Single-cell transcriptomics (scRNA-seq) has revolutionized the understanding of the spatial architecture of tissue structure and function. Advancing the “transcript-centric” view of scRNA-seq ...analyses is presently restricted by the limited resolution of proteomics and genome-wide techniques to analyze post-translational modifications. Here, by combining spatial cell sorting with transcriptomics and quantitative proteomics/phosphoproteomics, we established the spatially resolved proteome landscape of the liver endothelium, yielding deep mechanistic insight into zonated vascular signaling mechanisms. Phosphorylation of receptor tyrosine kinases was detected preferentially in the central vein area, resulting in an atypical enrichment of tyrosine phosphorylation. Prototypic biological validation identified Tie receptor signaling as a selective and specific regulator of vascular Wnt activity orchestrating angiocrine signaling, thereby controlling hepatocyte function during liver regeneration. Taken together, the study has yielded fundamental insight into the spatial organization of liver endothelial cell signaling. Spatial sorting may be employed as a universally adaptable strategy for multiomic analyses of scRNA-seq-defined cellular (sub)-populations.
Display omitted
•ScRNA-seq-guided spatial sort enables multiomic dissection of the liver vasculature•Liver sinusoidal endothelial cells have a hybrid vascular-lymphatic phenotype•Tyrosine phosphorylation of endothelial cell molecules is enriched on central vein•Endothelial Tie1 shapes hepatic Wnt signal zonation and promotes liver regeneration
Inverso, Shi et al. generate a multiomic encyclopedia of liver endothelial cells (L-ECs) with spatial resolution of transcriptome, proteome, and phosphoproteome. The study provides insight into liver vascular zonation and a template for scRNA-seq-data-guided spatial proteome and phosphoproteome analyses.
Mass customization logistics service mode provides a new way to maintain the sustainable cooperative relationship between customers and integrators. One of the key factors to maintain the sustainable ...development of logistics service supply chain under MC mode is to locate a suitable customer order decoupling point (CODP) location. This paper investigates the problem of CODP in the logistics service supply chain based on the fuzzy set theory under the mass customization mode. With the help of a fuzzy QFD method and a new service quality function that we constructed, this paper quantifies the quality of a logistics service when the LSI selects a different CODP. Then, the fuzzy set of the high-quality logistics service and the fuzzy set of the satisfactory delivery time are built. Based on those two new fuzzy sets, this paper builds a new fuzzy programming model on CODP positioning. The solving methods of this model under different conditions are given. Finally, the influence of some important parameters on the optimal CODP position is studied by sensitivity analysis on a specific numerical case.
We investigate the performance and stability of a platinum (Pt) supported on indium tin oxide (ITO) electrocatalyst. ITO was synthesized using the co-precipitation method and uniform particles with a ...B.E.T. surface area larger than 40m2/g were obtained. Pt was dispersed onto the ITO by colloidal deposition followed by reduction with formaldehyde. Previous rotating disk electrode (RDE) work has shown that Pt/ITO possesses high activity and stability for the oxygen reduction reaction (ORR). However, this catalyst exhibited very poor performance and stability in an operating polymer electrolyte fuel cell (PEFC) membrane electrode assembly (MEA). For H2/O2 PEFC operation at 80°C and 75% relative humidity (RH), the current density obtained at 0.55V was only 90mA/cm2, and the maximum current density was only 150–160mA/cm2, when Pt/ITO was used at the cathode, whereas a limiting current density of 3900mA/cm2 was readily obtained using a benchmark Pt/C catalyst under identical conditions. The low performance with Pt/ITO was primarily due to the high overall cell resistance of the MEA (>400mOhmcm2). X-ray photoelectron spectroscopy (XPS) was employed to investigate the degradation of the Pt/ITO catalyst in the PEFC electrode during operation. The deconvolution of the indium 3d XPS peak revealed the presence of two peaks: the first was assigned to indium oxide in ITO (at 445.6eV), while the second was assigned to indium hydroxide (at 446.6eV). We observed an increase in surface hydroxide concentration (compared to pristine Pt/ITO) after the Pt/ITO catalyst was used either at the cathode or anode of an operating PEFC. We postulate that the surface hydroxides form a passivating layer that increases the electrode resistance and undermines PEFC performance.
•Contrary to RDE results, low performance was observed with a Pt/ITO electrocatalyst in an operating PEFC.•X-ray photoelectron spectroscopy (XPS) revealed Pt/ITO catalyst degradation during PEFC operation.•Indium 3d XPS peak deconvolution revealed the degradation mechanism, namely the formation of surface indium hydroxides.•We postulate that the surface hydroxides form a passivating layer that increases the electrode resistance and undermines PEFC performance.
The gas diffusion layer (GDL), as a key component of proton exchange membrane fuel cells (PEMFCs), plays a crucial role in PEMFC’s polarization performance, particularly in mass transport properties ...at high current densities. To elucidate the correlation between GDLs’ structure and their mass transport properties, a limiting current test with the H2 molecular probe was established and employed to investigate three representative GDLs with and without the microporous layer (MPL). By varying humidity and back pressure, the mass transport resistance of three GDLs was measured in an operating fuel cell, and an elaborate analysis of H2 transport was conducted. The results showed that the transport resistance (RDM) of GDLs was affected by the thickness and pore size distribution of the macroporous substrate (MPS) and the MPL. In the process of gas transport, the smaller pore size and thicker MPL increase the force of gas on the pore wall, resulting in an increase in transmission resistance. Through further calculation and analysis, the total transport resistance can be divided into pressure-related resistance (RP) and pressure-independent resistance (RNP). RP mainly originates from the transport resistance in both MPLs and the substrate layers of GDLs, exhibiting a linear relationship to the pressure; RNP mainly originates from the transport resistance in the MPLs. 29BC with thick MPL shows the largest RNP, and T060 without MPL shows the RNP = 0. This methodology enables in situ measurements of mass transport resistances for gas diffusion media, which can be easily applied for developing and deploying PEMFCs.
Multiple morphological abnormalities of the sperm flagella (MMAF) is a subtype of severe asthenoteratozoospermia with poorly understood genetic etiology. SPAG6 is a core axonemal component that plays ...a critical role in the formation of cilia and sperm flagella. Previous studies have reported that mutations in SPAG6 cause primary ciliary dyskinesia (PCD), but the association between SPAG6 gene variants and the MMAF phenotype has not yet been described.
We performed whole-exome sequencing (WES) in two unrelated Han Chinese men with MMAF. Sanger sequencing was used to validate the candidate variants. Routine semen analysis was carried out according to the WHO guidelines (5
Edition). Sperm morphology was assessed using modified Papanicolaou staining. Scanning and transmission electron microscopy (S/TEM) was performed to observe the ultrastructural defects of the sperm flagella. Western blot analysis and immunofluorescence (IF) of spermatozoa were performed to examine the expression of SPAG6 protein. Assisted fertilization with intracytoplasmic sperm injection (ICSI) was applied.
Two homozygous SPAG6 variants were identified by WES and Sanger validation in two patients with MMAF phenotype (F1 II-1: c.308C > A, p. A103D; F2 II-1: c. 585delA, p. K196Sfs*6). Semen analysis showed progressive rates of less than 1%, and most of the spermatozoa presented MMAF by Papanicolaou staining. TEM revealed that the overall axonemal ultrastructure was disrupted and primarily presented an abnormal "9 + 0" configuration. No other PCD-related symptoms were found on physical examination and medical consultations, as well as lung CT screening. The level of SPAG6 protein was significantly decreased in the spermatozoa, and IF analysis revealed that SPAG6 staining was extremely weak and discontinuous in the sperm flagella of the two patients. Notably, F1 II-1 and his wife conceived successfully after undergoing ICSI.
Our research provides new evidence for a potential correlation between SPAG6 variants and the MMAF phenotype.
Using a variety of in situ techniques, we tracked the structural stability and concomitantly the electrocatalytic oxygen reduction reaction (ORR) of platinum nanoparticles on ruthenium–titanium mixed ...oxide (RTO) supports during electrochemical accelerated stress tests, mimicking fuel cell operating conditions. High-energy X-ray diffraction (HE-XRD) offered insights in the evolution of the morphology and structure of RTO-supported Pt nanoparticles during potential cycling. The changes of the atomic composition were tracked in situ using scanning flow cell measurements coupled to inductively coupled plasma mass spectrometry (SFC-ICP-MS). We excluded Pt agglomeration, particle growth, dissolution, or detachment as cause for the observed losses in catalytic ORR activity. Instead, we argue that Pt surface poisoning is the most likely cause of the observed catalytic rate decrease. Data suggest that the gradual growth of a thin oxide layer on the Pt nanoparticles due to strong metal–support interaction (SMSI) is the most plausible reason for the suppressed catalytic activity. We discuss the implications of the identified catalyst degradation pathway, which appear to be specific for oxide supports. Our conclusions offer previously unaddressed aspects related to oxide-supported metal particle electrocatalysts frequently deployed in fuel cells, electrolyzers, or metal–air batteries.
DNA mismatch repair (MMR) is a highly conserved pathway that corrects DNA replication errors, the loss of which is attributed to the development of various types of cancers. Although well ...characterized, MMR factors remain to be identified. As a 3'-5' exonuclease and endonuclease, meiotic recombination 11 homolog A (MRE11A) is implicated in multiple DNA repair pathways. However, the role of MRE11A in MMR is unclear.
Initially, short-term and long-term survival assays were used to measure the cells' sensitivity to N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Meanwhile, the level of apoptosis was also determined by flow cytometry after MNNG treatment. Western blotting and immunofluorescence assays were used to evaluate the DNA damage within one cell cycle after MNNG treatment. Next, a GFP-heteroduplex repair assay and microsatellite stability test were used to measure the MMR activities in cells. To investigate the mechanisms, western blotting, the GFP-heteroduplex repair assay, and chromatin immunoprecipitation were used.
We show that knockdown of MRE11A increased the sensitivity of HeLa cells to MNNG treatment, as well as the MNNG-induced DNA damage and apoptosis, implying a potential role of MRE11 in MMR. Moreover, we found that MRE11A was largely recruited to chromatin and negatively regulated the DNA damage signals within the first cell cycle after MNNG treatment. We also showed that knockdown of MRE11A increased, while overexpressing MRE11A decreased, MMR activity in HeLa cells, suggesting that MRE11A negatively regulates MMR activity. Furthermore, we show that recruitment of MRE11A to chromatin requires MLH1 and that MRE11A competes with PMS2 for binding to MLH1. This decreases PMS2 levels in whole cells and on chromatin, and consequently comprises MMR activity.
Our findings reveal that MRE11A is a negative regulator of human MMR.
Non-obstructive azoospermia (NOA) is the most severe type of male infertility, affecting 1% of men worldwide. Most of its etiologies remain idiopathic. Although genetic studies have identified dozens ...of NOA genes, monogenic mutations can also account for a small proportion of idiopathic NOA cases. Hence, this genetic study was conducted to explore the causes of monogenic variants of NOA in a cohort of Chinese patients.
Following the screening using chromosomal karyotyping, Y chromosome microdeletion analyses, and sex hormone assessments, subsequent whole-exome sequencing analysis was performed in 55 unrelated idiopathic NOA patients with male infertility to explore potential deleterious variants associated with spermatogenesis. We also performed Sanger sequencing to demonstrate the variants. Testicular biopsy or microsurgical testicular sperm extraction was also performed to confirm the diagnosis of NOA and identify spermatozoa. Hematoxylin and eosin staining was performed to assess the histopathology of spermatogenesis.
Abnormal testicular pathological phenotypes included Sertoli cell-only syndrome, maturation arrest, and hypospermatogenesis. Using bioinformatics analysis, we detected novel variants in two recessive genes, FANCA (NM_000135, c.3263C > T, c.1729C > G) and SYCE1 (NM_001143763, c.689_690del); one X-linked gene, TEX11 (NM_031276, c.466A > G, c.559_560del); and two dominant genes, DMRT1 (NM_021951, c.425C > T, c.340G > A) and PLK4 (NM_001190799, c.2785A > G), in eight patients, which corresponded to 14.55% (8/55) of the patients.
This study presented some novel variants of known pathogenic genes for NOA. Further, it expanded the variant spectrum of NOA patients, which might advance clinical genetic counseling in the future.
PDF
