Structural basis for urate recognition and apigenin inhibition of human GLUT9.

Urate, the physiological form of uric acid and a potent antioxidant in serum, plays a pivotal role in scavenging reactive oxygen species. Yet excessive accumulation of urate, known as hyperuricemia, is the primary risk factor for the development of gout. The high-capacity urate transporter GLUT9 represents a promising target for gout treatment. Here, we present cryo-electron microscopy structures of human GLUT9 in complex with urate or its inhibitor apigenin at overall resolutions of 3.5 Å and 3.3 Å, respectively. In both structures, GLUT9 exhibits an inward open conformation, wherein the substrate binding pocket faces the intracellular side. These structures unveil the molecular basis for GLUT9's substrate preference of urate over glucose, and show that apigenin acts as a competitive inhibitor by occupying the substrate binding site. Our findings provide critical information for the development of specific inhibitors targeting GLUT9 as potential therapeutics for gout and hyperuricemia.

Plasticity in ventral pallidal cholinergic neuron-derived circuits contributes to comorbid chronic pain-like and depression-like behaviour in male mice.

Nucleus- and cell-specific interrogation of individual basal forebrain (BF) cholinergic circuits is crucial for refining targets to treat comorbid chronic pain-like and depression-like behaviour. As the ventral pallidum (VP) in the BF regulates pain perception and emotions, we aim to address the role of VP-derived cholinergic circuits in hyperalgesia and depression-like behaviour in chronic pain mouse model. In male mice, VP cholinergic neurons innervate local non-cholinergic neurons and modulate downstream basolateral amygdala (BLA) neurons through nicotinic acetylcholine receptors. These cholinergic circuits are mobilized by pain-like stimuli and become hyperactive during persistent pain. Acute stimulation of VP cholinergic neurons and the VP-BLA cholinergic projection reduces pain threshold in naïve mice whereas inhibition of the circuits elevated pain threshold in pain-like states. Multi-day repetitive modulation of the VP-BLA cholinergic pathway regulates depression-like behaviour in persistent pain. Therefore, VP-derived cholinergic circuits are implicated in comorbid hyperalgesia and depression-like behaviour in chronic pain mouse model.

Differential remodeling of subthalamic projections to basal ganglia output nuclei and locomotor deficits in 6-OHDA-induced hemiparkinsonian mice.

The subthalamic nucleus (STN) controls basal ganglia outputs via the substantia nigra pars reticulata (SNr) and the globus pallidus internus (GPi). However, the synaptic properties of these projections and their roles in motor control remain unclear. We show that the STN-SNr and STN-GPi projections differ markedly in magnitude and activity-dependent plasticity despite the existence of collateral STN neurons projecting to both the SNr and GPi. Stimulation of either STN projection reduces locomotion; in contrast, inhibition of either the STN-SNr projection or collateral STN neurons facilitates locomotion. In 6-OHDA-hemiparkinsonian mice, the STN-SNr projection is dramatically attenuated, but the STN-GPi projection is robustly enhanced; apomorphine inhibition of the STN-GPi projection through D2 receptors is significantly augmented and improves locomotion. Optogenetic inhibition of either the STN-SNr or STN-GPi projection improves parkinsonian bradykinesia. These results suggest that the STN-GPi and STN-SNr projections are differentially involved in motor control in physiological and parkinsonian conditions.

Three gorges dam shifts estuarine heavy metal risk through suspended sediment gradation.

Damming alters downstream sediment supply relationships and erosion in the estuarine delta. Given that sediment grainsize serves as a key parameter for the ability to adsorb heavy metals from water, the assessment of estuarine heavy metal risk needs to get connected initially. Hence, fine suspended sediment (<63 µm) in the Yangtze River estuary (YRE) was divided into four grainsize fractions to simulate the surface suspended sediment concentration (SSC) and grainsize composition before and after the completion of the Three Gorges Dam (TGD). Representative months were selected for flood peak reduction (October) and runoff compensation in the dry season (March) to maximize the scheduling impact of the TGD on runoff and riverine sediment input to the YRE. An improved Water Quality Index (WQI) approach was proposed to assess the combined risk alteration of five heavy metals in six estuarine sensitive targets due to TGD-induced sediment characteristics. The results demonstrated that TGD significantly but tardily reduced the SSC and the proportion of fine sediment in the YRE, decreasing the risk of heavy metals resuspension. Seasonally, the total SSC became higher in the flood season than in the dry season during post-TGD period. However, the fine SSC in the flood season was averaged only 59.7% of that in the dry season due to the pronounced grainsize coarsening effect. As the significant reduction in fine SSC overcomes the increase in heavy metal content per unit of SS, the integrated resuspension risk declined significantly, particularly for Pb and Cr. Spatially, the risk reduction for sensitive targets near the turbidity maximum zone (TMZ) is 8.4 times greater than for inner river channel. However, undiminished anthropogenic metal inputs to the YRE signified greater pressures on the depositional environment.

The fate of microplastics in estuary: A quantitative simulation approach.

Microplastics pollution is an emerging environmental concern. However, there are almost no MPs numerical simulation studies in the Yangtze Estuary which is considered as the largest plastic export in the world and quantitative simulation is not carried out in the existing models. Therefore, completing quantitative simulation and exploring different patterns of MPs transport are the main objectives of this study. In addition, the concentration distribution and risk of MPs are also analyzed. Mass-Number method is proposed to quantitatively simulate microplastics concentration in Feb. and May with errors of less than 18%. Compared with sediment flocculation and settling transport, independent floating transport is more susceptible to surface currents resulting in increased beaching and more inhomogeneous concentration distribution. Meanwhile, under the influence of current, local topography and salt wedge, the MPs perform linear motion and clockwise spiral motion inside and outside the estuary and rapidly form a "hot spot" on the southeastern part of Chongming Island and 57% to 90% of MPs are beached or settled inside the estuary, especially on the north shore. Therefore, MPs risk in some sensitive targets should be concerned according to risk assessment results. Our results break the space-time limit and explore the fate of MPs in the Yangtze Estuary and provide new idea and concern of MPs numerical simulation.

How the Yangtze River transports microplastic to the east China sea.

Land transportation in the Yangtze River basin is an important source of microplastics in the East China Sea, so it is significant to clarify the source, fate and river-sea transition of microplastics. Taking the Yangtze River as the study area, the interpolation method was used to analyze the monthly changes of the microplastic load in the estuary, the input-output model was used to estimate the flux of microplastics into the sea, and the inflow process of microplastics was studied through correlation analysis. The results showed that: (1) The load of microplastics in the Yangtze River estuary varied with season, reaching the maximum in October, with a monthly load of 3.91 Gg; (2) The total amount of microplastics entering the sea in the Yangtze River basin was higher than the medium level, which was 7.02 Gg. Among them, tributary input was the most important source, accounting for 62.9%. Non-point and point sources were further subdivided into 11 categories, with the largest proportion of microplastics generated during vehicle trip. Spatially speaking, the microplastics transported in the midstream accounted for the largest proportion, accounting for 55.56%; (3) microplastics had a strong correlation with COD and TP, indicating that the inflow process of microplastics was similar to that of traditional pollutants, which were river retention, wastewater treatment plant removal, water consumption removal and inflow to the East China Sea. Although the proportion of the last one was only 8.05%, the ecological risk was still not negligible due to the huge amount.

How climate change and land-use evolution relates to the non-point source pollution in a typical watershed of China.

The water quality of Le 'an River Watershed (LRW) is crucial to the water environmental safety of Poyang Lake, especially the concentration of nitrogen and phosphorus. The effect of climate and land use change on watershed water quality has always been under the attention of local managers. More importantly, the lack of detailed studies on climate and land use impact on river water quality has prevented sustainable water security management in the LRW. Therefore, this study aimed to quantify the weight of climate and land use on nutrient loss in the LRW, respectively. We divided the historical period (1990-2020) into six scenarios and a baseline scenario. TN and TP losses in the watershed were simulated using Soil and Water Assessment Tool (SWAT), and the weight of climate and land use were quantified in overall, by period, and by region. The results showed that the weight of climate was greatly higher than land use with values around 90%. However, the weight of land use had a positive cumulative effect in a certain period, and its influence could not be neglected. The climate in all scenarios led to a reduction in nutrient loss, while land use was found to slightly increase the nutrient loss yield. In addition to, unique regional topographic features, urbanization rates, and climatic conditions could cause spatial heterogeneity in the climatic and land use weights.

Comparative structural analysis of human Nav1.1 and Nav1.5 reveals mutational hotspots for sodium channelopathies.

Among the nine subtypes of human voltage-gated sodium (Nav) channels, the brain and cardiac isoforms, Nav1.1 and Nav1.5, each carry more than 400 missense mutations respectively associated with epilepsy and cardiac disorders. High-resolution structures are required for structure-function relationship dissection of the disease variants. We report the cryo-EM structures of the full-length human Nav1.1-ß4 complex at 3.3 Å resolution here and the Nav1.5-E1784K variant in the accompanying paper. Up to 341 and 261 disease-related missense mutations in Nav1.1 and Nav1.5, respectively, are resolved. Comparative structural analysis reveals several clusters of disease mutations that are common to both Nav1.1 and Nav1.5. Among these, the majority of mutations on the extracellular loops above the pore domain and the supporting segments for the selectivity filter may impair structural integrity, while those on the pore domain and the voltage-sensing domains mostly interfere with electromechanical coupling and fast inactivation. Our systematic structural delineation of these mutations provides important insight into their pathogenic mechanism, which will facilitate the development of precise therapeutic interventions against various sodium channelopathies.

Molecular characterization and complement activating functional analysis of a new collectin(TfCol-11) from Trachidermus fasciatus.

The complement system is a crucial component of the innate immune system that links innate and adaptive immunity. CL-11, a protein similar to Mannose-binding lectin (MBL), plays significant role in the innate immune system in mammals and fish, serving as an initiator of the lectin pathway of complement activation. In this study, a CL-11 homolog (TfCol-11) was identified in roughskin sculpin (Trachidermus fasciatus), and its expression and role in immune responses were characterized. The open reading frame of TfCol-11 is 795 bp long, encoding a 264 amino acid polypeptide. The deduced amino acid sequence of this protein is highly homologous to sequences in other teleosts, and is similar to vertebrate CL-11, containing a canonical collagen-like region, a carbohydrate recognition domain, and a neck region. Recombinant TfCol-11 purified from Escherichia coli(E.coli) was able to bind to different microbes in a Ca2+-independent manner. Meanwhile, a 993 bp-long of partial MASP cDNA with a 96 bp 5' untranslated region (UTR) was also cloned from roughskin sculpin, containing 299 amino acids and consisting of three domains (CUB-EGF-CUB). qRT-PCR indicated that TfCol-11 and MASP mRNAs were predominately co-expressed in the liver. The temporal expression of TfCol-11 and MASP were both drastically up-regulated in the liver, skin, and blood by LPS challenge. Recombinant TfCol-11 purified from E.coli BL21(DE3) was able to agglutinate some bacteria in a Ca2+-dependent manner. In addition, an in vitro pull-down experiment demonstrated that TfCol-11 was able to bind to MASP, and in vivo experiments showed that TfCol-11 was associated with increased membrane attack complex (MAC) levels. It is therefore possible that TfCol-11 may plays a role in activating the complement system and in the defense against invading microorganisms in roughskin sculpin.