Crystal structure of a tetrameric RNA G-quadruplex formed by hexanucleotide repeat expansions of C9orf72 in ALS/FTD.

The abnormal GGGGCC hexanucleotide repeat expansions (HREs) in C9orf72 cause the fatal neurodegenerative diseases including amyotrophic lateral sclerosis and frontotemporal dementia. The transcribed RNA HREs, short for r(G4C2)n, can form toxic RNA foci which sequestrate RNA binding proteins and impair RNA processing, ultimately leading to neurodegeneration. Here, we determined the crystal structure of r(G4C2)2, which folds into a parallel tetrameric G-quadruplex composed of two four-layer dimeric G-quadruplex via 5'-to-5' stacking in coordination with a K+ ion. Notably, the two C bases locate at 3'- end stack on the outer G-tetrad with the assistance of two additional K+ ions. The high-resolution structure reported here lays a foundation in understanding the mechanism of neurological toxicity of RNA HREs. Furthermore, the atomic details provide a structural basis for the development of potential therapeutic agents against the fatal neurodegenerative diseases ALS/FTD.

PRDX1 Interfering Peptide Disrupts Amino Acids 70-90 of PRDX1 to Inhibit the TLR4/NF-κB Signaling Pathway and Attenuate Neuroinflammation and Ischemic Brain Injury.

Ischemic stroke ranks among the leading causes of death and disability in humans and is accompanied by motor and cognitive impairment. However, the precise mechanisms underlying injury after stroke and effective treatment strategies require further investigation. Peroxiredoxin-1 (PRDX1) triggers an extensive inflammatory cascade that plays a pivotal role in the pathology of ischemic stroke, resulting in severe brain damage from activated microglia. In the present study, we used molecular dynamics simulation and nuclear magnetic resonance to detect the interaction between PRDX1 and a specific interfering peptide. We used behavioral, morphological, and molecular experimental methods to demonstrate the effect of PRDX1-peptide on cerebral ischemia-reperfusion (I/R) in mice and to investigate the related mechanism. We found that PRDX1-peptide bound specifically to PRDX1 and improved motor and cognitive functions in I/R mice. In addition, pretreatment with PRDX1-peptide reduced the infarct area and decreased the number of apoptotic cells in the penumbra. Furthermore, PRDX1-peptide inhibited microglial activation and downregulated proinflammatory cytokines including IL-1ß, IL-6, and TNF-α through inhibition of the TLR4/NF-κB signaling pathway, thereby attenuating ischemic brain injury. Our findings clarify the precise mechanism underlying PRDX1-induced inflammation after ischemic stroke and suggest that the PRDX1-peptide can significantly alleviate the postischemic inflammatory response by interfering with PRDX1 amino acids 70-90 and thereby inhibiting the TLR4/NF-κB signaling pathway. Our study provides a theoretical basis for a new therapeutic strategy to treat ischemic stroke.

Boundary sample-based class-weighted semi-supervised learning for malignant tumor classification of medical imaging.

Medical image classification plays a pivotal role within the field of medicine. Existing models predominantly rely on supervised learning methods, which necessitate large volumes of labeled data for effective training. However, acquiring and annotating medical image data is both an expensive and time-consuming endeavor. In contrast, semi-supervised learning methods offer a promising approach by harnessing limited labeled data alongside abundant unlabeled data to enhance the performance of medical image classification. Nonetheless, current methods often encounter confirmation bias due to noise inherent in self-generated pseudo-labels and the presence of boundary samples from different classes. To overcome these challenges, this study introduces a novel framework known as boundary sample-based class-weighted semi-supervised learning (BSCSSL) for medical image classification. Our method aims to alleviate the impact of intra- and inter-class boundary samples derived from unlabeled data. Specifically, we address reliable confidential data and inter-class boundary samples separately through the utilization of an inter-class boundary sample mining module. Additionally, we implement an intra-class boundary sample weighting mechanism to extract class-aware features specific to intra-class boundary samples. Rather than discarding such intra-class boundary samples outright, our approach acknowledges their intrinsic value despite the difficulty associated with accurate classification, as they contribute significantly to model prediction. Experimental results on widely recognized medical image datasets demonstrate the superiority of our proposed BSCSSL method over existing semi-supervised learning approaches. By enhancing the accuracy and robustness of medical image classification, our BSCSSL approach yields considerable implications for advancing medical diagnosis and future research endeavors.

Ghost fishing efficiency in swimming crab (Portunus trituberculatus) pot fishery.

Abandoned, lost, or otherwise discarded fishing gear (ALDFG) is a global challenge that negatively affects marine environment through plastic pollution and continued capture of marine animals, so-called "ghost fishing". In different pot fisheries, ghost fishing related to ALDFG is of concern, including pot fishery targeting swimming crab (Portunus trituberculatus). This study quantified the ghost fishing efficiency by comparing it to the catch efficiency of actively fished pots of the commercial fishery. The results showed that the ghost fishing affects both target and bycatch species. On average, the ghost fishing pots captured 12.53 % (confidence intervals: 10.45 %-15.00 %) undersized crab and 15.70 % (confidence intervals: 12.08 %-20.74 %) legal-sized crab compared to the actively fished pots. Few individuals of several bycatch species were also captured by ghost fishing pots. The results of this study emphasized the need to develop new management strategies for reducing marine pollution by ALDFG and associated negative effects in this pot fishery.

Effect of gillnet mesh size on the capture probability and capture patterns in the Asian paddle crab (Charybdis japonica) fishery.

In the Asian paddle crab (Charybdis japonica) gillnet fishery in the Yellow Sea, China, the minimum mesh size (MMS) regulation has been of a major importance due to high bycatch rates of undersized crabs. In this study, we evaluated how gillnet mesh size can affect the capture probability of C. japonica and capture patterns in this fishery by comparing the performance of gillnets with four different mesh sizes (60, 70, 80, and 90 mm). Our results showed that changes in gillnet mesh size significantly affect the capture probability of different sizes of crabs. Specifically, increased mesh size decreased the capture probability of undersized crabs and their fraction in the catches decreased from 64 % to 24 % when mesh size was increased from 60 mm to 90 mm. In contrast, gillnets with larger mesh sizes significantly improved the capture probability of legal-sized crabs. Moreover, no significant differences were observed for the species catch composition between gillnets of different mesh sizes. Based on these results, we recommend 90 mm as the MMS in gillnets to improve sustainability in C. japonica fishery.

The N-terminal region of Cdc6 specifically recognizes human DNA G-quadruplex.

Guanine (G)-rich nucleic acid sequences can form diverse G-quadruplex structures located in functionally significant genome regions, exerting regulatory control over essential biological processes, including DNA replication in vivo. During the initiation of DNA replication, Cdc6 is recruited by the origin recognition complex (ORC) to target specific chromosomal DNA sequences. This study reveals that human Cdc6 interacts with G-quadruplex structure through a distinct region within the N-terminal intrinsically disordered region (IDR), encompassing residues 7-20. The binding region assumes a hook-type conformation, as elucidated by the NMR solution structure in complex with htel21T18. Significantly, mutagenesis and in vivo investigations confirm the highly specific nature of Cdc6's recognition of G-quadruplex. This research enhances our understanding of the fundamental mechanism governing the interaction between G-quadruplex and the N-terminal IDR region of Cdc6, shedding light on the intricate regulation of DNA replication processes.

TAT-W61 peptide attenuates neuronal injury through blocking the binding of S100b to the V-domain of Rage during ischemic stroke.

Ischemic stroke is a devastative nervous system disease associated with high mortality and morbidity rates. Unfortunately, no clinically effective neuroprotective drugs are available now. In ischemic stroke, S100 calcium-binding protein b (S100b) binds to receptor for advanced glycation end products (Rage), leading to the neurological injury. Therefore, disruption of the interaction between S100B and Rage can rescue neuronal cells. Here, we designed a peptide, termed TAT-W61, derived from the V domain of Rage which can recognize S100b. Intriguingly, TAT-W61 can reduce the inflammatory caused by ischemic stroke through the direct binding to S100b. The further investigation demonstrated that TAT-W61 can improve pathological infarct volume and reduce the apoptotic rate. Particularly, TAT-W61 significantly improved the learning ability, memory, and motor dysfunction of the mouse in the ischemic stroke model. Our study provides a mechanistic insight into the abnormal expression of S100b and Rage in ischemic stroke and yields an invaluable candidate for the development of drugs in tackling ischemic stroke. KEY MESSAGES: S100b expression is higher in ischemic stroke, in association with a high expression of many genes, especially of Rage. S100b is directly bound to the V-domain of Rage. Blocking the binding of S100b to Rage improves the injury after ischemic stroke.

Applying an ensemble of small models in predicting habitat suitability of invasive M. sallei along the southern coast of China.

A false mussel Mytilopsis sallei has caused serious ecological and economic losses after invading in China. In this research, we first assessed the niche differentiation between its native range and invasive range in China and then predicted the habitat suitability along the southern coast of China under present and future climatic circumstances. Distance to shore and water depth were the first two important factors in affecting the distribution of M. sallei, followed by minimum chlorophyll concentration and salinity. The niche of M. sallei shows significant expansion and unfilling. The ensemble of small models can account for few occurrences and presents high predictive performance. A general reduction and northward movement of suitable areas were found in the southern coast of China in the future. This study furnished significant insights regarding the areas under invasive risks, and provided valuable information for preventing the further invasion of M. sallei in China.

Comparison of physical properties and fishing performance between biodegradable PLA and conventional PA trammel nets in grey mullet (Mugil cephalus) and red-lip mullet (Liza haematocheila) fishery.

Marine plastic pollution and continuous capture of marine animals, so-called "ghost fishing", by abandoned, lost, or otherwise discarded fishing gear (ALDFG) are global concerns. This study investigated whether biodegradable polylactic acid (PLA) monofilaments can be used to replace conventionally used non-biodegradable polyamide (PA) in trammel net fishery for limiting ALDFG associated effects. It evaluated the physical properties of PLA and PA monofilaments and compared fishing performance of PLA and PA trammel nets in a commercial mullet fishery in the Yellow Sea, China. Although PA monofilament exhibited superior physical properties, no significant differences in catch efficiency between PA and PLA trammel nets were observed. Fish of both species were mainly captured by pocketing which can further explain observed similar catch efficiency. These initial results suggest a potential for applying biodegradable materials in trammel net fisheries. Therefore, further long-term testing is encouraged to investigate whether this promising performance is persistent over long-term.

An essential Noc3p dimerization cycle mediates ORC double-hexamer formation in replication licensing.

Replication licensing, a prerequisite of DNA replication, helps to ensure once-per-cell-cycle genome duplication. Some DNA replication-initiation proteins are sequentially loaded onto replication origins to form pre-replicative complexes (pre-RCs). ORC and Noc3p bind replication origins throughout the cell cycle, providing a platform for pre-RC assembly. We previously reported that cell cycle-dependent ORC dimerization is essential for the chromatin loading of the symmetric MCM double-hexamers. Here, we used Saccharomyces cerevisiae separation-of-function NOC3 mutants to confirm the separable roles of Noc3p in DNA replication and ribosome biogenesis. We also show that an essential and cell cycle-dependent Noc3p dimerization cycle regulates the ORC dimerization cycle. Noc3p dimerizes at the M-to-G1 transition and de-dimerizes in S-phase. The Noc3p dimerization cycle coupled with the ORC dimerization cycle enables replication licensing, protects nascent sister replication origins after replication initiation, and prevents re-replication. This study has revealed a new mechanism of replication licensing and elucidated the molecular mechanism of Noc3p as a mediator of ORC dimerization in pre-RC formation.

Selective C9orf72 G-Quadruplex-Binding Small Molecules Ameliorate Pathological Signatures of ALS/FTD Models.

The G-quadruplex (G4) forming C9orf72 GGGGCC (G4C2) expanded hexanucleotide repeat (EHR) is the predominant genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Developing selective G4-binding ligands is challenging due to the conformational polymorphism and similarity of G4 structures. We identified three first-in-class marine natural products, chrexanthomycin A (cA), chrexanthomycin B (cB), and chrexanthomycin C (cC), with remarkable bioactivities. Thereinto, cA shows the highest permeability and lowest cytotoxicity to live cells. NMR titration experiments and in silico analysis demonstrate that cA, cB, and cC selectively bind to DNA and RNA G4C2 G4s. Notably, cA and cC dramatically reduce G4C2 EHR-caused cell death, diminish G4C2 RNA foci in (G4C2)29-expressing Neuro2a cells, and significantly eliminate ROS in HT22 cells. In (G4C2)29-expressing Drosophila, cA and cC significantly rescue eye degeneration and improve locomotor deficits. Overall, our findings reveal that cA and cC are potential therapeutic agents deserving further clinical study.

Cryo-EM structure of human hexameric MCM2-7 complex.

The central step in the initiation of eukaryotic DNA replication is the loading of the minichromosome maintenance 2-7 (MCM2-7) complex, the core of the replicative DNA helicase, onto chromatin at replication origin. Here, we reported the cryo-EM structure of endogenous human single hexameric MCM2-7 complex with a resolution at 4.4 Å, typically an open-ring hexamer with a gap between Mcm2 and Mcm5. Strikingly, further analysis revealed that human MCM2-7 can self-associate to form a loose double hexamer which potentially implies a novel mechanism underlying the MCM2-7 loading in eukaryote. The high-resolution cryo-EM structure of human MCM2-7 is critical for understanding the molecular mechanisms governing human DNA replication, especially the MCM2-7 chromatin loading and pre-replicative complex assembly.

Improving size selectivity of round pot for Charybdis japonica by configuring escape vents in the Yellow Sea, China.

Sustainable development of the important economic species, Asian paddle crab (Charybdis japonica), has attracted attention in the coastal waters of the Yellow Sea, China. The commonly used round pots are almost nonselective, resulting in severe bycatch of juveniles. In this study, we explored a method to improve the size selectivity for C. japonica by mounting escape vents on the side panels of each pot. The selectivity of pots with escape vent sizes of 70 mm × 20 mm, 70 mm × 25 mm, 70 mm × 30 mm, and 70 mm × 35 mm was tested using a catch comparison method. The estimated minimum landing size (MLS) of carapace height (27 mm), according to the regulated MLS of carapace length (50 mm), was used as a reference point to explain the results. Significant increases in the size of crabs caught by pots were found with the enlargement of escape vent size (Kruskal-Wallis test, P < 0.01). The pots with 70 mm × 20 mm, 70 mm × 25 mm and 70 mm × 30 mm escape vents released nearly 50%, 75% and 95% of undersized individuals, respectively, and these three types of pots retained approximately 90% of legal-sized individuals compared with the control pots without escape vents. The pots with 70 mm × 35 mm escape vents released nearly all undersized individuals, but they also released most legal-sized individuals. Pots with an escape vent size of 70 mm × 30 mm were recommended for the sustainable development of C. japonica in the Yellow Sea of China. The results of this study reiterate the importance of carapace height for determining the size selectivity, which can serve as a reference to formulate management regulations in the coastal waters of the Yellow Sea, China.

Estimating the impact of climate change on the potential distribution of Indo-Pacific humpback dolphins with species distribution model.

As IUCN critically vulnerable species,the Indo-Pacific humpback dolphins (Sousa chinensis) have attracted great public attention in recent years. The threats of human disturbance and environmental pollution to this population have been documented extensively. However, research on the sensitivity of this species to climate change is lacking. To understand the effect of climate change on the potential distribution of Sousa chinensis, we developed a weighted ensemble model based on 82 occurrence records and six predictor variables (e.g., ocean depth, distance to shore, mean temperature, salinity, ice thickness, and current velocity). According to the true skill statistic (TSS) and the area under the receiver operating characteristic curve (AUC), our ensemble model presented higher prediction precision than most of the single-algorithm models. It also indicated that ocean depth and distance to shore were the most important predictors in shaping the distribution patterns. The projections for the 2050s and 2100s from our ensemble model indicated a severe adverse impact of climate change on the Sousa chinensis habitat. Over 75% and 80% of the suitable habitat in the present day will be lost in all representative concentration pathway emission scenarios (RCPS) in the 2050s and 2100s, respectively. With the increased numbers of records of stranding and deaths of Sousa chinensis in recent years, strict management regulations and conservation plans are urgent to safeguard the current suitable habitats. Due to habitat contraction and poleward shift in the future, adaptive management strategies, including designing new reserves and adjusting the location and range of reserves according to the geographical distribution of Sousa chinensis, should be formulated to minimize the impacts of climate change on this species.

Penetration of Arsenic and Deactivation of a Honeycomb V2O5-WO3/TiO2 Catalyst in a Glass Furnace.

Deactivation of honeycomb V2O5-WO3/TiO2 catalysts by arsenic has been studied widely in coal-fired power plants but rarely in glass furnaces. In this paper, deactivated catalysts that had been used for more than 4000 h were analyzed. We maintained the catalysts in their original monolith shape to retain their adhered substance and used appropriate methods to strip the substance layer by layer. With various characterization techniques, it was determined that the adhered substance was composed almost entirely of Na2SO4 and CaSO4. We also quantified the penetration depth of arsenic visually, which was more than 370 µm. A three-stage penetration and deactivation process induced by arsenic was proposed. It was pointed out that molten and volatile As2O3 played a key role in the deactivation process, while substances in the solid state had little impact on the deep bulk of the catalyst. In this study, we proposed an integrated deactivation process consisting of adhesion, penetration, and deactivation in a honeycomb V2O5-WO3/TiO2 catalyst by arsenic in a glass furnace. Finally, we also provided guidance on alleviating the deactivation caused by arsenic. The key is to convert molten and volatile As2O3 to solid-state substances before it contacts the catalyst.

Crystal structure of parallel G-quadruplex formed by the two-repeat ALS- and FTD-related GGGGCC sequence.

The hexanucleotide repeat expansion, GGGGCC (G4C2), within the first intron of the C9orf72 gene is known to be the most common genetic cause of both amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). The G4C2 repeat expansions, either DNA or RNA, are able to form G-quadruplexes which induce toxicity leading to ALS/FTD. Herein, we report a novel crystal structure of d(G4C2)2 that self-associates to form an eight-layer parallel tetrameric G-quadruplex. Two d(G4C2)2 associate together as a parallel dimeric G-quadruplex which folds into a tetramer via 5'-to-5' arrangements. Each dimer consists of four G-tetrads connected by two CC propeller loops. Especially, the 3'-end cytosines protrude out and form C·C+•C·C+/ C·C•C·C+ quadruple base pair or C•C·C+ triple base pair stacking on the dimeric block. Our work sheds light on the G-quadruplexes adopted by d(G4C2) and yields the invaluable structural details for the development of small molecules to tackle neurodegenerative diseases, ALS and FTD.

Structural basis of DNA replication origin recognition by human Orc6 protein binding with DNA.

The six-subunit origin recognition complex (ORC), a DNA replication initiator, defines the localization of the origins of replication in eukaryotes. The Orc6 subunit is the smallest and the least conserved among ORC subunits. It is required for DNA replication and essential for viability in all species. Orc6 in metazoans carries a structural homology with transcription factor TFIIB and can bind DNA on its own. Here, we report a solution structure of the full-length human Orc6 (HsOrc6) alone and in a complex with DNA. We further showed that human Orc6 is composed of three independent domains: N-terminal, middle and C-terminal (HsOrc6-N, HsOrc6-M and HsOrc6-C). We also identified a distinct DNA-binding domain of human Orc6, named as HsOrc6-DBD. The detailed analysis of the structure revealed novel amino acid clusters important for the interaction with DNA. Alterations of these amino acids abolish DNA-binding ability of Orc6 and result in reduced levels of DNA replication. We propose that Orc6 is a DNA-binding subunit of human/metazoan ORC and may play roles in targeting, positioning and assembling the functional ORC at the origins.

An Essential and Cell-Cycle-Dependent ORC Dimerization Cycle Regulates Eukaryotic Chromosomal DNA Replication.

Eukaryotic DNA replication licensing is a prerequisite for, and plays a role in, regulating genome duplication that occurs exactly once per cell cycle. ORC (origin recognition complex) binds to and marks replication origins throughout the cell cycle and loads other replication-initiation proteins onto replication origins to form pre-replicative complexes (pre-RCs), completing replication licensing. However, how an asymmetric single-heterohexameric ORC structure loads the symmetric MCM (minichromosome maintenance) double hexamers is controversial, and importantly, it remains unknown when and how ORC proteins associate with the newly replicated origins to protect them from invasion by histones. Here, we report an essential and cell-cycle-dependent ORC "dimerization cycle" that plays three fundamental roles in the regulation of DNA replication: providing a symmetric platform to load the symmetric pre-RCs, marking and protecting the nascent sister replication origins for the next licensing, and playing a crucial role to prevent origin re-licensing within the same cell cycle.

Exploring spatial nonstationary environmental effects on Yellow Perch distribution in Lake Erie.

BACKGROUND: Global regression models under an implicit assumption of spatial stationarity were commonly applied to estimate the environmental effects on aquatic species distribution. However, the relationships between species distribution and environmental variables may change among spatial locations, especially at large spatial scales with complicated habitat. Local regression models are appropriate supplementary tools to explore species-environment relationships at finer scales. METHOD: We applied geographically weighted regression (GWR) models on Yellow Perch in Lake Erie to estimate spatially-varying environmental effects on the presence probabilities of this species. Outputs from GWR were compared with those from generalized additive models (GAMs) in exploring the Yellow Perch distribution. Local regression coefficients from the GWR were mapped to visualize spatially-varying species-environment relationships. K-means cluster analyses based on the t-values of GWR local regression coefficients were used to characterize the distinct zones of ecological relationships. RESULTS: Geographically weighted regression resulted in a significant improvement over the GAM in goodness-of-fit and accuracy of model prediction. Results from the GWR revealed the magnitude and direction of environmental effects on Yellow Perch distribution changed among spatial locations. Consistent species-environment relationships were found in the west and east basins for adults. The different kinds of species-environment relationships found in the central management unit (MU) implied the variation of relationships at a scale finer than the MU. CONCLUSIONS: This study draws attention to the importance of accounting for spatial nonstationarity in exploring species-environment relationships. The GWR results can provide support for identification of unique stocks and potential refinement of the current jurisdictional MU structure toward more ecologically relevant MUs for the sustainable management of Yellow Perch in Lake Erie.