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Background: Gefitinib is an important drug used to treat Non-Small Cell Lung Cancer (NSCLC) with EGFR activating mutations, but drug resistance restricts its clinical application. In this present study, combined Jin Fu Kang Decoction (JFKD) and gefitinib showed specific cytotoxicity to gefitinib-resistant cancer cells (PC-9/gef). Objective: This study aimed to decipher the molecular mechanism of the JFKD on drug resistance when used together with Gefitinib and to find the contributing bio-active substance(s) in JFKD based on the putative mechanism. Methods: To investigate the combined effect of gefitinib and JFKD, in vitro experiments were conducted on the established gefitinib-resistant PC-9 subclone, while in vivo experiments were conducted on the BALB/c nude mice with PC-9/gef xenografts. Western blot was used to evaluate the protein expression, and Ultra-Performance Liquid Chromatography (UPLC) coupled with quadrupole time-offlight Mass Spectrometry (MS) was used to detect the bio-active compounds of JFKD. Results: The expression of the PTEN-relevant protein p-EGFR, p-Akt in vitro was inhibited more when combined JKFD and gefitinib were used, whereas the activities of PDCD4 and PTEN were increased; remarkably, in vivo experiments showed enhanced tumor growth inhibition when treated with this combination. Due to this combination, the effect on the gefitinib-resistant cell line, one of the JFKD-induced anti-cancer mechanisms, was found. To link the putative mechanism and the anticancer compounds in JFKD, 14 saponins and flavonoids were detected. Conclusion

Gastric cancer (GC) is a global health issue that lacks effective treatment options. Afatinib is a tyrosine kinase inhibitor (TKI) that has shown promising results in the treatment of GC. However, resistance to afatinib is inevitable and hampers its clinical application. To date, there is limited knowledge regarding the mechanisms underlying the resistance of GC cells to afatinib. This study aimed to identify novel factors that may contribute to the resistance of GC cells to afatinib. We found that upregulation of calmodulin 2 (CALM2), a member of the CALM family, confers resistance to afatinib in GC cells. Knockdown of CALM2 can overcome the resistance to afatinib by promoting mitochondrial apoptosis in a caspase-dependent manner. Mechanistically, it was found that the downregulation of CALM2 led to the upregulation of the FoxO3a/Puma axis. Inhibition of either FoxO3a or Puma abrogated the effects of CALM2 downregulation in GC cells. In addition, we revealed that CALM2 knockdown inhibited Akt signaling, which is responsible for blocking the FoxO3a/Puma axis. Altogether, our results indicated that CALM2 could be considered a potential target to overcome the resistance of GC cells to afatinib.

The programed bimodal photoelectrochemical (PEC)-sensing platform based on DNA structural switching induced by targets binding to aptamers was innovatively designed for the simultaneous detection of mucin 1 (MUC1) and microRNA 21 (miRNA-21). To promote excellent current intensity as well as enhance the sensitivity of aptasensors, the evenly distributed WO3/Fe2O3 heterojunction was prepared as a transducer material, notably reducing the background signal response and extending the absorption of light. The multifunctional paper-based biocathode was assembled to provide a visual colorimetric assay. When introducing the integrated signal probe (ISP) composed of signal probe 1 (sP1) and signal probe 2 (sP2) on paper-based working units modified with gold nanoparticles (AuNPs), recognition sites of two targets were formed. In the presence of MUC1 protein, both sP1 and the target on the working unit were released into the corresponding colorimetric unit because of the DNA specific recognition. The horseradish peroxidase-streptavidin (HRP-SA) carried by free sP1 could oxidize 3,3',5,5'-tetramethylbenzidine (TMB) to turn a blue-colored oxidized TMB (oxTMB) in the presence of hydrogen peroxide (H2O2), which ultimately gained a higher photocurrent signal. Furthermore, miRNA-21 was modified on another working unit by binding with sP2, leading to changes in the current signal and thus enabling real-time detection of analytes with the assistance of a digital multimeter. The PEC aptasensor offered a wide dynamic range of 10 fg·mL-1-100 ng mL-1 for MUC1 and 0.1 pM-10 nM for miRNA-21, with a low detection limit of 3.4 fg·mL-1 and 36 fM, respectively. It laid the foundation for synchronous detection of multiple analytes and initiated a new way for the enhancement in modern next-generation disease diagnosis.

Spinal muscular atrophy (SMA) is a devastating genetic neuromuscular disease. Diffuse neuropathology has been reported in SMA patients and mouse models, however, functional changes in brain regions have not been studied. In the SMNΔ7 mouse model, we identified three types of differences in neuronal function in the cerebellum and motor cortex from two age groups: P7–9 (P7) and P11–14 (P11). Microelectrode array studies revealed significantly lower spontaneous firing and network activity in the cerebellum of SMA mice in both age groups, but it was more profound in the P11 group. In the motor cortex, however, neural activity was not different in either age group. Whole-cell patch-clamp was used to study the function of output neurons in both brain regions. In cerebellar Purkinje cells (PCs) of SMA mice, the input resistance was larger at P7, while capacitance was smaller at P11. In the motor cortex, no difference was observed in the passive membrane properties of layer V pyramidal neurons (PN5s). The action potential threshold of both types of output neurons was depolarized in the P11 group. We also observed lower spontaneous excitatory and inhibitory synaptic activity in PN5s and PCs respectively from P11 SMA mice. Overall, these differences suggest functional alterations in the neural network in these motor regions that change during development. Our results also suggest that neuronal dysfunction in these brain regions may contribute to the pathology of SMA. Comprehensive treatment strategies may consider motor regions outside of the spinal cord for better outcomes.

The neural circuits that support human cognition are a topic of enduring interest. Yet, there are limited tools available to map brain circuits in the human and nonhuman primate brain. We harnessed high-resolution diffusion MR tractography, anatomic, and transcriptomic data from individuals of either sex to investigate the evolution and development of frontal cortex circuitry. We applied machine learning to RNA sequencing data to find corresponding ages between humans and macaques and to compare the development of circuits across species. We transcriptionally defined neural circuits by testing for associations between gene expression and white matter maturation. We then considered transcriptional and structural growth to test whether frontal cortex circuit maturation is unusually extended in humans relative to other species. We also considered gene expression and high-resolution diffusion MR tractography of adult brains to test for cross-species variation in frontal cortex circuits. We found that frontal cortex circuitry development is extended in primates, and concomitant with an expansion in corticocortical pathways compared with mice in adulthood. Importantly, we found that these parameters varied relatively little across humans and studied primates. These data identify a surprising collection of conserved features in frontal cortex circuits across humans and Old World monkeys. Our work demonstrates that integrating transcriptional and structural data across temporal dimensions is a robust approach to trace the evolution of brain pathways in primates.SIGNIFICANCE STATEMENTDiffusion MR tractography is an exciting method to explore pathways, but there are uncertainties in the accuracy of reconstructed tracts. We broaden the repertoire of toolkits to enhance our ability to trace human brain pathways from diffusion MR tractography. Our integrative approach finds corresponding ages across species and transcriptionally defines neural circuits. We used this information to test for variation in circuit maturation across species and found a surprising constellation of similar features in frontal cortex neural circuits across humans and primates. Integrating across scales of biological organization expands the repertoire of tools available to study pathways in primates, which opens new avenues to study pathways in health and diseases of the human brain.

Traction and quantitative detection of trace amount of target cells inside of biochip system in real-time has been a challenge for biomedical and clinical researchers. In this manuscript, we report fabrication of a photoelectrochemical platform that has integrated both biometric recognition and signal acquisition through microfabrication technology. In this chip, a ternary ZnO/CdTe/Bi nanorod array is fabricated, which significantly extends the absorption wavelength from the UV to the visible and even near-infrared regions for both photocarrier generation and surface plasmon resonance, ultimately achieving the amplification of initial photocurrent responses. The artificially designed aptamers with amino groups are assembled on the surface of the outermost Bi nanoparticles, which are used as signal probes due to the specific recognition to the nasopharyngeal carcinoma 5-8F cell. We demonstrate that different concentration of 5-8F cells is captured by aptamers, and the signal changes accordingly with the amount of the cells that have been trapped. As a result, the proposed biochip demonstrates rapid response in a wide linear range of 10

The axon initial segment (AIS) is a critical locus of control of action potential (AP) generation and neuronal information synthesis. Concussive traumatic brain injury gives rise to diffuse axotomy, and the majority of neocortical axonal injury arises at the AIS. Consequently, concussive traumatic brain injury might profoundly disrupt the functional specialization of this region. To investigate this hypothesis, one and two days after mild central fluid percussion injury in Thy1-YFP-H mice, we recorded high-resolution APs from axotomized and adjacent intact layer 5 pyramidal neurons and applied a second derivative (2°) analysis to measure the AIS- and soma-regional contributions to the AP upstroke. All layer 5 pyramidal neurons recorded from sham animals manifested two stark 2° peaks separated by a negative intervening slope. In contrast, within injured mice, we discovered a subset of axotomized layer 5 pyramidal neurons in which the AIS-regional 2° peak was abolished, a functional perturbation associated with diminished excitability, axonal sprouting and distention of the AIS as assessed by staining for ankyrin-G. Our analysis revealed an additional subpopulation of both axotomized and intact layer 5 pyramidal neurons that manifested a melding together of the AIS- and soma-regional 2° peaks, suggesting a more subtle aberration of sodium channel function and/or translocation of the AIS initiation zone closer to the soma. When these experiments were repeated in animals in which cyclophilin-D was knocked out, these effects were ameliorated, suggesting that trauma-induced AIS functional perturbation is associated with mitochondrial calcium dysregulation.

Objective. Oral leukoplakia (OLK) is the most common precancerous lesion in the oral cavity. This study aimed to explore key biomarkers for monitoring OLK for early diagnosis of oral squamous cell carcinoma (OSCC) and screen small-molecule drugs for the prevention of OSCC. Method. The Gene Expression Omnibus (GEO) database was explored to extract two microarray datasets, namely, GSE85195 and GSE25099. The data of the normal group, OLK group, and OSCC group were analyzed by weighted gene coexpression network analysis (WGCNA) to identify the most significant gene module and differentially expressed genes (DEGs). The intersection genes were extracted as the key genes of OLK carcinogenesis. Subsequently, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were analyzed in the module. Connectivity Map and molecular docking were used to screen small-molecule drugs. The diagnostic values of four key genes were identified and verified in the GSE26549 dataset. Results. WGCNA obtained the red module (r = −0.91, p < 0.05 ) with the strongest correlation with cancerous phenotype. GO enrichment analysis showed 60 pathways, including 28 biological processes, 11 cell components, and 21 molecular functions, and KEGG enrichment analysis showed 4 pathways ( p < 0.05 ). In the differential expression analysis, there was no intersection between the upregulated genes and the red module genes. However, the intersection of the downregulated genes and the red module genes yielded 4 key genes: dopachrome tautomerase (DCT), keratin 3 (KRT3), keratin 76 (KRT76), and FAM3 metabolic regulation signal molecule B (FAM3B). The area under the curve of the diagnostic model constructed by these four genes was 0.963 (CI = 0.913–1.000). The sensitivity was 0.933, and the specificity was 0.923. The diagnostic model was successfully verified in GSE26549 (AUC = 0.745, CI = 0.638–0.851). Compared with the diagnostic models of the previous studies, the diagnostic efficiency of this model was the highest. The small-molecule drugs, selumetinib and benidipine, were selected according to the gene expression profile and showed binding activity when docking with the above molecules. Conclusions. This study provides new targets and drugs for OLK. These targets could be used as the key diagnostic molecules for long-term follow-up of OLK. The small-molecule drugs selumetinib and benidipine could be used for the prevention and treatment of OSCC.

Cardiovascular and cerebrovascular diseases are a serious threat to human health and increase the annual death ratio at a considerable pace. This is not uncommon even among teenagers and martial arts athletes. Due to the increased risk associated with strenuous exercise in the context of a quiescent cardiac abnormality, athletes have a higher rate of heart attack and stroke than their nonathletic colleagues. The mortality rate due to this disease is extremely high, which needs to be controlled at the initial stages. At present, the recognition and analysis of ECG signals is still an issue and requires an expert to analyse it and identify its hidden patterns. The most challenging aspect of ECG signal classification is the irregularities in the signals, which are critical for detecting patient status. Each heartbeat is made up of a variety of action impulse waveforms produced by various cardiac tissues. Classification of heartbeats is difficult because waveforms vary from person to person and are identified by certain features. At present, the automatic identification of ECG signals still requires manual design, which has low accuracy and cannot be widely used in clinical practice. This study proposes an intelligent system based on deep learning and machine learning methods to classify and diagnose ECG signals to improve their classification and recognition accuracy. It improves the detection ability of martial arts athletes’ arrhythmia disease and obtains accurate arrhythmia diagnosis information. MIT-BIH arrhythmia dataset has been used for the experimental analysis. The performance of the proposed scheme is evaluated with the help of various performance measures. We conduct comprehensive experiments, and the results show that the algorithms used in this paper are robust.

Lung cancer remains the leading cause of cancer-related death worldwide, and the high incidence rates are worrisome. Exosomes are a class of extracellular vesicles secreted by most cells, including RNAs, proteins and lipids. Exosomes can mediate cell-to-cell communication in both physiologic and pathologic processes. Accumulated evidences show that cancer-derived exosomes aid in the recruitment and reprogramming of constituents correlated with tumor microenvironment. Furthermore, exosome-based clinical trials have been completed in advanced lung cancer patients. In this review, we discuss the roles of exosomes in a lung cancer microenvironment, such as its participation in lung cancer initiation, progression and metastasis as well as being involved in angiogenesis, epithelial-mesenchymal transition (EMT), immune escape, and drug resistance. In addition, we focus on the potential of exosomes as diagnostic and prognostic biomarkers in lung cancer, as well as the challenges faced by and advantages of exosomes as drug delivery vehicles and in exosome-based immunotherapy.

The programed bimodal photoelectrochemical (PEC)-sensing platform based on DNA structural switching induced by targets binding to aptamers was innovatively designed for the simultaneous detection of mucin 1 (MUC1) and microRNA 21 (miRNA-21). To promote excellent current intensity as well as enhance the sensitivity of aptasensors, the evenly distributed WO

Objective: We aimed to clarify the significance of PET-CT for detecting occult lymph node metastasis and for affecting prognosis in early-stage tongue squamous cell carcinoma (SCC).Methods: Patients with surgically treated primary cT1-2N0 tongue SCC who agreed to undergo a preoperative PET-CT scan were prospectively enrolled. The primary study outcomes were occult neck lymph node metastasis and locoregional control (LRC). The Kaplan-Meier method was used to analyze the LRC rate, and then the factors that were significant in the Kaplan-Meier method were assessed in the Cox model to determine the independent factors.Results: A total of 135 patients were included, and the median maximum standardized uptake value (SUV max) of the primary tumor was 9.0. When analyzing the PET-CT results, 18 patients were recognized as having neck lymph node metastasis, and 12 patients were proven to have pathologic lymph nodes. A total of 117 patients did not have neck lymph node metastasis reported by PET-CT, and five patients were proven to have pathologic lymph nodes. The sensitivity and specificity of PET-CT for predicting occult metastasis were 70.6 and 94.9%, respectively. In patients with an SUV max ≤ 9.0, the 5-year LRC rate was 95%; in patients with an SUV max >9.0, the 5-year LRC rate was 85%, and the difference was significant. Further Cox model analyses confirmed the independence of the SUV max for predicting LRC.Conclusion: PET-CT has a high specificity for predicting occult lymph node metastasis, and an SUV max >9.0 is significantly associated with worse LRC in cT1-2N0 tongue SCC.

An effective “on–off–on” photoelectrochemical (PEC)/visual sensing system based on cleaning-switchable lab-on-paper device was designed to achieve ultrasensitive detection of analytes. The first amplified “signal-on” PEC state was gained by CdS quantum dots sensitized leaf-shape ZnO (CdS QDs/leaf-shape ZnO) structure, which was assembled on reduced graphene oxide (rGO) modified paper electrode. Then Au-modified prism-anchored octahedral CeO2 nanoparticles (Au@PO-CeO2 NPs), as an efficient signal quencher, were immobilized on the CdS QDs/leaf-shape ZnO with the assistance of DNA hybridization, resulting in a noticeable photocurrent response decrement with the “signal-off” PEC state. With the addition of analytes, the quencher Au@PO-CeO2 NPs were immediately released from the sensing surface and robust PEC response was recovered to the signal-on state again. Meanwhile, the disengaged quencher in electrolyte solution flowed to the colorimetric detection area of lab-on-paper device and catalyzed oxidation of ...

Objective— The genetic contribution to coronary artery disease (CAD) remains largely unclear. We combined genetic screening with functional characterizations to identify novel loci and candidate genes for CAD. Approach and Results— We performed genome-wide screening followed by multicenter validation in 8 cohorts consisting of 21 828 participants of Han ethnicity and identified 3 novel intragenic SNPs (single nucleotide polymorphisms), rs9486729 ( SCML4 [Scm polycomb group protein-like 4]; odds ratio, 1.25; 95% CI, 1.17–1.34; P =3.51×10 −11 ), rs17165136 ( THSD7A [thrombospondin type 1 domain-containing 7A]; odds ratio 1.28; 95% CI, 1.21–1.35; P −25 ), and rs852787 ( DAB1 [disabled-1]; odds ratio, 1.29; 95% CI, 1.21–1.38; P =2.02×10 −14 ), associated with CAD with genome-wide significance. The risk allele of rs9486729 and protective allele of rs17165136 were associated with the decreased expression of their host genes, SCML4 and THSD7A , respectively, whereas rs852787 did not have transcriptional effects on any gene. Knockdown of SCML4 activated endothelial cells by increasing the expression of IL-6 , E-selectin , and ICAM and weakened their antiapoptotic activity, whereas the knockdown of THSD7A had little effect on these endothelial cell functions but attenuated monocyte adhesion via decreasing the expression of ICAM , L-selectin , and ITGB2 . We further showed that inhibiting the expression of SCML4 exacerbated endothelial dysfunction and vascular remodeling in a rat model with partial carotid ligation. Conclusions— We identify 3 novel loci associated with CAD and show that 2 genes, SCML4 and THSD7A , make functional contributions to atherosclerosis. How rs852787 and its host gene DAB1 are linked to CAD needs further studies.

The detection of cardiac markers with high sensitivity and specificity is exceedingly essential for the early diagnosis and warning of cardiovascular disease. In this work, we developed a microcavity-based sandwich immune chip to realize the ultrasensitive detection of cardiac troponin I (cTnI) and creatine kinase isoenzyme MB (CK-MB) by surface-enhanced Raman spectroscopy (SERS) method. Herein, polystyrene microspheres modified with gold nanoparticles (Au NPs) were fixed on the substrate via polydopamine, which was used as an immune capture chip. 5,5′-Dithio bis-(2-nitrobenzoic acid) (DTNB) and 4-mercaptobenzoic acid (4MBA) labelled Au NPs served as immune signal probes for cTnI and CK-MB, respectively. Light confinement effect of polystyrene (PS) microcavities and localized surface plasmon resonance (LSPR) near the Au NPs synergistically strengthened the electromagnetic field and promoted the interaction between light and matter, which enhanced SERS performance with an enhancement factor of 1.95 × 1012 for SERS sensing. The immunoassay results demonstrated the sensitive quantitative detection with the limit of detection (LOD) of 3.16 pg/mL and 4.27 pg/mL for cTnI and CK-MB, separately. The method also had good specificity, reproducibility and stability. It could be expected that our proposed microcavity-based SERS chip immunoassay technique would be considered as an effective diagnostic tool for ultrasensitive detection of cardiac biomarkers.

Inspired by the design of folding greeting cards and tissue drawing covers, a photoelectrochemical (PEC) lab-on-paper device with a controllable fluid separator, producing both reaction zone and detection zone, was explored for ultrasensitive detection of adenosine 5′-triphosphate (ATP) via mimic peroxidase-transfer enhancement of photocurrent response. To realize it, the DNA1, aptamer, and DNA2 as well as the mimic peroxidase of G-quadruplex/hemin modified Au nanocubes were linked on the graphene oxide-functionalized reaction zone via the DNA hybridization. Meanwhile, three-dimensional CuO nanoflowers (CuO NFs) as a photoactive material with outstanding electron transfer ability and absorption of light were grown in situ on the detection zone, providing a PEC active interface. Besides, an innovative fluid separator was elaborately designed by assembling a strip of paper with a hydrophilic channel, providing an effective way to bridge the gap between the two zones with a controllable drawing way, which could successfully avoid the signal interference caused by modifying biomolecules layer by layer on photosensitive materials. In the presence of ATP, the G-quadruplex/hemin modified in the reaction zone was dissociated due to the specific recognition of ATP with aptamer and released into the detection zone with the assistance of controllable fluid separator. The free G-quadruplex/hemin could catalyze hydrogen peroxide to generate oxygen for the consumption of photo-induced electrons from CuO NFs, which could further promote the electron-hole carriers separation efficiency, and eventually resulting in the enhancement of PEC signal. The proposed PEC lab-on-paper device could be employed for specific detection of ATP in the range from 5.0 to 3.0 × 103 nM with a detection limit of 2.1 nM.

A renewable photoelectrochemical/synchronous visualized sensing platform with two functional cells, named sensing and electrochromic cells, was developed for ultrasensitive detection of carcinoembryonic antigen (CEA). For achieving a boosted photocurrent intensity, the Ni:FeOOH/BiVO4 nanocomposites were engineered and served as the photoanode materials, which were then combined with the cathode material Prussian blue (PB) to form the sensing cell. Through the sandwich-type immunoreaction and enzymatic cascade amplification, the carried GOx could catalyse the decomposition of glucose to produce H2O2, leading to improvement of electron transfer efficiency of Ni:FeOOH/BiVO4 under the light irradiation and accompanying by the PB reduced to Prussian white (PW). By combining digital multimeter (DMM)-joined circuit to collect photocurrents with PB electrochromic indicator, ultrasensitive and naked eye visual detection of CEA was realized synchronously. On the electrochromic cell, laccase as the biocathode enabled PW back to the original PB state through the biocatalyzing oxygen reduction of the laccase, gaining an updatable sensing platform. Under the optimal conditions, the bimodal biosensor shows acceptable dynamic working range for CEA from 1 pg·mL−1 to 90 ng·mL−1 with a detection limit (LOD) of 0.35 pg mL−1. Meanwhile, it also has great potential applications in real sample analysis.

An effective "on-off-on" photoelectrochemical (PEC)/visual sensing system based on cleaning-switchable lab-on-paper device was designed to achieve ultrasensitive detection of analytes. The first amplified "signal-on" PEC state was gained by CdS quantum dots sensitized leaf-shape ZnO (CdS QDs/leaf-shape ZnO) structure, which was assembled on reduced graphene oxide (rGO) modified paper electrode. Then Au-modified prism-anchored octahedral CeO

Activity-dependent changes in the properties of the motor system underlie the necessary adjustments in its responsiveness on the basis of the environmental and developmental demands of the organism. Although plastic changes in the properties of the spinal cord have historically been neglected because of the archaic belief that the spinal cord is constituted by a hardwired network that simply relays information to muscles, plenty of evidence has been accumulated showing that synapses impinging on spinal motoneurons undergo short- and long-term plasticity. In the brain, brief changes in the activity level of the network have been shown to be paralleled by changes in the intrinsic excitability of the neurons and are suggested to either reinforce or stabilize the changes at the synaptic level. However, rapid activity-dependent changes in the intrinsic properties of spinal motoneurons have never been reported. In this study, we show that in neonatal mice the intrinsic excitability of spinal motoneurons is depressed after relatively brief but sustained changes in the spinal cord network activity. Using electrophysiological techniques together with specific pharmacological blockers of KCNQ/Kv7 channels, we demonstrate their involvement in the reduction of the intrinsic excitability of spinal motoneurons. This action results from an increased M-current, the product of the activation of KCNQ/Kv7 channels, which leads to a hyperpolarization of the resting membrane potential and a decrease in the input resistance of spinal motoneurons. Computer simulations showed that specific up-regulations in KCNQ/Kv7 channels functions lead to a modulation of the intrinsic excitability of spinal motoneurons as observed experimentally. These results indicate that KCNQ/Kv7 channels play a fundamental role in the activity-dependent modulation of the excitability of spinal motoneurons.

Five yeast strains were tested for their ability to remove T-2 toxin from yeast extract peptone dextrose medium. The ability of Saccharomyces cerevisiae 1221 (SC1221) was the strongest among five strains, and 36.50 ± 1.40% of T-2 toxin was removed after incubation at 30C for 96 h. The mode of removal was physical binding rather than biotransformation. The total amount of T-2 toxin released from viable Saccharomyces cerevisiae 1221 cell–toxin complexes was 37.39 ± 0.48% of total bound T-2 toxin, respectively, after three times of washing with phosphate buffered saline, which was 8.17 ± 0.75% after incubation with simulated gastric fluid at 37C for 4 h, and which was 13.22 ± 0.61% after incubation with simulated intestinal fluid at 37C for 4 h, respectively. Practical Applications T-2 toxin is the most toxic trichothecene, which is commonly found in cereal grains, animal feed and human food produced from contaminated grains. The best solution for decontamination should be detoxification by microorganisms, giving a possibility for removal of mycotoxins under mild conditions, without using harmful chemicals and significant losses in nutritive value and palatability of decontaminated food and feed. Yeasts have immense potential as tools in tackling the problem of mycotoxins in cereal-based foods and in animal feeds. The objective of this work was to investigate the effect of food-grade yeast strains on the removal of T-2 toxin in vitro. Results of this study indicated that the strain of Saccharomyces cerevisiae 1221 (SC1221) has a strong ability to remove T-2 toxin. Therefore, this strain could be potentially applied in food and feed industry for detoxification of T-2 toxin.

Significant grain refinement in cast metals can be achieved through the application of electric currents during the solidification process. The present paper investigates the distribution of electric currents on the grain size of solidified Al-7wt.%Si alloy under the application of electric current with constant parameters flowing through two parallel electrodes into the melt within a cylindrical mould. The distribution of electric current was controlled by applying an electrical insulation material coating, boron nitride (NB), to the sidewall of the electrodes. Experimental results showed that the employment of these insulated electrodes can reduce grain size in comparison with the reference case of electrodes without BN coating. Flow measurements were performed in Ga-20wt.%In-12wt.%Sn liquid metal. Higher intensity forced flow occurred when the sidewall of the electrodes was insulated. In order to understand the underlying mechanism behind the stronger forced flow, corresponding numerical simulations were performed to reveal the distributions of the electric current, magnetic field, Lorentz force, and the resultant forced flow. The results achieved indicate that the mechanism of grain refinement driven by electric current is dendrite fragmentation induced by forced flow. In addition, a novel approach to enhance the grain refinement without additional input of current energy was developed.

The genetic contribution to coronary artery disease (CAD) remains largely unclear. We combined genetic screening with functional characterizations to identify novel loci and candidate genes for CAD.We performed genome-wide screening followed by multicenter validation in 8 cohorts consisting of 21 828 participants of Han ethnicity and identified 3 novel intragenic SNPs (single nucleotide polymorphisms), rs9486729 (We identify 3 novel loci associated with CAD and show that 2 genes

The axon initial segment (AIS) is the site of action potential (AP) initiation, thus a crucial regulator of neuronal activity. In excitatory pyramidal neurons, the high density of voltage-gated sodium channels (NaV1.6) at the distal AIS regulates AP initiation. A surrogate AIS marker, ankyrin-G (ankG) is a structural protein regulating neuronal functional via clustering voltage-gated ion channels. In neuronal circuits, changes in presynaptic input can alter postsynaptic output via AIS structural-functional plasticity. Recently, we showed experimental mild traumatic brain injury (mTBI) evokes neocortical circuit disruption via diffuse axonal injury (DAI) of excitatory and inhibitory neuronal systems. A key finding was that mTBI-induced neocortical electrophysiological changes involved non-DAI/ intact excitatory pyramidal neurons consistent with AIS-specific alterations. In the current study we employed Thy1-yellow fluorescent protein (YFP)-H mice to test if mTBI induces AIS structural and/or functional plasticity within intact pyramidal neurons 2 days after mTBI. We used confocal microscopy to assess intact YFP+ pyramidal neurons in layer 5 of primary somatosensory barrel field (S1BF), whose axons were continuous from the soma of origin to the subcortical white matter (SCWM). YFP+ axonal traces were superimposed on ankG and NaV1.6 immunofluorescent profiles to determine AIS position and length. We found that while mTBI had no effect on ankG start position, the length significantly decreased from the distal end, consistent with the site of AP initiation at the AIS. However, NaV1.6 structure did not change after mTBI, suggesting uncoupling from ankG. Parallel quantitative analysis of presynaptic inhibitory terminals along the postsynaptic perisomatic domain of these same intact YFP+ excitatory pyramidal neurons revealed a significant decrease in GABAergic bouton density. Also within this non-DAI population, patch-clamp recordings of intact YFP+ pyramidal neurons showed AP acceleration decreased 2 days post-mTBI, consistent with AIS functional plasticity. Simulations of realistic pyramidal neuron computational models using experimentally determined AIS lengths showed a subtle decrease is NaV1.6 density is sufficient to attenuate AP acceleration. Collectively, these findings highlight the complexity of mTBI-induced neocortical circuit disruption, involving changes in extrinsic/presynaptic inhibitory perisomatic input interfaced with intrinsic/postsynaptic intact excitatory neuron AIS output.

Very long chain fatty acids (VLCFAs) are essential lipid components in many plants. 3-Ketoacyl-CoA synthase (KCS) catalyzes the condensation reaction to form 3-ketoacyl-CoA in VLCFA synthesis. AtELO4 has been reported to be involved in VLCFA synthesis, functioning as a KCS in Arabidopsis. However, no studies on other three AtELO members have been reported. Here, we initially found by real-time PCR in Arabidopsis thaliana (L.) Heynh. that AtELO1, AtELO3, and AtELO4 displayed characteristic expression patterns, but AtELO2 was nearly expressed in any organ. Then the transient expression of ELO-like-eGFP fusions in Arabidopsis green leaf protoplasts showed that AtELO1, AtELO3, and AtELO4 were localized in the endoplasmic reticulum (ER), where VLCFA synthesis took place. Finally, we found that the contents of all fatty acids were decreased by 10–20% in seeds of atelo1 T-DNA insertion mutants. In seeds of Pro35S:AtELO1 plants, the levels of all remaining components, except C20:0 and C20:3, were significantly increased. Taken together, our study revealed biological functions of AtELO members and might lay the foundation for further genetic manipulations to generate oil crops with the high oil content.

Altered GABA-mediated inhibition is proposed to play a role in the pathogenesis of epilepsy. Previous studies have demonstrated a loss of somatostatin-containing GABAergic interneurons innervating granule cells in epileptic animals. However, the reorganization of synapses between interneurons and granule cells has not been investigated. We studied synapse organization in an animal model of temporal lobe epilepsy (TLE) using continuous hippocampal stimulation. The distribution of axon terminals and inhibitory synapses on granule cell dendrites was studied using a combination of immunohistochemistry and pre-embedding electron microscopy techniques. A whole-cell patch-clamp technique was applied to study the functional changes in GABAergic input from different interneurons. In epileptic animals, the density of cholecystokinin (CCK)-immunoreactive (IR) fibers and α2 subunit containing GABAA receptors in the inner molecular layer of the dentate gyrus was reduced. Quantitative immuno-electron microscopy study revealed that the ratio of CCK-containing symmetric synapses to the total symmetric synapses was reduced. The frequency of GABAergic synaptic currents (sIPSC) was decreased and their amplitude was increased. The inhibitory effect of the activation of cannabinoid 1 (CB1) receptors was also reduced in epileptic animals. Isolation of CCK- and parvalbumin (PV)-containing GABAergic inputs by N- and P/Q-type calcium channel blockers respectively suggested that GABA release from CCK-containing interneurons was selectively reduced in epileptic rats. This study found that there was a loss of CCK-containing GABAergic synapses to granule cells both morphologically and functionally. These studies add to our understanding of the mechanisms that contribute to altering GABAergic inhibition of granule cells in TLE.

Fatty acyl thioesterases control the termination of intraplastidial fatty acid synthesis by hydrolyzing fatty acyl-ACP complexes. The fatty acyl thioesterase A (FATA) gene family in Arabidopsis comprises two members, i.e., FATA1 and FATA2. Previous studies have shown that FATAs display high specificity for unsaturated fatty acids. However, the expression pattern and individual roles of these two FATA genes remains unknown. In this study, we initially studied the expression patterns of FATA1 and FATA2 in various organs of Arabidopsis and we found that FATA1 was expressed at low level in all organs examined and FATA2 was detected in all organs examined, with especially high accumulation in siliques. The transient expression of a FATA2-eGFP fusion in Arabidopsis green leaf protoplasts showed that FATA2 was localized in chloroplasts. A T-DNA insertion mutant line of FATA2 (named fata2) was obtained and used for phenotypic observation. Semiquantitative RT-PCR assay showed that the expression level of FATA2 decreased significantly in fata2 compared with that in wild type. Furthermore, fata2 mutants produced longer siliques with more seeds, whereas seed size was slightly smaller than that of wild type. Compositional analysis of seed oil revealed that, except for a subtly decreased C24:0 and unchanged C22:0 level, all other fatty acids were increased by between 10 and 60 % in fata2 dry seeds compared with those in wild-type. Taken together, our results indicate that FATA2 plays important roles in lipid metabolism in seeds and in silique development in Arabidopsis thaliana.