroom-fabricated counterparts, the wave displacements and resonant frequencies of the different fabricated devices were directly measured through scanning laser Doppler vibrometry. Finally, to exhibit the capability of the aerosol jet printed devices for lab-on-a-chip applications, we successfully conducted acoustic streaming and particle concentration experiments. Overall, we demonstrated a novel solution-based, direct-write, single-step, cleanroom-free additive manufacturing technique to rapidly develop SAW microfluidic devices that shows viability for applications in the fields of biology, chemistry, engineering, and medicine.

Three new compounds from the seeds of Sinapis alba L.

Three undescribed compounds, 1-(4-hydroxyphenyl)-3-((4-hydroxyphenyl)thio) -2-propanone (I), 4'-hydroxybenzyl 4-(4''- hydroxybenzyloxy) benzyl thioether (II) and 3-(4'-hydroxybenzyl)-6-hydroxy-quinazolinone (III) were isolated from the seeds of Sinapis alba L. The structures of all compounds were elucidated based on NMR and MS analysis, along with the comparison with published data. The potential targets of compounds I-III were identified by virtual screening. The potential inhibitory effects of these compounds on protein kinase C theta type were compared by molecular docking. The docking score of compound III was the highest.

Poly(2-vinylpyridine)/MCM-41 Composites with Micropores and Switchable Mesopores for the Removal of Cr(VI).

Porous structure design and reversible regulation of pore size during adsorption-desorption are crucial to the removal of pollutants in water such as Cr(VI). In this paper, micropores and switchable mesopores were constructed on MCM-41 to further improve adsorption-desorption performance of Cr(VI) via the confinement effect of micropores and opening and closing of mesopores. 2-Vinylpyridine was introduced and polymerized into the pores and on the pore mouth of MCM41 modified by C═C group (AM41) under the irradiation of ultraviolet light. The obtained samples (PM41) possessed mesopores (2.73 nm) and micropores (1.36 nm), where mesopores could open or close under different pH and micropores showed the confinement effect because their pore size is close to Cr(VI) diameter (0.87 nm). Compared with MCM-41, the introduction of poly(2-vinylpyridine) enhanced obviously its adsorptive ability and it trapped most of the Cr(VI) (99%) in solution, 12 times higher than that of the parent sample. The change of pore size is favorable to the cycle performance, and after 3 times recycling, the removal rate of Cr(VI) by PM41-20 remained above 88%. Langmuir isotherm showed a better data correlation than the Freundlich model. Cr(VI) in solution was removed by electrostatic interaction between the pyridine group and Cr(VI) and the confinement effect from micropores.

Non-targeted Profiling of Sea Buckthorn Fruit Oil Fingerprints from 3 Regions and Study on Its Antioxidant Activity.

This study investigated the chemical and volatile characteristics of sea buckthorn fruits from three different regions in China. The chemical composition of the volatile oil was determined by using a non-targeted gas chromatography and mass spectrometry (GC/MS) method and the differences in chemical composition among the three producing areas were compared by heatmap providing a visual basis for researchers. A total of 93 compounds were identified, including 52 compounds from the Northeast China, 51 from the Xinjiang region, and 37 from Inner Mongolia region. Then, the in vitro antioxidant activity of sea buckthorn fruit oil was measured using DPPH, ABTS, and SOD inhibition tests, and the results showed that sea buckthorn fruit oil in northeast China was the strongest antioxidant, followed by Inner Mongolia and Xinjiang. The results of the CCK-8 experiment indicated that within the tested concentration, there is no cell cytotoxicity of the essential oil in human umbilical vein endothelial cells (HUVECs) cells. The results could supply reference to distinguish sea buckthorn fruit from different production areas and, meanwhile, clarify the activity and safety of sea buckthorn oil.

Isorhynchophylline improves lipid metabolism disorder by mediating a circadian rhythm gene Bmal1 in spontaneously hypertensive rat.

Hypertension is a progressive metabolic disease characterized by circadian regulation of lipid metabolism disorder. Identifying specific lipid components and maintaining circadian homeostasis of lipid metabolism might be a promising therapeutic strategy for hypertension. Isorhynchophylline (IRP) can regulate lipid metabolism; however, the underlying mechanism of IRP in improving lipid metabolism rhythm disorder is still unclear. The lipid circadian biomarkers and abnormal metabolic pathways intervened by IRP were investigated using diurnal lipidomic research methods. The 24-h circadian changes in mRNA and protein expression levels of circadian genes, including Bmal1, Clock, Cry1, Cry2, Per1, and Per2, and lipid metabolism-related factors (PPARα and LPL) were determined using RT-PCR and western blot analyses, respectively. The underlying mechanisms were intensively investigated by inhibiting Bmal1. Molecular docking and drug affinity responsive target stability analyses were performed to assess the binding affinity of IRP and Bmal1. IRP treatment could effectively improve 24-h blood pressure, ameliorate the lipid metabolic rhythm disorder, reverse the expression levels of circadian rhythm genes, and regulate lipid metabolism-related genes (PPARα and LPL) by mediating Bmal1. This study highlighted the potential effects of IRP in maintaining the circadian homeostasis of lipid metabolism and the treatment of hypertension.