SP6 controls human cytotrophoblast fate decisions and trophoblast stem cell establishment by targeting MSX2 regulatory elements.

The commitment and differentiation of human placental progenitor cytotrophoblast (CT) cells are crucial for a successful pregnancy, but the underlying mechanism remains poorly understood. Here, we identified the transcription factor (TF), specificity protein 6 (SP6), as a human species-specific trophoblast lineage TF expressed in human placental CT cells. Using pluripotent stem cells as a model, we demonstrated that SP6 controls CT generation and the establishment of trophoblast stem cells (TSCs) and identified msh homeobox 2 (MSX2) as the downstream effector in these events. Mechanistically, we showed that SP6 interacts with histone acetyltransferase P300 to alter the landscape of H3K27ac at targeted regulatory elements, thereby favoring transcriptional activation and facilitating CT cell fate decisions and TSC maintenance. Our results established SP6 as a regulator of the human trophoblast lineage and implied its role in placental development and the pathogenies of placental diseases.

The analysis on groundwater storage variations from GRACE/GRACE-FO in recent 20 years driven by influencing factors and prediction in Shandong Province, China.

Monitoring and predicting the regional groundwater storage (GWS) fluctuation is an essential support for effectively managing water resources. Therefore, taking Shandong Province as an example, the data from Gravity Recovery and Climate Experiment (GRACE) and GRACE Follow-On (GRACE-FO) is used to invert GWS fluctuation from January 2003 to December 2022 together with Watergap Global Hydrological Model (WGHM), in-situ groundwater volume and level data. The spatio-temporal characteristics are decomposed using Independent Components Analysis (ICA), and the impact factors, such as precipitation and human activities, which are also analyzed. To predict the short-time changes of GWS, the Support Vector Machines (SVM) is adopted together with three commonly used methods Long Short-Term Memory (LSTM), Singular Spectrum Analysis (SSA), Auto-Regressive Moving Average Model (ARMA), as the comparison. The results show that: (1) The loss intensity of western GWS is significantly greater than those in coastal areas. From 2003 to 2006, GWS increased sharply; during 2007 to 2014, there exists a loss rate - 5.80 ± 2.28 mm/a of GWS; the linear trend of GWS change is - 5.39 ± 3.65 mm/a from 2015 to 2022, may be mainly due to the effect of South-to-North Water Diversion Project. The correlation coefficient between GRACE and WGHM is 0.67, which is consistent with in-situ groundwater volume and level. (2) The GWS has higher positive correlation with monthly Global Precipitation Climatology Project (GPCP) considering time delay after moving average, which has the similar energy spectrum depending on Continuous Wavelet Transform (CWT) method. In addition, the influencing facotrs on annual GWS fluctuation are analyzed, the correlation coefficient between GWS and in-situ data including the consumption of groundwater mining, farmland irrigation is 0.80, 0.71, respectively. (3) For the GWS prediction, SVM method is adopted to analyze, three training samples with 180, 204 and 228 months are established with the goodness-of-fit all higher than 0.97. The correlation coefficients are 0.56, 0.75, 0.68; RMSE is 5.26, 4.42, 5.65 mm; NSE is 0.28, 0.43, 0.36, respectively. The performance of SVM model is better than the other methods for the short-term prediction.

Aromatic pentaamide macrocycles bind anions with high affinity for transport across biomembranes.

The convergent positioning of functional groups in biomacromolecules leads to good binding, catalytic and transport capabilities. Synthetic frameworks capable of convergently locking functional groups with minimized conformational uncertainty-leading to similar properties-are highly desirable but rare. Here we report C5-symmetric aromatic pentaamide macrocycles synthesized in one pot from the corresponding monomers. Their crystal structures reveal a star-shaped, fully constrained backbone that causes ten alternating NH/CH hydrogen-bond donors and five large amide dipoles to orient towards the centre of the macrocycle. With a highly electropositive cavity in a high-energy unbound state, the macrocycles bind anions in a 1:1 stoichiometry in solution, with high affinity for halides and very high affinity for oxoanions. We demonstrate that such macrocycles are able to transport anions across lipid bilayers with a high chloride selectivity and restore the depleted airway surface liquid of cystic fibrosis airway cell cultures.

High-affinity single and double helical pseudofoldaxanes with cationic guests.

Two aromatic oligoamides, the 8-residue H8 and 16-residue H16, that adopt stable, cavity-containing helical conformations were examined for their complexation of a rodlike dicationic guest, octyl viologen (OV2+) and para-bis(trimethylammonium)benzene (TB2+). Studies based on 1D and 2D 1H NMR, isothermal titration calorimetry (ITC), and X-ray crystallography demonstrated that H8 and H16 wraps around two OV2+ ions as a double helix and a single helix, respectively, resulting in 2 : 2 and 1 : 2 complexes. Compared to H8, the longer H16 binds the OV2+ ions with much higher binding affinity and with extraordinary negative cooperativity. In contrast to its 1 : 2 binding with OV2+, the binding of helix H16 with the bulkier guest TB2+ shows a 1 : 1 ratio. Host H16 also selectively binds OV2+ in the presence of TB2+. This novel host-guest system features pairwise placement of the otherwise strongly repulsive OV2+ ions in the same cavity, strong negative cooperativity, and mutual adaptability of hosts and guests. The resultant complexes are highly stable [2]-, [3]-, and [4]pseudo-foldaxanes with few known precedents.

Reconstructing a long-term water storage-based drought index in the Yangtze River Basin.

Drought is a prolonged dry period in the natural climate cycle, and is one of the most costly weather events. The Gravity Recovery and Climate Experiment (GRACE) derived terrestrial water storage anomalies (TWSA) have been widely used to assess drought severity. However, the relatively short cover period of GRACE and GRACE Follow-On limit our knowledge about the characterization and evolution of drought over decades time scale. This study proposes a standardized GRACE reconstructed TWSA index (SGRTI) to assess the drought severity based on a statistical reconstruction method calibrated by GRACE observations. Results show that the SGRTI correlates well with 6-month scale SPI and SPEI, with correlation coefficients reaching 0.79 and 0.81 in the YRB from 1981 to 2019. Soil moisture can capture drought condition like the SGRTI, while cannot further reflect deeper water storage depletion. The SGRTI is also comparable to the SRI and in-situ water level. As a case study for the Yangtze River Basin, its three sub-basins experience more frequent droughts, shorter drought duration, and lower severity drought, as identified by SGRTI during 1992-2019 relative to 1963-1991. The presented SGRTI in this study can provide a valuable supplement to the drought index before the GRACE era.

Single ultrasound-guided thoracic paravertebral block with a large volume of anesthetic for microwave ablation of lung tumors.

Objective: To compare single ultrasound-guided thoracic paravertebral block (TPVB) using a large volume of anesthetic with local anesthesia (LA) in computed tomography (CT)-guided pulmonary microwave ablation. Subjects and methods: Eighty patients who underwent CT-guided microwave ablation of pulmonary tumors were randomly divided into the TPVB group and the LA group. Patients of the TPVB group were anesthetized with a single injection of a large volume (40 ml) of 0.375% ropivacaine injection at T4, and those of the LA group had local infiltration by the surgeon at the puncture site, and emergency rescue with propofol injection was administered when the patient could not tolerate pain in either group. The following variables were recorded in both groups: general conditions; volume of propofol injection for emergency rescue during ablation; visual analog scale (VAS) scores during ablation and at 0, 2, 12, and 24 h after ablation; the need to use analgesics for rescue within 24 h after ablation; number of ablations; number of punctures performed by the surgeon; patient's movements during puncturing; and puncturing-associated complications. Results: Compared with the TPVB group, the amount of emergency use of propofol injection was significantly more in the LA group (P < 0.05). There were no significant differences in the VAS scores recorded intraoperatively and at 0, 2, 12, and 24 h after ablation between the two groups (P > 0.05). There was a significant difference in the patient's movements upon puncturing between the two groups (P < 0.05), but there were no significant differences in the numbers of punctures and ablations between the two groups (P > 0.05). The number of patients using analgesics within 24 h after the operation was also more in the LA group than in the TPVB group, and the difference between the two groups was statistically significant (P < 0.05). Conclusion: Single ultrasound-guided TPVB with a large volume of anesthetic offers effective analgesia for microwave ablation of lung tumors, helping the patient cooperate with the operating surgeon to reduce injury from multiple lung punctures. Further studies are recommended to validate these findings.

Ultra-Tight Host-Guest Binding with Exceptionally Strong Positive Cooperativity.

Cooperativity plays a critical role in self-assembly and molecular recognition. A rigid aromatic oligoamide macrocycle with a cyclodirectional backbone binds with DABCO-based cationic guests in a 2 : 1 ratio in high affinities (Ktotal ≈1013  M-2 ) in the highly polar DMF. The host-guest binding also exhibits exceptionally strong positive cooperativity quantified by interaction factors α that are among the largest for synthetic host-guest systems. The unusually strong positive cooperativity, revealed by isothermal titration calorimetry (ITC) and fully corroborated by mass spectrometry, NMR and computational studies, is driven by guest-induced stacking of the macrocycles and stabilization from the alkyl end chains of the guests, interactions that appear upon binding the second macrocycle. With its tight binding driven by extraordinary positive cooperativity, this host-guest system provides a tunable platform for studying molecular interactions and for constructing stable supramolecular assemblies.

Cavity-containing aromatic oligoamide foldamers and macrocycles: progress and future perspectives.

As a major class of foldamers, aromatic oligoamide foldamers have attracted intense interest. The rigidity of aromatic residues and amide linkages allows the development of foldamers with readily predictable, stable conformations. Aromatic oligoamide foldamers having backbones fully constrained by intramolecular hydrogen bonds have attracted wide attention. Depending on their lengths, such foldamers adopt crescent or helical conformations with highly negative inner cavities. Cyclizing the backbone of the aromatic oligoamides affords the corresponding macrocycles which are characterised by persistent shapes and non-deformable inner cavities. With their defined, inner cavities, such aromatic oligoamide foldamers and macrocycles have served as hosts for cationic and polar guests, and as transmembrane channels for transporting ions and molecules. Recent synthetic progress resulted in the construction of multi-turn hollow helices that offer three-dimensional inner pores with adjustable depth. Reducing the number of backbone-constraining hydrogen bonds leads to oligoamides which, with their partially constrained backbones, undergo either solvent- or guest-dependent folding. One class of such aromatic olgioamide foldamders, which offer multiple backbone amide NH groups as hydrogen-bond donors, are designed to bind anions with adjustable affinities.

Controlling Water Flow through a Synthetic Nanopore with Permeable Cations.

There is presently intense interest in the development of synthetic nanopores that recapitulate the functional properties of biological water channels for a wide range of applications. To date, all known synthetic water channels have a hydrophobic lumen, and while many exhibit a comparable rate of water transport as biological water channels, there is presently no rationally designed system with the ability to regulate water transport, a critical property of many natural water channels. Here, we describe a self-assembling nanopore consisting of stacked macrocyclic molecules with a hybrid hydrophilic/hydrophobic lumen exhibiting water transport that can be regulated by alkali metal ions. Stopped-flow kinetic assays reveal a non-monotonic-dependence of transport on cation size as well as a strikingly broad range of water flow, from essentially none in the presence of the sodium ion to as high a flow as that of the biological water channel, aquaporin 1, in the absence of the cations. All-atom molecular dynamics simulations show that the mechanism underlying the observed sensitivity is the binding of cations to defined sites within this hybrid pore, which perturbs water flow through the channel. Thus, beyond revealing insights into factors that can modulate a high-flux water transport through sub-nm pores, the obtained results provide a proof-of-concept for the rational design of next-generation, controllable synthetic water channels.

Stable pseudo[3]rotaxanes with strong positive binding cooperativity based on shape-persistent aromatic oligoamide macrocycles.

New aromatic oligoamide macrocycles with C3-symmetry bind a bipyridinium guest (G) to form compact pseudo[3]rotaxanes involving interesting enthalpic and entropic contributions. The observed high stabilities and strong positive binding cooperativity are found in few other host-guest systems.

Dipropinonates of Sugar Alcohols as Water-Soluble, Nontoxic CPAs for DMSO-Free Cell Cryopreservation.

Sorbitol, mannitol, xylitol, and erythritol, four readily available sugar alcohols with poor or no membrane permeability, are converted into their corresponding dipropionates by acylating their primary hydroxyl groups. With enhanced membrane permeability, these diesters are expected to permeate the cell membranes and, upon their hydrolysis, release the corresponding sugar alcohols inside the cells. NIH-3T3 cells incubated with these diesters before being frozen at -80 °C exhibited considerably higher total recovery over those incubated with the free sugar alcohols or media only. Among the four diesters, those of sorbitol, especially mannitol, showed cryoprotective effects comparable to that shown by 5% DMSO. This work has demonstrated the feasibility of converting readily available, naturally occurring compounds into membrane-permeable derivatives that serve as water-soluble, nontoxic alternatives to DMSO.

Precipitation Changes in the Three Gorges Reservoir Area and the Relationship with Water Level Change.

As the largest hydroelectric project worldwide, previous studies indicate that the Three Gorges Dam (TGD) affects the local climate because of the changes of hydrological cycle caused by the impounding and draining of the TGD. However, previous studies do not analyze the long-term precipitation changes before and after the impoundment, and the variation characteristics of local precipitation remain elusive. In this study, we use precipitation anomaly data derived from the CN05.1 precipitation dataset between 1988 and 2017 to trace the changes of precipitation before and after the construction of the TGD (i.e., 1988-2002 and 2003-2017), in the Three Gorges Reservoir Area (TGRA). Results showed that the annual and dry season precipitation anomaly in the TGRA presented an increasing trend, and the precipitation anomaly showed a slight decrease during the flood season. After the impoundment of TGD, the precipitation concentration degree in the TGRA decreased, indicating that the precipitation became increasingly uniform, and the precipitation concentration period insignificantly increased. A resonance phenomenon between the monthly average water level and precipitation anomaly occurred in the TGRA after 2011 and showed a positive correlation. Our findings revealed the change of local precipitation characteristics before and after the impoundment of TGD and showed strong evidence that this change had a close relationship with the water level.

Major Factors for the Persistent Folding of Hybrid α, ß, γ-Hybrid Peptides Into Hairpins.

Factors responsible for the persistent adoption of hairpin conformations by hybrid oligopeptides, each having a central ß/α dipeptide segment flanked by aromatic γ-amino acid (γAr) residues, are probed. Our recent studies revealed that tetrapeptide 1 and 2, having central dipeptide segments consisting of ß-alanine (ß-Ala) and glycine (Gly), and L-ß-homophenylalanine (L-ß-homoPhe) and Gly residues, respectively, that are flanked by γAr residues, fold into well-defined, expanded ß-turns with doubly H-bonded γAr residues. Replacing the γAr residues of 1 and 2 with L-Val and L-Leu residues results in tetrapetides 1 ' and 2 ' that fail to fold into defined conformations, which confirms the decisive role played by the H-bonded γAr residues in the promoting folding of 1 and 2. Attaching L-Val and L-Leu residues to the termini of 1 affords hexapeptide 1a. With an additional H-bond between its L-Val and L-Leu residues, peptide 1a folds into a hairpin with higher stability than that of 1, indicating that the expanded ß-turn can nucleate and stabilize ß-hairpin with longer ß-strands. Attaching L-Val and L-Leu residues to the termini of 2 affords hexapeptide 2a. Substituting the L-ß-homoPhe residue of 2a with a D-ß-homoPhe residue gives hexapeptide 2b. Surprisingly, hexapeptide 2a fold into a hairpin showing the similar stability as those of tetrapeptides 1 and 2. Hexapeptide 2b, with its combination of a D-ß-homoPhe residue and the L-Val/L-Leu pair, fold into a hairpin that is significantly more stable than the other hybrid peptides, demonstrating that a combination of hetero-chirality between the ß-amino acid residue of the dipeptide loop and the α-amino acid residues of the ß-strands enhances the stability of the resultant ß-hairpin.

Multiturn Hollow Helices: Synthesis and Folding of Long Aromatic Oligoamides.

Aromatic oligoamides adopting helical conformations are synthesized by coupling carboxyl-terminated basic units having two, four, and eight residues to amine-terminated oligomer precursors. Coupling yields show no noticeable reduction with the size of the basic units or the final product. One- and two-dimensional NMR spectroscopy and computational studies demonstrate the reliable helical folding of these oligomers. The X-ray structure of 16mer 7 reveals a compact, multiturn helix having a 9 Å inner pore.

Ultrasensitive liposome-based assay for the quantification of fundamental ion channel properties.

One of the most widely used approaches to characterize transmembrane ion transport through nanoscale synthetic or biological channels is a straightforward, liposome-based assay that monitors changes in ionic flux across the vesicle membrane using pH- or ion-sensitive dyes. However, failure to account for the precise experimental conditions, in particular the complete ionic composition on either side of the membrane and the inherent permeability of ions through the lipid bilayer itself, can prevent quantifications and lead to fundamentally incorrect conclusions. Here we present a quantitative model for this assay based on the Goldman-Hodgkin-Katz flux theory, which enables accurate measurements and identification of optimal conditions for the determination of ion channel permeability and selectivity. Based on our model, the detection sensitivity of channel permeability is improved by two orders of magnitude over the commonly used experimental conditions. Further, rather than obtaining qualitative preferences of ion selectivity as is typical, we determine quantitative values of these parameters under rigorously controlled conditions even when the experimental results would otherwise imply (without our model) incorrect behavior. We anticipate that this simply employed ultrasensitive assay will find wide application in the quantitative characterization of synthetic or biological ion channels.

Spatio-Temporal Variations in Groundwater Revealed by GRACE and Its Driving Factors in the Huang-Huai-Hai Plain, China.

The Huang-Huai-Hai (3H) Plain is the major crop-producing region in China. Due to the long-term overexploitation of groundwater for irrigation, the groundwater funnel is constantly expanding and the scarcity of water resources is prominent in this region. In this study, Gravity Recovery and Climate Experiment (GRACE) and hydrological models were used to estimate the spatial-temporal changes of groundwater storage (GWS) and the driving factors of GWS variations were discussed in the 3H Plain. The results showed that GRACE-based GWS was depleted at a rate of -1.14 ± 0.89 cm/y in the 3H Plain during 2003 to 2015. The maximum negative anomaly occurred in spring due to agricultural irrigation activities. Spatially, the loss of GWS in the Haihe River Basin is more serious than that in the Huaihe River Basin, presenting a decreasing trend from south to north. Conversely, the blue water footprint (WFblue) of wheat exhibited an increasing trend from south to north. During the drought years of 2006, 2013, and 2014, more groundwater was extracted to offset the surface water shortage, leading to an accelerated decline in GWS. This study demonstrated that GWS depletion in the 3H Plain is well explained by reduced precipitation and groundwater abstraction due to anthropogenic irrigation activities.

Reverse Turn Foldamers: An Expanded ß-Turn Motif Reinforced by Double Hydrogen Bonds.

Hybrid tetrapeptides sharing a backbone with a central α/ß-dipeptide segment flanked by aromatic γ-amino acid residues fold into the same hairpin conformation with an expanded ß-turn. This hairpin/ß-turn motif is general for accommodating different α- and ß-amino acid residues. Replacing glycine with other α-amino acid residues has an insignificant influence on or slightly decreases the stabilities of the folded conformations; substituting ß-alanine with other ß-amino acid residues enhances the stabilities of the folded structures.

Self-Assembly and Molecular Recognition in Water: Tubular Stacking and Guest-Templated Discrete Assembly of Water-Soluble, Shape-Persistent Macrocycles.

Supramolecular chemistry in aqueous media is an area with great fundamental and practical significance. To examine the role of multiple noncovalent interactions in controlled assembling and binding behavior in water, the self-association of five water-soluble hexakis(m-phenylene ethynylene) (m-PE) macrocycles, along with the molecular recognition behavior of the resultant assemblies, is investigated with UV-vis, fluorescence, CD, and NMR spectroscopy, mass spectrometry, and computational studies. In contrast to their different extents of self-aggregation in organic solvents, all five macrocycles remain aggregated in water at concentrations down to the micromolar (µM) range. CD spectroscopy reveals that 1-F6 and 1-H6, two macrocycles carrying chiral side chains and capable of H-bonded self-association, assemble into tubular stacks. The tubular stacks serve as supramolecular hosts in water, as exemplified by the interaction of macrocycles 1-H6 and 2-H6 and guests G1 through G4, each having a rod-like oligo(p-phenylene ethynylene) (p-PE) segment flanked by two hydrophilic chains. Fluorescence and 1H NMR spectroscopy revealed the formation of kinetically stable, discrete assemblies upon mixing 2-H6 and a guest. The binding stoichiometry, determined with fluorescence, 1H NMR, and ESI-MS, reveals that the discrete assemblies are novel pseudorotaxanes, each containing a pair of identical guest molecules encased by a tubular stack. The two guest molecules define the number of macrocyclic molecules that comprise the host, which curbs the "infinite" stack growth, resulting in a tubular stack with a cylindrical pore tailoring the length of the p-PE segment of the bound guests. Each complex is stabilized by the action of multiple noncovalent forces including aromatic stacking, side-chain H-bonding, and van der Waals interactions. Thus, the interplay of multiple noncovalent forces aligns the molecules of macrocycles 1 and 2 into tubular stacks with cylindrical inner pores that, upon binding rod-like guests, lead to tight, discrete, and well-ordered tubular assemblies that are unprecedented in water.

Bridging Terrestrial Water Storage Anomaly During GRACE/GRACE-FO Gap Using SSA Method: A Case Study in China.

The terrestrial water storage anomaly (TWSA) gap between the Gravity Recovery and Climate Experiment (GRACE) and its follow-on mission (GRACE-FO) is now a significant issue for scientific research in high-resolution time-variable gravity fields. This paper proposes the use of singular spectrum analysis (SSA) to predict the TWSA derived from GRACE. We designed a case study in six regions in China (North China Plain (NCP), Southwest China (SWC), Three-River Headwaters Region (TRHR), Tianshan Mountains Region (TSMR), Heihe River Basin (HRB), and Lishui and Wenzhou area (LSWZ)) using GRACE RL06 data from January 2003 to August 2016 for inversion, which were compared with Center for Space Research (CSR), Helmholtz-Centre Potsdam-German Research Centre for Geosciences (GFZ), Jet Propulsion Laboratory (JPL)'s Mascon (Mass Concentration) RL05, and JPL's Mascon RL06. We evaluated the accuracy of SSA prediction on different temporal scales based on the correlation coefficient (R), Nash-Sutcliffe efficiency (NSE), and root mean square error (RMSE), which were compared with that of an auto-regressive and moving average (ARMA) model. The TWSA from September 2016 to May 2019 were predicted using SSA, which was verified using Mascon RL06, the Global Land Data Assimilation System model, and GRACE-FO results. The results show that: (1) TWSA derived from GRACE agreed well with Mascon in most regions, with the highest consistency with Mascon RL06 and (2) prediction accuracy of GRACE in TRHR and SWC was higher. SSA reconstruction improved R, NSE, and RMSE compared with those of ARMA. The R values for predicting TWS in the six regions using the SSA method were 0.34-0.98, which was better than those for ARMA (0.26-0.97), and the RMSE values were 0.03-5.55 cm, which were better than the 2.29-5.11 cm RMSE for ARMA as a whole. (3) The SSA method produced better predictions for obvious periodic and trending characteristics in the TWSA in most regions, whereas the detailed signal could not be effectively predicted. (4) The predicted TWSA from September 2016 to May 2019 were basically consistent with Global Land Data Assimilation System (GLDAS) results, and the predicted TWSA during June 2018 to May 2019 agreed well with GRACE-FO results. The research method in this paper provides a reference for bridging the gap in the TWSA between GRACE and GRACE-FO.