Identification and Functional Studies of Arabidopsis thaliana Ubc13-interacting E3 Ubiquitin Ligases

In eukaryotic organisms, polyubiquitination is the modification of a protein with polymerized ubiquitin (Ub) chain. This process is well known for its function in targeting proteins for degradation by the 26S proteasome. However, a polyUb chain assembled through the lysine 63 residue of the Ub moiety (Lys63-linked polyubiquitination) has been shown to play a signaling role rather than targeting proteins for degradation. In plants, the functions of Lys63-linked polyubiquitination are currently not well understood. Ub-protein ligase (E3) catalyzes the last step in the ubiquitination reactions, and to a large extent it also determines the substrate specificity of protein ubiquitination. In order to study the roles of Lys63-linked polyubiquitination in plants, two E3s of Arabidopsis thaliana, proteins encoded by AtCHIP and At1g74370 (tentatively named E3-A1), were chosen for functional studies, since they interacted with AtUbc13A protein. Sequence analysis showed that AtCHIP is the only member in the family. A T-DNA insertion mutant line (Atchip-1) was obtained to study the AtCHIP gene knock-out effect. The mutant line was grown in normal conditions and further tested in a variety of conditions: hormonal treatments, osmotic stress, seed deterioration, high temperature stress, high-intensity light stress, oxidative stress and DNA damaging stress. However, no clear difference was observed between the mutant and wild type plants based on the several parameters measured. Sequence analysis of E3-A1 indicated two closely related proteins, tentatively named E3-A2 and E3-A3. As E3-A1 and E3-A2 appeared to share more sequence similarity, RNA interference (RNAi) transformants, with the level of transcripts for either of the two E3-A genes reduced by over 90% were generated. Selected RNAi mutant lines for E3-A1 and E3-A2 were crossed with each other, and double RNAi mutants were obtained. However, no distinct phenotype was detected under normal, high-sucrose or hormonal conditions for either single or double RNAi lines. Although various assays did not reveal a significant phenotype in the mutants in this study, the materials generated and the assays used will benefit a wider range of phenotypic survey in the future

Layer Construction of Three-Dimensional Z2 Monopole Charge Nodal Line Semimetals and prediction of the abundant candidate materials

The interplay between symmetry and topology led to the concept of symmetry-protected topological states, including all non-interacting and weakly interacting topological quantum states. Among them, recently proposed nodal line semimetal states with space-time inversion ($\mathcal{PT}$) symmetry which are classified by the Stiefel-Whitney characteristic class associated with real vector bundles and can carry a nontrivial $\mathbb{Z}_2$ monopole charge have attracted widespread attention. However, we know less about such 3D $\mathbb{Z}_2$ nodal line semimetals and do not know how to construct them. In this work, we first extend the layer construction previously used to construct topological insulating states to topological semimetallic systems. We construct 3D $\mathbb{Z}_2$ nodal line semimetals by stacking of 2D $\mathcal{PT}$-symmetric Dirac semimetals via nonsymmorphic symmetries. Based on our construction scheme, effective model and combined with first-principles calculations, we predict two types of candidate electronic materials for $\mathbb{Z}_2$ nodal line semimetals, namely 14 Si and Ge structures and 108 transition metal dichalcogenides $MX_2$ ($M$=Cr, Mo, W, $X$=S, Se, Te). Our theoretical construction scheme can be directly applied to metamaterials and circuit systems. Our work not only greatly enriches the candidate materials and deepens the understanding of $\mathbb{Z}_2$ nodal line semimetal states but also significantly extends the application scope of layer construction

3D carbon allotropes: Topological quantum materials with obstructed atomic insulating phases, multiple bulk-boundary correspondences, and real topology

The study of topological phases with unconventional bulk-boundary correspondences and nontrivial real Chern number has garnered significant attention in the topological states of matter. Using the first-principle calculations and theoretical analysis, we perform a high-throughput material screening of the 3D obstructed atomic insulators (OAIs) and 3D real Chern insulators (RCIs) based on the Samara Carbon Allotrope Database (SACADA). Results show that 422 out of 703 3D carbon allotropes are 3D OAIs with multiple bulk-boundary correspondences, including 2D obstructed surface states (OSSs) and 1D hinge states, which are in one dimension and two dimensions lower than the 3D bulk, respectively. The 2D OSSs in these OAIs can be modified when subjected to appropriate boundaries, which benefits the investigation of surface engineering and the development of efficient topological catalysts. These 422 OAIs, which have 2D and 1D boundary states, are excellent platforms for multi-dimensional topological boundaries research. Remarkably, 138 of 422 OAIs are also 3D RCIs, which show a nontrivial real topology in the protection of spacetime inversion symmetry. Our work not only provides a comprehensive list of 3D carbon-based OAIs and RCIs, but also guides their application in various aspects based on multiple bulk-boundary correspondences and real topological phases

Screening and Characterization of Potential Antioxidant Probiotics Isolated from the Gut of Hybrid Grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂)

Some symbiotic probiotics have antioxidant activities and could improve the antioxidant capacity of the host. There is still no report on the screening of host-derived antioxidant probiotics for grouper farming. In this study, 369 out of 583 isolates were screened from the gut of hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus♂) based on their non-hemolytic characteristics. Subsequent preliminary screening with 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) scavenging rate assay resulted in 36 potential antioxidant isolates. After comprehensive evaluation with nine different antioxidant assays (DPPH scavenging rate, 2,2’-azino-bis (3-ethylbezothiazoline)-6-sulfonic acid radical scavenging rate, iron ion reducing ability, reducing activity, O2-· scavenging rate,·OH scavenging rate, ferrous ion chelating rate, hydrogen peroxide tolerance, oxygen-free radical scavenging ability, et al.), 10 isolates with strong antioxidant abilities were screened from 36 potential antioxidant isolates. Then some other probiotic properties, such as simulated gastrointestinal fluid tolerance, adhesion, digestive enzyme activity, and antibacterial activity of the 10 selected isolates were evaluated. All 10 isolates were also identified using the molecular method. Finally, Vibrio rhodolitus GO 91 and Shewanella corallii GO 310, as representatives of the two genera resulting from the identification of the 10 isolates, and with the best overall probiotic properties, were selected from the 10 isolates. Isolates GO 91 and GO 310 were further tested for their safety performances. Antibiotic sensitivity tests showed that GO91 and GO310 were sensitive to many commonly used aquaculture antibiotics. The in vivo challenge test of GO 91 and GO 310 didn’t cause any disease symptoms or death in hybrid grouper. Therefore, isolates GO 91 and GO 310 showed great potential to be used as probiotics in hybrid grouper farming

Ultrasound-Stimulated Microbubbles Enhance Radiosensitization of Nasopharyngeal Carcinoma

Background/Aims: Recent studies indicate that therapies targeting the vasculature can significantly sensitize tumors to radiation. Ultrasound-stimulated microbubbles (USMBs) are regarded as a promising radiosensitizer. In this study, we investigated the effect of USMBs on the sensitivity of nasopharyngeal carcinoma (NPC) to radiation. Methods: Human NPC (CNE-2) cells and human umbilical vein endothelial cells (HUVECs) were exposed to radiation (0, 2, and 8 Gy) alone or in combination with USMBs. Cell viability and apoptosis were measured with the MTT assay and flow cytometry, respectively. The angiogenic activity of HUVECs was detected using matrigel tubule formation. The in vitro effects induced by these treatments were confirmed in vivo with xenograft models of CNE-2 cells in nude mice by examining vascular integrity using color Doppler flow imaging and cell survival using immunohistochemistry. Additionally, the in vivo and in vitro expressions of angiotensin II (ANG II) and its receptor (AT1R) were detected by immunohistochemistry and western blotting, respectively. With CNE-2 cells and HUVECs transfected with control, ANG II, or AT1R, perindopril (an inhibitor of angiotensin-converting enzyme) and candesartan (an inhibitor of AT1R) were used to verify the role of ANG II and AT1R in the radiosensitivity of tumor and endothelial cells by USMBs, by determining cell viability and apoptosis and angiogenic activity. Results: In the NPC xenografts, USMBs slightly reduced blood flow and CD34 expression, increased tumor cell death and ANG II and AT1R expression, and significantly enhanced the effects of radiation. With CNE-2 cells and HUVECs, the USMBs further enhanced the inhibition of tumor cell viability and endothelial tubule formation and further enhanced the increase in ANG II and AT1R due to radiation. Furthermore, perindopril and candesartan significantly enhanced the inhibitory effect of radiation and USMBs on tumor cell growth and angiogenesis in vitro. Conclusions: We have demonstrated for the first time that USMB exposure can significantly enhance the destructive effect on NPC of radiation, and this effect might be further increased by ANG II and AT1R inhibition. Our findings suggest that USMBs can be used as a promising sensitizer of radiotherapy to treat NPC, and the clinical effect might be increased by ANG II and AT1R inhibition

Cross-cultural adaptation and validation of the PedsQL™ stem cell transplant module in China: A methodological and cross-sectional study

BackgroundHematopoietic stem cell transplantation (HSCT), as a mature technology, has significantly improved the survival rate of children. However, there lack efficient scales to assess the quality of life (QoL) of children with HSCT in China, which has important implications in the care of this population. This study aimed to translate the original English Pediatric Quality of Life Inventory™ (PedsQL™) Stem Cell Transplant Module into a Chinese mandarin version, and evaluate its reliability.MethodsChildren of ages 2–18 years who had received HSCT at Children's Hospital of Nanjing Medical University and Children's Hospital of Fudan University were recruited. Children or their parents were asked to fill the PedsQL™ 4.0 Generic Core Scales, PedsQL™ Stem Cell Transplant Module, and PedsQL™ Family Information Form. Feasibility was evaluated by completion rate and the percentage of missing items, reliability by the internal consistency and test-retest reliability, and validity by factor analysis and correlation analysis between the scores of total scale and each dimension.ResultsA total of 120 children (mean age 6.37, SD = 3.674) and some parents were included. A low percentage of items were missed in returned reports. Cronbach's alpha coefficient reached 0.70 in the majority of dimensions of both child self-report and parent proxy-report. Test-retest reliability was 0.685 in parents' forms and 0.765 in child's forms. Eight factors were extracted, with a cumulative contribution rate of 74.54%. The correlation between PedsQL™ 4.0 and Transplant Module was 0.748 for children self-report and 0.808 for parent proxy-report.ConclusionsThis study provides evidence that the Chinese mandarin version of the PedsQL™ Stem Cell Transplant is feasible, reliable and valid in evaluating the QoL of Chinese children after HSCT

AAGAB is an assembly chaperone regulating AP1 and AP2 clathrin adaptors

Multimeric cargo adaptors such as AP2 play central roles in intracellular membrane trafficking. We recently discovered that the assembly of the AP2 adaptor complex, a key player in clathrin-mediated endocytosis, is a highly organized process controlled by alpha- and gamma-adaptin-binding protein (AAGAB, also known as p34). In this study, we demonstrate that besides AP2, AAGAB also regulates the assembly of AP1, a cargo adaptor involved in clathrin-mediated transport between the trans-Golgi network and the endosome. However, AAGAB is not involved in the formation of other adaptor complexes, including AP3. AAGAB promotes AP1 assembly by binding and stabilizing the &gamma; and &sigma; subunits of AP1, and its mutation abolishes AP1 assembly and disrupts AP1-mediated cargo trafficking. Comparative proteomic analyses indicate that AAGAB mutation massively alters surface protein homeostasis, and its loss-of-function phenotypes reflect the synergistic effects of AP1 and AP2 deficiency. Taken together, these findings establish AAGAB as an assembly chaperone for both AP1 and AP2 adaptors and pave the way for understanding the pathogenesis of AAGAB-linked diseases. &nbsp;</p

Gut microbiome is associated with metabolic syndrome accompanied by elevated gamma-glutamyl transpeptidase in men

It is predicted that by 2035, metabolic syndrome (MS) will be found in nearly more than half of our adult population, seriously affecting the health of our body. MS is usually accompanied by the occurrence of abnormal liver enzymes, such as elevated gamma-glutamyl transpeptidase (GGT). More and more studies have shown that the gut microbiota is involved in MS; however, the correlation between gut microbiota and MS with elevated GGT has not been studied comprehensively. Especially, there are few reports about its role in the physical examination of the population of men with MS and elevated GGT. By using the whole-genome shotgun sequencing technology, we conducted a genome-wide association study of the gut microbiome in 66 participants diagnosed as having MS accompanied by high levels of GGT (case group) and 66 participants with only MS and normal GGT level (control group). We found that the number of gut microbial species was reduced in participants in the case group compared to that of the control group. The overall microbial composition between the two groups is of significant difference. The gut microbiota in the case group is characterized by increased levels of “harmful bacteria” such as Megamonas hypermegale, Megamonas funiformis, Megamonas unclassified, Klebsiella pneumoniae, and Fusobacterium mortiferum and decreased levels of “beneficial bacteria” such as Faecalibacterium prausnitzii, Eubacterium eligens, Bifidobacterium longum, Bifidobacterium pseudocatenulatum, Bacteroides dorei, and Alistipes putredinis. Moreover, the pathways of POLYAMSYN-PWY, ARG+POLYAMINE-SYN, PWY-6305, and GOLPDLCAT-PWY were also increased in the case group, which may play a role in the elevation of GGT by producing amine, polyamine, putrescine, and endogenous alcohol. Taken together, there are apparent changes in the composition of the gut microbiome in men with MS and abnormal GGT levels, and it is high time to discover specific gut microbiome as a potential therapeutic target in that population. More in-depth studies of relevant mechanism could offer some new methods for the treatment of MS with elevated GGT