Biology subjects

Moore, S. J.

Publications and source records attributed to Moore, S. J..

At least 19 recordsLinked to original sources

bioRxiv2026

Protocol for Single-Cell Patch-Proteomics: identify specific proteins associated with electrophysiological properties from individual neurons

Single-cell patch-proteomics is a novel and powerful technique combining electrophysiological and quantitative proteomic characterization from individually sampled neurons. Following whole-cell patch clamp characterization of neuronal excitability, we collect intracellular contents and, using the latest advancements in Liquid Chromatography and Mass Spectrometry, identify and quantify peptides and their corresponding proteins. By cell-matched one-to-one associations between electrophysiological properties and protein abundance, this approach provides a means to elucidate molecular mechanisms of neuronal function on individual neuron basis.

neuroscience↗

bioRxiv2026

Quantifying the impact of experimental hut design on intervention evaluation outcomes and predicted reductions in vectorial capacity

Experimental hut trials (EHTs) are WHO-recommended for the entomological evaluation of insecticide-treated nets (ITNs), but several hut designs are in operational use, and structural differences between them may confound efficacy predictions and limit cross-site comparability. We developed a Bayesian hierarchical framework comprising a host-seeking model, which jointly estimates biting deterrence and preprandial mortality while accounting for night-to-night variation and overdispersion, and a postprandial mortality model, which expresses hut and net effects as hazard ratios through a complementary log-log link. We applied it to a comparative trial of four hut designs (East African, West African, Ifakara and Rapley) conducted at a single site in Tanzania, evaluating eight ITNs when new and after twenty washes. Posterior estimates parameterise a vectorial capacity framework to predict reductions in transmission potential. Hut design influenced baseline mosquito behaviour and all three modes of action. Relative to the Rapley reference, baseline feeding rates were substantially lower in the East African and West African huts and closer to Rapley in the Ifakara hut. Preprandial mortality was amplified in the Ifakara hut. Comparing to previous analysis provides evidence that combining mortality before and after feeding into a single endpoint does not reliably reflect impact, supporting the decomposition of entomological outcomes into separate modes of action. Expressing modes of action as mechanism-specific parameters allows the estimates to be carried directly into transmission models. For every net, the predicted reduction in vectorial capacity was greatest in the Ifakara hut and smallest in the West African and Rapley huts. The effect of hut design on predicted impact exceeded that of washing the nets twenty times. Results indicate that the hut design under which trial data were collected should be considered when forecasting population-level effect.

ecology↗

bioRxiv2026

Multi-site phenotypic characterisation of Kisumu colonies for comparison of insecticide susceptibility between testing centres

Testing of insecticides and insecticide-based vector control tools relies on the use of bioassays whereby insects are exposed and mortality, or another relevant entomological endpoint, is measured. To be able to compare results between products and across time and space standard laboratory strains are used. Insecticide-susceptible strains are often used as comparators in studies involving insecticide-resistant populations. Kisumu is a long-established, widely-used insecticide-susceptible strain of Anopheles gambiae. In this study, 16 facilities across different countries exposed their Kisumu colonies aged 3-5 days to four insecticides (permethrin, alpha-cypermethrin, DDT and pirimiphos-methyl) at a range of concentrations in WHO tube assays to assess susceptibility. Insecticide-treated filter papers were made at the Liverpool School of Tropical Medicine and sent to testing centres, along with a standard protocol and testing conditions requirements, to standardise testing as much as possible. A first observation was that DDT results were affected by the use of Risella oil, the WHO-recommended carrier oil but immiscible in acetone, leading to uneven coating of papers and increased variability in bioassay results (40x variance between colonies). Bioassay data generated by the testing facilities suggest a wide range of susceptibility to different insecticides between colonies of Kisumu: LC50 values between colonies differed, though not significantly, by up to 4x for permethrin, 6x for pirimiphos-methyl, and 15x for alpha-cypermethrin. Some colonies are classified as insecticide resistant based on the WHO definition of <98% mortality on exposure to a discriminating concentration. The assumption that Kisumu is a standard susceptible strain, consistent between facilities is challenged, which has implications when comparing results between studies. We propose genotypic analysis of the colonies to identify signs of resistance markers indicative of contamination from resistant strains or wild mosquitoes, and generation of guidance to support the maintenance and monitoring of susceptible mosquito strains.

zoology↗

bioRxiv2026

Cell-type-specific ATF6α programs regulate epithelial mitochondrial homeostasis and pericyte remodeling during physiological and exposure-accelerated lung aging

Proteostasis declines with lung aging, while the role of the Unfolded Protein Response (UPR) in lung aging and age-associated pulmonary diseases remains understudied. We investigated how deficiency in the UPR sensor ATF6 affects physiological and smoke exposure-accelerated lung aging. ATF6 -deficient mice exhibited accelerated alveolar simplification, a sign of lung parenchymal aging, which was exacerbated by smoking. Nevertheless, small airway vascular fibrotic remodeling, a prominent smoking-induced pathology, was not evident in smoke-exposed ATF6 -deficient mice. Mechanistically, these divergent phenotypes arose from cell-type-specific ATF6 programs. In alveolar epithelial type 2 cells (AEC2s), the facultative progenitors of the lung parenchyma, ATF6 maintained mitochondrial bioenergetics and sustained efficient redifferentiation into alveolar epithelial type 1 cells (AEC1s). In lung pericytes, ATF6 promoted extravasation, redifferentiation into myofibroblast-like cells, and production of collagens 1 and 3. These findings identify ATF6 as a cell-type-specific regulator of differentiation programs during lung aging and highlight the need to study ATF6 under defined physiological and pathological contexts before therapeutically targeting this pathway.

cell biology↗

bioRxiv2026

Multi-centre laboratory study to determine discriminating concentrations for broflanilide and isocycloseram resistance monitoring in mosquitoes

1In 2024-2025 a multi-centre study involving seven international laboratories was conducted with the support of the World Health Organization (WHO). The aim of the study was to establish and validate discriminating concentrations (DCs) in WHO bottle bioassays for monitoring susceptibility to broflanilide and isocycloseram in Anopheles gambiae s.s., An. funestus, An. stephensi and Aedes aegypti. The following values are recommended for adoption as DCs for broflanilide: 10 {micro}g/bottle for An. gambiae and Ae. aegypti, 15 {micro}g/bottle for An. funestus, and 25 {micro}g/bottle for An. stephensi. The recommended DCs for isocycloseram are 15 {micro}g/bottle for Ae. aegypti, 30 {micro}g/bottle for An. gambiae, 50 {micro}g/bottle for An. stephensi, and 60 {micro}g/bottle for An. funestus. Based on the experiences of conducting this study, which represents the application of a generic protocol for establishing discriminating concentrations produced by the WHO, technical recommendations are made on the generation and analysis of DC data for insecticides in future.

zoology↗

bioRxiv2026

Acarbose improves cognitive function in a mouse model of normal aging but not Alzheimer's disease

Structured AbstractO_ST_ABSINTRODUCTIONC_ST_ABSDeclines in function occur in both "normal" aging (in the absence of disease) and age-related pathological contexts, like Alzheimers disease (AD). Whether "anti-aging" interventions (that extend lifespan) also promote cognitive function in aging and AD remains unexplored. METHODSWe assessed the effect of acarbose (1000 ppm from 4 months of age) on spatial learning and memory using the Morris water maze in young adult (6 mo), mid-aged (12 mo), or aged (24 mo) cohorts of normal aging (Ntg-HET3) and AD-relevant (5xFAD-HET3) genetically heterogeneous mice. RESULTSIn mid-aged and aged Ntg-HET3 mice, acarbose treatment resulted in performance equivalent to young adults. Conversely, acarbose failed to ameliorate age-related deficits in 5xFAD-HET3 mice. DISCUSSIONThis work demonstrates that anti-aging interventions can also promote cognitive longevity in normal aging. Further, it reinforces that AD is not simply accelerated aging and requires therapies beyond anti-aging interventions that target its unique molecular and cellular drivers.

neuroscience↗

bioRxiv2026

A liquid handling platform for standardised quantification of cell-free enzymatic activity encoded by antimicrobial resistance genes

Antimicrobial resistance (AMR) is a growing global threat to human health, and rapid methods for characterising emerging antimicrobial resistance genes (ARGs) are needed. Here, we develop a semi-automated workflow using cell-free gene expression (CFE) systems to measure the activity of two ARGs encoded on plasmid DNA that produce rifampicin-inactivating and gentamicin-inactivating enzymes. We validated the use of a small benchtop Myra liquid handling system compared to manual pipetting, with no statistical differences observed. After optimising the pre-incubation time of ARGs and dispensing protocol, expression of aac(3)-IIa increased the half-maximal inhibition concentration (IC50) of gentamicin by over 150-fold, while arr-3 increased the IC50 of rifampicin by approximately 20-fold compared to controls. Future work could extend this platform to characterise novel ARGs identified through genomic surveillance or rapidly profile activity of new or derivative antibiotics. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=87 SRC="FIGDIR/small/720151v1_ufig1.gif" ALT="Figure 1"> View larger version (20K): org.highwire.dtl.DTLVardef@50bbdcorg.highwire.dtl.DTLVardef@5d98e4org.highwire.dtl.DTLVardef@e0a3fforg.highwire.dtl.DTLVardef@2ef12a_HPS_FORMAT_FIGEXP M_FIG C_FIG

synthetic biology↗

bioRxiv2026

Estimating mosquito bionomics parameters with a hierarchical Bayesian model

BackgroundThe malaria transmission potential and the vulnerability of Anopheles mosquitoes to different vector control methods depend, among other factors, on the endophily, endophagy, anthropophagy and survival of each species. Local information on these bionomic parameters is generally unavailable. MethodsTo address this, we estimated species-specific values of these parameters using an augmented version of the global database of bionomics data by Massey et al. (2016). We applied inclusion and exclusion criteria to select eligible studies with relevant experimental designs that minimise bias from collection methods for parous, sac, endophagy, and endophily rates as well as for the resting duration. For the human blood index (HBI), we separated data from indoor and outdoor collections. We fitted hierarchical Bayesian models with levels based on Anopheles taxonomy to estimate these quantities. Based on the estimated bionomics, we quantified the expected vectorial capacity reduction after the introduction of a pyrethroid-pyrrole insecticide-treated net (ITN) for 57 Anopheles species. ResultsWe identified 26 eligible studies for endophagy and 61 for the parous rate, leading to a Bayesian posterior average for the Anopheles genus of 42% (95% credible interval: 18-70) and 55% (32-77) respectively. HBI values widely varied depending on the location of collection, except for some species showing strong anthropophilic behaviours. Resting duration was estimated to be 2.1 days (1.2 - 4.8) at the genus level. Few studies were available to estimate the sac and endophily rates, which prevented us from deriving precise estimates for the whole Anopheles genus. Our estimates of the vectorial capacity reduction following the introduction of a pyrrole-pyrethroid ITN ranged between 48% and 76% across species, highlighting the important differences among mosquito species in vulnerability to vector control interventions. ConclusionThis work demonstrates how data from both Anopheles species complexes and individual species can be leveraged to generate species-specific estimates of bionomic parameters, capturing the local characteristics and behaviour of malaria vectors. The dataset is readily updatable as new data become available. However, more frequent and standardised field surveys are still needed to accurately characterise local vector behaviour.

zoology↗

bioRxiv2025

Genomic Evidence for a Novel Introduction and Intra-host Diversity of DENV-2 in Dar es Salaam, Tanzania

Dengue virus (DENV) poses a growing risk in Tanzania, yet its diversity in mosquitoes is poorly understood. Using Nanopore sequencing, we recovered full coding sequences from six DENV-2 positive mosquito pools collected in Dar es Salaam outside outbreak periods. Phylogenetic analysis placed the sequences in a distinct monophyletic clade within genotype II, separate from strains linked to Tanzanias 2014 outbreak. Instead, they clustered with Asian lineages, showing closest relatedness to DENV-2 from Kenya (2013) and India (2014), with divergence estimated around 2010. Variant profiling identified 212 low-frequency intra-host variants, largely nonsynonymous changes in NS3 and NS5. These results suggest an unrecognized introduction of genotype II now circulating silently in local mosquito populations. Our findings highlight the value of genomic surveillance in vectors for early detection of arboviral threats, even when human cases are absent.

genomics↗

bioRxiv2025

Controlling malaria mosquito reproduction via the octopamine beta2 receptor

Mosquito reproduction in a broad sense involves multiple steps from acoustic recognition of mating partners to egg hatching. We show that the octopamine receptor AgOct{beta}2R controls different aspects of this process in a sexually dimorphic manner. AgOct{beta}2R knockout males present auditory defects that impair their ability to inseminate females, whilst knockout females are sterile. These phenotypes suggest AgOct{beta}2R as a target to impair mosquito reproduction at multiple levels. We test the reproductive effects of the insecticide amitraz, an AgOct{beta}2R agonist, showing that amitraz exposure reduces insemination in the lab but not in the field and has no effects on female sterility, excluding its applicability as a mating disruptor. Pharmacological assays reveal that AgOct{beta}2R sensitivity to amitraz is reduced compared to other arthropods, but its responses can be altered by modifying residues in the binding pocket. Together, our results establish AgOct{beta}2R as a promising target to disrupt mosquito reproduction but emphasize the necessity of developing new tools to exploit this approach.

neuroscience↗

bioRxiv2024

Field evaluation of a volatile pyrethroid spatial repellent and etofenprox-treated clothing for outdoor protection against forest malaria vectors in Cambodia

Cambodias goal to eliminate malaria by 2025 is challenged by persisting transmission in the countrys forest and forest fringe areas. People living in, or traveling to the forest, are exposed to malaria vector bites during the day due to Anopheles daytime biting; and during the night, due to low bed net use and open sleeping structures. Volatile pyrethroid spatial repellents (VPSRs), and insecticide treated clothing (ITC) may help address these gaps in protection. In this field study the authors evaluated the outdoor application of one passive, transfluthrin-based VPSR, four etofenprox-ITCs paired with a picaridin topical repellent, and a combination of VPSR and ITC against wild Anopheles landing in Cambodia. Mathematical modeling was also used to predict the reduction of vectorial capacity of these interventions. A 7x7 Latin-square (6 interventions and one control) was conducted over 49 collection nights in seven temporary, open structures in a forest in Mondulkiri Province, Cambodia. Pairs of participants conducted human landing catches (HLCs) from 18h00 to 06h00, with each collector conducting collections for six hours. A randomly selected subset of collected Anopheles were identified to species using molecular methods. The rate ratio of each intervention compared to the control on Anopheles landings was estimated using a mixed-effect negative binomial regression with intervention, structure, and collector-pair as fixed-effects, and with collection date and structure-night as random effects. The modeling assessment aims to predict the relative reduction in vectoral capacity. Initial calculations involved establishing a "baseline scenario" without intervention, utilizing biometric parameters for Anopheles dirus. Various scenarios accounting for intervention coverage and adherence were then considered. The study aims to update parameters using field study estimates for wild Anopheles, incorporating multiple semi-field estimates for interventions and accounting for the variability and uncertainty in parameter values. Of the total 8,294 Anopheles specimens collected, 15% (n=1,242) of specimens were confirmed to species or species group via PCR. Fifteen species were confirmed; Anopheles dirus Form A was predominant (n=429), followed by Anopheles maculatus (n=189), and Anopheles minimus (n=60). All six interventions reduced Anopheles landing substantially; protective efficacies ranged between 61% (95% confidence interval (CI): 48 - 71%) (etofenprox-ITC, washed) and 95% (95% CI: 93 - 96%) (combined VPSR and unwashed etofenprox-ITC). Finally, the modelling assessment demonstrates significant reductions in vectoral capacity, with the highest impact observed for the combined ITC and VPSR as well as the VPSR used alone, although effectiveness decreases with intervention aging, and variability exists in the magnitude of predicted reductions due to differences in experimental conditions. These transfluthrin-based VPSR and etofenprox ITC interventions have the potential to reduce outdoor and daytime Anopheles biting by providing substantial protection against Anopheles landing. One or more of these tools may play a valuable role in the push for elimination in Cambodia and the Greater Mekong Subregion if programs can achieve effective coverage.

ecology↗

bioRxiv2023

Inference for entomological semi-field experiments: Fitting a mathematical model assessing personal and community protection of vector-control interventions

The effectiveness of vector-control tools is often assessed by experiments as a reduction in mosquito landings using human landing catches (HLCs). However, HLCs alone only quantify a single characteristic and therefore do not provide information on the overall impacts of the intervention product. Using data from a recent semi-field study which used time-stratified HLCs, aspiration of non-landing mosquitoes, and blood feeding, we suggest a Bayesian inference approach for fitting such data to a stochastic model. This model considers both personal protection, through a reduction in biting, and community protection, from mosquito mortality and disarming (prolonged inhibition of blood feeding). Parameter estimates are then used to predict the reduction of vectorial capacity induced by etofenpox-treated clothing, picaridin topical repellents, transfluthrin spatial repellents and metofluthrin spatial repellents, as well as combined interventions for Plasmodium falciparum malaria in Anopleles minimus. Overall, all interventions had both personal and community effects, preventing biting and killing or disarming mosquitoes. This led to large estimated reductions in the vectorial capacity, with substantial impact even at low coverage. As the interventions aged, fewer mosquitoes were killed; however the impact of some interventions changed from killing to disarming mosquitoes. Overall, this inference method allows for additional modes of action, rather than just reduction in biting, to be parameterised and highlights the tools assessed as promising malaria interventions.

systems biology↗

bioRxiv2023

Sub-lethal exposure to chlorfenapyr reduces the probability of developing Plasmodium falciparum parasites in surviving Anopheles mosquitoes

Pyrethroid resistance in the key malaria vectors threatens the success of pyrethroid-treated nets. To overcome pyrethroid resistance, Interceptor(R) G2 (IG2), a first-in-class dual insecticidal net that combines alpha-cypermethrin with chlorfenapyr was developed. Chlorfenapyr is a pro-insecticide, requiring bio-activation by oxidative metabolism within the insects mitochondria, constituting a mode of action preventing cross-resistance to pyrethroids. Recent epidemiological trials conducted in Benin and Tanzania confirm IG2s public health value in areas with pyrethroid-resistant Anopheles mosquitoes. As chlorfenapyr might also interfere with the metabolic mechanism of the Plasmodium parasite, we hypothesised that chlorfenapyr may provide additional transmission-reducing effects even if a mosquito survives a sub-lethal dose. Therefore, we tested the effect of chlorfenapyr netting to reduce Plasmodium falciparum transmission using a modified WHO tunnel test with a dose yielding sub-lethal effects. Pyrethroid-resistant Anopheles gambiae s.s. with established mixed-function oxidases and Vgsc-L995F knockdown resistance alleles were exposed to untreated netting and netting treated with 200 mg/m3 chlorfenapyr for 8 hours overnight and then fed on gametocytemic blood meals from naturally infected individuals. Prevalence and intensity of oocysts and sporozoites were determined on day 8 and day 16 after feeding. Both prevalence and intensity of P. falciparum infection in the surviving mosquitoes were substantially reduced in the chlorfenapyr-exposed mosquitoes compared to untreated nets. The odds ratios in the prevalence of oocysts and sporozoites were 0.33 (95% confidence interval; 95% CI: 0.23-0.46) and 0.43 (95% CI: 0.25-0.73), respectively, while only the incidence rate ratio for oocysts was 0.30 (95% CI: 0.22-0.41). We demonstrated that sub-lethal exposure of pyrethroid-resistant mosquitoes to chlorfenapyr substantially reduces the proportion of infected mosquitoes and the intensity of the P. falciparum infection. This will likely also contribute to the reduction of malaria in communities beyond the direct killing of mosquitoes. Author summaryMalaria remains a serious problem in many tropical and sub-tropical areas, affecting the welfare and health of many individuals. Since 2016, malaria has increased and the emergence of mosquitoes that are resistant to different classes of insecticides used in vector control tools may have contributed to some of this increase. Therefore, insecticides with a different mode of action are required to manage vector resistance to insecticides used for public health vector control. One of the main resistance mechanisms is metabolic resistance where mosquitoes upregulate detoxification enzymes to break down insecticides. Chlorfenapyr is a pyrrole-pro-insecticide that is metabolised by these detoxification enzymes from chlorfenapyr to tralopyril that disrupts mitochondrial function in mosquitoes. We therefore hypothesized that the metabolites of chlorfenapyr may also have an effect on Plasmodia since they, too possess mitochondria and this could reduce the development of Plasmodium in mosquitoes that survived a sub-lethal dose of chlorfenapyr. In this study we established and evaluated a modified WHO tunnel assay to investigate the effect of chlorfenapyr in Plasmodium-infected Anopheles mosquitoes. In this bioassay, we found that chlorfenapyr substantially reduces the proportion of Plasmodium-infected mosquitoes at doses sub-lethal to mosquitoes. Our findings demonstrate that chlorfenapyr provides additional benefits beyond mosquito killing although the mechanism of action requires further elucidation.

cell biology↗

bioRxiv2023

A cell-free strategy for profiling intracellular antibiotic sensitivity and resistance

Antimicrobial resistance (AMR) is a pandemic spread across multiple priority infectious disease threats. While the cell envelope plays a key role in AMR, this also makes it challenging to study how antibiotics function inside the cell. Herein, we present a Klebsiella pneumoniae cell-free gene expression (CFE) platform for the rapid profiling of intracellular antibiotic sensitivity and resistance. This cell-free approach provides the unique macromolecular and metabolite components from this microbe, which include multiple antibiotic targets from transcription, translation, and metabolic processes. First, we compare the K. pneumoniae CFE system to whole cell antimicrobial assays. We find that several antibiotic classes show higher sensitivity in the CFE system, suggesting limitations in antibiotic transport in the whole cell assay. Next, we evolved K. pneumoniae strains with resistance to specific antibiotics and use whole genome sequencing analysis for genotyping. As an exemplary case, we show that a single RNA polymerase beta subunit variant H526L (also frequently found in multidrug resistant Mycobacterium tuberculosis) confers a 58-fold increase in CFE resistance to rifampicin. Overall, we describe a safe (i.e., non-living, non-pathogenic) platform suitable for studying an infectious disease model in a Containment Level 1 laboratory. Our CFE strategy is generalisable to laboratory and clinical K. pneumoniae strains and provides a new experimental tool to profile intracellular AMR variants. In conclusion, our CFE tool provides a significant advance towards understanding AMR and complements wider infectious disease studies.

microbiology↗

bioRxiv2022

A novel beta-adrenergic like octopamine receptor modulates the audition of malaria mosquitoes and serves as insecticide target

Hearing is an essential sense in the life cycle of malaria mosquitoes. Within large swarms formed transiently at dusk, mosquitoes acoustically recognize their mating partners by their wingbeats. Indeed, malaria mosquitoes only respond to the flight tones of mating partners during swarm time. This phenomenon implies a sophisticated context- and time-dependent modulation of mosquito audition, the mechanisms of which are still largely unknown. Using transcriptomics, we identify a complex network of candidate neuromodulators regulating mosquito hearing. Among them, octopamine stands out as regulator of the auditory performance during swarm time. To explore octopamines roles in mosquito hearing, we carried out an in-depth analysis of octopamine-mediated effects on auditory function. We found that octopamine affects the properties of the mosquito ear on multiple levels: it modulates the tuning and stiffness of the flagellar sound receiver and it controls the erection of antennal fibrillae in males. We found that two different receptors are driving octopamines auditory roles, including a novel beta octopamine receptor. We also demonstrate that the octopaminergic auditory control system can be targeted by insecticides. Our findings identify octopamine signalling as a key component of hearing and mating partner detection in malaria mosquitoes, and as a potential novel target for mosquito control.

neuroscience↗

bioRxiv2022

Self-organizing Single-Rosette Brain Organoids from Human Pluripotent Stem Cells

The field of brain organoid research is complicated by morphological variability with multiple neural rosette structures per organoid. We have developed a new human brain organoid technique that generates self-organizing, single-rosette spheroids (SOSRS) with reproducible size, cortical-like lamination, and cell diversity. Rather than patterning a 3-dimensional embryoid body, we initiate brain organoid formation from a 2-dimensional monolayer of human pluripotent stem cells (hPSCs) that is patterned with small molecules into neuroepithelium and differentiated to cells of the developing dorsal cerebral cortex. This approach recapitulates the 2D to 3D transition from neural plate to neural tube that occurs during neurodevelopment. The vast majority of monolayer fragments form spheres with a single central lumen and consistent growth rates. Over time, the SOSRS develop appropriately ordered lamination consistent with six cortical layers by immunocytochemistry and single cell RNA-sequencing. The reproducibility of this method has allowed us to demonstrate robust structural phenotypes arising from chemical teratogen exposure or when modeling a genetic neurodevelopmental epileptic disorder. This platform should advance studies of human cortical development, brain disorder mechanisms, and precision therapies. SUMMARY STATEMENTSimple procedure for generating reproducible single rosette cortical brain organoids used to identify robust structural phenotypes with neuroteratogen exposure and in a genetic neurodevelopmental disease model.

neuroscience↗

bioRxiv2021

A curcumin direct protein (DiPro) biosensor for cell-free prototyping

In synthetic biology, biosensors are routinely coupled to a gene expression cascade for detecting small molecules and physical signals. We posit that an alternative direct protein (DiPro) biosensor mechanism, could provide a new opportunity for rapid detection of specific chemicals. Herein, we reveal a fluorescent curcumin DiPro biosensor, based on the Escherichia coli double bond reductase (EcCurA) as a detection system. We characterise the EcCurA DiPro biosensor and propose enhanced curcumin fluorescence is generated through {pi}-{pi} stacking between protein and ligand. Using a cell-free synthetic biology approach, we use the EcCurA DiPro biosensor to fine tune 10 reaction parameters (cofactor, substrate, and enzyme levels) for cell-free biosynthesis, assisted through acoustic liquid handling robotics. Overall, we increase EcCurA-curcumin fluorescence by 80-fold. We speculate that a generic DiPro biosensor fluorescence mechanism can be further exploited for a wider range of chemicals that share intrinsic fluorescence and have a suitable binding protein.

synthetic biology↗

bioRxiv2021

Flight-tone mediated circadian control of audibility in mating swarms of Anopheles mosquitoes

Mating swarms of malaria mosquitoes form every day at sunset throughout the tropical world, they typically last less than 30 minutes. Activity patterns must thus be highly synchronized between the sexes. Moreover, males must be able to identify the few sporadically entering females by detecting the females faint flight tones. We here show that the Anopheles circadian clock ensures a tight synchrony of male and female activity and -importantly - also retunes the males acoustic detection system: by lifting their own flight tones at dusk, males actively enhance the audibility of females. The reported phenomenon of harmonic convergence is a random by-product of the mosquitoes flight tone variance. Intriguingly, flight tones of individual mosquitoes occupy narrow -partly non-overlapping-frequency ranges, suggesting that the audibility of individual females varies across males. One Sentence SummaryMale mosquitoes sharpen their hearing at sunset.

neuroscience↗