Neuromodulatory Compensation of Cortical Neural Activity on Electrodeposited Pt/Ir Modified Microelectrode Arrays for Temperature Transients.

Temperature has a profound influence on various neuromodulation processes and has emerged as a focal point. However, the effects of acute environmental temperature fluctuations on cultured cortical networks have been inadequately elucidated. To bridge this gap, we have developed a brain-on-a-chip platform integrating cortical networks and electrodeposited Pt/Ir modified microelectrode arrays (MEAs) with 3D-printed bear-shaped triple chambers, facilitating control of temperature transients. This innovative system administers thermal stimuli while concurrently monitoring neuronal activity, including spikes and local field potentials, from 60 microelectrodes (diameter: 30 µm; impedance: 9.34 ± 1.37 kΩ; and phase delay: -45.26 ± 2.85°). Temperature transitions of approximately ±10 °C/s were applied to cortical networks on MEAs via in situ perfusion within the triple chambers. Subsequently, we examined the spatiotemporal dynamics of the brain-on-a-chip under temperature regulation at both the group level (neuronal population) and their interactions (network dynamics) and the individual level (cellular activity). Specifically, we found that after the temperature reduction neurons enhanced the overall information transmission efficiency of the network through synchronous firing to compensate for the decreased efficiency of single-cell level information transmission, in contrast to temperature elevation. By leveraging the integration of high-performance MEAs with perfusion chambers, this investigation provides a comprehensive understanding of the impact of temperature on the spatiotemporal dynamics of neural networks, thereby facilitating future exploration of the intricate interplay between temperature and brain function.

Cellulose-Encapsulated Composite Electrolyte Design: Toward Chemically and Mechanically Enhanced Solid-Sodium Batteries.

Sulfide- and halide-based ceramic ionic conductors exhibit comparable ionic conductivity with liquid electrolytes and are candidates for high-energy- and high-power-density all-solid-state batteries. These materials, however, are inherently brittle, making them unfavorable for applications. Here, we report a mechanically enhanced composite Na+ conductor that contains 92.5 wt % of sodium thioantimonate (Na3SbS4, NSS) and 7.5 wt % of sodium carboxymethyl cellulose (CMC); the latter serves as the binder and an electrochemically inert encapsulation layer. The ceramic and binder constituents were integrated at the particle level, providing ceramic NSS-level Na+ conductivity in the NSS-CMC composite. The more than 5-fold decrease of electrolyte thickness obtained in NSS-CMC composite provided a 5-fold increase in Na+ conductance compared to NSS ceramic pellets. As a result of the CMC encapsulation, this NSS-CMC composite shows increased moisture resistivity and electrochemical stability, which significantly promotes the cycling performance of NSS-based solid-state batteries. This work demonstrates a well-controlled, orthogonal process of ceramic-rich, composite electrolyte processing: independent streams for ceramic particle formation along with binder encapsulation in a solvent-assisted environment. This work also provides insights into the interplay among the solvent, the polymeric binder, and the ceramic particles in composite electrolyte synthesis and implies the critical importance of identifying the appropriate solvent/binder system for precise control of this complicated process.

Dual-Component Interlayer Enables Uniform Lithium Deposition and Dendrite Suppression for Solid-State Batteries.

ß-Lithium thiophosphate (LPS) exhibits high Li+ conductivity and has been identified as a promising ceramic electrolyte for safe and high-energy-density all-solid-state batteries. Integrating LPS into solid-state lithium (Li) batteries would enable the use of a Li electrode with the highest deliverable capacity. However, LPS-based batteries operate at a limited current density before short-circuiting, posing a major challenge for the development of application-relevant batteries. In this work, we designed a dual-component interfacial protective layer called LiSn-LiN that forms in situ between the Li electrode and LPS electrolyte. The LiSn component, Li22Sn5, exhibits enhanced Li diffusivity compared with the metallic lithium and facilitates a more uniform lithium deposition across the electrode surface, thus eliminating Li dendrite formation. Meanwhile, the LiN component, Li3N, shows enhanced mechanical stiffness compared with LPS and functions to suppress dendrite penetration. This chemically robust LiSn-LiN interlayer provides a more than doubled deliverable critical current density compared to systems without interfacial protection. Through combined XPS and XAFS analyses, we determined the local structure and the formation kinetics of the key functional Li22Sn5 phase formed via the electrochemical reduction of a Sn3N4 precursor. This work demonstrates an example of the structural-specific design of a protective interlayer with a desired function - dendrite suppression. The structure of a functional protective layer for a given solid-state battery should be tailored based on the given battery configuration and its unique interfacial chemistry.

Investigating Communication Dynamics in Neuronal Network using 3D Gold Microelectrode Arrays.

Although in vitro neuronal network models hold great potential for advancing neuroscience research, with the capacity to provide fundamental insights into mechanisms underlying neuronal functions, the dynamics of cell communication within such networks remain poorly understood. Here, we develop a customizable, polymer modified three-dimensional gold microelectrode array with sufficient stability for high signal-to-noise, long-term, neuronal recording of cultured networks. By using directed spatial and temporal patterns of electrical stimulation of cells to explore synaptic-based communication, we monitored cell network dynamics over 3 weeks, quantifying communication capability using correlation heatmaps and mutual information networks. Analysis of synaptic delay and signal speed between cells enabled us to establish a communication connectivity model. We anticipate that our discoveries of the dynamic changes in communication across the neuronal network will provide a valuable tool for future studies in understanding health and disease as well as in developing effective platforms for evaluating therapies.

SWCNTs/PEDOT:PSS nanocomposites-modified microelectrode arrays for revealing locking relations between burst and local field potential in cultured cortical networks.

Burst and local field potential (LFP) are fundamental components of brain activity, representing fast and slow rhythms, respectively. Understanding the intricate relationship between burst and LFP is crucial for deciphering the underlying mechanisms of brain dynamics. In this study, we fabricated high-performance microelectrode arrays (MEAs) using the SWCNTs/PEDOT:PSS nanocomposites, which exhibited favorable electrical properties (low impedance: 12.8 ± 2.44 kΩ) and minimal phase delay (-11.96 ± 1.64°). These MEAs enabled precise exploration of the burst-LFP interaction in cultured cortical networks. After a 14-day period of culture, we used the MEAs to monitor electrophysiological activities and revealed a time-locking relationship between burst and LFP, indicating the maturation of the neural network. To further investigate this relationship, we modulated burst firing patterns by treating the neural culture with increasing concentrations of glycine. The results indicated that glycine effectively altered burst firing patterns, with both duration and spike count increasing as the concentration rose. This was accompanied by an enhanced level of time-locking between burst and LFP but a decrease in synchrony among neurons. This study not only highlighted the pivotal role of SWCNTs/PEDOT:PSS-modified MEAs in elucidating the interaction between burst and LFP, bridging the gap between slow and fast brain rhythms in vitro but also provides valuable insights into the potential therapeutic strategies targeting neurological disorders associated with abnormal rhythm generation.

Comparative transcriptome analysis reveals growth and molecular pathway of body color regulation in turbot (Scophthalmus maximus) exposed to different light spectrum.

Fish body color changes play vital roles in adapting to ecological light environment and influencing market value. However, the initial mechanisms governing the changes remain unknown. Here, we scrutinized the impact of light spectrum on turbot (Scophthalmus maximus) body coloration, exposing them to red, blue, and full light spectra from embryo to 90 days post hatch. Transcriptome and quantitative real-time PCR (qRT-PCR) analyses were employed to elucidate underlying biological processes. The results showed that red light induced dimorphism in turbot juvenile skin pigmentation: some exhibited black coloration (Red_Black_Surface, R_B_S), while others displayed lighter skin (Red_White_Bottom, R_W_B), with red light leading to reduced skin lightness (L*) and body weight, particularly in R_B_S group. Transcriptomic and qRT-PCR analyses showcased upregulated gene expressions related to melanin synthesis in R_B_S individuals, notably tyrosinase (tyr), tyrosinase-related protein 1 (tyrp1), and dopachrome tautomerase (dct), alongside solute carrier family 24 member 5 (slc24a5) and oculocutaneous albinism type II (oca2) as pivotal regulators. Nervous system emerged as a critical mediator in spectral environment-driven color regulation. N-methyl d-aspartate (NMDA) glutamate receptor, and calcium signaling pathway emerged as pivotal links intertwining spectral conditions, neural signal transduction, and color regulation. The individual differences in NMDA glutamate receptor expression and subsequent neural excitability seemed responsible for dichromatic body coloration in red light-expose juveniles. This study provides new insights into the comprehending of fish adaptation to environment and methods for fish body color regulation and could potentially help enhance the economic benefit of fish farming industry.

2D Hybrid Perovskites Employing an Organic Cation Paired with a Neutral Molecule.

Two-dimensional (2D) hybrid perovskites harness the chemical and structural versatility of organic compounds. Here, we explore 2D perovskites that incorporate both a first organic component, a primary ammonium cation, and a second neutral organic module. Through the experimental examination of 42 organic pairs with a range of functional groups and organic backbones, we identify five crystallization scenarios that occur upon mixing. Only one leads to the cointercalation of the organic modules with distinct and extended interlayer spacing, which is observed with the aid of X-ray diffraction (XRD) pattern analysis combined with cross-sectional transmission electron microscopy (TEM) and elemental analysis. We present a picture in which complementary pairs, capable of forming intermolecular bonds, cocrystallize with multiple structural arrangements. These arrangements are a function of the ratio of organic content, annealing temperature, and substrate surface characteristics. We highlight how noncovalent bonds, particularly hydrogen and halogen bonding, enable the influence over the organic sublattice in hybrid halide perovskites.

Using Human-Induced Pluripotent Stem Cell Derived Neurons on Microelectrode Arrays to Model Neurological Disease: A Review.

In situ physiological signals of in vitro neural disease models are essential for studying pathogenesis and drug screening. Currently, an increasing number of in vitro neural disease models are established using human-induced pluripotent stem cell (hiPSC) derived neurons (hiPSC-DNs) to overcome interspecific gene expression differences. Microelectrode arrays (MEAs) can be readily interfaced with two-dimensional (2D), and more recently, three-dimensional (3D) neural stem cell-derived in vitro models of the human brain to monitor their physiological activity in real time. Therefore, MEAs are emerging and useful tools to model neurological disorders and disease in vitro using human iPSCs. This is enabling a real-time window into neuronal signaling at the network scale from patient derived. This paper provides a comprehensive review of MEA's role in analyzing neural disease models established by hiPSC-DNs. It covers the significance of MEA fabrication, surface structure and modification schemes for hiPSC-DNs culturing and signal detection. Additionally, this review discusses advances in the development and use of MEA technology to study in vitro neural disease models, including epilepsy, autism spectrum developmental disorder (ASD), and others established using hiPSC-DNs. The paper also highlights the application of MEAs combined with hiPSC-DNs in detecting in vitro neurotoxic substances. Finally, the future development and outlook of multifunctional and integrated devices for in vitro medical diagnostics and treatment are discussed.

Exploring retinal ganglion cells encoding to multi-modal stimulation using 3D microelectrodes arrays.

Microelectrode arrays (MEA) are extensively utilized in encoding studies of retinal ganglion cells (RGCs) due to their capacity for simultaneous recording of neural activity across multiple channels. However, conventional planar MEAs face limitations in studying RGCs due to poor coupling between electrodes and RGCs, resulting in low signal-to-noise ratio (SNR) and limited recording sensitivity. To overcome these challenges, we employed photolithography, electroplating, and other processes to fabricate a 3D MEA based on the planar MEA platform. The 3D MEA exhibited several improvements compared to planar MEA, including lower impedance (8.73 ± 1.66 kΩ) and phase delay (-15.11° ± 1.27°), as well as higher charge storage capacity (CSC = 10.16 ± 0.81 mC/cm2), cathodic charge storage capacity (CSCc = 7.10 ± 0.55 mC/cm2), and SNR (SNR = 8.91 ± 0.57). Leveraging the advanced 3D MEA, we investigated the encoding characteristics of RGCs under multi-modal stimulation. Optical, electrical, and chemical stimulation were applied as sensory inputs, and distinct response patterns and response times of RGCs were detected, as well as variations in rate encoding and temporal encoding. Specifically, electrical stimulation elicited more effective RGC firing, while optical stimulation enhanced RGC synchrony. These findings hold promise for advancing the field of neural encoding.

Recent Progress and Perspectives on Neural Chip Platforms Integrating PDMS-Based Microfluidic Devices and Microelectrode Arrays.

Recent years have witnessed a spurt of progress in the application of the encoding and decoding of neural activities to drug screening, diseases diagnosis, and brain-computer interactions. To overcome the constraints of the complexity of the brain and the ethical considerations of in vivo research, neural chip platforms integrating microfluidic devices and microelectrode arrays have been raised, which can not only customize growth paths for neurons in vitro but also monitor and modulate the specialized neural networks grown on chips. Therefore, this article reviews the developmental history of chip platforms integrating microfluidic devices and microelectrode arrays. First, we review the design and application of advanced microelectrode arrays and microfluidic devices. After, we introduce the fabrication process of neural chip platforms. Finally, we highlight the recent progress on this type of chip platform as a research tool in the field of brain science and neuroscience, focusing on neuropharmacology, neurological diseases, and simplified brain models. This is a detailed and comprehensive review of neural chip platforms. This work aims to fulfill the following three goals: (1) summarize the latest design patterns and fabrication schemes of such platforms, providing a reference for the development of other new platforms; (2) generalize several important applications of chip platforms in the field of neurology, which will attract the attention of scientists in the field; and (3) propose the developmental direction of neural chip platforms integrating microfluidic devices and microelectrode arrays.

PPy/SWCNTs-Modified Microelectrode Array for Learning and Memory Model Construction through Electrical Stimulation and Detection of In Vitro Hippocampal Neuronal Network.

The learning and memory functions of the brain remain unclear, which are in urgent need for the detection of both a single cell signal with high spatiotemporal resolution and network activities with high throughput. Here, an in vitro microelectrode array (MEA) was fabricated and further modified with polypyrrole/carboxylated single-walled carbon nanotubes (PPy/SWCNTs) nanocomposites as the interface between biological and electronic systems. The deposition of the nanocomposites significantly improved the performance of microelectrodes including low impedance (60.3 ± 28.8 k Ω), small phase delay (-32.8 ± 4.4°), and good biocompatibility. Then the modified MEA was used to apply learning training and test on hippocampal neuronal network cultured for 21 days through electrical stimulation, and multichannel electrophysiological signals were recorded simultaneously. During the process of learning training, the stimulus/response ratio of the hippocampal learning population gradually increased and the response time gradually decreased. After training, the mean spikes in burst, number of bursts, and mean burst duration increased by 53%, 191%, and 52%, respectively, and the correlation of neurons in the network was significantly enhanced from 0.45 ± 0.002 to 0.78 ± 0.002. In addition, the neuronal network basically retained these characteristics for at least 5 h. These results indicated that we have successfully constructed a learning and memory model of hippocampal neurons on the in vitro MEA, contributing to understanding learning and memory based on synaptic plasticity. The proposed PPy/SWCNTs-modified in vitro MEA will provide a promising platform for the exploration of learning and memory mechanism and their applications in vitro.

Nanomaterial-based microelectrode arrays for in vitro bidirectional brain-computer interfaces: a review.

A bidirectional in vitro brain-computer interface (BCI) directly connects isolated brain cells with the surrounding environment, reads neural signals and inputs modulatory instructions. As a noninvasive BCI, it has clear advantages in understanding and exploiting advanced brain function due to the simplified structure and high controllability of ex vivo neural networks. However, the core of ex vivo BCIs, microelectrode arrays (MEAs), urgently need improvements in the strength of signal detection, precision of neural modulation and biocompatibility. Notably, nanomaterial-based MEAs cater to all the requirements by converging the multilevel neural signals and simultaneously applying stimuli at an excellent spatiotemporal resolution, as well as supporting long-term cultivation of neurons. This is enabled by the advantageous electrochemical characteristics of nanomaterials, such as their active atomic reactivity and outstanding charge conduction efficiency, improving the performance of MEAs. Here, we review the fabrication of nanomaterial-based MEAs applied to bidirectional in vitro BCIs from an interdisciplinary perspective. We also consider the decoding and coding of neural activity through the interface and highlight the various usages of MEAs coupled with the dissociated neural cultures to benefit future developments of BCIs.

Dietary ingredient change induces a transient MyD88-dependent mucosal enteric glial cell response and promotes obesity.

OBJECTIVE: Consumption of a modern Western-type high-fat low-fiber diet increases the risk of obesity. However, how a host responds to such a diet, especially during the early period of dietary transition from a previous low-fat and fiber-rich diet, remains poorly explored. METHODS: Wild-type C57BL/6 mice were fed a normal chow diet or a high-fat diet. Enteric glial cell (EGC) activation was detected through quantitative real-time PCR (qRT-PCR), immunoblotting and immunohistology analysis. Fluorocitrate or genetic deletion of glial fibrillary acidic protein (GFAP)-positive glial-intrinsic myeloid differentiation factor 88 (Myd88) was used to inhibit EGC activation, and the effect of a high-fat diet on obesity was further investigated. The role of MYD88-dependent sensing of commensal products in adipocyte was observed to analyze the effect of obesity. RESULTS: A dietary shift from a normal chow diet to a high-fat diet in mice induced a transient early-phase emergence of a GFAP-positive EGC network in the lamina propria of the ileum, accompanied with an increase in glial-derived neurotrophic factor production. Inhibition of glial cell activity blocked this response. GFAP-positive glial Myd88 knockout mice gained less body weight after high-fat diet (HFD) feeding than littermate controls. In contrast, adipocyte deletion of Myd88 in mice had no effect on weight gain but instead exacerbated glucose intolerance. Furthermore, short-term fluorocitrate intervention during HFD feeding attenuated body weight gain. CONCLUSIONS: Our findings indicate that EGCs are early responders to intestinal ecosystem changes and the GFAP-positive glial Myd88 signaling participates in regulating obesity.

Degradation at the Na3SbS4/Anode Interface in an Operating All-Solid-State Sodium Battery.

All-solid-state sodium batteries utilize earth-abundant elements and are sustainable systems for large-scale energy storage and electric transportation. Replacing flammable carbonate-based electrolytes with solid-state ionic conductors promotes battery safety. Using solid-state electrolytes (SEs) also eliminates the need for packing when fabricating tandem cells, potentially enabling further enhanced energy density. Na3SbS4, a Na+ conductor, remains stable in dry air and shows high Na+ conductivity (σ ≈ 1.0 × 10-3 S/cm) and is thus a promising SE for applications in sodium batteries. However, upon repeated electrochemical cycling, Na3SbS4-containing Na batteries exhibit decaying capacity and limited cycle life, which is likely associated with the decomposition of Na3SbS4 at the electrode/electrolyte interface. This work presents an in-depth analysis of the decomposition chemistry occurring at the Na3SbS4/anode interface using combined in situ Raman and post-mortem characterization. The results indicate that the SbS43- counterion is electrochemically reduced when experiencing Na+ reduction potentials, and this reduction chemistry likely follows multiple pathways. The observed reduction products include SbS33-, the Sb2S74- dimer, the NaSb binary phase, and Na2S. We also observed the irreversibility of the decomposition and, as a consequence, the accumulation of the degradation products over cycles. Also notable is the heterogeneity of this degradation chemistry across the interface. Through the spectroelectrochemical characterizations, we reveal the possible mechanisms of the Na3SbS4 decomposition at the solid electrolyte/anode interface in an operating device.

FAM96A is essential for maintaining organismal energy balance and adipose tissue homeostasis in mice.

The iron (Fe) metabolism plays important role in regulating systemic metabolism and obesity development. The Fe inside cells can form iron-sulfur (Fe-S) clusters, which are usually assembled into target proteins with the help of a conserved cluster assembly machinery. Family with sequence similarity 96A (FAM96A; also designated CIAO2A) is a cytosolic Fe-S assembly protein involved in the regulation of cellular Fe homeostasis. However, the biological function of FAM96A in vivo is still incompletely defined. Here, we tested the role of FAM96A in regulating organismal Fe metabolism, which is relevant to obesity and adipose tissue homeostasis. We found that in mice genetically lacking FAM96A globally, intracellular Fe homeostasis was interrupted in both white and brown adipocytes, but the systemic Fe level was normal. FAM96A deficiency led to adipocyte hypertrophy and organismal energy expenditure reduction even under nonobesogenic normal chow diet-fed conditions. Mechanistically, FAM96A deficiency promoted mechanistic target of rapamycin (mTOR) signaling in adipocytes, leading to an elevation of de novo lipogenesis and, therefore, fat mass accumulation. Furthermore, it also caused mitochondrial defects, including defects in mitochondrial number, ultrastructure, redox activity, and metabolic function in brown adipocytes, which are known to be critical for the control of energy balance. Moreover, adipocyte-selective FAM96A knockout partially phenocopied global FAM96A deficiency with adipocyte hypertrophy and organismal energy expenditure defects but the mice were resistant to high-fat diet-induced weight gain. Thus, FAM96A in adipocytes may autonomously act as a critical gatekeeper of organismal energy balance by coupling Fe metabolism to adipose tissue homeostasis.

Grid cell remapping under three-dimensional object and social landmarks detected by implantable microelectrode arrays for the medial entorhinal cortex.

Grid cells with stable hexagonal firing patterns in the medial entorhinal cortex (MEC) carry the vital function of serving as a metric for the surrounding environment. Whether this mechanism processes only spatial information or involves nonspatial information remains elusive. Here, we fabricated an MEC-shaped microelectrode array (MEA) to detect the variation in neural spikes and local field potentials of the MEC when rats forage in a square enclosure with a planar, three-dimensional object and social landmarks in sequence. The results showed that grid cells exhibited rate remapping under social conditions in which spike firing fields closer to the social landmark had a higher firing rate. Furthermore, global remapping showed that hexagonal firing patterns were rotated and scaled when the planar landmark was replaced with object and social landmarks. In addition, when grid cells were activated, the local field potentials were dominated by the theta band (5-8 Hz), and spike phase locking was observed at troughs of theta oscillations. Our results suggest the pattern separation mechanism of grid cells in which the spatial firing structure and firing rate respond to spatial and social information, respectively, which may provide new insights into how the brain creates a cognitive map.

A tyrosine catabolic intermediate 4-hydroxyphenylpyruate attenuates murine endotoxic shock by blocking NLRP3 inflammasome activation.

The metabolic alterations of amino acid metabolism are closely associated with inflammatory response. However, relatively little is known about the roles of phenylalanine (Phe)/tyrosine (Tyr) catabolites during inflammation. Nitisinone (NTBC) is an orphan drug used to treat hereditary tyrosinemia type I potentially by changing Phe/Tyr metabolic flow. In this study, we used NTBC as a tool to investigate the potential role of the Phe/Tyr catabolic pathway in inflammatory responses. We found that NTBC was effective in tempering the bacterial endotoxin lipopolysaccharide (LPS)-induced septic shock in mice. Mechanistically, the protective effect was related to the accumulation of a Phe/Tyr catabolic intermediate, 4-hydroxyphenylpyruvate (4-HPP), induced by the NTBC treatment. 4-HPP could inhibit NLRP3 inflammasome priming and activation processes and therefore reduce IL-1ß release and pyroptosis. Like NTBC, 4-HPP was also effective in attenuating endotoxic shock in mice. Our results suggest the Phe/Tyr catabolic pathway as a potential immunoregulatory hub that may be exploited therapeutically to alleviate inflammation.

A Neural Sensor with a Nanocomposite Interface for the Study of Spike Characteristics of Hippocampal Neurons under Learning Training.

Both the cellular- and population-level properties of involved neurons are essential for unveiling the learning and memory functions of the brain. To give equal attention to these two aspects, neural sensors based on microelectrode arrays (MEAs) have been in the limelight due to their noninvasive detection and regulation capabilities. Here, we fabricated a neural sensor using carboxylated graphene/3,4-ethylenedioxythiophene:polystyrenesulfonate (cGO/PEDOT:PSS), which is effective in sensing and monitoring neuronal electrophysiological activity in vitro for a long time. The cGO/PEDOT:PSS-modified microelectrodes exhibited a lower electrochemical impedance (7.26 ± 0.29 kΩ), higher charge storage capacity (7.53 ± 0.34 mC/cm2), and improved charge injection (3.11 ± 0.25 mC/cm2). In addition, their performance was maintained after 2 to 4 weeks of long-term cell culture and 50,000 stimulation pulses. During neural network training, the sensors were able to induce learning function in hippocampal neurons through precise electrical stimulation and simultaneously detect changes in neural activity at multiple levels. At the cellular level, not only were three kinds of transient responses to electrical stimulation sensed, but electrical stimulation was also found to affect inhibitory neurons more than excitatory neurons. As for the population level, changes in connectivity and firing synchrony were identified. The cGO/PEDOT:PSS-based neural sensor offers an excellent tool in brain function development and neurological disease treatment.

Recent progress and perspectives of continuous in vivo testing device.

Devices for continuous in-vivo testing (CIVT) can detect target substances in real time, thus providing a valuable window into a patient's condition, their response to therapeutics, metabolic activities, and neurotransmitter transmission in the brain. Therefore, CIVT devices have received increased attention because they are expected to greatly assist disease diagnosis and treatment and research on human pathogenesis. However, CIVT has been achieved for only a few markers, and it remains challenging to detect many key markers. Therefore, it is important to summarize the key technologies and methodologies of CIVT, and to examine the direction of future development of CIVT. We review recent progress in the development of CIVT devices, with consideration of the structure of these devices, principles governing continuous detection, and nanomaterials used for electrode modification. This detailed and comprehensive review of CIVT devices serves three purposes: (1) to summarize the advantages and disadvantages of existing devices, (2) to provide a reference for development of CIVT equipment to detect additional important markers, and (3) to discuss future prospects with emphasis on problems that must be overcome for further development of CIVT equipment. This review aims to promote progress in research on CIVT devices and contribute to future innovation in personalized medical treatments.

New Evidence for the Existence of Two Kiss/Kissr Systems in a Flatfish Species, the Turbot (Scophthalmus maximus), and Stimulatory Effects on Gonadotropin Gene Expression.

Seasonal reproduction is generally controlled by the hypothalamus-pituitary-gonadal (HPG) axis in fish. Previous studies have demonstrated that the kisspeptin (Kiss)/kisspeptin receptor (Kissr) system, a positive regulator of the HPG axis, mediates the responses to environmental cues. Turbot (Scophthalmus maximus), a representative species of Pleuronectiformes, is one of the most commercially important fish species cultured in Europe and North China. However, the mechanisms by which the Kiss/Kissr system regulates the reproductive axis of turbot according to seasonal changes, especially photoperiod, have not been clearly characterized. In the current study, the cDNA sequences of kiss2/kissr2, along with kiss1/kissr3 which was thought to be lost in flatfish species, were cloned and functionally characterized. The kiss1, kiss2, and kissr3 transcripts were highly detected in the brain and gonad, while kissr2 mRNA was only abundantly expressed in the brain. Moreover, kiss/kissr mRNAs were further examined in various brain areas of both sexes. The kiss1, kissr2, kissr3 mRNAs were highly expressed in the mesencephalon, while a substantial degree of kiss2 transcripts were observed in the hypothalamus. During annual reproductive cycle, both kiss and kissr transcript levels declined significantly from the immature to mature stages and increased at the degeneration stage in the brains of both sexes, especially in the mesencephalon and hypothalamus. The ovarian kiss1, kiss2, and kissr2 mRNA levels were highest at the vitellogenic stage (mature stage), while expression of kissr3 was highest at the immature stage. The testicular kiss and kissr transcripts were highest in the immature and degeneration stages, and lowest at the mature stage. In addition, intraperitoneal injection of Kiss1-10 and Kiss2-10 significantly stimulated mRNA levels of pituitary lhß, fhsß, and gthα. In summary, two Kiss/Kissr systems were firstly proven in a flatfish species of turbot, and it has a positive involvement in controlling the reproduction of the Kiss/Kissr system in turbot. The results will provide preliminary information regarding how the Kiss/Kissr system controls seasonal reproduction in turbot broodstock.