Hyper-production of porcine contagious pleuropneumonia subunit vaccine proteins in Escherichia coli by developing a bicistronic T7 expression system.

The ApxII toxin and the outer membrane lipoprotein (Oml) of Actinobacillus pleuropneumoniae are important vaccine antigens against porcine contagious pleuropneumonia (PCP), a prevalent infectious disease affecting the swine industry worldwide. Previous studies have reported the recombinant expression of ApxII and Oml in Escherichia coli; however, their yields were not satisfactory. Here, we aimed to enhance the production of ApxII and Oml by constructing a bicistronic expression system based on the widely used T7 promoter. To create efficient T7 bicistronic expression cassettes, 16 different fore-cistron sequences were introduced downstream of the T7 promoter. The expression of three vaccine antigens Oml1, Oml7, and ApxII in the four strongest bicistronic vectors were enhanced compared to the monocistronic control. Further optimization of the fermentation conditions in micro-well plates (MWP) led to improved production. Finally, the production yields reached unprecedented levels of 2.43 g L-1 of Oml1, 2.59 g L-1 of Oml7, and 1.21 g L-1 of ApxII, in a 5 L bioreactor. These three antigens also demonstrated well-protective immunity against A. pleuropneumoniae infection. In conclusion, this study establishes an efficient bicistronic T7 expression system that can be used to express recombinant proteins in E. coli and achieves the hyper-production of PCP vaccine proteins.

Contactless Monitoring of Respiratory Rate and Breathing Absence from Head Movements Using an RGB Camera.

Contactless vital sign monitoring is more demanding for long-term, continuous, and unobtrusive measurements. Camera-based respiratory monitoring is receiving growing interest with advanced video technologies and computational power. The volume variations of the lungs for airflow changes create a periodic movement of the torso, but identifying the torso is more challenging than face detection in a video. In this paper, we present a unique approach to monitoring respiratory rate (RR) and breathing absence by leveraging head movements alone from an RGB video because respiratory motion also influences the head. Besides our novel RR estimation, an independent algorithm for breathing absence detection using signal feature extraction and machine learning techniques identifies an apnea event and improves overall RR estimation accuracy. The proposed approach was evaluated using videos from 30 healthy subjects who performed various breathing tasks. The breathing absence detector had 0.87 F1 score, 0.9 sensitivity, and 0.85 specificity. The accuracy of spontaneous breathing rate estimation increased from 2.46 to 1.91 bpm MAE and 3.54 to 2.7 bpm RMSE when combining the breathing absence result with the estimated RR.Clinical relevance- Our contactless respiratory monitoring can utilize a consumer RGB camera to offer a significant benefit in continuous monitoring of neonatal monitoring, sleep monitoring, telemedicine or telehealth, home fitness with mild physical movement, and emotion detection in the clinic and remote locations.

Enhanced Contactless Heart Rate Monitoring Using Camera with Motion Artifact Removal During Physical Activities.

Contactless monitoring of heart rate (HR) can improve passive and continuous tracking of cardiovascular activities and overall people's health. Remote photoplethysmography (rPPG) using a camera eliminates the need for a wearable device. rPPG-based HR has shown promising results to be accurate and comparable to conventional methods such as contact PPG. Most experiments use stationary subjects while motion is known to affect the accuracy of remote PPG. In this paper, a novel methodology is introduced to enhance the accuracy and reliability of HR monitoring based on rPPG in the presence of physical activities like Yoga. This method quickly and accurately tracks HR and analyzes head motion to exclude unreliable data within short windows of rPPG signals. The method was tested with smartphone video data collected from 60 subjects when they are doing activities with varying levels of movement. Results show that our method without motion removal improves the accuracy of the HR readings by 0.7 bpm, reaching 3.57 bpm on average for a 30-sec-window. The accuracy is further improved by another 1.3 bpm after removing the motion artifacts, and reaches 2.29 bpm.Clinical relevance- The enhancement of HR readings from shorter rPPG signal with motion tolerance during physical activities can ultimately help with a more reliable HR tracking of people in uncontrolled settings like home which is a critical step towards remote health-care or wellness tracking.

Power Optimized Smartwatch-Earbuds Multimodal System for Monitoring Activities of Daily Living.

Activities of daily living is an important entity to monitor for promoting healthy lifestyle for chronic disease patients, children and the healthy population. This paper presents a smartwatch and earbuds inertial sensors based multi-modal power efficient end-to-end mobile system for continuous, passive and accurate detection of broad daily activity classes. We collected various posture, stationary and moving activity data from 40 diverse subjects using earbuds and smartwatch and develop the novel power optimized end-to-end operational system consisting of i) optimized device sampling rates and Bluetooth packet transfer rates, ii) data buffering mechanism, iii) background services, and iv) optimized model size, and demonstrating 93% macro recall score in detecting various activities. Our power optimized solution uses 80%, 40% and 33.33% less battery power for the smartphone, smartwatch, and earbuds respectively, compared to a power agnostic system with an estimated continuous no-charging run time of 50 hours, 16.67 hours, and 25 hours for the smartphone, smartwatch, and earbuds respectively.Clinical relevance- The end-to-end power optimized activity detection system presented in this paper will assist practicing clinicians toward treatment of various chronic disease patients (e.g. diabetes, hypertension, heart disease and obesity) by long-term, continuous monitoring of their lifestyle and sedentary behavior.

RRDetection: Respiration Rate Estimation Using Earbuds During Physical Activities.

In modern times, earbuds have become both popular and essential accessories for people to use with a wide range of devices in their everyday lives. Moreover, the respiration rate is a crucial vital sign that is sensitive to various pathological conditions. Many earbuds now come equipped with multiple sensing capabilities, including inertial and acoustic sensors. These sensors can be used by researchers to passively monitor users' vital signs, such as respiration rates. While current earbud-based breath rate estimation algorithms mostly focus on resting conditions, recent studies have demonstrated that respiration rates during physical activities can predict cardio-respiratory fitness for healthy individuals and pulmonary conditions for respiratory patients. To address this gap, we propose a novel algorithm called RRDetection that leverages the motion sensors in ordinary earbuds to detect respiration rates during light to moderate physical activities.

Estimating SpO2 with Deep Oxygen Desaturations from Facial Video Under Various Lighting Conditions: A Feasibility Study.

This paper presents a feasibility study to collect data, process signals, and validate accuracy of peripheral oxygen saturation (SpO2) estimation from facial video in various lighting conditions. We collected facial videos using RGB camera, without auto-tuning, from subjects when they were breathing through a mouth tube with their nose clipped. The videos were record under four lighting conditions: warm color temperature and normal brightness, neutral color temperature and normal brightness, cool color temperature and normal brightness, neutral color temperature and dim brightness. The air inhaled by the subjects was manually controlled to gradually induce hypoxemia and lower subjects' SpO2 to as low as 81%. We first extracted the remote photoplethysmogram (rPPG) signals from the videos. We applied the principle of pulse oximetry and extracted the ratio of ratios (RoR) for two color combinations: Red/Blue and Red/Green. Next, we assessed SpO2 estimation accuracy against the ground truth, a Transfer Standard Pulse Oximeter. We have achieved an RMSE of 1.93% and a PCC of 0.97 under the warm color temperature and normal brightness lighting condition using leave-one-subject-out cross validation between two subjects. The results have demonstrated the feasibility to estimate SpO2 remotely and accurately using consumer level RGB camera with suitable camera configuration and lighting condition.Clinical Relevance- This work demonstrates that SpO2 can be estimated accurately using an RGB camera without auto-tuning and under warm color temperature, enabling continuous SpO2 monitoring applications that require noncontact sensing.

ApoE enhances mitochondrial metabolism via microRNA-142a/146a-regulated circuits that suppress hematopoiesis and inflammation in hyperlipidemia.

Apolipoprotein E (ApoE) is recognized for its pleiotropic properties that suppress inflammation. We report that ApoE serves as a metabolic rheostat that regulates microRNA control of glycolytic and mitochondrial activity in myeloid cells and hematopoietic stem and progenitor cells (HSPCs). ApoE expression in myeloid cells increases microRNA-146a, which reduces nuclear factor κB (NF-κB)-driven GLUT1 expression and glycolytic activity. In contrast, ApoE expression reduces microRNA-142a, which increases carnitine palmitoyltransferase 1a (CPT1A) expression, fatty acid oxidation, and oxidative phosphorylation. Improved mitochondrial metabolism by ApoE expression causes an enrichment of tricarboxylic acid (TCA) cycle metabolites and nicotinamide adenine dinucleotide (NAD+) in macrophages. The study of mice with conditional ApoE expression supports the capacity of ApoE to foster microRNA-controlled immunometabolism. Modulation of microRNA-146a and -142a in the hematopoietic system of hyperlipidemic mice using RNA mimics and antagonists, respectively, improves mitochondrial metabolism, which suppresses inflammation and hematopoiesis. Our findings unveil microRNA regulatory circuits, controlled by ApoE, that exert metabolic control over hematopoiesis and inflammation in hyperlipidemia.

High-throughput process development from gene cloning to protein production.

In the post-genomic era, the demand for faster and more efficient protein production has increased, both in public laboratories and industry. In addition, with the expansion of protein sequences in databases, the range of possible enzymes of interest for a given application is also increasing. Faced with peer competition, budgetary, and time constraints, companies and laboratories must find ways to develop a robust manufacturing process for recombinant protein production. In this review, we explore high-throughput technologies for recombinant protein expression and present a holistic high-throughput process development strategy that spans from genes to proteins. We discuss the challenges that come with this task, the limitations of previous studies, and future research directions.

ApoE expression in macrophages communicates immunometabolic signaling that controls hyperlipidemia-driven hematopoiesis & inflammation via extracellular vesicles.

While apolipoprotein E (apoE) expression by myeloid cells is recognized to control inflammation, whether such benefits can be communicated via extracellular vesicles is not known. Through the study of extracellular vesicles produced by macrophages derived from the bone marrow of Wildtype (WT-BMDM-EV) and ApoE deficient (EKO-BMDM-EV) mice, we uncovered a critical role for apoE expression in regulating their cell signaling properties. WT-BMDM-EV communicated anti-inflammatory properties to recipient myeloid cells by increasing cellular levels of apoE and miR-146a-5p, that reduced NF-κB signalling. They also downregulated cellular levels of miR-142a-3p, resulting in increased levels of its target carnitine palmitoyl transferase 1A (CPT1A) which improved fatty acid oxidation (FAO) and oxidative phosphorylation (OxPHOS) in recipient cells. Such favorable metabolic polarization enhanced cell-surface MerTK levels and the phagocytic uptake of apoptotic cells. In contrast, EKO-BMDM-EV exerted opposite effects by reducing cellular levels of apoE and miR-146a-5p, which increased NF-κB-driven GLUT1-mediated glucose uptake, aerobic glycolysis, and oxidative stress. Furthermore, EKO-BMDM-EV increased cellular miR-142a-3p levels, which reduced CPT1A levels and impaired FAO and OxPHOS in recipient myeloid cells. When cultured with naïve CD4+ T lymphocytes, EKO-BMDM-EV drove their activation and proliferation, and fostered their transition to a Th1 phenotype. While infusions of WT-BMDM-EV into hyperlipidemic mice resolved inflammation, infusions of EKO-BMDM-EV increased hematopoiesis and drove inflammatory responses in myeloid cells and T lymphocytes. ApoE-dependent immunometabolic signaling by macrophage extracellular vesicles was dependent on transcriptional axes controlled by miR-146a-5p and miR-142a-3p that could be reproduced by infusing miR-146a mimics & miR-142a antagonists into hyperlipidemic apoE-deficient mice. Together, our findings unveil a novel property for apoE expression in macrophages that modulates the immunometabolic regulatory properties of their secreted extracellular vesicles.

Flavonoids from Seabuckthorn (Hippophae rhamnoides L.) restore CUMS-induced depressive disorder and regulate the gut microbiota in mice.

Seabuckthorn (Hippophae rhamnoides L.), which is enriched with flavonoids, including isorhamnetin, quercetin and kaempferol, is a representative example of "medicine food homology" targeting several diseases. Major depressive disorders seriously threaten mental health worldwide and may even lead to death. Chronic unpredictable mild stress (CUMS)-induced depressive-like symptoms in mice are usually considered as the highest similarity to the situation in humans. Herein, we determined the potential functions of the flavonoid-enriched fraction from Seabuckthorn, which was named SBF, in treating major depressive disorder in mice. In the CUMS-induced mouse model, the intake of SBF reversed their depressive behaviors and relieved the CUMS-disturbed levels of neurotrophins, neurotransmitters, stress-related hormones, and inflammation-related cytokines. Additionally, the treatment of depressive mice with SBF showed ability to regulate the gut microbiota, especially in decreasing the abundance of Lactobacillaceae, while increasing the abundance of Lachnospiraceae at the family level. The results suggest the beneficial effects of Seabuckthorn flavonoids in functioning as a health food supplement to treat major depressive disorders.

Leaderless Bicistronic Design for Precise and Reliable Control of Gene Expression in Corynebacterium Glutamicum.

In synthetic biology, the precise control of gene expression is challenging due to the limited orthogonality of expression elements. Here, to address this issue and improve the reusability of genetic elements, we developed a bicistronic expression cassette in Corynebacterium glutamicum based on a leaderless promoter lacking a 5'UTR. The created leaderless bicistronic design (BCD) significantly improved the orthogonality of expression elements across different genes of interest. We also explored the importance of the fore-cistron and SD motif in maintaining the strength of leaderless BCDs. Additionally, we established a library containing 55,901 fore-cistrons and demonstrated that the regulatory range of gene expression in leaderless BCDs can be broader by modifying the fore-cistron sequence. This study provides a novel synthetic biology tool based on leaderless BCD for fine-tuning gene expression in C. glutamicum using fore-cistrons. Moreover, the strategy developed here can also be applied to improve the performance of other leaderless promoters in other bacteria.

Differential regulation of BIRC2 and BIRC3 expression by inflammatory cytokines and glucocorticoids in pulmonary epithelial cells.

Roles for the baculoviral inhibitor of apoptosis repeat-containing (BIRC) genes, BIRC2 and BIRC3, may include signaling to the inflammatory transcription factor, nuclear factor-κB (NF-κB) and protection from cell death. However, distinct functions for each BIRC are not well-delineated. Given roles for the epithelium in barrier function and host defence, BIRC2 and BIRC3 expression was characterized in pulmonary epithelial cell lines and primary human bronchial epithelial cells (pHBECs) grown as undifferentiated cells in submersion culture (SC) or as highly differentiated cells at air-liquid interface (ALI). In A549 cells, interleukin-1ß (IL1B) and tumor necrosis factor α (TNF) induced BIRC3 mRNA (~20-50-fold), with maximal protein expression from 6-24 h. Similar effects occurred in BEAS-2B and Calu-3 cells, as well as SC and ALI pHBECs. BIRC2 protein was readily detected in unstimulated cells, but was not markedly modulated by IL1B or TNF. Glucocorticoids (dexamethasone, budesonide) modestly increased BIRC3 mRNA and protein, but showed little effect on BIRC2 expression. In A549 cells, BIRC3 mRNA induced by IL1B was unchanged by glucocorticoids and showed supra-additivity with TNF-plus-glucocorticoid. Supra-additivity was also evident for IL1B-plus-budesonide induced-BIRC3 in SC and ALI pHBECs. Using A549 cells, IL1B- and TNF-induced BIRC3 expression, and to a lesser extent, BIRC2, was prevented by NF-κB inhibition. Glucocorticoid-induced BIRC3 expression was prevented by silencing and antagonism of the glucocorticoid receptor. Whereas TNF, but not IL1B, induced degradation of basal BIRC2 and BIRC3 protein, IL1B- and TNF-induced BIRC3 protein remained stable. Differential regulation by cytokines and glucocorticoids shows BIRC2 protein expression to be consistent with roles in rapid signaling events, whereas cytokine-induced BIRC3 may be more important in later effects. While TNF-induced degradation of both BIRCs may restrict their activity, cytokine-enhanced BIRC3 expression could prime for its function. Finally, shielding from glucocorticoid repression, or further enhancement by glucocorticoid, may indicate a key protective role for BIRC3.

Flavonoids from Seabuckthorn (Hippophae rhamnoides L.) mimic neurotrophic functions in inducing neurite outgrowth in cultured neurons: Signaling via PI3K/Akt and ERK pathways.

BACKGROUND: Various brain disorders, including neurodegenerative diseases and major depressive disorders, threaten an increasing number of patients. Seabuckthorn, a fruit from Hippophae rhamnoides L., is an example of "medicine food homology". The fruit has enriched flavonoids that reported to have benefits in treating cognitive disorders. However, the studies on potential functions of Seabuckthorn and/or its flavonoid-enriched fraction in treating neurodegenerative disorders are limited. PURPOSE: This study aimed to determine the ability and mechanism of the flavonoid-enriched fraction of Seabuckthorn (named as SBF) in mimicking the neurotrophic functions in inducing neurite outgrowth of cultured neurons. METHODS: Cultured PC12 cell line, SH-SY5Y cell line and primary neurons (cortical and hippocampal neurons isolated from E17-19 SD rat embryos) were the employed models to evaluate SBF in inducing neurite outgrowth by comparing to the effects of NGF and BDNF. Immuno-fluorescence staining was applied to identify the morphological change during the neuronal differentiation. Luciferase assay was utilized for analyzing the transcriptional regulation of neurofilaments and cAMP/CREB-mediated gene. Western blot assay was conducted to demonstrate the expressions of neurofilaments and phosphorylated proteins. RESULTS: The application of SBF induced neuronal cell differentiation, and this differentiating activation was blocked by the inhibitors of PI3K/Akt and ERK pathways. Additionally, SBF showed synergy with neurotrophic factors in stimulating the neurite outgrowth of cultured neurons. Moreover, the major flavonoids within SBF, i.e., isorhamnetin, quercetin and kaempferol, could account for the neurotrophic activities of SBF. CONCLUSION: Seabuckthorn flavonoids mimicked neurotrophic functions in inducing neuronal cell differentiation via activating PI3K/Akt and ERK pathways. The results suggest the beneficial functions of Seabuckthorn as a potential health food supplement in treating various brain disorders, e.g., neurodegenerative diseases.

The neurotrophic activities of brain-derived neurotrophic factor are potentiated by binding with apigenin, a common flavone in vegetables, in stimulating the receptor signaling.

AIMS: We aimed to identify the neurotrophic activities of apigenin (4',5,7-trihydroxyflavone) via its coordination with brain-derived neurotrophic factor (BNDF) and an elevated signaling of tyrosine kinase receptor B (Trk B receptor). METHODS: The direct binding of apigenin to BDNF was validated by ultrafiltration and biacore assay. Neurogenesis, triggered by apigenin and/or BDNF, was determined in cultured SH-SY5Y cells and rat cortical neurons. The amyloid-beta (Aß)25-35 -induced cellular stress was revealed by propidium iodide staining, mitochondrial membrane potential, bioenergetic analysis, and formation of reactive oxygen species levels. Activation of Trk B signaling was tested by western blotting. RESULTS: Apigenin and BDNF synergistically maintained the cell viability and promoted neurite outgrowth of cultured neurons. In addition, the BDNF-induced neurogenesis of cultured neurons was markedly potentiated by applied apigenin, including the induced expressions of neurofilaments, PSD-95 and synaptotagmin. Moreover, the synergy of apigenin and BDNF alleviated the (Aß)25-35 -induced cytotoxicity and mitochondrial dysfunction. The synergy could be accounted by phosphorylation of Trk B receptor, and which was fully blocked by a Trk inhibitor K252a. CONCLUSION: Apigenin potentiates the neurotrophic activities of BDNF through direct binding, which may serve as a possible treatment for its curative efficiency in neurodegenerative diseases and depression.

The ethanolic extract of peanut shell attenuates the depressive-like behaviors of mice through modulation of inflammation and gut microbiota.

Peanut shell is an agricultural byproduct being wasted on a large scale, which is in urgent need to be recycled. To fully utilize its pharmacological ingredients, e.g. luteolin, eriodyctiol, and 5,7-dihydroxychromone, we evaluated the curative effect of ethanol extract deriving from peanut shell (PSE) in treating chronic unpredictable mild stress (CUMS)-induced depressive mice. The chronic stress lasted for 10 weeks, and PSE at 100-900 mg/kg/day was gavaged to mice in the last 2 weeks of modeling. The depressive behaviors were assessed by analyses of sucrose preference, tail suspension, and forced swimming. The brain injury was demonstrated by Hematoxylin and Eosin (H&E), Nissl body, and TdT-mediated dUTP nick end labeling (TUNEL) stainings in the mouse hippocampus. Biochemical indicators were analyzed, including levels of neurotrophic factors, neurotransmitters, stress hormones, and inflammatory mediators. The feces were collected for the 16S rDNA sequencing of gut microbiome. Administration of PSE improved the sucrose water consumption of depressive mice, while it decreased the immobile time in tail suspension and forced swimming tests. Meanwhile, the anti-depressive effect of PSE was supported by ameliorated histochemical staining, increased levels of neurotrophic factors and neurotransmitters, as well as down-regulated stress hormones. Furthermore, the treatment of PSE was able to mitigate the levels of inflammatory cytokines in brain, serum, and small intestine. Besides, the tight junction proteins, e.g., occludin and ZO-1, of gut showed elevated expressions, which coincided with the elevated abundance and diversity of gut microbiota upon PSE treatment. This study validated the therapeutic efficacy of PSE in fighting against depression, as well as its modulatory action on inflammation and gut microbiota, which promoted the recycling of this agricultural waste to be health supplements of added value.

Deep Multivariate Domain Translation for Device Invariant Pulmonary Patient Identification from Cough and Speech Sounds.

Pulmonary audio sensing from cough and speech sounds in commodity mobile and wearable devices is increasingly used for remote pulmonary patient monitoring, home healthcare, and automated disease analysis. Patient identification is important for such applications to ensure system accuracy and integrity, and thus avoiding errors and misdiagnosis. Widespread usage and deployment of such patient identification models across various devices are challenging due to domain shift of acoustic features because of device heterogeneity. Because of this phenomenon, a patient identification model developed using audio data collected with one type of device is not usable when deployed in another type of device, which is a concern for model portability and general usability. This paper presents a framework utilizing a multivariate deep neural network regressor as a feature translator between source device and target device domains to reduce the effect of domain shift for better model portability. Extensive and empirical experiments of our translation framework consisting of two different human sound (speech and cough) based pulmonary patient identification tasks using audio data collected from 91 real patients demonstrate that it can recover up to 64.8% of lost accuracy due to domain shift across two common and widely used mobile and wearable devices: smartphone and smartwatch. Clinical Relevance- The methods presented in this paper will enable efficient and easy portability of pulmonary patient identification models from cough and speech across various mobile and wearable devices used by a patient.

Remote Photoplethysmography and Heart Rate Estimation by Dynamic Region of Interest Tracking.

Remote photoplethysmography (PPG) estimates vital signs by measuring changes in the reflected light from the human skin. Compared to traditional PPG techniques, remote PPG enables contactless measurement at a reduced cost. In this paper, we propose a novel method to extract remote PPG signals and heart rate from videos. We propose an algorithm to dynamically track regions of interest (ROIs) and combine the signals from all ROIs based on signal qualities. To maintain a stable frame rate and accuracy, we propose a dynamic down-sampling approach, which makes our system robust to the different video resolutions and user-camera distances. We also propose the strategy of adaptive measurement time to estimate HR, which can achieve comparable accuracy in HR estimation while reducing the average measurement time. To test the accuracy of the proposed system, we have collected data from 30 subjects with facial masks. Experimental results show that the proposed system can achieve 3.0 bpm mean absolute error in HR estimation.

Motion- Based Respiratory Rate Estimation with Motion Artifact Removal Using Video of Face and Upper Body.

Respiratory rate (RR) is a significant indicator of health conditions. Remote contactless measurement of RR is gaining popularity with recent respiratory tract infection awareness. Among various methods of contactless RR measurement, a video of an individual can be used to obtain an instantaneous RR. In this paper, we introduce an RR estimation based on the subtle motion of the head or upper chest captured on an RGB camera. Motion-based respiratory monitoring allows us to acquire RR from individuals with partial face coverings, such as glasses or a face mask. However, motion-based RR estimation is vulnerable to the subject's voluntary movement. In this work, adaptive selection between face and chest regions plus a motion artifact removal technique enables us to obtain a much cleaner respiratory signal from the video recordings. The average mean absolute error (MAE) for controlled and natural breathing is 1.95 BPM using head motion only and 1.28 BPM using chest motion only. Our results demonstrate the possibility of continuous monitoring of breathing rate in real-time with any personal device equipped with an RGB camera, such as a laptop or a smartphone.

A Herbal Mixture of Sesami Semen Nigrum and Longan Arillus Induces Neurite Outgrowth in Cultured Neurons and Shows Anti-Depression in Chronic Mild Stress-Induced Rats.

Medicinal food homology is referring to a group of food itself being considered as herbal medicine without a boundary of usage. Under the guidance of this food/medicine principle, the current study aims to develop anti-depressant from this food/medicine catalog. The herbal mixture of Sesami Semen Nigrum and Longan Arillus was evaluated in cultured PC12 rat pheochromocytoma cells, rat primary cortical neurons, and in chronic mild stress (CMS)-induced depressive rat model. The combination of two ethanolic extracts of Sesami Semen Nigrum and Longan Arillus in 1 : 1 ratio mimicked the function of nerve growth factor (NGF) and synergistically induced neurite outgrowth of PC12 cells. Besides, the expression and phosphorylation of tropomyosin receptor kinase A (TrkA) of the cultured cells were also elevated. This neurotrophic activity of herbal mixture was further supported by the increased expressions of biomarkers for neurogenesis and synaptogenesis in cortical neurons. Moreover, the depressed rats were soothed by the intake of herbal mixture, showing improved performance in behavior tests, as well as reversed levels of neurotransmitters and neurotrophic factors. Our results provide a new way to make full use of the current food/medicine resources, as to accelerate the development of therapeutics for depression.

Hypersecretion of OmlA antigen in Corynebacterium glutamicum through high-throughput based development process.

Outer membrane lipoprotein A (OmlA) is a vaccine antigen against porcine contagious pleuropneumonia (PCP), a disease severely affecting the swine industry. Here, we aimed to systematically potentiate the secretory production of OmlA in Corynebacterium glutamicum (C. glutamicum), a widely used microorganism in the food industry, by establishing a holistic development process based on our high-throughput culture platform. The expression patterns, expression element combinations, medium composition, and induction conditions were comprehensively screened or optimized in microwell plates (MWPs), followed by fermentation parameter optimization in a 4 × 1 L parallel fermentation system (CUBER4). An unprecedented yield of 1.01 g/L OmlA was ultimately achieved in a 5-L bioreactor following the scaling-up strategy of fixed oxygen mass transfer coefficient (kLa), and the produced OmlA antigen showed well-protective immunity against Actinobacillus pleuropneumoniae challenge. This result provides a rapid and reliable pipeline to achieve the hyper-production of OmlA, and possibly other recombinant vaccines, in C. glutamicum. KEY POINTS: • Established a holistic development process and applied it to potentiate the secretion of OmlA. • The secretion of OmlA reached an unprecedented yield of 1.01 g/L. • The recombinant OmlA antigen induced efficient protective immunity.