Chaperoning shape-shifting tau in disease.

Many neurodegenerative diseases, including Alzheimer's, originate from the conversion of proteins into pathogenic conformations. The microtubule-associated protein tau converts into ß-sheet-rich amyloid conformations, which underlie pathology in over 25 related tauopathies. Structural studies of tau amyloid fibrils isolated from human tauopathy tissues have revealed that tau adopts diverse structural polymorphs, each linked to a different disease. Molecular chaperones play central roles in regulating tau function and amyloid assembly in disease. New data supports the model that chaperones selectively recognize different conformations of tau to limit the accumulation of proteotoxic species. The challenge now is to understand how chaperones influence disease processes across different tauopathies, which will help guide the development of novel conformation-specific diagnostic and therapeutic strategies.

Self-Priming Cyclic Amplification Accelerating CRISPR Sensor for Sensitive and Specific MicroRNA Analysis with No Background.

In this work, we demonstrate for the first time the application of the phosphorothioated-terminal hairpin formation and self-priming extension (PS-THSP) reaction for miRNA assays. A self-priming amplification accelerating CRISPR sensor was well-established for sensitive and specific miRNA detection by integrating the PS-THSP reaction and CRISPR/Cas12a system. The sensor consists of three steps: (1) the formation of a complete PS-THSP template in the presence of target miRNA and ligase; (2) the exponential isothermal amplification of the PS-THSP reaction under the action of DNA polymerase; (3) the activation of the CRISPR/Cas12a fluorescence system to generate signals. We used miR-21 as a model target. The sensor can achieve sensitive detection of miR-21 without the involvement of any primers, and the special design of the CRISPR proto-spacer neighbor motif (PAM) sequence effectively avoids the interference of the background signal. In addition, the sensor can not only identify single-base mutant homologous sequences but also show stable performance in complex biological matrices. We have successfully used this sensor to accurately analyze miR-21 in different cell lines and real clinical samples, demonstrating its great potential in clinical diagnosis.

Versatile GO/ANFs Aerogel for Highly Efficient Solar-Powered Water Purification in Wide Environments.

Solar-driven interfacial evaporation is a very promising choice for producing clean water. Despite the considerable investigation of pure NaCl brine purification, solar-driven complex water purification, such as real-world seawater desalination as well as domestic and industrial wastewater treatment, has rarely been investigated, mainly due to its compositions being much more complicated than NaCl brine. Herein, we developed a graphene oxide/aramid nanofiber (GO/ANFs) aerogel by a freeze-drying process. The GO/ANFs aerogel combined opened porous microchannels, superhydrophilicity, anti-oil-fouling capacity, enhanced broad-spectrum light absorption (more than 92%), and good solar/heat management. These integrated properties enabled the GO/ANFs aerogel to be an advanced solar interfacial evaporator for efficient freshwater production with the characteristics of localized heat conversion, quick water transport, and salt crystallization inhibition, and the rate of steam production rate was as high as 2.25 kg m-2 h-1 upon exposure to 1 solar irradiation. Importantly, the high-water-vapor generation rate was maintained even under complicated conditions, including real-world seawater, dye water, emulsions, and corrosive liquid environments. Considering its promising adaptability to a wide range of environments, this work hopes to inspire the development of brine desalination, wastewater purification, clean water production, and solar energy utilization.

Mechanical Robust GO/PVA Hydrogel for Strong and Recyclable Adhesion in Air, Underwater, and Underoil Environments.

Adhesive hydrogels are considered to be promising interfacial adhesive materials for various applications; however, their adhesive strength is significantly reduced when immersed in liquid environments (water and oil) due to obstruction of the liquid layer or swelling in liquid, and they could not always be reused when the failure of the adhesive performance occurred. Herein, a graphite oxide/poly(vinyl alcohol) (GO/PVA) hydrogel with strong adhesion in air and under liquid environments was developed by rationally regulating the interactions of water and dimethyl sulfoxide (DMSO) in the binary liquid system. The strong interaction between water and DMSO allowed the water layer of the GO/PVA hydrogel on the hydrogel surface to act as a shield to repel oil in air, under water, and even when immersed in oil, and it also endowed the obtained hydrogel with antiswelling property when immersed in water and oil. Importantly, the GO/PVA hydrogel could serve as an advanced adhesive to firmly bond different substrates in air, under water, and under oil, and interestingly, its dry and wet adhesive performance was repeatable and recyclable. This work is expected to be an important addition to the field of adhesive soft materials.

Long Non-Coding RNA Detection Based on Multi-Probe-Induced Rolling Circle Amplification for Hepatocellular Carcinoma Early Diagnosis.

Long non-coding RNAs (lncRNAs) played vital roles in physiological and pathological conditions. Consistent results from cell experiments, animal experiments, and clinical studies suggested that lncRNA HULC was an oncogenic lncRNA serving as a potential diagnostic and prognostic marker of hepatocellular carcinoma. In this study, we developed a fluorescent biosensor for lncRNA HULC detection based on rolling circle amplification (RCA) induced by multi-primer probes. Multiple primer probes can not only combine with lncRNA to break its secondary structure, which was conducive to lncRNA captured by Y-shaped probes, but also trigger multiple RCA reactions to achieve signal amplification and the goal of sensitive detection of lncRNA. Compared to previous detection methods, in this scheme, we took advantage of the long sequence characteristics of lncRNA to make it a carrier that can bind multiple primers to initiate RCA. This newly designed biosensor provided a linear range from 1 pM to 100 nM with a detection limit of 0.06 pM. This method can provide a new idea for the application of isothermal amplification in detecting lncRNA. Furthermore, the application of the biosensor in liver cancer cell lines and whole blood samples from hepatocellular carcinomatosis patients also confirmed that the method had good selectivity and sensitivity to lncRNA HULC. This method offered a new way for transforming specific lncRNA into clinical application for diagnosis, prognosis, or predicting treatment response.

Highly Sensitive Aptasensor for Detecting Cancerous Exosomes Based on Clover-like Gold Nanoclusters.

Exosome-based liquid biopsy technologies play an increasingly prominent role in tumor diagnosis. However, the simple and sensitive method for counting exosomes still faces considerable challenges. In this work, the CD63 aptamer-modified DNA tetrahedrons on the gold electrode were used as recognition elements for the specific capture of exosomes. Partially complementary DNA probes act as bridges linking trapped exosomes and three AuNP-DNA signal probes. This clover-like structure can tackle the recognition and sensitivity issues arising from the undesired AuNP aggregation event. When cancerous exosomes are present in the system, the high accumulation of methylene blue molecules from DNA-AuNP nanocomposites on the surface of the electrode leads to an intense current signal. According to the results, the aptasensor responds to MCF-7 cell-derived exosomes in the concentration range from 1.0 × 103 to 1.0 × 108 particles·µL-1, with the detection limit of 158 particles·µL-1. Furthermore, the aptasensor has been extended to serum samples from breast cancer patients and exhibited excellent specificity. To sum it up, the aptasensor is sensitive, straightforward, less expensive, and fully capable of receiving widespread application in clinics for tumor monitoring.

Simple and Rapid Way to a Multifunctionally Conductive Hydrogel for Wearable Strain Sensors.

Conductive hydrogels have gained increasing attention in the field of wearable smart devices. However, it remains a big challenge to develop a multifunctionally conductive hydrogel in a rapid and facile way. Herein, a conductive tannic acid-iron/poly (acrylic acid) hydrogel was synthesized within 30 s at ambient temperature by the tannic acid-iron (TA@Fe3+)-mediated dynamic catalytic system. The TA@Fe3+ dynamic redox autocatalytic pair could efficiently activate the ammonium persulfate to initiate the free-radical polymerization, allowing the gelation to occur easily and rapidly. The resulting hydrogel exhibited enhanced stretchability (3560%), conductivity (33.58 S/m), and strain sensitivity (gauge factor = 2.11). When damaged, it could be self-healed through the dynamic and reversible coordination bonds between the Fe3+ and COO- groups in the hydrogel network. Interestingly, the resulting hydrogel could act as a strain sensor to monitor various human motions including the huge movement of deformations (knuckle, wrist) and subtle motions (smiling, breathing) in real time due to its enhanced self-adhesion, good conductivity, and improved strain sensitivity. Also, the obtained hydrogel exhibited efficient electromagnetic interference (EMI) shielding performance with an EMI shielding effectiveness value of 24.5 dB in the X-band (8.2-12.4 GHz). Additionally, it displayed antibacterial properties, with the help of the activity of TA.

An integrated supervision framework to safeguard the urban river water quality supported by ICT and models.

Models and information and communication technology (ICT) can assist in the effective supervision of urban receiving water bodies and drainage systems. Single model-based decision tools, e.g., water quality models and the pollution source identification (PSI) method, have been widely reported in this field. However, a systematic pathway for environmental decision support system (EDSS) construction by integrating advanced single techniques has rarely been reported, impeding engineering applications. This paper presents an integrated supervision framework (UrbanWQEWIS) involving monitoring-early warning-source identification-emergency disposal to safeguard the urban water quality, where the data, model, equipment and knowledge are smoothly and logically linked. The generic architecture, all-in-one equipment and three key model components are introduced. A pilot EDSS is developed and deployed in the Maozhou River, China, with the assistance of environmental Internet of Things (IoT) technology. These key model components are successfully validated via in situ monitoring data and dye tracing experiments. In particular, fluorescence fingerprint-based qualitative PSI and Bayesian-based quantitative PSI methods are effectively coupled, which can largely reduce system costs and enhance flexibility. The presented supervision framework delivers a state-of-the-art management tool in the digital water era. The proposed technical pathway of EDSS development provides a valuable reference for other regions.

Acoustic Trauma Causes Cochlear Pericyte-to-Myofibroblast-Like Cell Transformation and Vascular Degeneration, and Transplantation of New Pericytes Prevents Vascular Atrophy.

Acoustic trauma disrupts cochlear blood flow and damages sensory hair cells. Damage and regression of capillaries after acoustic trauma have long been observed, but the underlying mechanism of pathology has not been understood. We show herein that loud sound causes change of phenotype from neural/glial antigen 2 positive/α-smooth muscle actin negative to neural/glial antigen 2 positive/α-smooth muscle actin positive in some pericytes (PCs) on strial capillaries that is strongly associated with up-regulation of transforming growth factor-ß1. The acoustic trauma also reduced capillary density and increased deposition of matrix proteins, particularly in the vicinity of transformed PCs. In a newly established in vitro three-dimensional endothelial cell (EC) and PC co-culture model, transformed PCs induced thicker capillary-like branches in ECs and increased collagen IV and laminin expression. Transplantation of exogenous PCs derived from neonatal day 10 mouse cochleae to acoustic traumatized cochleae, however, significantly attenuated the decreased vascular density in the stria. Transplantation of PCs pretransfected with adeno-associated virus 1-vascular endothelial growth factor-A165 under control of a hypoxia-response element markedly promotes vascular volume and blood flow, increased proliferation of PCs and ECs, and attenuated loud sound-caused loss in endocochlear potential and hearing. Our results indicate that loud sound-triggered PC transformation contributes to capillary wall thickening and regression, and young PC transplantation effectively rehabilitates the vascular regression and improves hearing.

Contactless treatment for scoliosis by electromagnetically controlled shape-memory alloy rods: a preliminary study in rabbits.

PURPOSE: To evaluate the safety and efficacy of a system aiming to correct scoliosis called "electromagnetically controlled shape-memory alloy rods" (EC-SMAR) used in a rabbit model. METHODS: We heat-treated shape-memory alloy (SMA) rods to achieve a transition temperature between 34 and 47 °C and a C-shape austenite phase. We then developed a water-cooled generator capable of generating an alternating magnetic field (100 kHz) for induction heating. We next studied the efficacy of this system in vitro and determined some parameters prior to proceeding with animal experiments. We then employed a rabbit model, in which we fixed a straight rod along the spinous processes intraoperatively, and conducted induction heating postoperatively every 4 days for 1 month, while performing periodic X-ray assessments. RESULTS: Significant kyphotic deformations with Cobb angles of about 45° (p < 0.01) were created in five rabbits, and no complications occurred throughout the experiment. The rabbits are still very much alive and do not show any signs of discomfort. CONCLUSIONS: This is the first system that can modulate spinal deformation in a gradual, contactless, noninvasive manner through electromagnetic induction heating applied to SMA alloy rods. Although this study dealt with healthy spines, it provides promising evidence that this device also has the capacity to correct human kyphosis and even scoliosis in the future. These slides can be retrieved under Electronic Supplementary Material.

Vision function of pseudophakic eyes with posterior capsular opacification under different speed and spatial frequency.

PURPOSE: To investigate the characteristics of dynamic visual acuity (DVA) and contrast sensitivity (CS) in pseudophakic patients with posterior capsular opacification (PCO). METHODS: Fifty-four eyes (36 patients) with PCO planned for laser capsulotomy were recruited. They underwent examinations of static visual acuity (SVA), DVA, CS and optical quality analysis (OQAS) before and one week after the laser treatment. Improvements in each index after laser treatment were analyzed. The visual quality of patients with good initial vision was studied separately. RESULTS: SVA, DVA and CS all significantly increased after capsulotomy (P < 0.05). Postoperative improvements in DVA were higher than in SVA, but they decreased when the speed increased. DVA at 15 dps gained the most improvement after capsulotomy. DVA at all analyzed speeds was significantly lower than SVA (P = 0.000). There was a significant speed-dependent decrease in DVA at lower speeds compared with higher speeds. The postoperative improvements in CS decreased when the spatial frequency was increased. The CS at the lower frequencies of 3 cpd and 6 cpd was the most improved after capsulotomy. CS was much lower at high frequencies (p < 0.05). There was a significant decrease in CS at higher spatial frequencies compared with lower frequencies. DVA improvements were correlated with CS improvements at medium spatial frequencies and with objective scattering index and Strehl ratio. The CS at all frequencies significantly improved for patients with good initial vision. CONCLUSION: PCO could impair dynamic vision function, but CS was a more sensitive indication of visual complaints in patients with slight PCO.

Icariin promotes osteogenic differentiation of bone marrow stromal cells and prevents bone loss in OVX mice via activating autophagy.

Osteoporosis (OP) results from the impaired function of endogenous bone marrow mesenchymal stem cells (BMSCs). Icariin (ICA) has shown potential osteoprotective effects. However, the molecular mechanism for the anabolic action of ICA remains largely unknown. The objective of the present study is to investigate whether ICA prevents bone loss by acting on BMSCs via affecting the level of autophagy after ovariectomy (OVX). The BMSCs were extracted from BALB/c mice treated with ICA, chloroquine (CQ, an autophagy inhibitor) or ICA + CQ. The OVX mice were injected with ICA, CQ, or ICA + CQ for 1 month. We performed Alizarin Red staining and alkaline phosphatase staining to detect osteogenic differentiation of BMSCs. Micro-CT, hematoxylin and eosin staining, Oil Red O staining, and tartrate-resistant acid phosphatase staining were used to assess the bone mass, lipid droplets and osteoclasts in femurs. Autophagy activity in BMSCs from different groups was evaluated by Western blot analysis. The osteogenic differentiation of BMSCs from OVX-induced OP mice was decreased. Treatment with ICA reduced bone loss and formation of osteoclasts and increased osteogenic differentiation of BMSCs in vitro and vivo. In addition, autophagy was enhanced in BMSCs of OVX mice treated with ICA. Our results indicate that ICA prevents OVX-induced bone loss possibly by strengthening the osteogenic differentiation of BMSCs via increasing autophagic activity.

The impact of age-related cataracts on colour perception, postoperative recovery and related spectra derived from test of hue perception.

BACKGROUND: Cataract patients were always excluded from studies on ageing of colour vision; thus, effect of age-related cataracts on deterioration of colour perception has not been analysed. In present study, impacts of age-related cataracts on colour discrimination, postoperative recovery and related spectra were investigated. METHODS: In this cohort study, thirty age-related cataract patients scheduled for binocular surgery and 30 elderly volunteers were enrolled. Colour discrimination under photopic (1000 lx) and mesopic (40 lx) conditions was evaluated with Farnsworth-Munsell 100-hue test. The total error score (TES) and partial error score (PES) were calculated. RESULTS: Preoperatively, the TES in the patient group was 129.7 ± 59.5 at 1000 lx and 194.6 ± 74.5 at 40 lx, exhibiting worse discrimination than the volunteer group (TES1000lux = 71.5 ± 37.5 and TES40lux = 113.1 ± 38.8, p ≤ 0.001). Inferior perception were detected in the yellow to green-yellow (Y-GY), green-yellow to green (GY-G), green to blue-green (G-BG) and blue-green to blue (BG-B) colour bands (p ≤ 0.003), corresponding to the 470 nm-580 nm range of the visible light spectrum. Under mesopic conditions, the impact expanded to all colour bands except for yellow-red to yellow (YR-Y). Postoperatively, the TES in the patient group were 80.4 ± 62.4 at 1000 lx and 112.0 ± 85.2 at 40 lx, which were lower than those of the preoperative phase (p ≤ 0.001) but similar to those of the volunteer group (p ≥ 0.505). Postoperative improvement occurred in the Y-GY, GY-G and G-BG colour bands (490 nm to 580 nm) at 1000 lx (p ≤ 0.001) and shifted to the Y-GY, GY-G, G-BG and BG-B colour bands (470 nm to 580 nm) at 40 lx (p ≤ 0.001). Deterioration of hue perception for decrement of illumination was detected in the red to yellow-red (R-YR), Y-GY, G-BG, BG-B, blue to purple-blue (B-PB) and red-purple to red (RP-R) colour bands (450 nm to 500 nm) in the volunteer group (p ≤ 0.002) and the R-YR, G-BG, BG-B, B-PB, PB-P and red-purple to red (RP-R) colour bands (from the short-wavelength end to 500 nm) in the patient group preoperatively (p ≤ 0.001). CONCLUSIONS: Phacoemulsification could effectively rebuild colour perception in patients with age-related cataract. The postoperative benefits were most significant in colour bands corresponding with spectrum from 470 nm to 580 nm.

Game theory-based visual tracking approach focusing on color and texture features.

It is difficult for a single-feature tracking algorithm to achieve strong robustness under a complex environment. To solve this problem, we proposed a multifeature fusion tracking algorithm that is based on game theory. By focusing on color and texture features as two gamers, this algorithm accomplishes tracking by using a mean shift iterative formula to search for the Nash equilibrium of the game. The contribution of different features is always keeping the state of optical balance, so that the algorithm can fully take advantage of feature fusion. According to the experiment results, this algorithm proves to possess good performance, especially under the condition of scene variation, target occlusion, and similar interference.

Cardioprotective effect of nicorandil against myocardial injury following cardiac arrest in swine.

INTRODUCTION: Nicorandil, a vasodilatory drug used to treat angina, was reported to protect against myocardial ischemia-reperfusion injury in various animal models. However, its cardioprotective action following cardiac arrest is unknown. We examined the cardioprotective effects of nicorandil in a porcine model of cardiac arrest and resuscitation. METHODS: Ventricular fibrillation was induced electrically for 4min in anesthetized domestic swine, followed by cardiopulmonary resuscitation. Sixteen successfully resuscitated animals were randomized to saline control (n=8) or nicorandil (n=8) groups. Nicorandil (150µg/kg) was administered by central intravenous injection at onset of restoration of spontaneous circulation (ROSC), followed by 3µg/kg/min infusion until reperfusion end. Sham-operated animals received surgery only (n=4). Hemodynamic parameters were monitored continuously. Blood samples were taken at baseline, 5, 30, 180, and 360min after ROSC. Left ventricular ejection fraction was assessed by echocardiography at baseline and 6h after ROSC. The animals were euthanized 6h after ROSC, and the cardiac tissue was removed for analysis. RESULTS: 6 h after ROSC, nicorandil had significantly improved all hemodynamic variables (all P<0.05) except the maximum rate of left ventricular pressure decline and heart rate (P>0.05) compared with the control group. Control animals showed elevated cardiac troponin I and lactate levels compared with sham animals, which were significantly decreased following nicorandil treatment (P<0.05). In the saline control group, the adenosine triphosphate (ATP) content was largely reduced but subsequently rescued by nicorandil (P<0.05). Histopathologic injury was reduced with nicorandil treatment. Nicorandil reduced cardiomyocyte apoptosis as evidenced by reduced terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL)-positive cells, decreased Bax and caspase-3 expression, and increased Bcl-2 expression in the myocardium (all P<0.05). CONCLUSION: Nicorandil exhibited cardioprotective effects on myocardial injury following cardiac arrest via improvement in post-resuscitation myocardial dysfunction and energy metabolism, reduction in myocardial histopathologic injury, and antiapoptotic effects.

Effect of thermal treatment on microcracking characteristics of granite under tensile condition based on bonded-particle model and moment tensor.

It is known that the heterogeneity caused by thermally induced micro-cracks and thermal stress can affect the mechanical behavior of granite. The laboratory-scale tests have the intrinsic limitation of non-repeatability and lack of effective methods to characterize the interaction effect between thermal micro-cracks and thermal stresses. In this study, we demonstrate how advancements in particle bonded model and moment tensor can help better understand the roles of high temperature in weakening granite and thermally induced cracking process in Brazilian test. Our results show that the types of micro-cracks (intergranular, intragranular, and transcrystalline ones) are related to their thermal expansion coefficients of mineralogical compositions. The intergranular tensile micro-cracks are predominant during the heating and heating-cooling processes. An obvious weakening of granite and non-central initiation is associated with the heterogeneity caused by the thermal damage and thermal stress. We also quantitatively evaluate the thermal damage based on orientation distribution, b-value, and nature of the sources, which gives a new microcracking perspective on tensile characteristics subjected to high temperature.

Extensive Analysis of Mulberry (Morus rubra L.) Polysaccharides with Different Maturities by Using Two-Step Extraction and LC/QqQ-MS.

A primary challenge of polysaccharide analysis is the need for comprehensive extraction and characterization methods. In this study, mulberry polysaccharides at different maturities were fully extracted through a two-step process involving ethylenediaminetetraacetic acid (EDTA) and sodium hydroxide (NaOH), and their structures were determined by a combination analysis of monosaccharides and glycosidic linkages based on liquid chromatography triple quadrupole mass spectrometry (LC/QqQ-MS). The results indicate mulberry polysaccharides mainly contain highly branched pectic polysaccharides, (1,3,6)-linked glucan, xylan, and xyloglucan, but the content of different portions varies at different maturity stages. HG decreases from 19.12 and 19.14% (green mulberry) to 9.80 and 6.08% (red mulberry) but increases to 17.83 and 11.83% as mulberry transitioned from red to black. In contrast, the contents of glucan showed opposite trends. When mulberry turns red to black, the RG-I arabinan chains decrease from 47.75 and 28.86% to 13.16 and 12.72%, while the galactan side chains increase from 1.18 and 1.91 to 8.3 and 6.49%, xylan and xyloglucan show an increase in content. Overall, the two-step extraction combined with LC/QqQ-MS provides a new strategy for extensive analysis of complex plant polysaccharides.

Polysaccharide structure evaluation of Ganoderma lucidum from different regions in China based on an innovative extraction strategy.

The polysaccharides and triterpenes are important functional components of Ganoderma lucidum, but traditional preparation process of G. lucidum functional components can only realize the preparation of single functional component, which has poor targeting and low efficiency. In this study, the existence state of the functional components of G. lucidum was revealed. Then, the single step extraction process for functional components was established, and the precise structure evaluation of polysaccharide and triterpenes was conducted based on the process. The results showed that preparation time required for this strategy is only one-sixth of the traditional one, and 50 % of raw materials can be saved. Structural analysis of the functional components revealed that triterpenes were mainly Ganoderic acid and Lucidenic acid, and the polysaccharide structure was mainly 1,3-glucan and 1,3,6-glucan. The establishment of single step extraction strategy and the evaluation of the fine structure of functional components improved the efficiency of preparation and result determination, and provided an important basis for the development and utilization of green and low-carbon G. lucidum and even edible fungi resources and human nutritional dietary improvement strategies.

Overcoming strength-ductility tradeoff with high pressure thermal treatment.

Conventional material processing approaches often achieve strengthening of materials at the cost of reduced ductility. Here, we show that high-pressure and high-temperature (HPHT) treatment can help overcome the strength-ductility trade-off in structural materials. We report an initially strong-yet-brittle eutectic high entropy alloy simultaneously doubling its strength to 1150 MPa and its tensile ductility to 36% after the HPHT treatment. Such strength-ductility synergy is attributed to the HPHT-induced formation of a hierarchically patterned microstructure with coherent interfaces, which promotes multiple deformation mechanisms, including dislocations, stacking faults, microbands and deformation twins, at multiple length scales. More importantly, the HPHT-induced microstructure helps relieve stress concentration at the interfaces, thereby arresting interfacial cracking commonly observed in traditional eutectic high entropy alloys. These findings suggest a new direction of research in employing HPHT techniques to help develop next generation structural materials.

Structure of 2-haloacid dehalogenase from Pseudomonas syringae pv. tomato DC3000.

2-Haloacid dehalogenases (2-HADs) catalyse the hydrolytic dehalogenation of 2-haloalkanoic acids, cleaving the carbon-halide bond at the C(α)-atom position and releasing a halogen atom. These enzymes are of interest for their potential use in bioremediation and in the synthesis of industrial chemicals. Here, the crystal structure of 2-HAD from Pseudomonas syringae pv. tomato DC3000 (ps-2-HAD) at 1.98 Å resolution solved using the single-wavelength anomalous dispersion method is reported. The ps-2-HAD molecule consists of two structurally distinct domains: the core domain and the subdomain. Enzymatic activity analysis of ps-2-HAD revealed its capacity to catalyse the dehalogenation of both L- and D-substrates; however, the structure of ps-2-HAD is completely different from that of DehI, which is the only DL-2-HAD enzyme that has been structurally characterized, but shows similar overall folding to L-HADs. Single mutations of four amino-acid residues at the putative active site showed that they are related to its enzymatic activity, yet three of them are nonconserved among HADs. These observations imply that ps-2-HAD has a novel active site and a unique catalytic behaviour compared with other HADs. This study provides a structural basis and biochemical evidence for further elucidation of the catalytic mechanism of 2-HAD.