Assessment of the Novel Anti-Seizure Potential of Validamycin A Using Zebrafish Epilepsy Model.

Epilepsy is a prevalent neurological disorder characterized by recurrent seizures. Validamycin A (VA) is an antibiotic fungicide that inhibits trehalase activity and is widely used for crop protection in agriculture. In this study, we identified a novel function of VA as a potential anti-seizure medication in a zebrafish epilepsy model. Electroencephalogram (EEG) analysis demonstrated that VA reduced pentylenetetrazol (PTZ)-induced seizures in the brains of larval and adult zebrafish. Moreover, VA reduced PTZ-induced irregular movement in a behavioral assessment of adult zebrafish. The developmental toxicity test showed no observable anatomical alteration when the zebrafish larvae were treated with VA up to 10 µM within the effective range. The median lethal dose of VA in adult zebrafish was > 14,000 mg/kg. These results imply that VA does not demonstrate observable toxicity in zebrafish at concentrations effective for generating anti-seizure activity in the EEG and alleviating abnormal behavior in the PTZ-induced epileptic model. Furthermore, the effectiveness of VA was comparable to that of valproic acid. These results indicate that VA may have a potentially safer anti-seizure profile than valproic acid, thus offering promising prospects for its application in agriculture and medicine.

Innovative Biofouling Control for Membrane Bioreactors in Cold Regions by Inducing Environmental Adaptation in Quorum-Quenching Bacteria.

Bacterial quorum quenching (QQ), whose mechanism involves the degradation of quorum-sensing signal molecules, is an effective strategy for controlling biofouling in membrane bioreactors (MBRs). However, MBRs operated at low temperatures, either due to cold climates or seasonal variations, exhibit severe deterioration in QQ efficiency. In this study, a modified culture method for Rhodococcus sp. BH4, a QQ bacterium, was developed to induce environmental adaptation in cold regions. BH4-L, which was prepared by the modified culture method, showed enhancement in QQ efficiency at low temperatures. The higher QQ efficiency obtained by employing BH4-L at 10 °C (compared with that obtained by employing BH4 at 10 °C) was attributed to the higher live/dead cell ratio in the BH4-L-entrapping beads. When BH4-L-entrapping beads were applied to lab-scale MBRs operated at low temperatures, membrane biofouling in MBRs at low temperatures was successfully mitigated because BH4-L could substantially reduce the concentration of signal molecules (N-acyl homoserine lactones) in the biocake. Employing BH4-L in QQ-MBRs could offer a novel solution to the problem of severe membrane biofouling in MBRs in cold regions.

Preparation of a mesoporous silica quorum quenching medium for wastewater treatment using a membrane bioreactor.

Various quorum quenching (QQ) media have been developed to mitigate membrane biofouling in a membrane bioreactor (MBR). However, most are expensive, unstable and easily trapped in hollow fibre membranes. Here, a sol-gel method was used to develop a mesoporous silica medium entrapping a QQ bacterial strain (Rhodococcus sp. BH4). The new silica QQ medium was able to remove quorum sensing signalling molecules via both adsorption (owing to their mesoporous hydrophobic structure) and decomposition with an enzyme (lactonase), preventing MBR biofouling without affecting the water quality. It also demonstrated a relatively long life span due to its non-biodegradability and its relatively small particle size (<1.0 mm), which makes it less likely to clog in a hollow fibre membrane module.

Smart Pack: Online Autonomous Object-Packing System Using RGB-D Sensor Data.

This paper proposes a novel online object-packing system which can measure the dimensions of every incoming object and calculate its desired position in a given container. Existing object-packing systems have the limitations of requiring the exact information of objects in advance or assuming them as boxes. Thus, this paper is mainly focused on the following two points: (1) Real-time calculation of the dimensions and orientation of an object; (2) Online optimization of the object's position in a container. The dimensions and orientation of the object are obtained using an RGB-D sensor when the object is picked by a manipulator and moved over a certain position. The optimal position of the object is calculated by recognizing the container's available space using another RGB-D sensor and minimizing the cost function that is formulated by the available space information and the optimization criteria inspired by the way people place things. The experimental results show that the proposed system successfully places the incoming various shaped objects in their proper positions.

Quorum sensing: an emerging link between temperature and membrane biofouling in membrane bioreactors.

Lab-scale membrane bioreactors (MBRs) were investigated at 12, 18, and 25 °C to identify the correlation between quorum sensing (QS) and biofouling at different temperatures. The lower the reactor temperature, the more severe the membrane biofouling measured in terms of the transmembrane pressure (TMP) during filtration. More extracellular polymeric substances (EPSs) that cause biofouling were produced at 18 °C than at 25 °C, particularly polysaccharides, closely associated with QS via the production of N-acyl homoserine lactone (AHL). However, at 12 °C, AHL production decreased, but the release of EPSs due to deflocculation increased the soluble EPS concentration. To confirm the temperature effect related to QS, bacteria producing AHL were isolated from MBR sludge and identified as Aeromonas sp., Leclercia sp., and Enterobacter sp. through a 16S rDNA sequencing analysis. Batch assays at 18 and 25 °C showed that there was a positive correlation between QS through AHL and biofilm formation in that temperature range.

Stopping Autoinducer-2 Chatter by Means of an Indigenous Bacterium ( Acinetobacter sp. DKY-1): A New Antibiofouling Strategy in a Membrane Bioreactor for Wastewater Treatment.

Bacterial quorum quenching (QQ) by means of degrading signaling molecules has been applied to antibiofouling strategies in a membrane bioreactor (MBR) for wastewater treatment. However, the target signaling molecules have been limited to N-acyl homoserine lactones participating in intraspecies quorum sensing. Here, an approach to disrupting autoinducer-2 (AI-2) signaling molecules participating in interspecies quorum sensing was pursued as a next-generation antibiofouling strategy in an MBR for wastewater treatment. We isolated an indigenous QQ bacterium ( Acinetobacter sp. DKY-1) that can attenuate the expression of the quorum-sensing (QS) response through the inactivation of an autoinducer-2 signaling molecule, 4,5-dihydroxy-2,3-pentanedione (DPD), among four kinds of autoinducer-2 QS bacteria. DKY-1 released AI-2 QQ compounds, which were verified to be hydrophilic with a molecular weight of <400 Da. The addition of DKY-1 entrapping beads into an MBR significantly decreased DPD concentration and remarkably reduced membrane biofouling. This new approach, combining molecular biology with wastewater engineering, could enlarge the range of QQ-MBR for antibiofouling and energy savings in the field of wastewater treatment.

Membrane biofouling behaviors at cold temperatures in pilot-scale hollow fiber membrane bioreactors with quorum quenching.

In this study, the seasonality of the biofouling behavior of pilot-scale membrane bioreactors (MBRs) run in parallel with vacant sheets and quorum quenching (QQ) sheets using real municipal wastewater was investigated. QQ media delayed fouling, but low temperatures caused severe biofouling. The greater amount of extracellular polymeric substances (EPSs) produced in cold weather was responsible for the faster biofouling of a membrane, even with QQ media. There were significant negative relationships between EPS levels and water temperature. Cold weather was detrimental to the degradation of quorum sensing signal molecules by QQ sheets, whose activity was restored with a higher dose of QQ bacteria. The QQ bacteria in the sheets experienced a slight loss in activity during the early stage of the field test, but survived in the pilot-scale MBR fed with real wastewater. There were no significant discrepancies in treatment efficiency among conventional, vacant, and QQ MBRs.

More Efficient Media Design for Enhanced Biofouling Control in a Membrane Bioreactor: Quorum Quenching Bacteria Entrapping Hollow Cylinder.

Recently, membrane bioreactors (MBRs) with quorum quenching (QQ) bacteria entrapping beads have been reported as a new paradigm in biofouling control because, unlike conventional post-biofilm control methods, bacterial QQ can inhibit biofilm formation through its combined effects of physical scouring of the membrane and inhibition of quorum sensing (QS). In this study, using a special reporter strain (Escherichia coli JB525), the interaction between QS signal molecules and quorum quenching bacteria entrapping beads (QQ-beads) was elucidated through visualization of the QS signal molecules within a QQ-bead using a fluorescence microscope. As a result, under the conditions considered in this study, the surface area of QQ-media was likely to be a dominant parameter in enhancing QQ activity over total mass of entrapped QQ bacteria because QQ bacteria located near the core of a QQ-bead were unable to display their QQ activities. On the basis of this information, a more efficient QQ-medium, a QQ hollow cylinder (QQ-HC), was designed and prepared. In batch experiments, QQ-HCs showed greater QQ activity than QQ-beads as a result of their higher surface area and enhanced physical washing effect because of their larger impact area against the membrane surface. Furthermore, it was shown that such advantages of QQ-HCs resulted in more effective mitigation of membrane fouling than from QQ-beads in lab-scale continuous MBRs.

Crossing the Border between Laboratory and Field: Bacterial Quorum Quenching for Anti-Biofouling Strategy in an MBR.

Quorum quenching (QQ) has recently been acknowledged to be a sustainable antifouling strategy and has been investigated widely using lab-scale membrane bioreactor (MBR) systems. This study attempted to bring this QQ-MBR closer to potential practical application. Two types of pilot-scale QQ-MBRs with QQ bacteria entrapping beads (QQ-beads) were installed and run at a wastewater treatment plant, feeding real municipal wastewater to test the systems' effectiveness for membrane fouling control and thus the amount of energy savings, even under harsh environmental conditions. The rate of transmembrane pressure (TMP) build-up was significantly mitigated in QQ-MBR compared to that in a conventional-MBR. Consequently, QQ-MBR can substantially reduce energy consumption by reducing coarse bubble aeration without compromising the effluent water quality. The addition of QQ-beads to a conventional MBR substantially affected the EPS concentrations, as well as microbial floc size in the mixed liquor. Furthermore, the QQ activity and mechanical stability of QQ-beads were well maintained for at least four months, indicating QQ-MBR has good potential for practical applications.

Fungal Quorum Quenching: A Paradigm Shift for Energy Savings in Membrane Bioreactor (MBR) for Wastewater Treatment.

In the last 30 years, the use of membrane bioreactors (MBRs) for advanced wastewater treatment and reuse have been expanded continuously, but they still suffer from excessive energy consumption resulting from the intrinsic problem of membrane biofouling. One of the major causes of biofouling in MBRs is bacterial quorum sensing (QS) via N-acylhomoserine lactones (AHLs) and/or autoinducer-2 (AI-2), enabling intra- and interspecies communications, respectively. In this study, we demonstrate that farnesol can substantially mitigate membrane biofouling in a MBR due to its quorum quenching (QQ) activity. When Candida albicans (a farnesol producing fungus) entrapping polymer beads (AEBs) were placed in the MBR, the rate of transmembrane pressure (TMP) rise-up was substantially decreased, even for lower aeration intensities. This finding corresponds to a specific aeration energy savings of approximately 40% (25% through the physical washing effect and a further 15% through the biological QQ effect of AEBs) compared to conventional MBRs without AEBs. A real-time RT-qPCR analysis revealed that farnesol secreted from C. albicans mitigated the biofilm formation in MBRs via the suppression of AI-2 QS. Successful control of biofouling and energy savings through fungal-to-bacterial QQ could be expanded to the plant scale for MBRs in wastewater treatment with economic feasibility.

Mirrored continuum and molecular scale simulations of the ignition of high-pressure phases of RDX.

We present a mirrored atomistic and continuum framework that is used to describe the ignition of energetic materials, and a high-pressure phase of RDX in particular. The continuum formulation uses meaningful averages of thermodynamic properties obtained from the atomistic simulation and a simplification of enormously complex reaction kinetics. In particular, components are identified based on molecular weight bin averages and our methodology assumes that both the averaged atomistic and continuum simulations are represented on the same time and length scales. The atomistic simulations of thermally initiated ignition of RDX are performed using reactive molecular dynamics (RMD). The continuum model is based on multi-component thermodynamics and uses a kinetics scheme that describes observed chemical changes of the averaged atomistic simulations. Thus the mirrored continuum simulations mimic the rapid change in pressure, temperature, and average molecular weight of species in the reactive mixture. This mirroring enables a new technique to simplify the chemistry obtained from reactive MD simulations while retaining the observed features and spatial and temporal scales from both the RMD and continuum model. The primary benefit of this approach is a potentially powerful, but familiar way to interpret the atomistic simulations and understand the chemical events and reaction rates. The approach is quite general and thus can provide a way to model chemistry based on atomistic simulations and extend the reach of those simulations.

Benefits of fungal-to-bacterial quorum quenching as anti-biofouling strategy in membrane bioreactors for wastewater treatment and water reuse.

This study addresses membrane biofouling in membrane bioreactors (MBRs) by exploring fungal-to-bacterial quorum quenching (QQ) strategies. While most research has been focused on bacterial-to-bacterial QQ tactics, this study identified fungal strain Vanrija sp. MS1, which is capable of degrading N-acyl-homoserine lactones (signaling molecules of Gram-negative bacteria). To determine the benefits of fungal over bacterial strains, after immobilization on fluidizing spherical beads in an MBR, MS1 significantly reduced the fouling rate by 1.8-fold compared to control MBR, decreased extracellular polymeric substance levels in the biofilm during MBR operation, and favorably changed microbial community and bacterial network, resulting in biofouling mitigation. It is noteworthy that, unlike Rhodococcus sp. BH4, MS1 enhanced QQ activity when switching from neutral to acidic conditions. These results suggest that MS1 has the potential for the effective treatment of acidic industrial wastewater sources such as semiconductor and secondary battery wastewater using MBRs.

Rapid deep learning-assisted predictive diagnostics for point-of-care testing.

Prominent techniques such as real-time polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and rapid kits are currently being explored to both enhance sensitivity and reduce assay time for diagnostic tests. Existing commercial molecular methods typically take several hours, while immunoassays can range from several hours to tens of minutes. Rapid diagnostics are crucial in Point-of-Care Testing (POCT). We propose an approach that integrates a time-series deep learning architecture and AI-based verification, for the enhanced result analysis of lateral flow assays. This approach is applicable to both infectious diseases and non-infectious biomarkers. In blind tests using clinical samples, our method achieved diagnostic times as short as 2 minutes, exceeding the accuracy of human analysis at 15 minutes. Furthermore, our technique significantly reduces assay time to just 1-2 minutes in the POCT setting. This advancement has the potential to greatly enhance POCT diagnostics, enabling both healthcare professionals and non-experts to make rapid, accurate decisions.

Membrane reciprocation and quorum quenching: An innovative combination for fouling control and energy saving in membrane bioreactors.

Membrane bioreactors (MBRs) play a crucial role in wastewater treatment, but they face considerable challenges due to fouling. To tackle this issue, innovative strategies are needed. This study investigated the effectiveness of membrane reciprocation and quorum quenching (QQ) to control fouling in MBRs. The study compared MBRs using membrane reciprocation (30 rpm) and QQ (injecting media containing 100 or 200 mg/L BH4) with conventional MBRs employing different air-scouring intensities. The results demonstrated that combining membrane reciprocation (30 rpm) with QQ (200 mg/L BH4) significantly extended the service time of MBRs, making it approximately six times longer than conventional methods. Moreover, this approach reduced physically reversible resistance. The reduction in signal molecules related to biofouling due to QQ showcased its critical role in controlling biofouling, even under high shear caused by membrane reciprocation. However, the impact of QQ on microbial community structure appeared relatively insignificant when compared to factors such as operation time, aeration intensity, and membrane reciprocation. By combining membrane reciprocation and QQ, the study achieved a remarkable 81 % energy saving compared to extensive aeration (103 s-1 in velocity gradient), in addition to the extended service time. Importantly, this combined antifouling approach did not negatively affect microbial characteristics and wastewater treatment, emphasizing its effectiveness in MBRs. Overall, the findings of this study offer valuable insights for developing synergistic fouling control strategies in MBRs, significantly improving the energy efficiency of the wastewater treatment process.

Microbial population dynamics and proteomics in membrane bioreactors with enzymatic quorum quenching.

Quorum sensing gives rise to biofilm formation on the membrane surface, which in turn causes a loss of water permeability in membrane bioreactors (MBRs) for wastewater treatment. Enzymatic quorum quenching was reported to successfully inhibit the formation of biofilm in MBRs through the decomposition of signal molecules, N-acyl homoserine lactones (AHLs). The aim of this study was to elucidate the mechanisms of quorum quenching in more detail in terms of microbial population dynamics and proteomics. Microbial communities in MBRs with and without a quorum quenching enzyme (acylase) were analyzed using pyrosequencing and compared with each other. In the quorum quenching MBR, the rate of transmembrane pressure (TMP) rise-up was delayed substantially, and the proportion of quorum sensing bacteria with AHL-like autoinducers (such as Enterobacter, Pseudomonas, and Acinetobacter) also decreased in the entire microbial community of mature biofilm in comparison to that in the control MBR. These factors were attributed to the lower production of extracellular polymeric substances (EPS), which are known to play a key role in the formation of biofilm. Proteomic analysis using the Enterobacter cancerogenus strain ATCC 35316 demonstrates the possible depression of protein expression related to microbial attachments to solid surfaces (outer membrane protein, flagellin) and the agglomeration of microorganisms (ATP synthase beta subunit) with the enzymatic quorum quenching. It has been argued that changes in the microbial population, EPS and proteins via enzymatic quorum quenching could inhibit the formation of biofilm, resulting in less biofouling in the quorum quenching MBR.

Does quorum quenching matter to microbial community dynamics in long-term membrane bioreactor operation?

Quorum quenching (QQ) has effectively prevented biofouling in membrane bioreactors (MBRs) employing isolated QQ bacterial strains. However, the influence of QQ on the microbial population still needs to be fully understood. This research aims to analyze the microbial population in MBRs over an extended period (>250 days) under different conditions, such as varying aeration intensities and doses of QQ bacteria, QQ media, and types of feed. Results show that no significant changes occurred in the structure and diversity of the microbial community in the mixed liquor and biofilm due to QQ treatment. Canonical correspondence analysis did reveal that the microbial communities were strongly influenced by feed types and phases. The microbial community composition varied between bacterial habitats (i.e., mixed liquor and biofilm), showing the two dominant phyla Proteobacteria and Bacteroidota in the former and Proteobacteria and Chloroflexi in the latter. The co-occurrence network analysis indicated that the biofilm (with 163 edges) in the MBR fed with real wastewater exhibited a more intricate network than the biofilm (with 53 edges) in the MBR fed with synthetic wastewater. With QQ, the biofilm exhibited more positive edges than negative ones. The phylogenetic investigation of communities showed that QQ barely affects functional gene-related quorum sensing (e.g., bacterial chemotaxis, motility proteins, and secretion) in mixed liquor but in biofilms at relatively large QQ doses (> 75 mg/L BH4). This research sheds light on the bacterial QQ's role in reducing MBR biofouling and provides crucial insights into its underlying mechanisms.

Sample-to-answer platform for the clinical evaluation of COVID-19 using a deep learning-assisted smartphone-based assay.

Since many lateral flow assays (LFA) are tested daily, the improvement in accuracy can greatly impact individual patient care and public health. However, current self-testing for COVID-19 detection suffers from low accuracy, mainly due to the LFA sensitivity and reading ambiguities. Here, we present deep learning-assisted smartphone-based LFA (SMARTAI-LFA) diagnostics to provide accurate decisions with higher sensitivity. Combining clinical data learning and two-step algorithms enables a cradle-free on-site assay with higher accuracy than the untrained individuals and human experts via blind tests of clinical data (n = 1500). We acquired 98% accuracy across 135 smartphone application-based clinical tests with different users/smartphones. Furthermore, with more low-titer tests, we observed that the accuracy of SMARTAI-LFA was maintained at over 99% while there was a significant decrease in human accuracy, indicating the reliable performance of SMARTAI-LFA. We envision a smartphone-based SMARTAI-LFA that allows continuously enhanced performance by adding clinical tests and satisfies the new criterion for digitalized real-time diagnostics.

Zebrafish EEG predicts the efficacy of antiepileptic drugs.

Background: Pharmacological evaluation of antiepileptic drugs (AEDs) using mammalian animals takes long time and is expensive. The zebrafish is a species commonly used to study brain functions, neurological diseases, and drug toxicity, and attracts more attention as an alternative animal model to substitute or supplement mammalian animals in drug development. Electroencephalogram (EEG) is a key indicator for diagnosing brain diseases such as epilepsy, by directly measuring the brain activity. We propose a novel method for pharmacological evaluation of AEDs based on EEG from adult zebrafish, which allows researchers to select more clinically valuable drugs at the early stage of AED screening. Methods: To evaluate the efficacy of AEDs, zebrafish EEG signals were measured after administering six AEDs (valproate acid, gabapentin, ethosuximide, oxcarbazepine, tiagabine, and topiramate) at various doses to pentylenetetrazol (PTZ)-induced seizure models. The change in seizure activity was investigated according to doses. The antiepileptic effect was determined by observing a significant decrease in at least one out of three indicators of the number, total duration, and mean duration of ictal events. Results: Using EEG signals from adult zebrafish, antiepileptic effects were observed with all six AEDs. Among them, antiepileptic effects depending on dose were confirmed with valproate acid, gabapentin, ethosuximide, and tiagabine. Moreover, the 50% effective doses (ED50) of valproate acid and tiagabine were determined based on zebrafish EEG for the first time, indicating that the quantitative inter-species comparison of the AED efficacy is possible between zebrafish and mammals such as rodents. Significance: The results show that zebrafish can be used to effectively and quantitatively evaluate the efficacy of AEDs based on EEG, the same method to evaluate antiepileptic effects in mammals, suggesting that the proposed method can contribute in reducing the cost and duration of search for AEDs and thus accelerate the drug development cycles.

Quorum quenching bacteria isolated from industrial wastewater sludge to control membrane biofouling.

N-acylhomoserine lactone (AHL)-based bacterial communication through quorum sensing (QS) is one of the main causes of biofouling. Although quorum quenching (QQ) has proven to be an effective strategy against biofouling in membrane bioreactors (MBRs) for municipal wastewater treatment, its applicability for industrial wastewater treatment has rarely been studied. This is the first study to isolate QQ strains from the activated sludge used to treat industrial wastewater containing toxic tetramethylammonium hydroxide (TMAH) and 1-methyl-2-pyrrolidinone. The two QQ strains from genus Bacillus (SDC-U1 and SDC-A8) survived and effectively degraded QS signals in the presence of TMAH. They also showed resistance to toxic byproducts of TMAH degradation such as ammonium and formaldehyde. They effectively reduced the biofilm formation of Pseudomonas aeruginosa PAO1 and mixed community of activated sludge. The strains isolated in this study thus have the potential to be employed to reduce membrane biofouling in MBRs during the treatment of TMAH-containing wastewater.

Layered Antibiofouling Composite Membrane for Quenching Bacterial Signaling.

Bacterial quorum quenching (QQ) media with various structures (e.g., bead, cylinder, hollow cylinder, and sheet), which impart biofouling mitigation in membrane bioreactors (MBRs), have been reported. However, there has been a continuous demand for membranes with QQ capability. Thus, herein, we report a novel double-layered membrane comprising an outer layer containing a QQ bacterium (BH4 strain) on the polysulfone hollow fiber membrane. The double-layered composite membrane significantly inhibits biofilm formation (i.e., the biofilm density decreases by ~58%), biopolymer accumulation (e.g., polysaccharide), and signal molecule concentration (which decreases by ~38%) on the membrane surface. The transmembrane pressure buildup to 50 kPa of the BH4-embedded membrane (17.8 h ± 1.1) is delayed by more than thrice (p < 0.05) of the control with no BH4 in the membrane's outer layer (5.5 h ± 0.8). This finding provides new insight into fabricating antibiofouling membranes with a self-regulating property against biofilm growth.