Pulmonary function testing dataset of pressure and flow, dynamic circumference, heart rate, and aeration monitoring.

Respiratory data was collected from 20 subjects, with an even sex distribution, in the low-risk clinical unit at the University of Canterbury. Ethical consent for this trial was granted by the University of Canterbury Human Research Ethics Committee (Ref: HREC 2023/30/LR-PS). Respiratory data were collected, for each subject, over three tests consisting of: 1) increasing set PEEP from a starting point of ZEEP using a CPAP machine; 2) test 1 repeated with two simulated apnoea's (breath holds) at each set PEEP; and 3) three forced expiratory manoeuvres at ZEEP. Data were collected using a custom pressure and flow sensor device, ECG, PPG, Garmin HRM Dual heartrate belt, and a Dräeger PulmoVista 500 Electrical Impedance Tomography (EIT) machine. Subject demographic data was also collected prior to the trial, in a questionnaire, with measurement equipment available. These data aim to inform the development of pulmonary mechanics models and titration algorithms.

Quantitative proteomics of the miR-301a/SOCS3/STAT3 axis reveals underlying autism and anxiety-like behavior.

Autism is a widespread neurodevelopmental disorder. Although the research on autism spectrum disorders has been increasing in the past decade, there is still no specific answer to its mechanism of action and treatment. As a pro-inflammatory microRNA, miR-301a is abnormally expressed in various psychiatric diseases including autism. Here, we show that miR-301a deletion and inhibition exhibited two distinct abnormal behavioral phenotypes in mice. We observed that miR-301a deletion in mice impaired learning/memory, and enhanced anxiety. On the contrary, miR-301a inhibition effectively reduced the maternal immune activation (MIA)-induced autism-like behaviors in mice. We further demonstrated that miR-301a bound to the 3'UTR region of the SOCS3, and that inhibition of miR-301a led to the upregulation of SOCS3 in hippocampus. The last result in the reduction of the inflammatory response by inhibiting phosphorylation of AKT and STAT3, and the expression level of IL-17A in poly(I:C)-induced autism-like features in mice. The obtained data revealed the miR-301a as a critical participant in partial behavior phenotypes, which may exhibit a divergent role between gene knockout and knockdown. Our findings ascertain that miR-301a negatively regulates SOCS3 in MIA-induced autism in mice and could present a new therapeutic target for ameliorating the behavioral abnormalities of autism.

Quantitative proteomics analysis identified new interacting proteins of JAL30 in Arabidopsis.

Jacalin-related lectins (JALs) are a unique group of plant lectins derived from the jacalin protein family, which play important roles in plant defense responses. JAL30/PBP1 (PYK10 binding protein 1) interacts with inactive PYK10, exerting negative regulatory control over the size of the PYK10 complex, which is formed and activated upon insect or pathogen invasion. However, the precise interplay between JAL30 and other components remains elusive. In this study, we found JAL30 as a nucleocytoplasmic protein, but no obvious phenotype was observed in jal30-1 single mutant. Through immunoprecipitation (IP) enrichment combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS), dozens of new JAL30 interacting proteins were found in addition to several reported ones. Gene Ontology (GO) analysis revealed that these interacting proteins were highly related to the wounding and bacterial stimuli, suggesting their potential involvement in the jasmonate (JA) response. Importantly, the expression of JAL30 was induced by MeJA treatment, further highlighting its relevance in plant defense mechanisms. A novel JAL30 interacting protein, ESM1, was identified and its interaction with JAL30 was confirmed by Co-immunoprecipitation. Moreover, ESM1 was found as an O-GlcNAcylated protein, suggesting that JAL30 may possess glycosylated protein binding ability, particularly in O-GlcNAcylated protein and peptide recognition. Overall, our study provides valuable insights into the interacting protein network and biological function of JAL30, demonstrates the interaction between JAL30 and ESM1, and uncovers the potential significance of JAL30 in plant defense system, potentially through its association with PYK10 complex or JA response. SIGNIFICANCE: The biological functions of lectin proteins, including defense responses, immunity responses, signal transduction, have been well studied. Lectin proteins were also utilized to enrich glycosylated proteins for their specific carbohydrates binding capability. Jacalin-related lectins (JALs) were found to involve in plant defense mechanism. However, it is not yet clear whether JALs could use for enrichment of glycosylated proteins. In this study, we used label-free quantification method to identify interacting proteins of JAL30. A novel interacting protein, ESM1, as an O-GlcNAcylated protein was found. ESM1 has been reported to take part in defense against insect herbivory. Therefore, our findings provided experimental evidence to confirm that JALs have potential to be developed as the bio-tools to enrich glycosylated proteins. Finally, our data not only illustrated the vital biological role of JALs in plants, but also verified unique function of JAL30 in recognizing O-GlcNAcylated proteins.

Respiratory monitoring dataset, with rapid expiratory occlusions, over increasing positive airway pressure ventilation.

Resting breathing data was collected from 80 smokers, vapers, asthmatics, and otherwise healthy people in the low-risk clinical unit at the University of Canterbury. Subjects were asked to breathe normally through a full-face mask connected to a Fisher and Paykel Healthcare SleepStyle SPSCAA CPAP device. PEEP (Positive End-Expiratory Pressure) support was increased from 4 to 12 cmH2O in 0.5 cmH2O increments. Data was also collected during resting breathing at ZEEP (0 cmH2O) before and after the PEEP trial. The trial was conducted under University of Canterbury Human Research Ethics Committee consent (Ref: HREC 2023/04/LR-PS). Data was collected by and Dräeger PulmoVista 500 EIT machine and a custom Venturi-based pressure and flow sensor device connected in series with the CPAP and full-face mask. The outlined dataset includes pressure, flow, volume, dynamic circumference (thoracic and abdominal, and cross-sectional aeration. Subject demographic data was self-reported using a questionnaire given prior to the trial.

Influence of a Structural Prior Mask on EIT Image Reconstruction.

Electrical Impedance Tomography (EIT) is a low-cost imaging method with promising results in visualizing ventilation distribution within the lungs. However, in clinical settings, the interpretability of EIT images is often limited by blurred anatomical alignment and reconstruction artifacts. Integrating structural priors into the EIT reconstruction process can enhance the interpretability of EIT images. In this contribution, we introduced a patient-specific structural prior mask into the EIT reconstruction process. Such prior mask ensures that only conductivity changes within the lung regions are reconstructed. With the aim to investigate the influence of the structural prior mask on the EIT images, we conducted numerical simulations in terms of four different ventilation status. EIT images were reconstructed with Gauss-Newton algorithm and discrete cosine transform-based EIT algorithm. We carried out quantitative analysis including the reconstruction error and figures of merit for the evaluation. The results show that the morphological structures of the lungs introduced by the prior mask are preserved in the EIT images, and the reconstruction artefacts are also limited. In conclusion, the incorporation of the structural prior mask enhances the interpretability of EIT images in clinical settings.Clinical relevance-The correct interpretation of an EIT image is crucial for a clinical diagnosis. This research demonstrates that a structural prior mask might have the potential to improve the interpretability of an EIT image, which facilitates the clinicians with a better understanding of EIT results.

Structural priors represented by discrete cosine transform improve EIT functional imaging.

Structural prior information can improve electrical impedance tomography (EIT) reconstruction. In this contribution, we introduce a discrete cosine transformation-based (DCT-based) EIT reconstruction algorithm to demonstrate a way to incorporate the structural prior with the EIT reconstruction process. Structural prior information is obtained from other available imaging methods, e.g., thorax-CT. The DCT-based approach creates a functional EIT image of regional lung ventilation while preserving the introduced structural information. This leads to an easier interpretation in clinical settings while maintaining the advantages of EIT in terms of bedside monitoring during mechanical ventilation. Structural priors introduced in the DCT-based approach are of two categories in terms of different levels of information included: a contour prior only differentiates lung and non-lung region, while a detail prior includes information, such as atelectasis, within the lung area. To demonstrate the increased interpretability of the EIT image through structural prior in the DCT-based approach, the DCT-based reconstructions were compared with reconstructions from a widely applied one-step Gauss-Newton solver with background prior and from the advanced GREIT algorithm. The comparisons were conducted both on simulation data and retrospective patient data. In the simulation, we used two sets of forward models to simulate different lung conditions. A contour prior and a detail prior were derived from simulation ground truth. With these two structural priors, the reconstructions from the DCT-based approach were compared with the reconstructions from both the one-step Gauss-Newton solver and the GREIT. The difference between the reconstructions and the simulation ground truth is calculated by the ℓ2-norm image difference. In retrospective patient data analysis, datasets from six lung disease patients were included. For each patient, a detail prior was derived from the patient's CT, respectively. The detail prior was used for the reconstructions using the DCT-based approach, which was compared with the reconstructions from the GREIT. The reconstructions from the DCT-based approach are more comprehensive and interpretable in terms of preserving the structure specified by the priors, both in simulation and retrospective patient data analysis. In simulation analysis, the ℓ2-norm image difference of the DCT-based approach with a contour prior decreased on average by 34% from GREIT and 49% from the Gauss-Newton solver with background prior; for reconstructions of the DCT-based approach with detail prior, on average the ℓ2-norm image difference is 53% less than GREIT and 63% less than the reconstruction with background prior. In retrospective patient data analysis, the reconstructions from both the DCT-based approach and GREIT can indicate the current patient status, but the DCT-based approach yields more interpretable results. However, it is worth noting that the preserved structure in the DCT-based approach is derived from another imaging method, not from the EIT measurement. If the structural prior is outdated or wrong, the result might be misleadingly interpreted, which induces false clinical conclusions. Further research in terms of evaluating the validity of the structural prior and detecting the outdated prior is necessary.

Effect of a Patient-Specific Structural Prior Mask on Electrical Impedance Tomography Image Reconstructions.

Electrical Impedance Tomography (EIT) is a low-cost imaging method which reconstructs two-dimensional cross-sectional images, visualising the impedance change within the thorax. However, the reconstruction of an EIT image is an ill-posed inverse problem. In addition, blurring, anatomical alignment, and reconstruction artefacts can hinder the interpretation of EIT images. In this contribution, we introduce a patient-specific structural prior mask into the EIT reconstruction process, with the aim of improving image interpretability. Such a prior mask ensures that only conductivity changes within the lung regions are reconstructed. To evaluate the influence of the introduced structural prior mask, we conducted numerical simulations with two scopes in terms of their different ventilation statuses and varying atelectasis scales. Quantitative analysis, including the reconstruction error and figures of merit, was applied in the evaluation procedure. The results show that the morphological structures of the lungs introduced by the mask are preserved in the EIT reconstructions and the reconstruction artefacts are decreased, reducing the reconstruction error by 25.9% and 17.7%, respectively, in the two EIT algorithms included in this contribution. The use of the structural prior mask conclusively improves the interpretability of the EIT images, which could facilitate better diagnosis and decision-making in clinical settings.

Microporous melamine-formaldehyde networks loaded on rice husks for dynamic removal of organic micropollutants.

The alteration of agricultural wastes into novel adsorbents can stimulate their scalability in realistic application, showing great economic and environmental advantages. Here, we proposed a strategy to engineer rice husk (RH) with microporous melamine-formaldehyde networks (MFNs) resins and the utilization for dynamic removal of organic micropollutants rapidly and efficiently. was pre-treated to acquire attractive surface and unique hierarchical porosity, endowing with surface functionalization and essential filtering properties. MFNs can be uniformly generated in-situ on the fully exposed cellulose backbones of the pre-treated RH. MFNs granules functionalized RH (RH@MFNs) exhibited high removal efficiencies over 90% within 30 min for the adsorption of hazardous organic compounds (e.g., phenolic and antibiotic micropollutants) in static tests. Experiment results and density functional theory (DFT) simulation revealed that the synergy of hydrogen bonding, π-πinteraction, and micropore preservation dominates the adsorption. Further dynamic adsorption experiments showed that the removal efficiency and equilibrium removal capacity towards bisphenol A by RH@MFNs packed bed up-flow column were 2.6 and 67 times higher than that of raw RH, respectively. The column adsorption fits well with the Thomas model and bed depth service time (BDST) kinetic model. The inherent macropores inside RH and the roughness caused by the spiky structures and mesopores outside RH, as well as the accumulated MFNs granules, can lead to local turbulence of water flow around RH@MFNs, enabling fast and efficient adsorption. This sustainable and cost-effective preparation of RH-based adsorbents sheds light on the rational design of biomass waste adsorbents for realistic wastewater.

Influence of hyperparameter on the Untrue Prior Detection in Discrete Transformation-based EIT Algorithm.

Incorporated with a structural prior, discrete cosine transformation (DCT) based electrical impedance tomog-raphy (EIT) algorithm can improve the interpretability of EIT images in clinical settings. However, this benefit comes with a risk of the untrue prior which yields a misleading result compromising clinical decision. The redistribution index is able to detect an untrue prior by analysing EIT reconstructions. In addition to structural priors, EIT reconstruction is also affected by the choice of hyperparameter A in DCT-based EIT algorithm. In this research, influence of hyperparameter on untrue prior detection is investigated in terms of simulation experiment. A series of simulation settings consisting of 30 different atelectasis scales was conducted, then reconstructed with 20 different hyperparameters, to investigate the behavior of redistribution index. The result shows, despite the fact that redistribution index is indeed influenced by the choice of the hyperparameter A, the detection of an untrue prior is not significantly affected. The untrue prior detection is rather stable regardless of the optimal hyperparameter. Clinical Relevance - Optimal hyperparameter is not always guaranteed in clinical settings. This research confirms that the untrue prior detection is not strongly influenced by the hyperparameter. An update of untrue priors incorporated into EIT approach will facilitate a better interpretation of EIT results and an accurate clinical decision.

Loss of schizophrenia-related miR-501-3p in mice impairs sociability and memory by enhancing mGluR5-mediated glutamatergic transmission.

Schizophrenia is a polygenetic disease, the heterogeneity of which is likely complicated by epigenetic modifications yet to be elucidated. Here, we performed transcriptomic analysis of peripheral blood RNA from monozygotic twins discordant for schizophrenia and identified a schizophrenia-associated down-regulated microRNA, miR-501-3p. We showed that the loss of miR-501-3p in germline knockout (KO) male mice resulted in dendritic structure defects, glutamatergic transmission enhancement, and sociability, memory, and sensorimotor gating disruptions, which were attenuated when miR-501 expression was conditionally restored in the nervous system. Combining the results of proteomic analyses with the known genes linked to schizophrenia revealed that metabotropic glutamate receptor 5 (mGluR5) was one of the miR-501-3p targets and was elevated in vivo upon loss of miR-501. Treatment with the mGluR5 negative allosteric modulator 3-2((-methyl-4-thiazolyl) ethynyl) pyridine or the N-methyl-d-aspartate receptor antagonist 2-amino-5-phosphonopentanoic acid ameliorated the deficits observed in Mir501-KO mice. The epigenetic and pathophysiological mechanism that links miR-501-3p to the modulation of glutamatergic transmission provides etiological implications for schizophrenia.

Differentiating Phenotypes of Coronavirus Disease-2019 Pneumonia by Electric Impedance Tomography.

Introduction: Coronavirus disease-2019 (COVID-19) pneumonia has different phenotypes. Selecting the patient individualized and optimal respirator settings for the ventilated patient is a challenging process. Electric impedance tomography (EIT) is a real-time, radiation-free functional imaging technique that can aid clinicians in differentiating the "low" (L-) and "high" (H-) phenotypes of COVID-19 pneumonia described previously. Methods: Two patients ("A" and "B") underwent a stepwise positive end-expiratory pressure (PEEP) recruitment by 3 cmH2O of steps from PEEP 10 to 25 and back to 10 cmH2O during a pressure control ventilation of 15 cmH2O. Recruitment maneuvers were performed under continuous EIT recording on a daily basis until patients required controlled ventilation mode. Results: Patients "A" and "B" had a 7- and 12-day long trial, respectively. At the daily baseline, patient "A" had significantly higher compliance: mean ± SD = 53 ± 7 vs. 38 ± 5 ml/cmH2O (p < 0.001) and a significantly higher physiological dead space according to the Bohr-Enghoff equation than patient "B": mean ± SD = 52 ± 4 vs. 45 ± 6% (p = 0.018). Following recruitment maneuvers, patient "A" had a significantly higher cumulative collapse ratio detected by EIT than patient "B": mean ± SD = 0.40 ± 0.08 vs. 0.29 ± 0.08 (p = 0.007). In patient "A," there was a significant linear regression between the cumulative collapse ratios at the end of the recruitment maneuvers (R 2 = 0.824, p = 0.005) by moving forward in days, while not for patient "B" (R 2 = 0.329, p = 0.5). Conclusion: Patient "B" was recognized as H-phenotype with high elastance, low compliance, higher recruitability, and low ventilation-to-perfusion ratio; meanwhile patient "A" was identified as the L-phenotype with low elastance, high compliance, and lower recruitability. Observation by EIT was not just able to differentiate the two phenotypes, but it also could follow the transition from L- to H-type within patient "A." Clinical Trial Registration: www.ClinicalTrials.gov, identifier: NCT04360837.

A Comparison of Epileptogenic Effect of Status Epilepticus Treated With Diazepam, Midazolam, and Pentobarbital in the Mouse Pilocarpine Model of Epilepsy.

Status epilepticus (SE) is a medical emergency associated with acute severe systemic damage and high mortality. Moreover, symptomatic SE is one of the highest risk factors for epileptogenesis. While the antiepileptic drugs (AEDs) are chosen in favor of acute control of SE, the potential short-term and long-term effects of such AEDs have been ignored in clinics. In this study, we hypothesized that AEDs that are used to control acute SE might affect the feasibility for the chronic development of epileptogenesis after SE. Therefore, we sought to compare the epileptogenic effects of SE that are terminated by three AEDs, i.e., diazepam, midazolam, and pentobarbital, which are widely used as first-line anti-SE AEDs. For this purpose, we used a mouse model of SE induced by intraperitoneal (i.p.) injection of lithium chloride (LiCl)-pilocarpine. The pilocarpine-induced SE was terminated with diazepam, midazolam, or pentobarbital. Then we compared short-term and long-term effects of SE with different AED treatments by examining SE-associated mortality and behavioral spontaneous recurrent seizures (SRSs) and by using magnetic resonance imaging (MRI) and immunohistochemistry to evaluate pathological and cellular alterations of mice in the different treatment groups. We found that i.p. injections of diazepam (5 mg/kg), midazolam (10 mg/kg), and pentobarbital (37.5 mg/kg) were able to terminate acute pilocarpine-SE effectively, while pentobarbital treatment showed less neuroprotective action against lethality in the short phase following SE. Long-term evaluation following SE revealed that SE treated with midazolam had resulted in relatively less behavioral SRS, less hippocampal atrophy, and milder neuronal loss and gliosis. Our data revealed an obvious advantage of midazolam vs. diazepam or pentobarbital in protecting the brain from epileptogenesis. Therefore, if midazolam provides as strong action to quench SE as other AEDs in clinics, midazolam should be the first choice of anti-SE AEDs as it provides additional benefits against epileptogenesis.

Nucleus accumbens shell modulates seizure propagation in a mouse temporal lobe epilepsy model.

Temporal lobe epilepsy (TLE) is the most common form of epilepsy with focal seizures which in some conditions can develop into secondarily generalized tonic-clonic seizures by the propagation of epileptic activities in the temporal lobe to other brain areas. The nucleus accumbens (NAc) has been suggested as a treatment target for TLE as accumulating evidence indicates that the NAc, especially its shell, participates in the process of epileptic seizures of patients and animal models with TLE. The majority of neurons in the NAc are GABAergic medium spiny neurons (MSNs) expressing dopamine receptor D1 (D1R) or dopamine receptor D2 (D2R). However, the direct evidence of the NAc shell participating in the propagation of TLE seizures is missing, and its cell type-specific modulatory roles in TLE seizures are unknown. In this study, we microinjected kainic acid into basolateral amygdala (BLA) to make a mouse model of TLE with initial focal seizures and secondarily generalized seizures (SGSs). We found that TLE seizures caused robust c-fos expression in the NAc shell and increased neuronal excitability of D1R-expressing MSN (D1R-MSN) and D2R-expressing MSN (D2R-MSN). Pharmacological inhibition of the NAc shell alleviated TLE seizures by reducing the number of SGSs and seizure stages. Cell-type-specific chemogenetic inhibition of either D1R-MSN or D2R-MSN showed similar effects with pharmacological inhibition of the NAc shell. Both pharmacological and cell-type-specific chemogenetic inhibition of the NAc shell did not alter the onset time of focal seizures. Collectively, these findings indicate that the NAc shell and its D1R-MSN or D2R-MSN mainly participate in the propagation and generalization of the TLE seizures.

Astroglial CB1 Cannabinoid Receptors Mediate CP 55,940-Induced Conditioned Place Aversion Through Cyclooxygenase-2 Signaling in Mice.

Cannabinoids (CBs), such as phytocannabinoids, synthetic CBs, and endogenous CBs, can be neuroprotective, rewarding, or aversive. The aversive effects of CBs may hinder their medical and recreational applications. It is unknown which type of CB receptors mediates the direct aversive effects of synthetic CB CP 55,940 which is an analog of Δ9-tetrahydrocannabinol, the major psychoactive component of marijuana. In this study, we address this question by taking the advantage of systematic type 1 CB receptor (CB1R) knockout mice and conditional reinstatement of this receptor only in astrocytes. We show that CP 55,940 at a concentration of 1 mg/kg induces conditioned place aversion (CPA) and the CPA effect of CP 55,940 is mediated by the astroglial CB1Rs. Inhibiting cyclooxygenase-2 (COX-2) eliminates CP 55,940-induced CPA in mice that only express CB1Rs in astrocytes. These findings conclude that CPA effect of CP 55,940 is mediated by the astroglial CB1Rs through COX-2 signaling, suggesting that selective COX-2 inhibition or precise isolation of astroglial CB1R activity may be the strategy for treating aversive response of medical and recreational administrations of marijuana.

Redistribution Index - Detection of an Outdated Prior Information in the Discrete Cosine Transformation-based EIT Algorithm.

The morphological prior information incorporated with the discrete cosine transformation (DCT) based electrical impedance tomography (EIT) algorithm can improve the interpretability of the EIT results in clinical settings. However, an outdated prior information can yield a misleading result compromising the accuracy of the clinical decisions. Detection of the outdated prior information is critical in the DCT-based EIT algorithm. In this contribution, a redistribution index calculated from the DCT approach result was proposed to quantify the possible error induced by the morphological prior information. Two simulations in terms of different atelectasis and collapse scales were conducted to evaluate the plausibility of the redistribution index. Thus, an experiential threshold for redistribution index was adopt as an indicator to the outdated prior in DCT-based EIT approach. A retrospective research was conducted with the seven-day patient monitor data as a proof-of-concept to verify plausibility and comparability of the redistribution index. From the evaluation, the redistribution index qualifies the function as an indicator for the outdated prior in the DCT-based EIT approach.Clinical relevance- This establishes an indicator to advice an update to the morphological prior information embedded in EIT approach, which lower the risk misleading interpretation of EIT results in mechanical ventilation monitoring.

The Conformationally Sensitive Spatial Distance Between the TM3-4 Loop and Transmembrane Segment 7 in the Glutamate Transporter Revealed by Paired-Cysteine Mutagenesis.

Excitatory amino acid transporters can maintain extracellular glutamate concentrations lower than neurotoxic levels by transferring neurotransmitters from the synaptic cleft into surrounding glial cells and neurons. Previous work regarding the structural studies of Glt Ph , Glt TK , excitatory amino acid transporter 1 (EAAT1), EAAT3 and alanine serine cysteine transporter 2 described the transport mechanism of the glutamate transporter in depth. However, much remains unknown about the role of the loop between transmembrane segment 3 and 4 during transport. To probe the function of this loop in the transport cycle, we engineered a pair of cysteine residues between the TM3-TM4 loop and TM7 in cysteine-less EAAT2. Here, we show that the oxidative cross-linking reagent CuPh inhibits transport activity of the paired mutant L149C/M414C, whereas DTT inhibits the effect of CuPh on transport activity of L149C/M414C. Additionally, we show that the effect of cross-linking in the mutant is due to the formation of the disulfide bond within the molecules of EAAT2. Further, L-glutamate or KCl protect, and D,L-threo-ß-benzyloxy-aspartate (TBOA) increases, CuPh-induced inhibition in the L149C/M414 mutant, suggesting that the L149C and M414C cysteines are closer or farther away in the outward- or inward-facing conformations, respectively. Together, our findings provide evidence that the distance between TM3-TM4 loop and TM7 alter when substrates are transported.

Application of genetic screening processor in screening neonatal glucose-6-phosphate dehydrogenase deficiency.

To evaluate the performance of genetic screening processor (GSP analyzer) in neonatal screening for glucose-6-phosphate dehydrogenase (G6PD)deficiency. The accuracy and precision of GSP analyzer was evaluated with the control materials from National Center for Clinical Laboratories and the low and high quality G6PD control kit (fluorescence analysis). GSP analyzer and semi-automatic fluorescence immunoanalyzer (1420 analyzer) were simultaneously used to detect 2622 neonatal screening samples and 41 confirmed samples to analyze the correlation and consistency of the test results; 78 floating samples and 78 non-floating samples were detected to compare the result. A total of 1 100 384 neonatal screening samples from January 2017 to December 2018 and 855 856 neonatal screening samples from January 2019 to December 2020 were detected with 1420 analyzer and GSP analyzer, respectively. Referring to the percentile method and the expert consensus, the new cut-off value of GSP analyzer for G6PD deficiency in screening was established. The relative bias of GSP analyzer in detecting G6PD was 0.71%-4.23%; the intra assay precision was 4.34%-4.91%, the inter assay precision was 0.85%-2.12%, and the total coefficient of variation was 5.44%-5.72%. There was a significant positive correlation between G6PD activity detected by GSP analyzer and 1420 analyzer (=0.740, <0.01). Forty-one clinical confirmed patients were identified by both 1420 analyzer and GSP analyzer (=0.945). The G6PD activity in floating dry blood spots detected by 1420 analyzer was significantly lower than that in non-floating dry blood spots (<0.05), but there was no significant difference in G6PD activity between floating and non-floating dry blood spots detected by GSP analyzer (>0.05). The sensitivities of GSP analyzer and 1420 analyzer in screening G6PD deficiency were both 100.00%, and the specificities were both more than 99.80%. Compared with 1420 analyzer, the positive predictive value, positive rate and prevalence of G6PD deficiency detected by GSP analyzer were increased, and the false positive rate was decreased (all <0.01). The new cut-off value was 26.1 U/dL for male and 29.1 U/dL for female according to the 99.1% percentile of the population. GSP analyzer has better detection performance with high automation, efficiency and throughput, which can be used in large-scale screening for neonatal G6PD deficiency.

MicroRNA 223 Targeting ATG16L1 Affects Microglial Autophagy in the Kainic Acid Model of Temporal Lobe Epilepsy.

This study aimed to explore whether microRNA (miR) 223 affects microglial autophagy by targeting autophagy-related 16-like 1 (ATG16L1) in the kainic acid (KA) model of temporal lobe epilepsy (TLE). The miRNA and mRNA expression levels were quantified using quantitative real-time polymerase chain reaction (qRT-PCR), and the protein expression was investigated using western blotting. A dual-luciferase reporter assay was used to test the direct interaction between miR 223 and ATG16L1. In situ hybridization was performed to measure the hippocampal expression of miR 223. We used immunofluorescence staining to assess the expression of ATG16L1 and microtubule-associated protein light chain 3 (LC3) in the murine hippocampal microglia. Inhibitor of miR 223 was utilized to investigate the role of miR 223 in TLE, and the epileptic activity was assessed using electroencephalography (EEG). The autophagosomes were observed by transmission electron microscopy. In patients with TLE, the murine KA model of TLE, and the KA-stimulated BV2 cells, miR 223, and sequestosome 1 (SQSTM1/P62) expressions were remarkably increased, whereas ATG16L1 and LC3 levels were significantly decreased. Using a dual-luciferase reporter assay, ATG16L1 was determined as a direct target of miR 223. Treatment with antagomir 223 alleviated epilepsy, prevented abnormalities in EEG recordings and increased the ATG16L1 and LC3 levels in KA-treated mice. Inhibition of miR 223 induced increased autophagy in BV2 cells upon Rapamycin stimulation. These findings show that miR 223 affects microglial autophagy via ATG16L1 in the KA model of TLE. The miR 223/ATG16L1 pathway may offer a new treatment option for TLE.

Optimal Brassinosteroid Levels Are Required for Soybean Growth and Mineral Nutrient Homeostasis.

Brassinosteroids (BRs) are steroid phytohormones that are known to regulate plant growth and nutrient uptake and distribution. However, how BRs regulate nutrient uptake and balance in legume species is not fully understood. Here, we show that optimal BR levels are required for soybean (Glycine max L.) seedling growth, as treatments with both 24-epicastasterone (24-epiCS) and the BR biosynthesis inhibitor propiconazole (PPZ) inhibit root growth, including primary root elongation and lateral root formation and elongation. Specifically, 24-epiCS and PPZ reduced the total phosphorus and potassium levels in the shoot and affected several minor nutrients, such as magnesium, iron, manganese, and molybdenum. A genome-wide transcriptome analysis identified 3774 and 4273 differentially expressed genes in the root tip after brassinolide and PPZ treatments, respectively. The gene ontology (GO) analysis suggested that genes related to "DNA-replication", "microtubule-based movement", and "plant-type cell wall organization" were highly responsive to the brassinolide and PPZ treatments. Furthermore, consistent with the effects on the nutrient concentrations, corresponding mineral transporters were found to be regulated by BR levels, including the GmPHT1s, GmKTs, GmVIT2, GmZIPs, and GmMOT1 genes. Our study demonstrates that optimal BR levels are important for growth and mineral nutrient homeostasis in soybean seedlings.