bioRxiv Science⌕ Search

Biology subjects

Yan, G.

Publications and source records attributed to Yan, G..

At least 19 recordsLinked to original sources

Local trait responses and aquatic microclimates increase projected habitat suitability for field-collected Anopheles stephensi in southern Ethiopia

BackgroundTrait-based models of mosquito environmental suitability commonly use global thermal performance curves compiled from laboratory studies. These curves may misrepresent suitability when local populations or larval habitats differ from globally synthesised expectations. This issue is particularly relevant for Anopheles stephensi, an urban malaria vector expanding in the Horn of Africa, where artificial aquatic habitats may generate microclimates poorly represented by global trait datasets or gridded air-temperature products. Methodology/Principal FindingsWe compared juvenile survival, development, and projected maximal population growth rate, rm, using locally measured data from field-collected Ethiopian An. stephensi and a global comparator combining pub-lished juvenile trait data. We fitted thermal performance curves for juvenile survival and development, used these curves to estimate temperature-dependent rm as a metric of habitat suitability, and projected rm across two matched temperature inputs: logger-measured larval habitat water temperature and ERA5-Land 2 m air temperature. Local juvenile trait responses produced higher projected rm than the global comparator across monitored habitats. Measured aquatic temperatures were warmer than ERA5-Land air temperatures, with a mean daily water-air offset of 3.26{whitebullet}C, and were more spatially heterogeneous among habitats. ERA5-Land resolved only two unique temperature series across the six monitored habitats. Sensitivity analyses showed that the Local-Global difference was robust for logger-measured water temperatures, whereas the ERA5-based difference was partly amplified by cooler temperatures below the local experimental range. Conclusions/SignificanceThis study provides the first locally derived juvenile thermal performance curves and temperature-dependent population growth estimates for field-collected An. stephensi in Africa, demonstrating that locally measured juvenile traits consistently predict higher habitat suitability than globally synthesised trait data. These find-ings highlight the importance of incorporating local mosquito trait data and aquatic microclimate measure-ments into predictive models to improve assessments of An. stephensi establishment, spread, and malaria risk in newly invaded urban environments. Author SummaryAnopheles stephensi is an invasive malaria mosquito that is rapidly spreading in urban areas of Africa. Pre-dicting where this species can establish is difficult because many models use mosquito temperature-response data from long-maintained laboratory colonies rather than from recently collected African populations. We measured juvenile survival and development in An. stephensi collected from Ethiopia, and compared pro-jections based on these local data with projections based on published laboratory-colony data. We also compared field-measured larval habitat water temperature with ERA5-Land air temperature. We found that local juvenile trait responses produced higher projected population growth than the global comparator across monitored habitats. Field-measured water temperatures were warmer and more variable among habitats than gridded air temperatures, which reduced habitat-level differences. These findings show that local mosquito biology and larval habitat microclimate measurements can change fine-scale estimates of suitability. This matters for surveillance and control because urban water containers may create suitable conditions that are poorly represented by broad climate datasets.

ecology↗

A Divergent Class of Arylamine N-Acetyltransferases Catalyzes Convergent Amidative Condensation of Polyketides in Manumycins Biosynthesis

Polyketides are prized for their structural complexity and therapeutic potential, yet the incorporation of amide bonds into their frameworks typically relies on linear, nonribosomal peptide synthetase (NRPS)-dependent assembly. The direct, convergent coupling of distinct polyketide chains via amide bond formation--an ideal strategy for combinatorial biosynthesis--has remained largely elusive. Here, we report the discovery of a novel family of arylamine N-acetyltransferases (NATs) from manumycin-type biosynthetic pathways that catalyze an unprecedented intermolecular amidative chain transfer/condensation between an acyl carrier protein (ACP)-tethered polyketide donor and a free polyketide acceptor. Biochemical, structural, and molecular dynamics studies reveal that the representative enzyme, ColC2, possesses a distinctive substrate-binding pocket that diverges from canonical arylamine NATs, conferring exceptional promiscuity toward diverse acyl donors and acceptors. We demonstrate the utility of this biocatalyst by coupling arylamines with either synthetic acyl-thioesters or polyketide synthase (PKS) machinery to generate a library of non-natural polyketide amides and manumycin derivatives. These findings establish a new paradigm for amide bond formation in polyketide biosynthesis and position arylamine NATs as powerful tools for the development of novel therapeutics through combinatorial synthesis.

synthetic biology↗

Anterior cingulate cortex projections to the amygdala in primates: topographic and layer-specific organization underlying emotion and mood regulation

Emotion and mood regulation critically depends on interactions between the anterior cingulate cortex (ACC) and the amygdala. However, the detailed architecture of ACC projections to their major targets, the basal (BA) and accessory (AcBA) basal nuclei of the amygdala, remains unclear. To address this issue, a combined retrograde and anterograde tracing with viral vectors were performed in macaques to map the projection patterns from pregenual (pgACC), subgenual (sgACC), and dorsal (dACC) subareas. Data revealed that ACC neurons projecting to the BA arose predominantly from the superficial layers (II/III) of all subareas and the deep layers (V/VI) of the sgACC, whereas ACC neurons projecting to the AcBA originated mainly in the deep layers of the sgACC and dACC. The present study defines the topographic and layer-specific organization of ACC-amygdala connectivity in primates and subserves to provide an anatomical basis for future causal and translational approaches, such as targeted interventions against ACC-related mood disorders. TeaserPrimate anterior cingulate cortex has topographic and layer-specific projections to amygdala that are involved in emotion and mood regulation.

neuroscience↗

Plasmodium vivax genome evolution for erythrocyte invasion in Duffy-negative infections from East Africa

Historically, Plasmodium vivax (Pv) malaria was rare in Africa due to the lack of Duffy antigen receptor for chemokines (DARC) expression on host erythrocytes, an essential receptor for Pv Duffy Binding Protein (PvDBP1)-mediated invasion. However, increased reports of Pv cases across Africa and among Duffy-negative individuals have led to the hypothesis that the parasites have evolved alternate invasion mechanisms that are DARC-independent. To investigate potential genetic adaptations underlying this phenomenon, we performed genome-wide single nucleotide polymorphisms (SNPs) analyses of 110 Pv isolates collected from Ethiopia, comprising 38 Duffy-negative and 72 Duffy-positive infections. Pv from Duffy positives exhibited markedly higher genetic variation (477,561 SNPs) than those from Duffy negatives (197,461 SNPs). Chromosomes 1, 2, 9, 10, and 12 harbored the highest SNPs densities, consistent across both host genotypes but elevated in Duffy-positive infections. Among 43 erythrocyte-binding gene candidates, tryptophan rich antigen 3 and 34 (TRAg3 and TRAg34) and members of the merozoite surface protein 3 (MSP3) family showed the greatest nucleotide diversity per kilobase, highlighting these loci as potential mediators of host-parasite interaction shifts. Signals of positive selection differed by host Duffy genotypes. In Duffy-positive Pv, adaptive signatures were observed in genes related to drug resistance (chloroquine resistance-associated protein CG1 and 26S proteasome regulatory subunit RPN2) and erythrocyte binding (MAEBL); whereas in Duffy negative Pv, positive selection was observed in genes linked to organellar maintenance and vesicle trafficking (plastid replication repair enzyme and the AP-5 complex subunit beta 1), implicating alternative metabolic or trafficking adaptations. Amino acid substitutions in invasion ligands (PvDBP1, PvEBP/DBP2, and PvRBP2b) were common in Duffy-positive Pv but largely absent in Duffy-negative ones, with almost half of the mutations located in critical binding domains. Overall, Pv isolates in Duffy-negative hosts displayed reduced genomic diversity yet retained high conservation across the genome, suggesting strong selective constraints and limited diversification. Phylogenetic comparison revealed that Ethiopian Pv clustered closely with other East African isolates, whereas Southeast Asian and South American Pv represented more distant lineages. These findings indicate that Pv circulating in Duffy-negative populations maintains a genetically conserved background, potentially reflecting stringent evolutionary bottlenecks and/or specialized host-parasite interactions required for invasion of Duffy-negative erythrocytes.

genomics↗

Early-evening biting by Anopheles stephensi in Southern Ethiopia: A Challenge for Bed Net Use as Vector Control in Africa

Anopheles stephensi is rapidly expanding across Africa, posing new challenges for malaria control. Its biting time patterns, however, remain poorly characterized, raising uncertainty about the effectiveness of bed nets against this invasive vector. To address this gap, we investigated diel biting activity, feeding propensity, and flight behavior using complementary behavioral assays on females reared from wild-caught larvae in Hawassa City, southern Ethiopia. Flight monitoring assays revealed that An. stephensi exhibited pronounced activity at dusk, beginning between 18:00 and 19:00 with the onset of scotophase, and little to no activity during the photophase. Blood-feeding propensity, defined as the proportion of mosquitoes taking a blood meal when offered, peaked during the early scotophase (18:00-22:00) at 33.3-51.7%, but was markedly reduced during daylight hours (0-16.7%). Human landing catches in large-cage enclosures confirmed this early evening activity: 83.5% of total landings occurred between 18:00 and 22:00 with a sharp peak at 18:00-19:00, corresponding to a mean biting rate of 17.8 bites per person per hour. These findings demonstrate that invasive An. stephensi primarily seek hosts and bite during the early evening, a time when people are often unprotected by bed nets. This behavior reduces the protective impact of conventional net-based interventions and underscores the need for African National Malaria Control Programs to deploy complementary measures such as spatial repellents and larval source management to mitigate early-evening transmission. Moreover, this study highlights the utility of integrated behavioral assays for estimating biting time, offering approaches that can be extended to other vector species across Africa.

zoology↗

Ultra-wide-field, deep, adaptive two-photon microscopy

Observing the activity patterns of large neural populations throughout the brain is essential for understanding brain function. However, capturing neural interactions across widely distributed brain regions from both superficial and deep cortical layers remains challenging with existing microscopy technologies. Here, we introduce a state-of-the-art two-photon microscopy system, ULTRA, capable of single-cell resolution imaging across an ultra-large field of view (FOV) exceeding 50 mm{superscript 2}, enabling deep and very wide field in vivo imaging. To demonstrate its capabilities, we conducted a series of experiments under multiple imaging conditions, successfully visualizing brain structures and neuronal activities spanning a spatial range of over 7 mm from superficial layers to depths of up to 900 m, while covering a volume of 45.24 mm3 in the mouse brain. This versatile imaging platform overcomes traditional spatial constraints, providing a powerful tool for comprehensive exploration of neuronal circuitry over extensive spatial scales with cellular resolution.

neuroscience↗

Rapid adaptive optics enabling near noninvasive high-resolution brain imaging in awake behaving mice

High-resolution imaging under physiological conditions is essential for studying biological mechanisms and disease processes. However, achieving this goal remains challenging due to optical aberrations and scattering from heterogeneous tissue structures, compounded by motion artifacts from awake animals. In this study, we developed a rapid and accurate adaptive optics system called multiplexing digital focus sensing and shaping (MD-FSS) for deep-tissue multiphoton microscopy. Under two-photon excitation, MD-FSS precisely measures the aberrated point spread function in approximately 0.1 s per measurement, effectively compensating for both aberrations and scattering to achieve subcellular resolution in deep tissue. Using MD-FSS integrated with two-photon microscopy, we achieved high-resolution brain imaging through thinned or optically cleared skull windows, two near noninvasive methods to access mouse brain, reaching depths up to 600 m below the pia in awake behaving mice. Our findings revealed significant differences in microglial functional states and microvascular circulation dynamics between awake and anesthetized conditions, highlighting the importance of studying brain function in awake mice through noninvasive methods. We captured functional imaging of fine neuronal structures at subcellular level in both somatosensory and visual cortices. Additionally, we demonstrated high-resolution imaging of microvascular structures and neurovascular coupling across multiple cortical regions and depths in the awake brain. Our work shows that MD-FSS robustly corrects tissue-induced aberrations and scattering through rapid PSF measurements, enabling near-noninvasive, high-resolution imaging in awake, behaving mice.

bioengineering↗

CellScope: High-Performance Cell Atlas Workflow with Tree-Structured Representation

Single-cell sequencing enables comprehensive profiling of individual cells, revealing cellular heterogeneity and function with unprecedented resolution. However, current analysis frameworks lack the ability to simultaneously explore and visualize cellular hierarchies at multiple biological levels. To address these limitations, we present CellScope, an innovative framework for constructing high-resolution cell atlases at multiple clustering levels. CellScope employs a two-step manifold fitting process for gene selection and noise reduction, followed by agglomerative clustering, and uniquely integrates UMAP visualization with hierarchical clustering to intuitively represent cellular relationships simultaneously at multiple levels--such as cell lineage, cell type, and cell subtype levels. Compared to established pipelines such as Seurat and Scanpy, CellScope comprehensively improves clustering performance, visualization clarity, computational efficiency, and algorithm interpretability, while reducing dependence on hyperparameters across a multitude of single-cell datasets. Most importantly, it can reveal new biological insights that other contemporary methods are unable to detect, thereby deepening our understanding of cellular heterogeneity and function, and potentially informing disease research.

bioinformatics↗

Mechanistic Study of palmatine in Regulating Pyroptosis in Sepsis Cells via Signaling Pathways

This study aims to investigate whether palmatine has a potential effect on sepsis-related pyroptosis in patients through relevant signaling pathways. Initially, the study explores the differential genes associated with pyroptosis in sepsis patients, along with their related enrichment processes, biological behaviors, and functional pathways. Patients with sepsis pyroptosis are then classified into subgroups based on pyroptosis-related genes to analyze the biological differences between these subtypes. Furthermore, the signaling pathways and biological functions associated with palmatine are studied to assess whether there are functional similarities or overlaps with the signaling pathways and biological behaviors observed in patients with sepsis pyroptosis. Finally, the findings are validated through wet-lab experiments. The results reveal that the signaling pathways related to palmatine partially overlap with those observed in sepsis pyroptosis patients, indicating that palmatine may influence sepsis pyroptosis through these pathways. Parmatine may improve the prognosis of sepsis by affecting the immune function of PRKACA, PTGS2, NLRP3, HSP90AA1 and PTPN22.

bioinformatics↗

Emergence of cooperation promoted by higher-order strategy updates

Cooperation is fundamental to human societies, and the interaction structure among individuals profoundly shapes its emergence and evolution. In real-world scenarios, cooperation prevails in multi-group (higher-order) populations, beyond just dyadic behaviors. Despite recent studies on group dilemmas in higher-order networks, the exploration of cooperation driven by higher-order strategy updates remains limited due to the intricacy and indivisibility of group-wise interactions. Here we investigate four categories of higher-order mechanisms for strategy updates in public goods games and establish their mathematical conditions for the emergence of cooperation. Such conditions uncover the impact of both higher-order strategy updates and network properties on evolutionary outcomes, notably highlighting the enhancement of cooperation by overlaps between groups. Interestingly, we discover that the group-mutual comparison update - selecting a high-fitness group and then imitating a random individual within this group - can prominently promote cooperation. Our analyses further unveil that, compared to pairwise interactions, higher-order strategy updates generally improve cooperation in most higher-order networks. These findings underscore the pivotal role of higher-order strategy updates in fostering collective cooperation in complex social systems. Author summaryHuman societies often organize and cooperate within social groups, where relatives, friends, neighbors, and colleagues influence behavior at both group and individual levels. Individuals may exhibit biased or neutral attitudes when selecting a neighboring group and a peer within it for imitation or comparison, a process termed as higher-order strategy update. These selection preferences originate from four personality types: aggressive, open-minded, myopic, and passive. This work demonstrates that the open-minded type - indiscriminately imitating a peer within a well-functioning group - significantly promotes cooperation. The mathematical framework proposed in this study deepens the understanding of how decision-making within higher-order structures affects the emergence and spread of cooperative behaviors.

evolutionary biology↗

Dietary manipulation of intestinal microbes prolongs survival in a mouse model of Hirschsprung disease

Enterocolitis is a common and potentially deadly manifestation of Hirschsprung disease (HSCR) but disease mechanisms remain poorly defined. Unexpectedly, we discovered that diet can dramatically affect the lifespan of a HSCR mouse model (Piebald lethal, sl/sl) where affected animals die from HAEC complications. In the sl/sl model, diet alters gut microbes and metabolites, leading to changes in colon epithelial gene expression and epithelial oxygen levels known to influence colitis severity. Our findings demonstrate unrecognized similarity between HAEC and other types of colitis and suggest dietary manipulation could be a valuable therapeutic strategy for people with HSCR. AbstractHirschsprung disease (HSCR) is a birth defect where enteric nervous system (ENS) is absent from distal bowel. Bowel lacking ENS fails to relax, causing partial obstruction. Affected children often have "Hirschsprung disease associated enterocolitis" (HAEC), which predisposes to sepsis. We discovered survival of Piebald lethal (sl/sl) mice, a well-established HSCR model with HAEC, is markedly altered by two distinct standard chow diets. A "Protective" diet increased fecal butyrate/isobutyrate and enhanced production of gut epithelial antimicrobial peptides in proximal colon. In contrast, "Detrimental" diet-fed sl/sl had abnormal appearing distal colon epithelium mitochondria, reduced epithelial mRNA involved in oxidative phosphorylation, and elevated epithelial oxygen that fostered growth of inflammation-associated Enterobacteriaceae. Accordingly, selective depletion of Enterobacteriaceae with sodium tungstate prolonged sl/sl survival. Our results provide the first strong evidence that diet modifies survival in a HSCR mouse model, without altering length of distal colon lacking ENS. HighlightsO_LITwo different standard mouse diets alter survival in the Piebald lethal (sl/sl) mouse model of Hirschsprung disease, without impacting extent of distal colon aganglionosis (the region lacking ENS). C_LIO_LIPiebald lethal mice fed the "Detrimental" diet had many changes in colon epithelial transcriptome including decreased mRNA for antimicrobial peptides and genes involved in oxidative phosphorylation. Detrimental diet fed sl/sl also had aberrant-appearing mitochondria in distal colon epithelium, with elevated epithelial oxygen that drives lethal Enterobacteriaceae overgrowth via aerobic respiration. C_LIO_LIElimination of Enterobacteriaceae with antibiotics or sodium tungstate improves survival of Piebald lethal fed the "Detrimental diet". C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=94 SRC="FIGDIR/small/637436v1_ufig1.gif" ALT="Figure 1"> View larger version (15K): org.highwire.dtl.DTLVardef@d95251org.highwire.dtl.DTLVardef@1ab58caorg.highwire.dtl.DTLVardef@5260b0org.highwire.dtl.DTLVardef@49ce42_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

Conformation-gated binding drives negative cooperativity in ATP:cob(I)alamin Adenosyltransferase for optimized cobalamin handling

Vitamin B12 (cobalamin) is a high-value yet scarce cofactor required for various metabolic processes, making its efficient handling important for maintaining metabolic homeostasis. While the involvement of ATP:cob(I)alamin adenosyltransferases (MMAB) in the synthesis, delivery, and repair of 5-deoxyadenosylcobalamin (AdoCbl) is well established, the kinetic mechanisms that regulate this process, particularly its negative cooperativity, remain poorly understood. Understanding these mechanisms is key to clarifying how MMAB efficiently uses AdoCbl, prevents resource wastage, and supports bacterial survival in nutrient-limited environments. Using single-molecule relative fluorescence (SRF) spectroscopy, we found that conformation-gated binding is the driving force behind MMABs preference for AdoCbl over hydroxocobalamin and is the underlying mechanism for negative cooperativity. This mechanism significantly slows down the binding of the second equivalent of AdoCbl, favoring the singly bound state. Our findings indicate that MMAB predominantly binds a single AdoCbl, optimizing the AdoCbl loading to methylmalonyl-CoA mutase. Additionally, our SRF approach also serves as a tool to explore other cofactor interactions, such as those between riboswitches and cobalamin derivatives, to provide insights into regulatory mechanisms of cobalamin sensing and gene regulation, which are crucial for bacterial adaptation to changing nutrient conditions. Significance StatementMMAB is important for B12-dependent propionate metabolism in bacteria. Our findings reveal that conformation-driven binding mechanism underlines the negative cooperativity of MMAB, as it favors the binding of the first AdoCbl while limiting further binding. The larger kon for the first site, combined with similar unbinding rates for both sites, could provide a solution for optimizing cobalamin handling and minimize unnecessary waste. Our single-molecule fluorescence approach offers a powerful tool for investigating other dynamic cofactor interactions, providing new insights into regulatory mechanisms in bacterial metabolism.

biophysics↗

Identification and characterization of ugpE required for the full virulence of Streptococcus suis

Streptococcus suis (S. suis) is an emerging zoonotic pathogen that threatens both animal and human health worldwide. UgpE is a protein subunit of the Ugp (uptake of glycerol phosphate) transporter system that is involved in glycerophospholipid synthesis in bacterial membranes. In this study, an ugpE deletion mutant was constructed and the effects of ugpE deletion on cell morphology, biofilm formation, and virulence were investigated. Deletion of ugpE did not affect bacterial growth but impaired cell chain formation and capsular synthesis by downregulating the mRNA levels of the capsular regulon genes cps-2B, cps-2C, and cps-2S. Deletion of ugpE also led to decreased tolerance to heat, oxidative, and acid-base stress. Crystal violet staining and scanning electron microscopy demonstrated ugpE negatively regulated biofilm formation in liquid culture and the rdar biofilm morphotype on agar plates. Moreover, ugpE deletion not only reduced hemolysin activity, survival in whole human blood, and anti-phagocytosis ability against porcine alveolar macrophages (PAM) but also enhanced bacterial adhesion and invasion of human cerebral microvascular endothelial cells (hCMEC/D3) by upregulating the expression of multiple genes associated with cell adhesion. In a mouse infection model, ugpE deletion significantly attenuated virulence and lowered the number of viable bacteria in the blood and major organs, as well as distribution of macrophages. In conclusion, this study identified UgpE as a novel virulence factor that plays a pivotal role in the regulation of virulence and biofilm formation of S. suis.

microbiology↗

Integration of binocular vision and motor state to promote prey pursuit

Most animals with two eyes combine the inputs to achieve binocular vision, which can serve numerous functions, and is particularly useful in hunting prey. However, the mechanisms by which visual information from the two eyes are combined remain largely unknown. Here, we designed a device to reversibly occlude the eyes of a head-fixed zebrafish larva, and used large-scale volumetric two-photon imaging to identify binocular neurons that respond to prey stimuli. We found these binocular prey-responsive neurons (bino-PRNs) were primarily located in three areas, the pretectum, thalamus, and nucleus isthmi. We then characterized the bino-PRNs functional properties, and found that their left and right eye receptive fields are offset to varying degrees, which would correspond to objects at naturalistic hunting distances for a larva with converged eyes. We also found that bino-PRNs have a significantly greater response in hunting trials, which could be the result of an eye convergence-related corollary discharge. We then optogenetically induced prey capture eye and tail movements, and found that this hunting command activates prey responsive neurons in the pretectum, thalamus, and nucleus isthmi. These findings indicate that bino-PRNs receive visual and motor input that would allow them to encode prey position in three dimensions.

neuroscience↗

Interpretable scRNA-seq Analysis with Intelligent Gene Selection

AO_SCPLOWBSTRACTC_SCPLOWSingle-cell RNA sequencing (scRNA-seq) data analysis faces multiple challenges, including high dimensionality, significant noise, and data loss. To effectively address these issues, we introduce AIGS, a robust and transparent single-cell analysis framework. AIGS utilizes an intelligent gene selection method that systematically identifies the most informative genes for clustering based on the normalized mutual information between pre-learned pseudo-labels and quantified genes. Additionally, AIGS incorporates a scale-invariant distance metric to assess cell-to-cell similarity, enhancing connections between homogenous cells and ensuring more accurate and robust results. Through comprehensive comparisons with state-of-the-art techniques, AIGS demonstrates superior performance in both clustering accuracy and multi-resolution visualization quality. Our in-depth analysis of clustering and visualization results further reveals that AIGS can uncover complex, stage-specific gene expression patterns during the same developmental cell stage.

bioinformatics↗

Opto-Myomatrix: μLED integrated microelectrode arrays for optogenetic activation and electrical recording in muscle tissue

ObjectiveOptogenetics is a valuable and widely-used technique that allows precise perturbations of selected groups of cells with high temporal and spatial resolution by using optical systems and genetic engineering technologies. This study aims to develop Opto-Myomatrix, a novel optogenetic tool for precise muscle fiber control and high-resolution electrophysiological recording. MethodBased on a flexible and biocompatible polymer substrate, the device incorporates an integrated {micro}LED that delivers light at 465 nm for optogenetic stimulation and 32 PEDOT:PSS-coated electrodes for electromyography (EMG) recording. We also added a reflector to improve optical power output. ResultsThe fabricated Opto-Myomatrix device achieves an optical output intensity as high as 129.46 mW/mm2 in the direction of interest, which is enhanced by nearly 100%. The PEDOT:PSS-coated electrodes exhibit 85% lower impedance than uncoated contacts, enabling high signal-to-noise EMG acquisition. We investigated heat dissipation characteristics of the {micro}LED through measurements and finite element simulations, confirming that temperature changes remain within safe limits. The Opto-Myomatrix device was implanted in transgenetic mice and successfully stimulated targeted jaw muscles, inducing movement while simultaneously capturing EMG signals. ConclusionThe Opto-Myomatrix effectively combines precise optical stimulation and high-quality EMG recording in a flexible and biocompatible device, focusing on optogenetic applications in muscle. SignificanceWhile optogenetic tools are well-established for brain and central nervous system (CNS) research, the development of Opto-Myomatrix addresses a critical gap by enabling precise muscle fiber control and high-resolution recording for advancing neuromuscular studies.

bioengineering↗

The dual resistance mechanism of CYP325G4 and CYP6AA9 in Culex pipiens pallens legs according to transcriptome and proteome analysis

Mosquitoes within the Culex pipiens complex play a crucial role in human disease transmission. Insecticides, especially pyrethroids, are used to control these vectors. Mosquito legs are the main entry point and barrier for insecticides to gain their neuronal targets. However, the resistance mechanism in mosquito legs is unclear. Herein, we employed transcriptomic analyses and isobaric tags for relative and absolute quantitation techniques to investigate the resistance mechanism, focusing on Cx. pipiens legs. We discovered 2346 differentially expressed genes (DEGs) between deltamethrin-resistant (DR) and deltamethrin-sensitive (DS) mosquito legs, including 41 cytochrome P450 genes. In the same comparison, we identified 228 differentially expressed proteins (DEPs), including six cytochrome P450 proteins). Combined transcriptome and proteome analysis revealed only two upregulated P450 genes, CYP325G4 and CYP6AA9. The main clusters of DEGs and DEPs were associated with metabolic processes, such as cytochrome P450-mediated metabolism of drugs and xenobiotics. Transcription analysis revealed high CYP325G4 and CYP6AA9 expression in the DR strain at 72 hours post-eclosion compared with that in the DS strain, particularly in the legs. Mosquitoes knocked down for CYP325G4 were more sensitive to deltamethrin than the controls. CYP325G4 knockdown reduced the expression of several chlorinated hydrocarbon (CHC)-related genes, which altered the cuticle thickness and structure. Conversely, CYP6AA9 knockdown increased CHC gene expression without altering cuticle thickness and structure. P450 activity analysis demonstrated that CYP325G4 and CYP6AA9 contributed to metabolic resistance in the midgut and legs. This study identified CYP325G4 as a novel mosquito deltamethrin resistance factor, being involved in both metabolic and cuticular resistance mechanisms. The previously identified CYP6AA9 was investigated for its involvement in metabolic resistance and potential cuticular resistance in mosquito legs. These findings enhance our comprehension of resistance mechanisms, identifying P450s as promising targets for the future management of mosquito vector resistance, and laying a theoretical groundwork for mosquito resistance management. Author SummaryCulex pipiens mosquitoes are the primary vector of the filamentous nematode, Wuchereria bancrofti and also involved in the transmission of other pathogens, such as West Nile virus (WNV), avian malarias, and avian pox virus. Insecticides, particularly pyrethroids, continue to be the primary method to control these significant vectors. Worryingly, resistance to insecticides has become widespread and is rapidly intensifying in Culex mosquitoes throughout China, posing a threat to the efficacy of insecticides. Legs are the main sites of contact with ITNs and sprayed insecticides, and the insecticides have to penetrate the leg cuticle to reach their targets.Therefore, the resistance mechanisms in mosquito legs deserve further investigation. Several reports have found a certain amount of P450 in insect legs. Unfortunately, none of the above reports have conducted further functional studies on P450s in the legs. Here, we have identified two P450 enzymes, CYP325G4 and CYP6AA9, through the integrated analysis of transcriptomics and proteomics. CYP325G4 enriched in the cuticle of resistant mosquitoes might possess a dual resistance mechanism involving metabolic resistance and cuticle resistance. CYP6AA9 was slightly different, possibly exerting metabolic resistance as its main function and also being involved in cuticle synthesis. Understanding the dual resistance mechanism of P450s in the metabolism of pyrethroid insecticides will have an important role in optimizing vector control strategies.

physiology↗

Insecticide resistance and its intensity in urban Anopheles arabiensis in Kisumu City, Western Kenya: Implications for malaria control in urban areas

BackgroundThe rise of insecticide resistance poses a growing challenge to the effectiveness of vector control tools, particularly in rural areas. However, the urban setting has received comparatively less focus despite its significance in attracting rural to urban migration. Unplanned urbanization, often overlooked, exacerbates insecticide resistance as Anopheles mosquitoes adapt to the polluted environments of rapidly expanding cities. This study aimed to assess the insecticide susceptibility status of malaria vectors and identify potential underlying mechanisms across three distinct ecological settings characterized by differing levels of urbanization in Kisumu County, Kenya. MethodsField-derived An. gambiae (s.l.) larvae collected from a long stretch of urban-to-rural continuum were phenotyped as either resistant or susceptible to six different insecticides using the World Health Organization (WHO) susceptibility test. Polymerase chain reaction (PCR) techniques were used to identify the species of the An. gambiae complex and screened for mutations at voltage-gated sodium channels (Vgsc-1014F, Vgsc-1014S, Vgsc-1575Y) and acetylcholinesterase Ace1-119S. Metabolic enzymes activities (non-specific {beta}-esterases and monooxygenases) were evaluated in mosquitoes not exposed to insecticides using microplate assays. Additionally, during larval sampling, a retrospective questionnaire survey was conducted to determine pesticide usage by the local inhabitants. ResultsAnopheles arabiensis dominated in urban (96.2%) and peri-urban (96.8%) areas, while An. gambiae (s.s.) was abundant in rural settings (82.7%). Urban mosquito populations showed high resistance intensity to deltamethrin (Mortality rate: 85.2% at 10x) and suspected resistance to Pirimiphos-methyl and bendiocarb while peri-urban and rural populations exhibited moderate resistance intensity to deltamethrin (mortality rate >98% at 10x). Preexposure of mosquitoes to a synergist piperonyl butoxide (PBO) significantly increased mortality rates: from 40.7% to 88.5% in urban, 51.9% to 90.3% in peri-urban, and 55.4% to 87.6% in rural populations for deltamethrin, and from 41.4% to 78.8% in urban, 43.7% to 90.7% in peri-urban, and 35% to 84.2% in rural populations for permethrin. In contrast, 100% mortality to chlorfenapyr and clothianidin was observed in all the populations tested. The prevalence of L1014F mutation was notably higher in urban An. arabiensis (0.22) unlike the peri-urban (0.11) and rural (0.14) populations while the L1014S mutation was more prevalent in rural An. gambiae (0.93). Additionally, urban An. arabiensis exhibited elevated levels of mixed function oxidases (0.8/mg protein) and non-specific esterases (2.12/mg protein) compared to peri-urban (0.57/mg protein and 1.5/mg protein, respectively) and rural populations (0.6/mg protein and 1.8/mg protein, respectively). Pyrethroids, apart from their use in public health through LLINs, were being highly used for agricultural purposes across all ecological settings (urban 38%, peri-urban 36% and rural 37%) followed by amidine group, with organophosphates, neonicotinoids and carbamates being of secondary importance. ConclusionThese findings show high resistance of An. arabiensis to insecticides commonly used for vector control, linked with increased levels of detoxification enzymes. The observed intensity of resistance underscores the pressing issue of insecticide resistance in urban areas, potentially compromising the effectiveness of vector control measures, especially pyrethroid-treated LLINs. Given the species unique behavior and ecology compared to An. gambiae, tailored vector control strategies are needed to address this concern in urban settings.

ecology↗