ABSTRACT
Optical nanoscopy of intact biological specimens has been transformed by recent advancements in hydrogel-based tissue clearing and expansion, enabling the imaging of cellular and subcellular structures with molecular contrast. However, existing high-resolution fluorescence microscopes have limited imaging depth, which prevents the study of whole-mount specimens without physical sectioning. To address this challenge, we developed "photochemical sectioning," a spatially precise, light-based sample sectioning process. By combining photochemical sectioning with volumetric lattice light-sheet imaging and petabyte-scale computation, we imaged and reconstructed axons and myelination sheaths across entire mouse olfactory bulbs at nanoscale resolution. An olfactory-bulb-wide analysis of myelinated and unmyelinated axons revealed distinctive patterns of axon degeneration and de-/dysmyelination in the neurodegenerative mouse, highlighting the potential for peta- to exabyte-scale super-resolution studies using this approach.
ABSTRACT
At each cell division, nanometer-scale motors and microtubules give rise to the micron-scale spindle. Many mitotic motors step helically around microtubules in vitro, and most are predicted to twist the spindle in a left-handed direction. However, the human spindle exhibits only slight global twist, raising the question of how these molecular torques are balanced. Here, we find that anaphase spindles in the epithelial cell line MCF10A have a high baseline twist, and we identify factors that both increase and decrease this twist. The midzone motors KIF4A and MKLP1 are together required for left-handed twist at anaphase, and we show that KIF4A generates left-handed torque in vitro. The actin cytoskeleton also contributes to left-handed twist, but dynein and its cortical recruitment factor LGN counteract it. Together, our work demonstrates that force generators regulate twist in opposite directions from both within and outside the spindle, preventing strong spindle twist during chromosome segregation.
Subject(s)
Anaphase , Kinesins , Microtubules , Spindle Apparatus , Humans , Spindle Apparatus/metabolism , Kinesins/metabolism , Kinesins/genetics , Microtubules/metabolism , Dyneins/metabolism , Dyneins/genetics , Torque , Chromosome Segregation , Actin Cytoskeleton/metabolism , Microtubule-Associated Proteins/metabolism , Microtubule-Associated Proteins/geneticsABSTRACT
Using biochar in agriculture to enhance soil carbon storage and productivity has been recognized as an effective means of carbon sequestration. However, the effects on crop yield and soil carbon and nitrogen can vary depending on environmental conditions, field management, and biochar conditions. Thus, we conducted a meta-analysis to identify the factors contributing to these inconsistencies. We found that biochar application significantly increased soil organic carbon (SOC), microbial biomass carbon (MBC), dissolved organic carbon (DOC), easily oxidized carbon (EOC), particulate organic carbon (POC), total nitrogen (TN), and the C:N ratio in topsoil (0-20 cm) and crop yields. Biochar was most effective in tropical regions, increasing SOC, Soil TN, and crop yield the most, with relatively moderate pyrolysis temperatures (550-650 °C) more conducive to SOC accumulation and relatively low pyrolysis temperatures (<350 °C) more conducive to increasing soil carbon components and crop yields. Biochar made from manure effectively increased soil carbon components and TN. Soil with low fertility (original SOC < 5 g kg-1; original TN < 0.6 g kg-1), coarse texture, and acidity (pH < 5.5) showed more effective results. However, biochar application rates should not be too high and should be combined with appropriate nitrogen fertilizer. And biochar application had long-term positive effects on soil carbon storage and crop yield. Overall, we recommend using small amounts of biochar with lower pyrolysis temperatures in soils with low fertility, coarse texture, and tropical regions for optimal economic and environmental benefits.
Subject(s)
Carbon , Soil , Charcoal/pharmacology , Agriculture/methods , Fertilizers , Nitrogen/analysisABSTRACT
The ridge-furrow rainfall harvesting system (RFRH) improved the water shortages, and reasonable fertilization can promote nutrient uptake and utilization of crops, leading to better yield in semi-arid regions. This holds significant practical significance for improving fertilization strategies and reducing the application of chemical fertilizers in semi-arid areas. This field study was conducted to investigate the effects of different fertilization rates on maize growth, fertilizer use efficiency, and grain yield under the ridge-furrow rainfall harvesting system during 2013-2016 in semiarid region of China. Therefore, a four-year localization field experiment was conducted with four fertilizer treatments: RN (N 0 kg hm-2, P2O5 0 kg hm-2), RL (N 150 kg hm-2, P2O5 75 kg hm-2), RM (N 300 kg hm-2, P2O5 150 kg hm-2), and RH (N 450 kg hm-2, P2O5 225 kg hm-2). The results showed that the total dry matter accumulation of maize increased with the fertilizer application rate. The nitrogen accumulation was highest under the RM treatment after harvest, average increase by 1.41% and 22.02% (P<0.05) compared to the RH and RL, respectively, whereas the phosphorus accumulation was increased with the fertilizer application rate. The nitrogen and phosphorus use efficiency both decreased gradually with the fertilization rate increased, where the maximum efficiency was observed under the RL. With the increase of fertilizer application rate, the maize grain yield initially increased and then decreased. Under linear fitting, the grain yield, biomass yield, hundred-kernel weight, and ear-grain number all showed a parabolic trend with the increase of fertilization rate. Based on comprehensive consideration, the recommended moderate fertilization rate (N 300 kg hm-2, P2O5 150 kg hm-2) is suitable for the ridge furrow rainfall harvesting system in semiarid region, and the fertilization rate can be appropriately reduced according to the rainfall.
ABSTRACT
How does wiring specificity of neural maps emerge during development? Formation of the adult Drosophila olfactory glomerular map begins with the patterning of projection neuron (PN) dendrites at the early pupal stage. To better understand the origin of wiring specificity of this map, we created genetic tools to systematically characterize dendrite patterning across development at PN type-specific resolution. We find that PNs use lineage and birth order combinatorially to build the initial dendritic map. Specifically, birth order directs dendrite targeting in rotating and binary manners for PNs of the anterodorsal and lateral lineages, respectively. Two-photon- and adaptive optical lattice light-sheet microscope-based time-lapse imaging reveals that PN dendrites initiate active targeting with direction-dependent branch stabilization on the timescale of seconds. Moreover, PNs that are used in both the larval and adult olfactory circuits prune their larval-specific dendrites and re-extend new dendrites simultaneously to facilitate timely olfactory map organization. Our work highlights the power and necessity of type-specific neuronal access and time-lapse imaging in identifying wiring mechanisms that underlie complex patterns of functional neural maps.
The brain's ability to sense, act and remember relies on the intricate network of connections between neurons. Organization of these connections into neural maps is critical for processing sensory information. For instance, different odors are represented by specific neurons in a part of the brain known as the olfactory bulb, allowing animals to distinguish between smells. Projection neurons in the olfactory bulb have extensions known as dendrites that receive signals from sensory neurons. Scientists have extensively used the olfactory map in adult fruit flies to study brain wiring because of the specific connections between their sensory and projection neurons. This has led to the discovery of similar wiring strategies in mammals. But how the olfactory map is formed during development is not fully understood. To investigate, Wong et al. built genetic tools to label specific types of olfactory projection neurons during the pupal stage of fruit fly development. This showed that a group of projection neurons directed their dendrites in a clockwise rotation pattern depending on the order in which they were born: the first-born neuron sent dendrites towards the top right of the antennal lobe (the fruit fly equivalent of the olfactory bulb), while the last-born sent dendrites towards the top left. Wong et al. also carried out high-resolution time-lapse imaging of live brains grown in the laboratory to determine how dendrites make wiring decisions. This revealed that projection neurons send dendrites in all directions, but preferentially stabilize those that extend in the direction which the neurons eventually target. Also, live imaging showed neurons could remove old dendrites (used in the larvae) and build new ones (to be used in the adult) simultaneously, allowing them to quickly create new circuits. These experiments demonstrate the value of imaging specific types of neurons to understand the mechanisms that assemble neural maps in the developing brain. Further work could use the genetic tools created by Wong et al. to study how wiring decisions are determined in this and other neural maps by specific genes, potentially yielding insights into neurological disorders associated with wiring defects.
Subject(s)
Drosophila Proteins , Olfactory Receptor Neurons , Animals , Drosophila melanogaster/genetics , Olfactory Pathways , Olfactory Receptor Neurons/physiology , Dendrites/physiology , Drosophila Proteins/genetics , Drosophila Proteins/metabolism , Time-Lapse Imaging , Drosophila/metabolismABSTRACT
Lattice light sheet microscopy excels at the noninvasive imaging of three-dimensional (3D) dynamic processes at high spatiotemporal resolution within cells and developing embryos. Recently, several papers have called into question the performance of lattice light sheets relative to the Gaussian sheets most common in light sheet microscopy. Here, we undertake a theoretical and experimental analysis of various forms of light sheet microscopy, which demonstrates and explains why lattice light sheets provide substantial improvements in resolution and photobleaching reduction. The analysis provides a procedure to select the correct light sheet for a desired experiment and specifies the processing that maximizes the use of all fluorescence generated within the light sheet excitation envelope for optimal resolution while minimizing image artifacts and photodamage. We also introduce a new type of "harmonic balanced" lattice light sheet that improves performance at all spatial frequencies within its 3D resolution limits and maintains this performance over lengthened propagation distances allowing for expanded fields of view.
ABSTRACT
China's urbanization has exceeded 64% and a large number of small and medium-sized cities are the key development areas in the new stage. In urban planning, it is very important to reveal the influence of environmental comfort on urban vitality to improve the life quality of residents in these towns. Thus, the study investigated the impact of environmental comfort on urban vitality using ordinary least squares regression in Wuxi County. Environmental comfort was assessed through a comprehensive analysis of a built-up area and urban vitality was represented by vitality intensity. In addition, the influence pathways were identified and model validation was verified. The conclusions are as follows: (1) Environmental comfort and urban vitality are distributed spatially similarly, and both gradually decline from the center to the periphery. It is high in the east and low in the west, high in the south and low in the north. (2) Population density, POI mixing degree, building density, and road network density have significant positive effects on urban vitality. Population density has the greatest impact on urban vitality. Building height, building age, and river buffer have significant negative effects on urban vitality. (3) The impact of comprehensive environmental comfort on urban vitality is positive, and in terms of time, the order of impact is afternoon > morning > evening. Finally, a method for assessing the impact of environmental comfort on urban vitality was constructed, and the promoting effect of environmental comfort improvements on the vitality was verified. These findings will fill the gap between urban physical space and social needs in planning practices and provide reference to improve vitality for urban planning in small and medium-sized cities.
Subject(s)
Urbanization , Humans , Cities , Urban Population , China , Population DensityABSTRACT
PIEZO1 channels play a critical role in numerous physiological processes by transducing diverse mechanical stimuli into electrical and chemical signals. Recent studies underscore the importance of endogenous PIEZO1 activity and localization in regulating mechanotransduction. To enable physiologically and clinically relevant human-based studies, we genetically engineered human induced pluripotent stem cells (hiPSCs) to express a HaloTag fused to endogenous PIEZO1. Combined with super-resolution imaging, our chemogenetic approach allows precise visualization of PIEZO1 in various cell types. Further, the PIEZO1-HaloTag hiPSC technology allows non-invasive monitoring of channel activity via Ca2+-sensitive HaloTag ligands, with temporal resolution approaching that of patch clamp electrophysiology. Using lightsheet imaging of hiPSC-derived neural organoids, we also achieve molecular scale PIEZO1 imaging in three-dimensional tissue samples. Our advances offer a novel platform for studying PIEZO1 mechanotransduction in human cells and tissues, with potential for elucidating disease mechanisms and development of targeted therapeutics.
ABSTRACT
At each cell division, nanometer-scale motors and microtubules give rise to the micron-scale spindle. Many mitotic motors step helically around microtubules in vitro, and most are predicted to twist the spindle in a left-handed direction. However, the human spindle exhibits only slight global twist, raising the question of how these molecular torques are balanced. Here, using lattice light sheet microscopy, we find that anaphase spindles in the epithelial cell line MCF10A have a high baseline twist, and we identify factors that both increase and decrease this twist. The midzone motors KIF4A and MKLP1 are redundantly required for left-handed twist at anaphase, and we show that KIF4A generates left-handed torque in vitro. The actin cytoskeleton also contributes to left-handed twist, but dynein and its cortical recruitment factor LGN counteract it. Together, our work demonstrates that force generators regulate twist in opposite directions from both within and outside the spindle, preventing strong spindle twist during chromosome segregation.
ABSTRACT
A combination of high-level ab initio calculations and anion photoelectron detachment (PD) measurements is reported for the UC, UC-, and UC+ molecules. To better compare the theoretical values with the experimental photoelectron spectrum (PES), a value of 1.493 eV for the adiabatic electron affinity (AEA) of UC was calculated at the Feller-Peterson-Dixon (FPD) level. The lowest vertical detachment energy (VDE) is predicted to be 1.500 eV compared to the experimental value of 1.487 ± 0.035 eV. A shoulder to lower energy in the experimental PD spectrum with the 355 nm laser can be assigned to a combination of low-lying excited states of UC- and excited vibrational states. The VDEs calculated for the low-lying excited electronic states of UC at the SO-CASPT2 level are consistent with the observed additional electron binding energies at 1.990, 2.112, 2.316, and 3.760 eV. Potential energy curves for the Ω states and the associated spectroscopic properties are also reported. Compared to UN and UN+, the bond dissociation energy (BDE) of UC (411.3 kJ/mol) is predicted to be considerably lower. The natural bond orbitals (NBO) calculations show that the UC0/+/- molecules have a bond order of 2.5 with their ground-state configuration arising from changes in the oxidation state of the U atom in terms of the 7s orbital occupation: UC (5f27s1), UC- (5f27s2), and UC+ (5f27s0). The behavior of the UN and UC sequence of molecules and anions differs from the corresponding sequences for UO and UF.
ABSTRACT
Mass spectrometric analysis of anionic products that result from interacting Ir- with H2O shows the efficient generation of [Ir(H2O)]- complexes and IrO- molecular anions. Anion photoelectron spectra of [Ir(H2O)]-, formed under various source conditions, exhibit spectral features that are due to three different forms of the complex: the solvated anion-molecule complex, Ir-(H2O), as well as the intermediates, [H-Ir-OH]- and [H2-Ir-O]-, where one and two O-H bonds have been broken, respectively. The measured and calculated vertical detachment energy values are in good agreement and, thus, support identification of all three types of isomers. The calculated reaction pathway shows that the overall reaction Ir- + H2O â IrO- + H2 is exothermic. Two minimum energy crossing points were found, which shuttle intermediates and products between singlet and triplet potential surfaces. This study presents the first example of water activation and splitting by single Ir- anions.
ABSTRACT
The results of calculations of the properties of the anion UN- including electron detachment are described, which further expand our knowledge of this diatomic molecule. High-level electronic structure calculations were conducted for the UN and UN- diatomic molecules and compared to photoelectron spectroscopy measurements. The low-lying Ω states were obtained using multireference CASPT2 including spin-orbit effects up to â¼20,000 cm-1. At the Feller-Peterson-Dixon (FPD) level, the adiabatic electron affinity (AEA) of UN is estimated to be 1.402 eV and the vertical detachment energy (VDE) is 1.423 eV. The assignment of the UN excited states shows good agreement with the experimental results with a VDE of 1.424 eV. An Ω = 4 ground state was obtained for UN- which is mainly associated with the 3H ΛS state. Thermochemical calculations estimate a bond dissociation energy (BDE) for UN- (U- + N) of 665.9 kJ/mol, â¼15% larger than that of UN and UN+. The NBO analysis reveals U-N triple bonds for the UN, UN-, and UN+ species.
ABSTRACT
The activation and transformation of H2O and CO2 mediated by electrons and single Pt atoms is demonstrated at the molecular level. The reaction mechanism is revealed by the synergy of mass spectrometry, photoelectron spectroscopy, and quantum chemical calculations. Specifically, a Pt atom captures an electron and activates H2O to form a H-Pt-OH- complex. This complex reacts with CO2via two different pathways to form formate, where CO2 is hydrogenated, or to form bicarbonate, where CO2 is carbonated. The overall formula of this reaction is identical to a typical electrochemical CO2 reduction reaction on a Pt electrode. Since the reactants are electrons and isolated, single atoms and molecules, we term this reaction a molecular-level electrochemical CO2 reduction reaction. Mechanistic analysis reveals that the negative charge distribution on the Pt-H and the -OH moieties in H-Pt-OH- is critical for the hydrogenation and carbonation of CO2. The realization of the molecular-level CO2 reduction reaction provides insights into the design of novel catalysts for the electrochemical conversion of CO2.
ABSTRACT
Understanding direct metal-metal bonding between actinide atoms has been an elusive goal in chemistry for years. We report for the first time the anion photoelectron spectrum of U2-. The threshold of the lowest electron binding energy (EBE) spectral band occurs at 1.0 eV, which corresponds to the electron affinity (EA) of U2, whereas the vertical detachment energy of U2- is found at EBE â¼ 1.2 eV. Electronic structure calculations on U2 and U2- were carried out with state-of-the-art theoretical methods. The computed values of EA(U2) and EA(U) and the difference between the computed dissociation energies of U2 and U2- are found to be internally consistent and consistent with experiment. Analysis of the bonds in U2 and U2- shows that while U2 has a formal quintuple bond, U2- has a quadruple bond, even if the effective bond orders differ only by 0.5 unit instead of one unit. The resulting experimental-computational synergy elucidates the nature of metal-metal bonding in U2 and U2-.
ABSTRACT
Elucidating the multifaceted processes of molecular activation and subsequent reactions gives a fundamental view into the development of iridium catalysts as they apply to fuels and propellants, for example, for spacecraft thrusters. Hydroxylamine, a component of the well-known hydroxylammonium nitrate (HAN) ionic liquid, is a safer alternative and mimics the chemistry and performance standards of hydrazine. The activation of hydroxylamine by anionic iridium clusters, Irn- (n = 1-5), depicts a part of the mechanism, where two hydrogen atoms are removed, likely as H2, and Irn(NOH)- clusters remain. The significant photoelectron spectral differences between these products and the bare clusters illustrate the substantial electronic changes imposed by the hydroxylamine fragment on the iridium clusters. In combination with DFT calculations, a preliminary reaction mechanism is proposed, identifying the possible intermediate steps leading to the formation of Ir(NOH)-.
ABSTRACT
The results of a combined experimental and computational study of the uranium atom are presented with the aim of determining its electron affinity. Experimentally, the electron affinity of uranium was measured via negative ion photoelectron spectroscopy of the uranium atomic anion, U-. Computationally, the electron affinities of both thorium and uranium were calculated by conducting relativistic coupled-cluster and multi-reference configuration interaction calculations. The experimentally determined value of the electron affinity of the uranium atom was determined to be 0.309 ± 0.025 eV. The computationally predicted electron affinity of uranium based on composite coupled cluster calculations and full four-component spin-orbit coupling was found to be 0.232 eV. Predominately due to a better convergence of the coupled cluster sequence for Th and Th-, the final calculated electron affinity of Th, 0.565 eV, was in much better agreement with the accurate experimental value of 0.608 eV. In both cases, the ground state of the anion corresponds to electron attachment to the 6d orbital.
ABSTRACT
A wide range of low oxidation state aluminum-containing cluster anions, LAln- (n = 1-14, L = N[Si(Me)3]2), were produced via reactions between aluminum cluster anions and hexamethyldisilazane (HMDS). These clusters were identified by mass spectrometry, with a few of them (n = 4, 6, and 7) further characterized by a synergy of anion photoelectron spectroscopy and density functional theory (DFT) based calculations. As compared to a previously reported method which reacts anionic aluminum hydrides with ligands, the direct reactions between aluminum cluster anions and ligands promise a more general synthetic scheme for preparing low oxidation state, ligated aluminum clusters over a large size range. Computations revealed structures in which a methyl-group of the ligand migrated onto the surface of the metal cluster, thereby resulting in "two metal-atom" insertion between Si-CH3 bond.
ABSTRACT
Artificial lakes that form during rapid urbanization often fail to achieve their desired functions, and gradually become eutrophic. Whether the external discharge or internal release of nutrients dominates the eutrophication of urban lakes has rarely been reported. In this study, a lake that had been formed during ten years of urbanization had become hyper-eutrophic. TP mainly contributed to the eutrophication and algal bloom in the lake. While the release potential of TP fluctuated, TN, particularly NH3-N, was constantly released from the sediment. Concentrations of anthropogenic metals (Pb, Cu and Cr) increased with the increasing depth of the sediment. Even for a lake that had formed rapidly in a short period, the internal phosphorus released from sediment was 1.9-times higher than that of the external discharge. The dominating contribution of internal pollution from sediment requires more attention to restore and manage these urban waters.
Subject(s)
Lakes , Urbanization , China , Environmental Monitoring , Eutrophication , Geologic Sediments , Nitrogen/analysis , Phosphorus/analysisABSTRACT
Mass spectrometric analysis of the anionic products of interaction between palladium hydride anions, PdH-, and carbon dioxide, CO2, in a reaction cell shows an efficient generation of the PdHCO2- intermediate and isolated formate product. Multiple isomers of the PdHCO2- intermediates are identified by a synergy between negative ion photoelectron spectroscopy and quantum-chemical calculations. It is shown that a direct mechanism, in which the H atom in PdH- directly activates and hydrogenates CO2, leads to the formation of the formate product. An indirect mechanism, on the other hand, leads to a stable HPdCO2- structure, where CO2 is chemisorbed onto the Pd atom.
ABSTRACT
Mass spectrometric analysis of the anionic products of interaction among Pt-, methane, and carbon dioxide shows that the methane activation complex, H3C-Pt-H-, reacts with CO2 to form [H3C-Pt-H(CO2)]-. Two hydrogenation and one C-C bond coupling products are identified as isomers of [H3C-Pt-H(CO2)]- by a synergy between anion photoelectron spectroscopy and quantum chemical calculations. Mechanistic study reveals that both CH4 and CO2 are activated by the anionic Pt atom and that the successive depletion of the negative charge on Pt drives the CO2 insertion into the Pt-H and Pt-C bonds of H3C-Pt-H-. This study represents the first example of the simultaneous functionalization of CH4 and CO2 mediated by single atomic anions.