Odorant-binding proteins (OBPs) in mosquitoes play central roles in chemosensory perception and are attractive targets for vector control strategies. However, some OBPs are also expressed in non-olfactory tissues, suggesting a pleiotropic function that has not been sufficiently investigated. The crystal structures of the pleiotropic AgamOBP9 of the Anopheles gambiae mosquito were determined in complex with three plant-derived bioactive molecules: the phenylpropanoids n-butyl cinnamate and methyl eugenol (ME), as well as the monoterpene p-menthane-3, 8-diol (PMD). Fluorescence competitive binding assays and binding free-energy calculations identified that n-butyl cinnamate and ME bind with Ki values in the micromolar range, in contrast to the weak affinity observed for PMD. Structural analysis revealed an extended internal cavity comprising two distinct ligand-binding regions. One region, located at the bottom of the cavity, accommodated all investigated ligands without undergoing significant conformational changes, suggesting a structurally preordered binding pocket. The second region, situated at the cavity entrance, binds MPD and PEG, both used as crystallization agents, and may therefore serve as a recognition site for molecules with diverse chemical features. In the AgamOBP9-butyl cinnamate-MPD complex, MPD binding to the entrance-site induces rearrangements of surrounding residues, including Arg8, Arg15, Tyr32, Lys33, and Trp35. These interactions appear to promote the convergence of helices α1, α2, and the α2-α3 connecting loop toward a more "closed" protein conformation. The conformational flexibility of AgamOBP9 at the entrance region was further supported by differential scanning calorimetry and molecular dynamics simulations, which suggested the presence of two independent thermodynamic domains within the protein. Structural superposition with the homologous AaegOBP22-linoleic acid complex indicated that the AgamOBP9 cavity could accommodate fatty acids or other long-chain molecules spanning both binding regions, implying potential functions beyond conventional olfactory signaling. These findings provide new insights into the molecular basis of ligand recognition by AgamOBP9, which may guide the discovery of novel OBP9-targeting ligands.

The cyclic monoterpene p-menthane-3,8-diol exhibits highly effective mosquito-repellent activity. Current industrial production of PMD depends on chemically synthesized citronellal as a precursor. To provide a green, sustainable alternative for citronellal supply, we engineered a Saccharomyces cerevisiae strain for efficient de novo biosynthesis of citronellal. By enhancing geraniol synthesis, systematically redesigning the endogenous oxidoreductase system, and implementing metabolic optimization strategies, a citronellal titer of 0.13 g/L was achieved in shake-flask cultivation. By integrating this microbial fermentation step with subsequent chemical catalysis, we established a biological-chemical method for PMD production, achieving a final PMD yield of 0.64 g/L. This work represents the first semisynthetic route from glucose to PMD via a biological-chemical method, offering a novel and sustainable paradigm for PMD manufacture.

Vector-borne diseases pose a rising global health challenge, necessitating the development of safe and effective pest protection agents. Here, we report a highly selective biocatalytic direct Prins cyclohydration for the synthesis of (1R)-cis-p-menthane-3,8-diol (PMD), a natural insect repellent with high efficacy. By strategically engineering squalene-hopene cyclases (SHCs), we achieved >96% diastereomeric excess, surpassing previous synthetic methods. Structural and mechanistic analyses suggest direct Prins cyclohydration and a precisely positioned water molecule within the enzyme's active pocket adjacent to the final carbocation that drives hydration and catalytic efficiency. Fine-tuning the biocatalytic setup enabled preparative scale production, without losing much product selectivity. Moreover, we demonstrate access to the other naturally occurring PMD isomers from (R)- and (S)-citronellal, as well as a one-pot cascade starting from E/Z-citral. This study paves the way for highly selective access to stereodefined terpene-derived repellents and establishes engineered squalene-hopene cyclases as a tool for direct asymmetric Prins cyclohydration.

Benchmarks are important tools for tracking the rapid advancements in large language model (LLM) capabilities. However, benchmarks are not keeping pace in difficulty: LLMs now achieve more than 90% accuracy on popular benchmarks such as Measuring Massive Multitask Language Understanding[1], limiting informed measurement of state-of-the-art LLM capabilities. Here, in response, we introduce Humanity's Last Exam (HLE), a multi-modal benchmark at the frontier of human knowledge, designed to be an expert-level closed-ended academic benchmark with broad subject coverage. HLE consists of 2,500 questions across dozens of subjects, including mathematics, humanities and the natural sciences. HLE is developed globally by subject-matter experts and consists of multiple-choice and short-answer questions suitable for automated grading. Each question has a known solution that is unambiguous and easily verifiable but cannot be quickly answered by internet retrieval. State-of-the-art LLMs demonstrate low accuracy and calibration on HLE, highlighting a marked gap between current LLM capabilities and the expert human frontier on closed-ended academic questions. To inform research and policymaking upon a clear understanding of model capabilities, we publicly release HLE at https://lastexam.ai .

This study systematically evaluated the synergistic effects of terpinen-4-ol (T4O) with p-menthane-3,8-diol (PMD) for enhanced mosquito repellency. Through screening 20 essential oils, we identified six candidates (cinnamon, eucalyptus, tea tree, litsea cubeba, moxa, peppermint) demonstrating >85% repellency. GC-MS analysis revealed 17 candidate compounds, with T4O exhibiting strong electrophysiological responses (0.031 ± 0.002 mV). Molecular docking confirmed its high binding affinity to mosquito odorant-binding proteins, suggesting its potential as an olfactory disruptor. In PMD-based binary formulations: TP2 (20% T4O) achieved 86.9 ± 1.8% spatial repellency, 30.6% higher than PMD alone; TP1 (10% T4O) provided 6.58 ± 0.38 h of skin protection, extending PMD's efficacy by 21.6%; TP5 (50% T4O) enhanced fumigation efficacy, reducing KT50 to 3.00 ± 0.32 min (78.7% decrease); TP3 (30% T4O) reduced LC50 by 17.2-25.0% against larval instars I-IV. This work demonstrates T4O's multidimensional enhancement of PMD's efficacy, offering an effective natural alternative to synthetic repellents.

Ixodes ricinus and Hyalomma excavatum are two widely dispersed vectors of pathogens, including those that cause Lyme disease and Crimean-Congo haemorrhagic fever in the human population. Recently developed, plant-derived, mosquito-repellent blends have shown promise against other vector clades, and this study assesses these blends as potential tick repellents. Blends of (+)-borneol, bornyl acetate, eugenol, isoeugenol and camphor were assessed in two formats: blends 3 and 4. Ticks were assessed using the moving object bioassay (Ixodes) or a dual-choice behavioural assay (Hyalomma). Both blends were compared against negative controls and four commercially available synthetic repellents: N,N-diethyl-3-methylbenzamide (DEET), 2-(2-hydroxyethyl)-1-methylpropylstyrene 1-piperidine carboxylate) (Picaridin), 3-(N-n-butyl-N-acetyl)-amino-propionic acid ethyl ester (IR3535) and p-menthane-3,8-diol (PMD). Results demonstrate the efficacies of blends 3 and 4; moreover, both were more effective than the commercial repellents (P < 0.05). Blend 3 was marginally more effective than blend 4, and differences in the repellent action were noted for each of the tick species, suggesting broad-spectrum vector-repellent activities, irrespective of life strategy. Overall, this work demonstrates the clear potential of blends 3 and 4 as tick repellents that offer an improved vector response over currently available commercial repellents. Furthermore, that the same repellent blends are capable of tick repellency in addition to mosquito repellency, offers the potential for widely dispersed usage across a range of integrated vector management strategies. © 2025 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

An in vitro repellent testing system for use with colony reared black flies, Simulium vittatum, is described. Postoviposition female S. vittatum were exposed to latex membranes treated with 15 μl of commercially available insect repellents every 2 h, up to 12 h. Repellents tested were the following: Repel® Plant-Based Lemon Eucalyptus Insect Repellent 2 (30% oil of lemon eucalyptus [OLE]); OFF!® Botanicals Insect Repellent IV (10% p-menthane-3,8-diol [PMD]); and Zevo™ On-Body (20% 3-[N-butyl-N-acetyl]-aminopropionic acid, ethyl ester, IR3535®). Untreated membranes served as control. The PMD and IR3535 had negative correlations between repellency rate and time (IR3535, m (slopes of mean repellencies over time) = -6.64; and PMD, m = -5.28), whereas OLE had none (m = 0). Statistical analysis demonstrated significance within all groups that included OLE or the control (P < 0.00), but none for groups consisting of PMD or IR3535 (P = 0.31).

Lethal lung developmental disorders (LLDDs), histologically classified as alveolar capillary dysplasia with misalignment of pulmonary veins (ACDMPV), congenital alveolar dysplasia (CAD), acinar dysplasia (AcDys), and primary pulmonary hypoplasia (PH), are rare diseases associated with high neonatal mortality due to refractory respiratory failure. Although ACDMPV mostly results from single nucleotide variants (SNVs) or copy-number variants (CNVs) involving FOXF1, AcDys, CAD, and PH are often associated with abnormalities within TBX4 or FGF10. These genes interact in the SHH-FOXF1 and TBX4-FGF10 signaling network and are known regulators of lung development. Recent studies conducted in TBX4-, FGF10-, or FOXF1-deficient LLDD lungs revealed decreased expression of TMEM100 at the transcriptomic and immunohistochemical levels. Here, we present four new patients with genetically and histopathologically confirmed LLDD, including ACDMPV (n = 2), AcDys (n = 1), and PH (n = 1), in whom we detected a heterozygous variants involving FOXF1 (n = 2) or TBX4 (n = 2). Additional immunohistochemical (TMEM100) and qPCR analyses (TMEM100, TBX4, FOXF1) performed in lung tissues of these newborns revealed a significant reduction in TMEM100, TBX4, and FOXF1 expression. Our results confirm previous findings indicating the possible involvement of TMEM100 in FOXF1-TBX4-FGF10 molecular signaling that, when disrupted, may lead to LLDD.

Mosquitoes threaten over half of the world's population through vectored diseases such as malaria, zika, yellow fever, dengue, and chikungunya. Mosquitoes have a highly developed olfactory system attuned to chemotaxis relating to host-seeking, mating, and oviposition behavior. In this study, we aimed to determine the spatial efficacy of 2 plant-based repellent blends (Blend3 and Blend4 that had previously been found to successfully repel Aedes, Anopheles and Culex mosquitoes in wind tunnel assays) in excluding Aedes aegypti from the window entry. A new cage system was developed for parallel "no-choice" and "choice" olfactometric assays. In the no-choice trial, Blends 3 and 4, as well as commercial products (N, N-diethyl-3-methylbenzamide, p-menthane-3,8-diol [PMD], 3-(N-n-butyl-N-acetyl)-amino-propionic acid ethyl ester, and 2-(2-hydroxyethyl)-1-methylpropylstyrene 1-piperidine carboxylate), were adsorbed into filter papers of different sizes and placed in a window created between 2 attached bug dorms. Then, the number of mosquitoes entering the window was counted through a 6-min period. In choice olfactometric assays, Blends 3, 4, and PMD were adsorbed into filter paper and the number of mosquitoes moving away from Blend 3 and PMD were compared. No-choice assays showed that Blend3 (P < 0.001) and Blend4 (P = 0.0012) were more repellent than the best commercial product PMD. Additionally, while Blend 4 was significantly more repellent than Blend 3 (P = 0.012) in the choice assay, overall, these 2 blends show promise as new repellents for the spatial exclusion of Aedes aegypti from window entry alone or as part of a "push-pull'' strategy.

Topical repellents provide protection against mosquito bites and their efficacy is often assessed using the arm-in-cage test. The arm-in-cage test estimates the repellent's protection time by exposing a repellent-treated forearm to host-seeking mosquitoes inside a cage at regular intervals until the first confirmed mosquito bite. However, the test does not reveal the repellents' behavioural mode of action. To understand how mosquitoes interact with topical repellents in the arm-in-cage test, we used a 3D infrared video camera system to track individual Aedes aegypti and Anopheles stephensi females during exposure to either a repellent-treated or an untreated forearm. The repellents tested were 20% (m/m) ethanolic solutions of N, N-diethyl-meta-toluamide, p-menthane-3,8-diol, icaridin and ethyl butylacetylaminopropionate. All four repellents substantially reduced the number of bites compared to an untreated forearm, while the flight trajectories indicate that the repellents do not prevent skin contact as the mosquitoes made multiple brief contacts with the treated forearm. We conclude that, in the context of the arm-in-cage test, topical repellents activate mosquitoes to disengage from the forearm with undirected displacements upon contact rather than being repelled at distance by volatile odorants.

Background/Objectives: In recent years, manufacturers have developed new low-profile stent grafts to allow endovascular treatment of abdominal aortic aneurysms (AAA) in patients with small access vessels. We evaluated the early and mid-term outcomes of the Incraft (Cordis Corp, Bridgewater, NJ, USA) ultra-low profile endograft implantation in a high-volume single center. Methods: Between 2014 and 2023, 133 consecutive endovascular aneurysm repair (EVAR) procedures performed using the Incraft endograft were recorded in a prospective database. Indications included infrarenal aortic aneurysms, common iliac aneurysms, and infrarenal penetrating aortic ulcers. Mid-term results were analyzed using the Kaplan-Meier method. Results: During the study period, 133 patients were treated with the Cordis Incraft endograft, in both elective and urgent settings. The Incraft graft was the first choice for patients with hostile iliac accesses, a feature characterizing at least one side in 90.2% of the patients in the study cohort. The immediate technical success rate was 78.2%. The intraoperative endoleak rate was 51.9% (20.3% type 1 A, 0.8% type 1 B, and 30.8% type 2 endoleak). Within 30 days, technical and clinical success rates were both 99.3%; all type 1A and 1B endoleaks were resolved at the 30-day follow-up CT-angiogram. After a mean follow-up of 35.4 months, the actuarial freedom from the re-intervention rate was 96.0%, 91.1%, and 84.0% at 1, 3, and 5 years, respectively. The iliac leg patency rate was 97.1%, 94.1%, and 93.1% at 1, 3, and 5 years, respectively. No statistically significant differences were observed between hostile and non-hostile access groups, nor between the groups with grade 1, grade 2, and grade 3 access hostility. Conclusions: The ultra-low profile Cordis Incraft endograft represents a valid option for the endovascular treatment of AAA in patients with hostile iliac accesses. The procedure can be performed with high rates of technical and clinical success at 30 days and the rates of iliac branch occlusion observed during the follow-up period appear acceptable in patients with poor aorto-iliac outflow.

Linear monoterpenes, versatile reaction biosubstrates, are bound and subsequently converted to various cyclic monomers and oligomers with excellent selectivity and efficiency, only in natural enzymes. We herein report bioinspired functions of synthetic polyaromatic cavities toward linear monoterpenes in the solution and solid states. The cavities are provided by polyaromatic coordination capsules, formed by the assembly of Pt(II) ions and bent bispyridine ligands with two anthracene panels. By using the capsule cavities, the selective binding of citronellal from mixtures with other monoterpenes and its preferential vapor binding over its derivatives are demonstrated in water and in the solid state, respectively. The capsule furthermore extracts p-menthane-3,8-diol, with high product- and stereoselectivity, from a reaction mixture obtained by the acid-catalyzed cyclization of citronellal in water. Thanks to the inner and outer polyaromatic cavities, the catalytic cyclization-dimerization of vaporized citronellal efficiently proceeds in the acid-loaded capsule solid and product/stereoselectively affords p-menthane-3,8-diol citronellal acetal (∼330% yield based on the capsule) under ambient conditions. The solid capsule reactor can be reused at least 5 times with enhanced conversion. The present study opens up a new approach toward mimicking terpene biosynthesis via synthetic polyaromatic cavities.