Anti-Inflammatory Effects of Chamaecyparis obtusa (Siebold & Zucc.) Endl. Leaf Extract Fermented by Ganoderma applanatum Mycelia.

Corticosteroids are commonly used anti-inflammatory agents. However, their prolonged use can lead to side effects. Therefore, the development of natural compounds with minimal side effects is necessary. This study was performed to investigate the anti-inflammatory effects and mechanisms of action of Chamaecyparis obtusa (Siebold & Zucc.) Endl. leaf (COL), bioconverted using Ganoderma applanatum (G. applanatum) in lipopolysaccharide (LPS)-induced RAW264.7 cells. The COL 70% EtOH extract fermented by G. applanatum (70COLGA) improved the high cytotoxicity of 70% EtOH extracts (70COL). When RAW264.7 cells were pre-treated with 100 and 200 µg/mL of 70COLGA for 2 h and then treated with LPS for 16 h, LPS induced the production of nitric oxide (NO), and the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) were significantly inhibited. When RAW264.7 cells were pre-treated with 100 and 200 µg/mL of 70COLGA for 2 h and then treated with LPS for 4 h, the phosphorylation of signal transducers and activators of transcription (STAT) was markedly decreased. In addition, 70COLGA markedly suppressed the production of the inflammatory cytokines interleukin (IL)-1ß and IL-6 in LPS-induced RAW264.7 cells. Analysis of pro-inflammatory molecules using cytokine arrays showed that macrophage inflammatory protein (MIP)-2, granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF) and IL-27 expressions were also suppressed by 200 µg/mL of 70COLGA in LPS-induced RAW264.7 cells. These results demonstrate that 70COLGA significantly prevented inflammatory responses by inhibiting the secretion of pro-inflammatory molecules in LPS-induced RAW264.7 cells. When RAW264.7 cells were pre-treated with 100 and 200 µg/mL of 70COLGA for 2 h and then treated with LPS-conditioned medium (LPS-CM) for 30 min, 70COLGA directly inhibited STAT activation. In summary, our findings suggest that 70COLGA has therapeutic potential for the treatment of inflammatory diseases.

Salmonella vector induces protective immunity against Lawsonia and Salmonella in murine model using prokaryotic expression system.

BACKGROUND: Lawsonia intracellularis is the causative agent of proliferative enteropathy and is associated with several outbreaks, causing substantial economic loss to the porcine industry. OBJECTIVES: In this study, we focused on demonstrating the protective effect in the mouse model through the immunological bases of two vaccine strains against porcine proliferative enteritis. METHODS: We used live-attenuated Salmonella Typhimurium (ST) secreting two selected immunogenic LI antigens (Lawsonia autotransporter A epitopes and flagellin [FliC]-peptidoglycan-associated lipoprotein-FliC) as the vaccine carrier. The constructs were cloned into a Salmonella expression vector (pJHL65) and transformed into the ST strain (JOL912). The expression of immunogenic proteins within Salmonella was evaluated via immunoblotting. RESULTS: Immunizing BALB/c mice orally and subcutaneously induced high levels of LI-specific systemic immunoglobulin G and mucosal secretory immunoglobulin A. In immunized mice, there was significant upregulation of interferon-γ and interleukin-4 cytokine mRNA and an increase in the subpopulations of cluster of differentiation (CD) 4+ and CD 8+ T lymphocytes upon splenocytes re-stimulation with LI antigens. We observed significant protection in C57BL/6 mice against challenge with 106.9 times the median tissue culture infectious dose of LI or 2 × 109 colony-forming units of the virulent ST strain. Immunizing mice with either individual vaccine strains or co-mixture inhibited bacterial proliferation, with a marked reduction in the percentage of mice shedding Lawsonia in their feces. CONCLUSIONS: Salmonella-mediated LI gene delivery induces robust humoral and cellular immune reactions, leading to significant protection against LI and salmonellosis.

Development of a Gait Feature-Based Model for Classifying Cognitive Disorders Using a Single Wearable Inertial Sensor.

BACKGROUND AND OBJECTIVES: Gait changes are potential markers of cognitive disorders (CDs). We developed a model for classifying older adults with CD from those with normal cognition using gait speed and variability captured from a wearable inertial sensor and compared its diagnostic performance for CD with that of the model using the Mini-Mental State Examination (MMSE). METHODS: We enrolled community-dwelling older adults with normal gait from the Korean Longitudinal Study on Cognitive Aging and Dementia and measured their gait features using a wearable inertial sensor placed at the center of body mass while they walked on a 14-m long walkway thrice at comfortable paces. We randomly split our entire dataset into the development (80%) and validation (20%) datasets. We developed a model for classifying CD using logistic regression analysis from the development dataset and validated it in the validation dataset. In both datasets, we compared the diagnostic performance of the model with that using the MMSE. We estimated optimal cutoff score of our model using receiver operator characteristic analysis. RESULTS: In total, 595 participants were enrolled, of which 101 of them experienced CD. Our model included both gait speed and temporal gait variability and exhibited good diagnostic performance for classifying CD from normal cognition in both the development (area under the receiver operator characteristic curve [AUC] = 0.788, 95% CI 0.748-0.823, p < 0.001) and validation datasets (AUC = 0.811, 95% CI 0.729-0.877, p < 0.001). Our model showed comparable diagnostic performance for CD with that of the model using the MMSE in both the development (difference in AUC = 0.026, standard error [SE] = 0.043, z statistic = 0.610, p = 0.542) and validation datasets (difference in AUC = 0.070, SE = 0.073, z statistic = 0.956, p = 0.330). The optimal cutoff score of the gait-based model was >-1.56. DISCUSSION: Our gait-based model using a wearable inertial sensor may be a promising diagnostic marker of CD in older adults. CLASSIFICATION OF EVIDENCE: This study provides Class III evidence that gait analysis can accurately distinguish older adults with CDs from healthy controls.

Blended Polymer Dry Electrodes for Reliable Electrocardiogram and Electromyogram Measurements and Their Eco-Friendly Disposal Led by Degradability in Hot Water.

To increase the human lifespan, healthcare monitoring devices that diagnose diseases and check body conditions have attracted considerable interest. Commercial AgCl-based wet electrodes with the advantages of high conductivity and strong adaptability to human skin are considered the most frequently used electrode material for healthcare monitoring. However, commercial AgCl-based wet electrodes, when exposed for a long period, cause an evaporation of organic solvents, which could reduce the signal-to-noise ratio of biosignals and stimulate human skin. In this context, we demonstrate a dry electrode for a poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS)-based blended polymer electrode using a combination of PEDOT:PSS, waterborne polyurethane (WPU) and ethylene glycol (EG) that could be reused for a long period of time to detect electrocardiography (ECG) and electromyography (EMG). Both ECG and EMG are reliably detected by the wireless real-time monitoring system. In particular, the proposed dry electrode detects biosignals without deterioration for over 2 weeks. Additionally, a double layer of a polyimide (PI) substrate and fluorinated polymer CYTOP induces the strong waterproof characteristics of external liquids for the proposed dry electrodes, having a low surface energy of 14.49 mN/m. In addition, the proposed electrode has excellent degradability in water; it dissolves in hot water at 60 °C.

Simple and cost-effective microfabrication of flexible and stretchable electronics for wearable multi-functional electrophysiological monitoring.

The fabrication of flexible and stretchable electronics is a critical requirement for the successful application of wearable healthcare devices. Although such flexible electronics have been commonly fabricated by microelectromechanical system (MEMS) technologies, they require a specialised equipment for vacuum deposition, photolithography, and wet and dry etching. A photolithography-free simple patterning method using a desktop plotter cutter has been proposed; however, the metal formation and electrode opening still rely on the MEMS technology. To address this issue, we demonstrate a simple, rapid, cost-effective, and a complete microfabrication process for flexible and stretchable sensor platforms encompassing conductor formation and patterning to encapsulate and open sensing windows, which only require an economic plotter cutter and readily available supplies. Despite its simplicity, the proposed process could stably create microscale features of 200 µm wide conductor lines and 1 mm window openings, which are in the useful range for various wearable applications. The feasibility of the simple fabrication of multi-functional sensors for various physiological monitoring applications was successfully demonstrated in electrochemical (glucose), electrical (electrocardiogram), mechanical (strain), and thermal (body temperature) modalities.

Evaluation methods for long-term reliability of polymer-based implantable biomedical devices.

Long-term reliability of implantable biomedical devices is a critical issue for their practical usefulness and successful translation into clinical application. Reliability is particularly of great concern for recently demonstrated devices based on new materials typically relying on polymeric thin films and microfabrication process. While reliability testing protocol has been well-established for traditional metallic packages, common evaluation methods for polymer-based microdevices has yet to be agreed upon, even though various testing methods have been proposed. This article is aiming to summarize the evaluation methods on long-term reliability of emerging biomedical implants based on polymeric thin-films in terms of their theories and implementation with specific focus on difference from the traditional metallic packages.

Antiviral Activities of Quercetin and Isoquercitrin Against Human Herpesviruses.

We previously reported that the ethyl acetate (EtOAc) fraction of a 70% ethanol extract of Elaeocarpus sylvestris (ESE) inhibits varicella-zoster virus (VZV) and human cytomegalovirus (HCMV) replication in vitro. PGG (1,2,3,4,6-penta-O-galloyl-ß-D-glucose) is a major chemical constituent of the EtOAc fraction of ESE that inhibits VZV but not HCMV replication. In this study, we comprehensively screened the chemical compounds identified in the EtOAc fraction of ESE for potential antiviral properties. Among the examined compounds, quercetin and isoquercitrin displayed potent antiviral activities against both VZV and HCMV with no significant cytotoxic effects. Both compounds strongly suppressed the expression of VZV and HCMV immediate-early (IE) genes. Our collective results indicated that, in addition to PGG, quercetin and isoquercitrin are bioactive compounds in the EtOAc fraction of ESE that effectively inhibit human herpesvirus replication.

Lactobacillus brevis strains from fermented aloe vera survive gastroduodenal environment and suppress common food borne enteropathogens.

Five novel Lactobacillus brevis strains were isolated from naturally fermented Aloe vera leaf flesh. Each strain was identified by Random Amplified Polymorphic DNA (RAPD) analysis and 16S rRNA sequence comparison. These strains were highly tolerant to acid, surviving in pH2.5 for up to 4 hours, and resistant to 5% bile salts at 37°C for 18 hours. Due to its tolerance to acid and bile salts, one strain passed through the gastric barrier and colonised the intestine after oral administration. All five strains inhibited the growth of many harmful enteropathogens without restraining most of normal commensals in the gut and hence named POAL (Probiotics Originating from Aloe Leaf) strains. Additionally, each strain exhibited discriminative resistance to a wide range of antibiotics. The L. brevis POAL strains, moreover, expressed high levels of the glutamate decarboxylase (GAD) gene which produces a beneficial neurotransmitter, γ-aminobutyric acid (GABA). These characteristics in all suggest that the novel L. brevis strains should be considered as potential food additives and resources for pharmaceutical research.