Protocol for adeno-associated virus-mediated optogenetic activation of olfactory output neurons in neonatal mice.

Optogenetic manipulation has proven a powerful tool for investigating the mechanisms underlying the function of neuronal networks, but implementing the technique on mammals during early development remains challenging. Here, we present a comprehensive workflow to specifically manipulate mitral/tufted cells (M/TCs), the output neurons in the olfactory circuit, mediated by adeno-associated virus (AAV) transduction and light stimulation in neonatal mice and monitor neuronal and network activity with in vivo electrophysiology. This method represents an efficient approach to elucidate functional brain development. For complete details on the use and execution of this protocol, please refer to Chen et al.1,2,3.

Preconfigured architecture of the developing mouse brain.

In the adult brain, structural and functional parameters, such as synaptic sizes and neuronal firing rates, follow right-skewed and heavy-tailed distributions. While this organization is thought to have significant implications, its development is still largely unknown. Here, we address this knowledge gap by investigating a large-scale dataset recorded from the prefrontal cortex and the olfactory bulb of mice aged 4-60 postnatal days. We show that firing rates and spike train interactions have a largely stable distribution shape throughout the first 60 postnatal days and that the prefrontal cortex displays a functional small-world architecture. Moreover, early brain activity exhibits an oligarchical organization, where high-firing neurons have hub-like properties. In a neural network model, we show that analogously right-skewed and heavy-tailed synaptic parameters are instrumental to consistently recapitulate the experimental data. Thus, functional and structural parameters in the developing brain are already extremely distributed, suggesting that this organization is preconfigured and not experience dependent.

Azonia-Naphthalene: A Cationic Hydrophilic Building Block for Stable N-Type Organic Mixed Ionic-Electronic Conductors.

Conjugated polymers that can efficiently transport both ionic and electronic charges have broad applications in next-generation optoelectronic, bioelectronic, and energy storage devices. To date, almost all the conjugated polymers have hydrophobic backbones, which impedes efficient ion diffusion/transport in aqueous media. Here, we design and synthesize a novel hydrophilic polymer building block, 4a-azonia-naphthalene (AN), drawing inspiration from biological systems. Because of the strong electron-withdrawing ability of AN, the AN-based polymers show typical n-type charge transport behaviors. We find that cationic aromatics exhibit strong cation-π interactions, leading to smaller π-π stacking distance, interesting ion diffusion behavior, and good morphology stability. Additionally, AN enhances the hydrophilicity and ionic-electronic coupling of the polymer, which can help to improve ion diffusion/injection speed, and operational stability of organic electrochemical transistors (OECTs). The integration of cationic building blocks will undoubtedly enrich the material library for high-performance n-type conjugated polymers.

Olfactory bulb activity shapes the development of entorhinal-hippocampal coupling and associated cognitive abilities.

The interplay between olfaction and higher cognitive processing has been documented in the adult brain; however, its development is poorly understood. In mice, shortly after birth, endogenous and stimulus-evoked activity in the olfactory bulb (OB) boosts the oscillatory entrainment of downstream lateral entorhinal cortex (LEC) and hippocampus (HP). However, it is unclear whether early OB activity has a long-lasting impact on entorhinal-hippocampal function and cognitive processing. Here, we chemogenetically silenced the synaptic outputs of mitral/tufted cells, the main projection neurons in the OB, during postnatal days 8-10. The transient manipulation leads to a long-lasting reduction of oscillatory coupling and weaker responsiveness to stimuli within developing entorhinal-hippocampal circuits accompanied by dendritic sparsification of LEC pyramidal neurons. Moreover, the transient silencing reduces the performance in behavioral tests involving entorhinal-hippocampal circuits later in life. Thus, neonatal OB activity is critical for the functional LEC-HP development and maturation of cognitive abilities.

Prediction model for day 3 embryo implantation potential based on metabolites in spent embryo culture medium.

BACKGROUND: Metabolites in spent embryo culture medium correlate with the embryo's viability. However, there is no widely accepted method using metabolite dada to predict successful implantation. We sought to combine metabolomic profiling of spent embryo culture medium and clinical variables to create an implantation prediction model as an adjunct to morphological screening of day 3 embryos. METHODS: This investigation was a prospective, nested case-control study. Forty-two day 3 embryos from 34 patients were transferred, and the spent embryo culture medium was collected. Twenty-two embryos implanted successfully, and the others failed. Metabolites in the medium relevant to implantation were detected and measured by Liquid Chromatography-Mass Spectrometry. Clinical signatures relevant to embryo implantation were subjected to univariate analysis to select candidates for a prediction model. Multivariate logistical regression of the clinical and metabolomic candidates was used to construct a prediction model for embryo implantation potential. RESULTS: The levels of 13 metabolites were significantly different between the successful and failed groups, among which five were most relevant and interpretable selected by Least Absolute Shrinkage and Selection Operator regression analysis. None of the clinical variables significantly affected day 3 embryo implantation. The most relevant and interpretable set of metabolites was used to construct a prediction model for day 3 embryo implantation potential with an accuracy of 0.88. CONCLUSIONS: Day 3 embryos'implantation potential could be noninvasively predicted by the spent embryo culture medium's metabolites measured by LC-MS. This approach may become a useful adjunct to morphological evaluation of day 3 embryos.

In-source fragmentation of nucleosides in electrospray ionization towards more sensitive and accurate nucleoside analysis.

Nucleosides have been found to suffer in-source fragmentation (ISF) in electrospray ionization mass spectrometry, which leads to reduced sensitivity and ambiguous identification. In this work, a combination of theoretical calculations and nuclear magnetic resonance analysis revealed the key role of protonation at N3 near the glycosidic bond during ISF. Therefore, an ultrasensitive liquid chromatography-tandem mass spectrometry system for 5-formylcytosine detection was developed with 300 fold signal enhancement. Also, we established a MS1-only platform for nucleoside profiling and successfully identified sixteen nucleosides in the total RNA of MCF-7 cells. Taking ISF into account, we can realize analysis with higher sensitivity and less ambiguity, not only for nucleosides, but for other molecules with similar protonation and fragmentation behaviors.

Ultrasensitive HPLC-MS Quantification of S-(2-Succino) Cysteine Based on Ethanol/Acetyl Chloride Derivatization in Fumarate Accumulation Cells.

Succination is a nonenzymatic and irreversible post-translational modification (PTM) with important biological significance, yielding S-(2-succino) cysteine (2SC) residue. This PTM is low in abundance and often requires a large amount of protein samples for 2SC quantification. In this work, an efficient quantification method based on ethanol/acetyl chloride chemical derivatization was developed. The three carboxyl groups of 2SC were all esterified to increase hydrophobicity, greatly improving its ionization efficiency. The sensitivity was increased by 112 times; the limit of detection was reduced to 0.885 fmol, and the protein usage was reduced by at least 10 times. The established method was used to detect the overall concentration of 2SC in fumarate accumulation cells quantitatively.

Preparation of Magnesium Hydroxide Flame Retardant from Hydromagnesite and Enhance the Flame Retardant Performance of EVA.

In this study, hydromagnesite, a rare natural hydrated alkaline magnesium carbonate, was used to synthesize magnesium hydroxide (MH) as a flame retardant for ethylene-vinyl acetate (EVA) to enhance its fire resistance and smoke suppression. Various concentrations of sodium hydroxide (NaOH) were used to alter the morphology and the flame-retardant efficiency of synthesized MH. EVA/MH composites were prepared through melt blending, and the influence of NaOH on the flame retardancy and mechanical properties was investigated by means of the limiting oxygen index (LOI), cone calorimeter test (CCT) and tensile test. The flame retardancy results demonstrated that composites exhibited remarkably improved flame retardant properties after introducing MH, reflected by an increase in the LOI value from 20% for neat EVA to roughly 38%. Additionally, the peak of heat release rate (pHRR), the total heat release (THR) and the peak of the smoke production rate for EVA3 were decreased by 37.6%, 20.7% and 44.4% compared with neat EVA, respectively. In the meantime, increasing char residues were also observed. The incorporation of different MH concentrations had a limited effect on the mechanical properties of the EVA/MH composites.

Simultaneous measurement of phenols by three-way fluorescence spectroscopy: A comparison of N-PLS/RBL, U-PLS/RBL and PARAFAC.

Phenol, o-cresol, p-cresol, catechol and resorcinol are five phenolic compounds with extremely similar structure. Their fluorescence spectra are hard to be analyzed because of the serious spectral overlaps between any two of the five phenolic components in the mixture system. In this experiment, multi-dimensional partial least-squares (N-PLS), unfolded partial least-squares (U-PLS) with residual bilinearization (RBL) and parallel factor analysis (PARAFAC) are employed to analyze the three-way fluorescence spectra aiming to achieve quantitative results. Meanwhile, a contrast of these three methods is given. The experiment results show that N-PLS/RBL and U-PLS/RBL algorithms are superior to PARARFAC in terms of analysis of highly overlapping three-way fluorescence spectra for concentration determination.

Developing a multispectral model for detection of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) changes in fish fillet using physarum network and genetic algorithm (PN-GA) method.

A multispectral model for the detection of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) changes in grass carp and salmon fillet was developed using physarum network and genetic algorithm (PN-GA) method for the first time. Partial least-squares regression (PLSR), multiple linear regressions (MLR), and principal component regression (PCR) algorithms were used to predict the DHA and EPA using optimal wavelengths selected by PN-GA. The MLR models showed the best DHA prediction results for both grass carp and salmon fillets, and also showed good prediction for EPA in grass carp fillet but poor prediction in salmon fillet. The MLR models were then applied for visualizing the spatial distribution of DHA and EPA changes in two fish fillets. The current results demonstrated that a developed multispectral imaging system could be feasibly constructed for DHA and EPA measurement in fish species with the optimal wavelengths selected by PN-GA method.

Study on Three-Dimensional Fluorescence Spectral Characteristics of Heterocyclic Pesticides in Different Environmental Conditions.

The impact analysis of different environments on the fluorescence emission spectrum of pesticides is critical in detecting the concentration of pesticides. In this paper, three kinds of pesticides, carbendazim, carbaryl and fuberidazole, were selected as the research objects. Under different environment, such as different pH values and the presence of different common anion or cation, three-dimensional fluorescence spectral emission (EEM) characteristic of pesticides were analyzed. The experimental results showed that the primary fluorescence peaks for three kinds of pesticides were at λex/λem=280/300, 310/340 and 280/335 nm (respectively); Carbendazim and fuberidazole had a secondary peak at 245/305 nm (PeakB) and 250/340 nm (PeakB). We can come to the conclusion that with the change of pH value, the characteristic of fluorescence emission of carbendazim and fuberidazole is similar. We can find that the fluorescence intensities of carbendazim and fuberidazole were enhanced with the declining of the solution acidity or alkalinity and the fluorescence intensity of carbaryl had not changed with the declining of the solution acidity, but it increased with the declining of the solution alkalinity; the fluorescence emission spectra of the three kinds of pesticides had good fluorescence characteristics with the scope of the pH varying from 6.16 to 7.4. Twelve common ions in water (CO2-3，SO2-4，NO-3，Cl-，HPO2-4，HCO-3，Mg2+，Zn2+，NH+4，Na+，Ca2+，K+) had no significant effect on fluorescence emission characteristics of carbendazim and fuberidazole. The fluorescence intensities were seriously influenced by Fe3+ and Cu2+. The results showed that the pesticides fluorescence intensities were decreased with the ion concentration increasing. It was necessary to consider the quenching effects on pesticides of Fe3+ and Cu2+for the analytic results. The obtained results provided the basic research for improving the accuracy of the heterocyclic pesticides measurement in water.

Suitability of hyperspectral imaging for rapid evaluation of thiobarbituric acid (TBA) value in grass carp (Ctenopharyngodon idella) fillet.

The suitability of hyperspectral imaging technique (400-1000 nm) was investigated to determine the thiobarbituric acid (TBA) value for monitoring lipid oxidation in fish fillets during cold storage at 4°C for 0, 2, 5, and 8 days. The PLSR calibration model was established with full spectral region between the spectral data extracted from the hyperspectral images and the reference TBA values and showed good performance for predicting TBA value with determination coefficients (R(2)P) of 0.8325 and root-mean-square errors of prediction (RMSEP) of 0.1172 mg MDA/kg flesh. Two simplified PLSR and MLR models were built and compared using the selected ten most important wavelengths. The optimised MLR model yielded satisfactory results with R(2)P of 0.8395 and RMSEP of 0.1147 mg MDA/kg flesh, which was used to visualise the TBA values distribution in fish fillets. The whole results confirmed that using hyperspectral imaging technique as a rapid and non-destructive tool is suitable for the determination of TBA values for monitoring lipid oxidation and evaluation of fish freshness.