Additional Interactions Research Articles

Rational Design for Enhancing Cellobiose Dehydrogenase Activity and Its Synergistic Role in Straw Degradation.

Addressing the demand for efficient biological degradation of straw, this study employed rational design coupled with structural biology and enzyme engineering techniques to enhance the catalytic activity of cellobiose dehydrogenase (PsCDH, CDH form Pycnoporus sanguineus). By predicting and modifying the active site and key amino acids of PsCDH, several CDH immobilized enzyme preparations with higher catalytic activities were successfully obtained. The excellent mutant T1 (C286Y/A461H/F464R) exhibited a 2.7-fold increase in enzyme activity compared to the wild type. Simulated calculations indicated that the enhancement of catalytic activity was primarily due to the formation of additional intermolecular interactions between CDH and the substrate, as well as the enlargement of the substrate pocket to reduce steric hindrance effects. Additionally, molecular dynamics simulation analysis revealed a potential correlation between structural stability and enzyme activity. When PsCDH was added to a multienzyme synergistic straw degradation system, the cellulose degradation rate increased by 1.84-fold. Moreover, mutant T1 further increased the degradation of lignocellulose in the mixed system. This study provides efficient enzyme sources and modification strategies for the high-efficiency biological conversion of straw and unconventional feedstock degradation, thereby possessing significant academic value and application prospects.

Tryptamine‐Derived Schiff Bases: Potent Antimicrobial Agents and Evaluation of Cytotoxicity, ADME and DNA Binding Properties

Inspired by the fact that the introduction of indole pharmacophore in organic scaffolds could enable interesting pharmacological properties, the series of novel tryptamine‐derived Schiff bases was synthetized. Tryptamine was used as a source of indole pattern, as well as an example of biogenic amines which chemical transformations lead to the compounds with prominent biological activities. The obtained results for antimicrobial activity against a range of bacterial and fungal strains and cytotoxic activities have revealed that Schiff base TSB4 combining the tryptamine and p‐nitro aryl patterns in the structure showed better antifungal activity at low concentrations than standard drug Fluconazole, while compound TSB6 with molecular scaffold composed from tryptamine and quinoline moieties showed certain cytotoxic effect on HCT‐116 cell line with a strongly expressed selectivity about healthy fibroblast cells, MRC‐5. For these two selected compounds, additional ADME analysis and DNA interactions were performed. to obtain better insight into their pharmacokinetics and determination of binding mode for DNA molecules. As results suggested, strong binding of examined compounds to CT‐DNA was observed, while the ADME screening showed that selected compounds possess suitable physicochemical properties for oral bioavailability and druglikeness.

In utero and childhood exposure to the great Chinese famine and risk of aging in adulthood

Background Early-life exposure to famine may influence the occurrence of chronic diseases and aging in midlife among those exposed. This study aims to explore the relationship between exposure to the Chinese Great Famine and aging in middle-aged individuals. Methods Participants born in 1963–1965 (unexposed), 1959–1961 (in utero exposure), and 1955–1957 (childhood exposure) from the Kailuan Study were included. Their biological age at 2010, 2014, and 2018 was investigated, and age acceleration (biological age minus actual age) was calculated to assess aging. Logistic regression analysis was employed to describe the relationship between famine exposure and the aging risk. Subgroup and sensitivity analysis were conducted to explore differences and stability in this relationship among different groups. Results A total of 17,543 participants were included in this study. Among them, 12,762 (72.7%) were male, and 4,781 (27.3%) were female, with 2,543 participants experiencing aging events. Compared to unexposed participants, those exposed during childhood and in utero exhibited a 1.69-fold (OR = 1.69, 95%CI: 1.53–1.87) and 1.22-fold (OR = 1.22, 95%CI: 1.08–1.37) increased risk of aging. Subgroup analysis revealed an interaction with income (P for interaction = 0.008), and additional interaction analysis suggested that increasing income could partially mitigate the detrimental effects of early-life famine exposure. Furthermore, experiencing famine in severely affected regions exacerbated the risk of aging (OR = 1.41, 95%CI: 1.21–1.63). Conclusion Exposure to famine in utero or during childhood may elevate the risk of midlife aging among exposed individuals, and these relationships are influenced by the severity of famine exposure. Increasing income may also help mitigate these effects.Trial registration: Kailuan study, ChiCTRTNRC11001489. Registered July 19, 2015 Retrospectively registered, https//www.chictr.org.cn/showprojEN.html?proj=8050&u_atoken=af46a0dee8d73f320bb5459ab7bbcfa9&u_asig=1a0c381017255295896468605e00cf .

Employee Insights into Organizational Justice and Job Performance: The Case of Insurance Companies

This paper explores employee insights into organizational justice and job performance. It also examines the association between employee insights into organizational justice and their personal traits and analyzes the association between employee insights into organizational justice and job performance. This study used descriptive statistics as well as correlation analysis. The survey data were gathered from 250 employees of ten insurance companies (including both life and non-life) in Kathmandu Valley. The results indicate a significant and positive association between employee insights into organizational justice and job performance. It indicates that the justice of management matters a lot in enhancing employee job performance at work. The study's findings also reveal a substantial association between all employees' demographic characteristics and their insights into organizational justice. It indicates that when employers treat their employees fairly, they do well on the job. To develop a climate of organizational justice, organizations must ensure that rewards and resources are distributed fairly. It should prioritize clear compensation policies, fair reward systems, and performance-based incentives. It must also prioritize the establishment of fair and consistent decision-making systems by creating standardized systems for evaluations, promotions, and disciplinary actions. In addition, fostering respectful and equitable interpersonal interactions can also help to build an environment of organizational justice. For this, organizations must train managers and supervisors on effective communication skills, emphasizing the importance of treating employees with dignity and respect, promoting open-door policies in which employees feel comfortable discussing concerns, and ensuring that managers provide clear and honest clarification for decisions. All of these activities ultimately increase employees' willingness to optimize their potential for higher levels of job performance.

Multivalent interactions between fully glycosylated influenza virus hemagglutinins mediated by glycans at distinct N-glycosylation sites

The hemagglutinin (HA) glycoprotein of influenza virus binds host cell receptors and mediates viral entry. Here we present cryo-EM structures of fully glycosylated HAs from H5N1 and H5N8 influenza viruses. We find that the H5N1 HA can form filaments that comprise two head-to-head HA trimers. Multivalent interactions between the two HA trimers are mediated by glycans attached to N158. The distal Sia1-Gal2-NAG3 sugar moiety of N158 interacts with the receptor binding site on the opposing HA trimer. Additional interactions are observed between NAG3 and residues K222 and K193. The H5N8 HA lacks the N158 glycosylation site and does not form the filamentous structure. However, the H5N8 HA exhibits an auto-inhibition conformation, where the receptor binding site is occupied by the glycan chain attached to residue N169 from a neighboring protomer. These structures represent native HA-glycan interactions, which may closely mimic the receptor-HA interactions on the cell surface.

A Survey on Intent-aware Recommender Systems

Many modern online services feature personalized recommendations. A central challenge when providing such recommendations is that the reason why an individual user accesses the service may change from visit to visit or even during an ongoing usage session. To be effective, a recommender system should therefore aim to take the users’ probable intent of using the service at a certain point in time into account. In recent years, researchers have thus started to address this challenge by incorporating intent-awareness into recommender systems. Correspondingly, a number of technical approaches were put forward, including diversification techniques, intent prediction models or latent intent modeling approaches. In this paper, we survey and categorize existing approaches to building the next generation of Intent-Aware Recommender Systems (IARS). Based on an analysis of current evaluation practices, we outline open gaps and possible future directions in this area, which in particular include the consideration of additional interaction signals and contextual information to further improve the effectiveness of such systems.

Rapid Coaggregation of Proteins Without Sequence Similarity: Possible Role of Conformational Complementarity.

Despite extensive research on the sequence-determined self-assembly of both pathogenic and nonpathogenic proteins, the question of how the sequence identity would influence the coassembly or cross-seeding of diverse proteins without distinct sequence similarity remains largely unanswered. Here, we demonstrate that the rapid coaggregation of proteins with negligible sequence similarity is fundamentally governed by preferred heteromeric interactions between their partially unfolded states via the gain of additional charge complementarity and hydrophobic interactions. The partial loss of intramolecular interactions and concurrent gain of non-native intrinsically disordered regions with sticky groups become crucial for both aggressive heteromeric primary nucleation and secondary nucleation events. The results signify the direct relevance of sequence-independent conformational cross-talk between diverse proteins to the foundational events required for the growth of biological multiprotein amyloid deposits.

Structural Study on Alkali‐Carboxylate Functionalized Late‐Transition Metal Bis(dithiocarbamate) Complexes

AbstractLithium, sodium, and potassium derivatives of amino‐acid derived bis(dithiocarbamate) complexes of bivalent nickel, palladium, platinum, and copper are readily available by treatment of the carboxylic‐acid functionalized complexes with the respective alkali metal hydroxide. Representative derivatives of the types A4[M(L1)2], A2[M(HL1)2], A[M(HL1)(H2L1)] (N‐dithioato‐iminodiacetate, L1), and A2[M(L2,3)2] (N‐dithioato‐l‐prolinate, L2, and ‐N‐benzylglycinate, L3; A=Li, Na, K; M=Ni, Pd, Pt, Cu) were isolated and characterized by spectroscopic and thermal methods. Six entries were accessible to single‐crystal X‐ray structure determination, revealing the presence of different coordination‐polymeric structures and hydrogen‐bonded assemblies comprising solvent‐separated cations in the solid state. While the transition metal always retains its all‐sulfur coordination with defined geometry, the alkali metals are structurally more flexible. The latter are usually coordinated by carboxylate and water, but additional interactions with sulfur donors are also relevant in some cases.

Identification of novel diarylpyrimidine derivatives as potent HIV-1 non-nucleoside reverse transcriptase inhibitors against wild-type and K103N mutant viruses

HIV-1 non-nucleoside reverse transcriptase inhibitors (NNRTIs) play a crucial role in combination antiretroviral therapy (cART). To further enhance their antiviral activity and anti-resistance properties, we developed a series of novel NNRTIs, by specifically targeting tolerant region I of the NNRTI binding pocket. Among them, compound 9t-2 displayed excellent anti-HIV-1 potency against wild-type and prevalent mutant strains with EC50 values between 0.0019 and 0.012 μM. This outperformed the positive drugs ETR, NVP and RPV. Aslo, ELISA results confirmed that these compounds can effectively inhibit the activity of HIV-1 RT. Molecular dynamics (MD) simulation studies indicated that the thiomorpholine-1,1-dioxide moiety of 9t-2 is capable of establishing additional interactions with residues P225, F227 and P236 in the tolerant region I, which contributed to its enhanced activity. Compound 9t-2 possessed negligible inhibitory effect on the five main CYP isoenzymes (IC50 > 10 μM), indicating a low potential for inducing CYP-mediated drug-drug interactions. In conclusion, compound 9t-2, with its enhanced anti-resistance properties, stands out as a promising lead compound for further optimization towards discovering the new generation of anti-HIV agents.

Addressing the Intracellular Vestibule of the Plasmodial Lactate Transporter PfFNT by p-Substituted Inhibitors Amplifies In Vitro Activity.

Inhibition of the lactate transporter PfFNT is a valid novel mode of action against malaria parasites. Current pyridine-substituted pentafluoro-3-hydroxy-pent-2-en-1-ones act as substrate analogs with submicromolar EC50 in vitro, and >99.7% activity in mice. The recently solved structure of a PfFNT-inhibitor complex visualized the binding mode. Here, we extended the inhibitor layout by series of amine- and anilide-linked pyridine p-substituents to generate additional interactions in the cytoplasmic vestibule. Virtual docking indicated hydrogen bonding to Tyr31 and Ser102. Fluorescence cross-correlation spectroscopy yielded respectively enhanced target affinity. Strikingly, the in vitro activity increased by 1 order of magnitude to 14.8 nM at negligible cytotoxicity. While p-amine substitutions were rapidly metabolized, the more stable p-acetanilide cleared 99.7% of parasites at 4 × 50 mg kg-1 in a mouse malaria model. Future stabilization of the p-substitution against metabolism may translate the gain in in vitro potency to the in vivo situation.

Nature of Topological Phase Transition of Kitaev Quantum Spin Liquids.

We investigate the nature of the topological quantum phase transition between the gapless and gapped Kitaev quantum spin liquid phases away from the exactly solvable point. The transition is driven by anisotropy of the Kitaev couplings. At the critical point, the two Dirac points of the gapless Majorana modes merge, resulting in the formation of a semi-Dirac point with quadratic and linear band touching directions. We derive an effective Gross-Neveu-Yukawa-type field theory that describes the topological phase transition in the presence of additional magnetic interactions. We obtain the infrared scaling form of the propagator of the dynamical Ising order parameter field and perform a renormalization-group analysis. The universality of the transition is found to be different from that of symmetry-breaking phase transitions of semi-Dirac electrons. However, as in the electronic case, the Majorana fermions acquire an anomalous dimension, indicative of the breakdown of the fractionalized quasiparticle description.

Surfactant-tolerance evolution of Bacillus clausii protease for enhancing activity and stability by reshaping the substrate access tunnel

Alkali proteases are crucial in numerous industries, especially in the laundry industry, but their inactivation by surfactants limits their effectiveness. This study employed substrate access tunnel engineering to improve the performance of WT bcPRO in surfactants. By modifying the key residues in the substrate pocket, the best variant N212S showed higher stability and activity in both AES and LAS. Molecular dynamics (MD) simulations provided insights into the enhanced stability and activity. The Asn212Ser mutation weakened the anti-correlation motion, increased the number of hydrogen bonds between amino acid residues, and made the protein structure more compact, contributing to its stability. Additionally, the mutation extended the substrate access tunnel and enabled additional interactions with the substrate, enhancing its catalytic activity in surfactants. This study demonstrates a strategy for reshaping the substrate access tunnel to improve protease stability and activity in surfactant environments, offering a promising protease candidate for the laundry industry.

Effect of repulsive interaction and initial velocity on collective motion process.

Self-propelled collective motion is a highly complex phenomenon, necessitating advanced practical and theoretical tools for comprehension. The significance of studying collective motion becomes apparent in its diverse applications. For instance, addressing evacuation challenges in scenarios with multiple agents can be achieved through an examination of collective motion. Research indicates that the transition of individuals (such as birds, fish, etc.) from a state of rest to equilibrium constitutes a phase transition. Our interest of the issue is to delve into the nature of this transitional phase and elucidate the parameters that shape it. Hence, the primary aim of this paper is to grasp the kinetic phase transition by examining how initial velocity and repulsive interactions impact the dynamics of the system. To gain insight into the complex behavior of multi-agent systems, we apply an extended version of the classical Vicsek model. This extension includes an additional interaction zone, the repulsive zone, where particles repel each other at close range to avoid collisions. Our study uses numerical simulations to explore the system's behavior under various conditions. The focus of this study is the impact of initial velocity on the collective movement of particles. The importance of this research lies in comprehending how velocity affects the overall movement. The conclusion we can draw from these results is that the initial velocity affects both the noise and the density. The novelty of the work is the transition phase, yet it lacks universal characteristics because the critical noise depends on the initial velocity system and the repulsion radius zone. Notably, the repulsion radius and particle density play pivotal roles in achieving a phase transition from one equilibrium state to another aligned equilibrium state.

Arg92Leu-cTnT Alters the cTnC-cTnI Interface Disrupting PKA-Mediated Relaxation.

Impaired left ventricular relaxation, high filling pressures, and dysregulation of Ca2+ homeostasis are common findings contributing to diastolic dysfunction in hypertrophic cardiomyopathy (HCM). Studies have shown that impaired relaxation is an early observation in the sarcomere-gene-positive preclinical HCM cohort, which suggests the potential involvement of myofilament regulators in relaxation. A molecular-level understanding of mechanism(s) at the level of the myofilament is lacking. We hypothesized that mutation-specific, allosterically mediated, changes to the cTnC (cardiac troponin C)-cTnI (cardiac troponin I) interface can account for the development of early-onset diastolic dysfunction via decreased PKA accessibility to cTnI. HCM mutations R92L-cTnT (cardiac troponin T; Arg92Leu) and Δ160E-cTnT (Glu160 deletion) were studied in vivo, in vitro, and in silico via 2-dimensional echocardiography, Western blotting, ex vivo hemodynamics, stopped-flow kinetics, time-resolved fluorescence resonance energy transfer, and molecular dynamics simulations. The HCM-causative mutations R92L-cTnT and Δ160E-cTnT result in different time-of-onset diastolic dysfunction. R92L-cTnT demonstrated early-onset diastolic dysfunction accompanied by a localized decrease in phosphorylation of cTnI. Constitutive phosphorylation of cTnI (cTnI-D23D24) was sufficient to recover diastolic function to non-Tg levels only for R92L-cTnT. Mutation-specific changes in Ca2+ dissociation rates associated with R92L-cTnT reconstituted with cTnI-D23D24 led us to investigate potential involvement of structural changes in the cTnC-cTnI interface as an explanation for these observations. We probed the interface via time-resolved fluorescence resonance energy transfer revealing a repositioning of the N-terminus of cTnI, closer to cTnC, and concomitant decreases in distance distributions at sites flanking the PKA consensus sequence. Implementing time-resolved fluorescence resonance energy transfer distances as constraints into our atomistic model identified additional electrostatic interactions at the consensus sequence. These data show that the early diastolic dysfunction observed in a subset of HCM is attributable to allosterically mediated structural changes at the cTnC-cTnI interface that impair accessibility of PKA, thereby blunting β-adrenergic responsiveness and identifying a potential molecular target for therapeutic intervention.

On the abundance and importance of AXXXA sequence motifs in globular proteins and their involvement in Cβ[sbnd]Cβ interaction

The AXXXA and GXXXG motifs are frequently observed in helices, especially in membrane proteins. The motif GXXXG is known to stabilize helix-helix association in membrane proteins via CαHO bonding. AXXXA sequence motif additionally stabilizes the folded state of proteins. We found 27,000 and 18,000 occurrences of AXXXA and GXXXG motifs in a non-redundant set of 6000 obligate homodimeric (OD) complexes. Interestingly, this is less pronounced in transient homodimers (TD) and heterodimers (HetD). On average each obligate homodimer contains four AXXXA motifs, it is 2 and 3.5 for HetD and TD, respectively. Focusing on the binding surface it is seen that 27 % of the ODs contain at least one AXXXA motif at the interface, whereas it is 17 % and 15 % for HetD and TD respectively. AXXXA predominantly stabilizes the OD quaternary structure via the side chain CβCβ interactions. This interaction is energetically favorable and is found to be a major driving force for OD quaternary structure stability. Cβ-Cβ interactions are observed ∼6 times higher than the known CαHO interaction for helix-helix stabilization. Two additional new interactions of CβO and OO are observed at the AXXXA containing interface regions. The occurrence of the motif gets drastically reduced if any of the terminal Ala residues are replaced by Gly. Our findings show the importance of AXXXA in providing stability to the quaternary structure through specific hydrophobic interactions and the specificity of the Ala residue at motif termini. The knowledge gained can be used for designing synthetic proteins of improved stability and for designing peptide-based therapeutics.

Distinguishing laser-induced dissociation pathways of O 2 molecule based on alignment-dependent dissociation spectra in intense laser fields

We experimentally investigate the dissociative single ionization process, O 2 → O + + O, of aligned O 2 molecule in intense laser fields. The yield of a vibrational structure in the kinetic energy release spectra is measured as a function of alignment angle. By quantitative comparison of the measured angle-dependent dissociation probability with the simulation of a classical model that considers ionization and thus additional interaction within the laser pulse, we are able to distinguish the dissociation pathway that contribute to the vibrational structure. It is found that for a relative low laser intensity, the vibrational structure are produced from the dissociation pathway of a 4Π u → f 4Π g − 1ω. As the laser intensity increases, the increasing of the population probability of higher vibrational states and the absorbtion probability of more photons makes another two dissociation pathways open and become dominant.

Grazing and precipitation addition interactions alleviate dominant species overgrowth and promote community productivity and biodiversity in a typical steppe

Grazing and precipitation are pivotal factors influencing the productivity and biodiversity of grassland ecosystems, largely through their effects on the growth and reproduction of dominant species. Approximately 50 % of terrestrial ecosystems are concurrently affected by grazing and precipitation addition (PA), yet the interactive effects of these factors remain underexplored. To elucidate the combined impacts of grazing and PA on the growth of dominant species and their influence on community structure and function, we initiated a four-year combined grazing and PA experiment based on a long term of grazing experiment in a typical steppe. The synergistic interaction between PA and grazing enhanced canopy diameter (CD), tiller density (TD), and seedling density (SD) in dominant species, while decreasing reproductive branch density (RB). Conversely, an antagonistic interaction increased plant height (PH) and TD but reduced SD. These responses suggest that dominant species adapt to combined grazing and PA pressures by shifting growth strategies towards lateral growth and asexual reproduction. The growth characteristics of dominant species exhibited four response patterns to grazing and PA interactions: full saturation, sufficient saturation, equal saturation, and deficit saturation, each with three corresponding thresholds: adaptation, optimum, and saturation points. Grazing decreased the precipitation response thresholds for PH, CD, RB, and population density, while increasing the optimal points for TD and SD. These changes in the growth of the dominant species resulted in a 33 % reduction in the aboveground biomass (AGB) of the community and triggered a 18 % increase in the coupling index between AGB and species richness within the community. Our findings highlight the role of dominant species in facilitating community adaptation to increased precipitation and rotational grazing, offering critical insights for developing sustainable grazing strategies under climate change.

Synergistic Effect between the APOE ε4 Allele with Genetic Variants of GSK3B and MAPT: Differential Profile between Refractory Epilepsy and Alzheimer Disease.

Temporal Lobe Epilepsy (TLE) is a chronic neurological disorder characterized by recurrent focal seizures originating in the temporal lobe. Despite the variety of antiseizure drugs currently available to treat TLE, about 30% of cases continue to have seizures. The etiology of TLE is complex and multifactorial. Increasing evidence indicates that Alzheimer's disease (AD) and drug-resistant TLE present common pathological features that may induce hyperexcitability, especially aberrant hyperphosphorylation of tau protein. Genetic polymorphic variants located in genes of the microtubule-associated protein tau (MAPT) and glycogen synthase kinase-3β (GSK3B) have been associated with the risk of developing AD. The APOE ε4 allele is a major genetic risk factor for AD. Likewise, a gene-dose-dependent effect of ε4 seems to influence TLE. The present study aimed to investigate whether the APOE ɛ4 allele and genetic variants located in the MAPT and GSK3B genes are associated with the risk of developing AD and drug-resistant TLE in a cohort of the Mexican population. A significant association with the APOE ε4 allele was observed in patients with AD and TLE. Additional genetic interactions were identified between this allele and variants of the MAPT and GSK3B genes.

Design and Synthesis of Novel Indole-Derived N-Methylcarbamoylguanidinyl Chitinase Inhibitors with Significantly Improved Insecticidal Activity.

Chitinases play an important role in the molting process of insects and are potential targets for the development of green insecticides. Based on the feature that the +1/+2 sites in OfChtI, OfChtII, and OfChi-h have tryptophan residues in mismatch-parallel position, a strategy to introduce indole scaffold into chitinase inhibitors was proposed, and multitarget chitinase inhibitors containing N-methylcarbamoylguanidinyl and indole scaffold were successfully synthesized. The inhibitory activity showed that compound 8u exhibited significant inhibitory activity against OfChtI, OfChtII, and OfChi-h, with IC50 values of 0.7, 0.79, and 0.58 μM, and Ki values of 0.05 ± 0.005, 0.065 ± 0.004, and 0.025 ± 0.006 μM, respectively. In vivo insecticidal activity showed that compounds 8a and 8g exhibited excellent insecticidal activity against Plutella xylostella and Mythimna separata, with LC50 values of 0.79 and 9.17 mg/L against P. xylostella, respectively, and 3.58 and 83.09 mg/L against M. separata, respectively, making them the most potent chitinase inhibitors with in vivo insecticidal activity discovered to date. The inhibition mechanism and binding free energy results suggested that N-methylcarbamoylguanidinyl binds to the -1 catalytic site, while additional interactions acquired by π-π stacking and hydrophobic interactions of the indole scaffold with tryptophan increase the binding affinity of the targets to chitinases. This work provides a new direction for the development of chitinase inhibitors with compounds 8a and 8g potentially serving as promising candidates for pesticide development.

Above-room-temperature intrinsic ferromagnetism in ultrathin van der Waals crystal Fe3+xGaTe2

Two-dimensional (2D) van der Waals (vdW) magnets are crucial for ultra-compact spintronics. However, so far, no vdW crystal has exhibited tunable above-room-temperature intrinsic ferromagnetism in the 2D ultrathin regime. Here, we report the tunable above-room-temperature intrinsic ferromagnetism in ultrathin vdW crystal Fe3+xGaTe2 (x = 0 and 0.3). By increasing the Fe content, the Curie temperature (TC) and room-temperature saturation magnetization of bulk Fe3+xGaTe2 crystals are enhanced from 354 to 376 K and 43.9 to 50.4 emu·g−1, respectively. Remarkably, the robust anomalous Hall effect in 3-nm Fe3.3GaTe2 indicates a record-high TC of 340 K and a large room-temperature perpendicular magnetic anisotropy energy of 6.6 × 105 J m−3, superior to other ultrathin vdW ferromagnets. First-principles calculations reveal the asymmetric density of states and an additional large spin exchange interaction in ultrathin Fe3+xGaTe2 responsible for robust intrinsic ferromagnetism and higher TC. This work opens a window for above-room-temperature ultrathin 2D magnets in vdW-integrated spintronics.