Optimization and evaluation of Atrina pectinata polysaccharides recovered by subcritical water extraction: A promising path to natural products.

In this study, the recovery of Atrina pectinata posterior adductor polysaccharides (APP-PS) using subcritical water extraction (SWE) was optimized by response surface methodology (RSM) and the physicochemical and biological properties of the recovered APP-PS were evaluated. The optimal extraction conditions, which resulted in a maximum yield of 55.58 ± 1.12 %, were temperature, 152.08 °C; extraction time, 10 min; solid-liquid ratio, 30 g/600 mL. The obtained APP-PS was found to be 88.05 ± 0.17 % total sugar. Fourier transform infrared (FT-IR) and Nuclear magnetic resonance (NMR) analyses confirmed the presence of the α-coordination of D-glucan in the polymer sample. The analysis of monosaccharide composition, along with thermogravimetric analysis, revealed the typical structure of the sample, composed of glucose alone. Total phenolic contents of APP-PS were measured as 5.47 ± 0.01 mg Gallic acid/g of dry sample and total flavonoids contents were determined to be 0.78 ± 0.06 mg Quercetin/g of dry sample. For biological activities, ABTS+, DPPH and FRAP antioxidant activities were measured to be 20.00 ± 0.71, 2.35 ± 0.05 and 4.02 ± 0.07 µg Trolox equivalent/100 g of dry sample, respectively. Additionally ACE inhibitory was confirmed to be 87.02 ± 0.47 %. These results showed that SWE is an effective method to recover biofunctional materials from marine organisms.

An experimental study on the performance of fire extinguishing system according to the agent charging condition and nozzle type.

In confined environments such as aircraft, an increase in mass impacts the overall system's performance, thus requiring sophisticated management. To verify whether the performance characteristics of fire extinguishing systems used in aircraft are satisfied, in this study was built a 1:1 scale test model. We examined the influence of the initial charge state and nozzles. Further, it measured the pressure inside the pipelines and vessels where multiple nozzles are installed to identify the flow and diffusivity characteristics of HFC-125 inside the pipelines and vessels. At a charging ratio of 54%, the initial pressure drop was smaller, and the lowest pressure before the bubble release point appeared 0.26 s later than when the charging ratio was 76%. The average pressure of each nozzle was 275.8 kPa higher under a charging ratio of 54% than 76% and increased further when the average concentration change was 54%, indicating that diffusivity increased. Although improvements occurred according to the charging ratio, the improvements according to the HFC-125 charging mass were more significant.

Exploring Bioactive Compounds in Brown Seaweeds Using Subcritical Water: A Comprehensive Analysis.

In this study, we characterized the bioactive properties of three important brown seaweed species, Sargassum thunbergii, Undaria pinnatifida, and Saccharina japonica, by subcritical water extraction (SWE), as these species are well known for their beneficial health effects. Their physiochemical properties, including potential antioxidant, antihypertensive, and α-glucosidase inhibitory activity, and the antibacterial activity of the hydroysates were also analyzed. The highest total phlorotannin, total sugar content, and reducing sugar content in the S. thunbergii hydrolysates were 38.82 ± 0.17 mg PGE/g, 116.66 ± 0.19 mg glucose/g dry sample, and 53.27 ± 1.57 mg glucose/g dry sample, respectively. The highest ABTS+ and DPPH antioxidant activities were obtained in the S. japonica hydrolysates (124.77 ± 2.47 and 46.35 ± 0.01 mg Trolox equivalent/g, respectively) and the highest FRAP activity was obtained in the S. thunbergii hydrolysates (34.47 ± 0.49 mg Trolox equivalent/g seaweed). In addition, the seaweed extracts showed antihypertensive (≤59.77 ± 0.14%) and α-glucosidase inhibitory activity (≤68.05 ± 1.15%), as well as activity against foodborne pathogens. The present findings provide evidence of the biological activity of brown seaweed extracts for potential application in the food, pharmaceutical, and cosmetic sectors.

Effect of airborne particle abrasion treatment of two types of 3D-printing resin materials for permanent restoration materials on flexural strength.

OBJECTIVES: This study aimed to assess the effects of airborne-particle abrasion (APA) on the flexural strength of two types of 3D-printing resins for permanent restoration. METHODS: Two types of 3D printing resins (urethane dimethacrylate oligomer; UDMA, ethoxylated bisphenol-A dimethacrylate; BEMA) constituting different components were printed. The specimen surfaces were subjected to APA using 50 and 110 µm alumina particles under different pressures. The three-point flexural strength was measured for each surface treatment group, and a Weibull analysis was performed. Surface characteristics were analyzed via surface roughness measurements and scanning electron microscopy. Dynamic mechanical analysis and nano-indentation measurements were limited to the control group. RESULTS: The three-point flexural strength according to the surface treatment was significantly lower in the UDMA group for large particle sizes and at high pressures; the BEMA group demonstrated low flexural strength for large particle sizes regardless of the pressure. After thermocycling, the flexural strengths of UDMA and BEMA significantly decreased in the group subjected to surface treatment. The Weibull modulus and characteristic strength of UDMA were higher than those of BEMA under different APA and thermocycling conditions. As the abrasion pressure and particle size increased, a porous surface formed, and the surface roughness increased. Compared with BEMA, UDMA featured a lower strain, greater strain recovery, and a negligible increase in modulus according to strain. SIGNIFICANCE: Thus, surface roughness increased with the sandblasting particle size and pressure of the 3D-printing resin. Hence, a suitable surface treatment method to improve adhesion can be determined by considering physical property changes.

Effect of Adhesion Conditions on the Shear Bond Strength of 3D Printing Resins after Thermocycling Used for Definitive Prosthesis.

Three-dimensional (3D) printing polymers such as urethane dimethacrylate (UDMA) and ethoxylated bisphenol A dimethacrylate (Bis-EMA) are typically used in definitive prosthesis and require surface treatments before bonding. However, surface treatment and adhesion conditions often affect long-term use. Herein, polymers were divided into Groups 1 and 2 for the UDMA and Bis-EMA components, respectively. The shear bond strength (SBS) between two types of 3D printing resins and resin cements was measured using Rely X Ultimate Cement and Rely X U200, according to adhesion conditions such as single bond universal (SBU) and airborne-particle abrasion (APA) treatments. Thermocycling was performed to evaluate the long-term stability. Sample surface changes were observed using a scanning electron microscope and surface roughness measuring instrument. The effect of interaction between the resin material and adhesion conditions on the SBS was analyzed via a two-way analysis of variance. The optimal adhesion condition for Group 1 was achieved when U200 was used after APA and SBU, whereas Group 2 was not significantly affected by the adhesion conditions. After thermocycling, the SBS significantly decreased in Group 1 without APA treatment and in the entire Group 2. Additionally, porosity, along with increased roughness, was observed on both material surfaces after APA.

Periplasmic Bacterial Biomineralization of Copper Sulfide Nanoparticles.

Metal sulfides are a common group of extracellular bacterial biominerals. However, only a few cases of intracellular biomineralization are reported in this group, mostly limited to greigite (Fe3 S4 ) in magnetotactic bacteria. Here, a previously unknown periplasmic biomineralization of copper sulfide produced by the magnetotactic bacterium Desulfamplus magnetovallimortis strain BW-1, a species known to mineralize greigite (Fe3 S4 ) and magnetite (Fe3 O4 ) in the cytoplasm is reported. BW-1 produces hundreds of spherical nanoparticles, composed of 1-2 nm substructures of a poorly crystalline hexagonal copper sulfide structure that remains in a thermodynamically unstable state. The particles appear to be surrounded by an organic matrix as found from staining and electron microscopy inspection. Differential proteomics suggests that periplasmic proteins, such as a DegP-like protein and a heavy metal-binding protein, could be involved in this biomineralization process. The unexpected periplasmic formation of copper sulfide nanoparticles in BW-1 reveals previously unknown possibilities for intracellular biomineralization that involves intriguing biological control and holds promise for biological metal recovery in times of copper shortage.

Microwave-assisted digestion method using diluted nitric acid and hydrogen peroxide for the determination of major and minor elements in milk samples by ICP-OES and ICP-MS.

A novel method for microwave-assisted digestion of milk samples using diluted HNO3and H2O2 with a single reaction chamber was developed for elemental analysis by ICP-based techniques. The optimal conditions for digestion were 0.25 g of sample mass, 6 mL of 0.1 molL-1HNO3and 2 mL of 30% H2O2 at 250 ℃ and 160 bar. The optimized procedure resulted in low residual carbon content and residual acidity of 260 mgL-1 and 0.06 mol L-1, respectively. The limits of detection ranged from 0.286Õ¸g g-1(Ca) to 82.990Õ¸g g-1(Fe). In addition, the proposed method was considered an excellent green analysis method with a final score of 87 based on the analytical Eco-Scale. Finally, the method was validated and applied to the determination of Al, As, Ba, Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Sb, Se, and Zn in milk samples from South Korea.

Diversity of Microbial Metal Sulfide Biomineralization.

Since the emergence of life on Earth, microorganisms have contributed to biogeochemical cycles. Sulfate-reducing bacteria are an example of widespread microorganisms that participate in the metal and sulfur cycles by biomineralization of biogenic metal sulfides. In this work, we review the microbial biomineralization of metal sulfide particles and summarize distinctive features from exemplary cases. We highlight that metal sulfide biomineralization is highly metal- and organism-specific. The properties of metal sulfide biominerals depend on the degree of cellular control and on environmental factors, such as pH, temperature, and concentration of metals. Moreover, biogenic macromolecules, including peptides and proteins, help cells control their extracellular and intracellular environments that regulate biomineralization. Accordingly, metal sulfide biominerals exhibit unique features when compared to abiotic minerals or biominerals produced by dead cell debris.

Effects of Short-term Mobile Application Use on Weight Reduction for Patients with Type 2 Diabetes.

BACKGROUND: The increasing prevalence of type 2 diabetes mellitus (T2DM) has led to a significant health burden. Technological advancements have highlighted the benefits of digital therapeutics for chronic diseases. In this study, we aimed to investigate the effects of a mobile application on weight reduction in patients with T2DM. METHODS: A total of 48 patients with T2DM was included in this single-center, randomized, controlled trial. In addition to conventional treatment, participants in the intervention group used a mobile application-based self-management system for diet, exercise, and medication adherence. The primary outcome of this study was weight change after 3 months of intervention, and secondary outcomes were metabolic parameters. RESULTS: After 12 weeks, no significant differences in body weight change were observed between the intervention and control groups (P=0.229). However, a significant difference was found in waist circumference (WC) between the two groups, wherein the control group showed an increase in WC (from 95.00±8.89 cm to 95.76±9.72 cm), while the intervention group showed a reduction (from 91.93±6.25 cm to 90.75±6.01 cm) with a significant time by group interaction (P=0.016). Additionally, participants with good compliance exhibited a more evident reduction in WC (P=0.037). However, no significant differences were found in other metabolic parameters between the two groups. CONCLUSION: Lifestyle modification using short-term mobile applications effectively reduced WC, especially in patients with good adherence to the application. However, weight reduction was not achieved.

Immunogenicity of an inactivated rabies vaccine for animals derived from the recombinant ERAGS strain.

PURPOSE: The aims of the present study were to evaluate the immunogenicity of an inactivated rabies vaccine based on the ERAGS strain. MATERIALS AND METHODS: The ERAGS virus propagated in Vero cells was inactivated with 3 mM binary ethylenimine for 8 hours. Three types of inactivated rabies vaccines were prepared to determine the minimum vaccine virus titers. Four further types of inactivated rabies vaccines were prepared by blending inactivated ERAGS with four different adjuvants; each vaccine was injected into mice, guinea pigs, and dogs to identify the optimal adjuvant. The immunogenicity of a Montanide (IMS) gel-adjuvanted vaccine was evaluated in cats, dogs, and cattle. Humoral immune responses were measured via a fluorescent antibody virus neutralization method and a blocking enzyme-linked immunosorbent assay. RESULTS: The minimum virus titer of the inactivated rabies vaccine was over 107.0 50% tissue culture infectious doses (TCID50 values)/mL. Of the four kinds of adjuvants, the IMS gel-adjuvanted vaccine induced the highest mean viral neutralizing antibody (VNA) titers of 6.24 and 2.36 IU/mL in guinea pigs and dogs, respectively, and was thus selected as the vaccine for the target animals. Cats, dogs, and cattle inoculated with the IMS gel-adjuvanted vaccine developed protective VNA titers ranging from 3.5 to 1.2 IU/mL at 4 weeks post-inoculation (WPI). CONCLUSION: Our data indicate that cats, dogs, and cattle inoculated with an inactivated rabies vaccine derived from the ERAGS strain developed protective immune responses that were maintained to 12 WPI.

Generation of a recombinant rabies virus expressing green fluorescent protein for a virus neutralization antibody assay.

BACKGROUND: Fluorescent antibody virus neutralization (FAVN) test is a standard assay for quantifying rabies virus-neutralizing antibody (VNA) in serum. However, a safer rabies virus (RABV) should be used in the FAVN assay. There is a need for a new method that is economical and time-saving by eliminating the immunostaining step. OBJECTIVES: We aimed to improve the traditional FAVN method by rescuing and characterizing a new recombinant RABV expressing green fluorescent protein (GFP). METHODS: A new recombinant RABV expressing GFP designated as ERAGS-GFP was rescued using a reverse genetic system. Immuno-fluorescence assay, peroxidase-linked assay, electron microscopy and reverse transcription polymerase chain reaction were performed to confirm the recombinant ERAGS-GFP virus as a RABV expressing the GFP gene. The safety of ERAGS-GFP was evaluated in 4-week-old mice. The rabies VNA titers were measured and compared with conventional FAVN and FAVN-GFP tests using VERO cells. RESULTS: The virus propagated in VERO cells was confirmed as RABV expressing GFP. The ERAGS-GFP showed the highest titer (108.0 TCID50/mL) in VERO cells at 5 days post-inoculation, and GFP expression persisted until passage 30. The body weight of 4-week-old mice inoculated intracranially with ERAGS-GFP continued to increase and the survival rate was 100%. In 62 dog sera, the FAVN-GFP result was significantly correlated with that of conventional FAVN (r = 0.95). CONCLUSIONS: We constructed ERAGS-GFP, which could replace the challenge virus standard-11 strain used in FAVN test.

Study on graphene oxide as a hole extraction layer for stable organic solar cells.

The development of an efficient and stable hole extraction layer (HEL) is crucial for commercializing organic solar cells (OSCs). Although a few candidates have been widely utilized as HELs for OSCs, the most appropriate material has been lacking. A few articles have recently reported graphene oxide (GO) as a well-working HEL that offers comparable performance to conventional HELs. However, a systematic study providing comprehensive insight into the GO-based OSC behavior is lacking. This article discusses broad topics, including the material properties, device efficiency, shelf lifetime, and impedance properties. We found that GO offers excellent properties, which are identical to those of conventional HELs, while the shelf lifetime shows a significant 6-fold increase. Furthermore, we discuss the significantly reduced space-charge limited region of an aged GO-based OSC compared with a PEDOT:PSS-based device, which is revealed to be a reason for the different shelf lifetime. We believe that the results will accelerate the development of GO as an HEL for OSCs and other optoelectronic devices.

Fast charging with high capacity for aluminum rechargeable batteries using organic additive in an ionic liquid electrolyte.

Aluminum-ion batteries have many advantages such as the natural abundance of aluminum, high theoretical capacity, and low cost. However, the ionic liquid commonly used as the electrolyte for aluminum-ion batteries has high viscosity, which hinders the migration of charge carriers. In this study, we used various organic solvents as additives for the ionic liquid electrolyte and investigated their effect on the battery performance. The electrolyte containing 45% (v/v) benzene had the best electrochemical properties, which led to a high specific capacity of 90 mA h g-1 at an extremely high current density of 5 A g-1.

Curvature Effect of a Phosphatidylethanolamine-Included Membrane on the Behavior of Cinnamycin on the Membrane.

The behavior of cinnamycin on a biomimetic membrane was studied with respect to the curvature of a phosphatidylethanolamine (PE)-included membrane with the adhesion measured using an atomic force microscope (AFM). The membrane was formed through vesicle fusion on the hydrophobic surface of silica spheres, which was used to define the curvature of the membrane. The hydrophobicity was generated by the reaction of alkyl-silane and analyzed with an X-ray photoelectron spectrometer. The cinnamycin, immobilized covalently to the AFM tip coated with 1-mercapto-1-undecanol that was observed to be inert to any adhesion to the membrane, showed that the adhesion became stronger with the increase in the curvature. The correlation between the adhesion and the curvature was linearly proportional. Since it was found that the cinnamycin was bound to a PE headgroup and the binding was enhanced by the interaction of the hydrophobic area located at one side of the cinnamycin, the linear proportionality seems to suggest that the interaction is related to the one dimensional orientation of the binding.

Kinetic and thermodynamic studies of cinnamycin specific-adsorption on PE-Included-Membranes using surface plasmon resonance.

The binding of the cinnamycin on the biomimetic membrane was studied with respect to time using the surface plasmon resonance(SPR). The membrane was composed of the inner layer tethered on the gold surface and the outer layer formed on the inner layer, which was at the desired ratio of dioleoylphosphatidylethanolamine(DOPE) to dioleoylphosphatidyl- choline(DOPC). On the bilayer, the cinnamycin solution was injected and showed different behavior of the binding with respect to time up on its concentration. For kinetic analysis, the behavior was converted to the coverage fraction with respect to time, which was ratio to the saturated response of 5 µM cinnamycin solution. The fraction change with respect to time was function of the available-site, which was eventually the subtraction of the fraction from one. With the fitting of the first order of the available site, the rate constant was acquired into 6∼7 × 10-3 s-1. Furthermore, the reciprocals of the fraction and the concentration were fitted with the Langmuir adsorption isotherm. From the fitting, the equilibrium constant was between 1 × 107 and 5 × 107 M-1.

Author Correction: The peptide AC 2 isolated from Bacillus-treated Trapa japonica fruit extract rescues DHT (dihydrotestosterone)-treated human dermal papilla cells and mediates mTORC1 signaling for autophagy and apoptosis suppression.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

NAB2-STAT6 fusion protein mediates cell proliferation and oncogenic progression via EGR-1 regulation.

Solitary fibrous tumors are rare mesenchymal tumors derived from soft tissues and vascular walls. NAB2-STAT6 fusion gene serves as a marker gene for this disease and consists of the truncated repressor domain of NGFI-A-Binding protein 2 (NAB2) and the intact activation domain of STAT6. In this study, we found that EGR-1 and the proliferation-related EGR-1 target gene IGF2 were upregulated in NIH-3T3 cells transfected with NAB2-STAT6. Additionally, p-Rb (Ser795) and cyclin D1 levels were upregulated, and cell proliferation was also enhanced. We identified that treatment with the IGF2 inhibitor reduced cell proliferation in NIH-3T3 cells transfected with NAB2-STAT6. The oncogenic progression was enhanced in NIH-3T3 cells transfected with NAB2-STAT6 compared with those transfected with the empty vector. Taken together, our study suggests that the NAB2-STAT6 fusion gene is associated with cell proliferation through EGR-1 transcriptional expression and IGF2 can be a drug target for the treatment of solitary fibrous tumors.

The peptide AC 2 isolated from Bacillus-treated Trapa japonica fruit extract rescues DHT (dihydrotestosterone)-treated human dermal papilla cells and mediates mTORC1 signaling for autophagy and apoptosis suppression.

The Trapa japonica fruit is a natural plant growing in ponds with its roots in the mud. It has long been used as a home remedy for many diseases; however, a major problem with this kind of natural extract is the multicomponents-multitargets for diseases. Such problems make it difficult to identify the mechanism of action. Another problem is quality control and consistency. The aim of this research was to isolate a single bioactive compound (peptide) derived from the Trapa japonica fruit. The research was conducted with various experimental techniques, such as fermentation and liquid chromatography, to isolate a peptide. We isolated the AC 2 peptide from Trapa japonica fruit and found it to be promising on human dermal papilla cells. Dihydrotestosterone (DHT) stresses human dermal papilla cells and is a major cause of hair loss resulting from hormones and environmental factors. The purpose of this research was to develop an understanding of the mechanism by which the AC 2 peptide rescues dihydrotestosterone (DHT)-treated human dermal papilla cells. We explored the effects of the AC 2 peptide on the cell biological functions of human dermal papilla cells (HDPs). HDPs were treated with the AC 2 peptide and DHT. Then, a cytotoxicity assay, flow cytometry, Western blot, immunoprecipitation, and 3D cell culture for immunohistochemistry were conducted to investigate the mTORC1 pathway and suppression of autophagy and apoptosis. In addition, we also synthesized the AC2 peptide as an alternative to the expensive and difficult isolation and purification procedures and confirmed its potential in biomedical applications. We also validated the effects of the synthetic AC2 peptide as well as the isolated and purified AC2 peptide and established their similarity. Although extensive research has been carried out on natural extracts, few single studies have isolated and separated a bioactive peptide (single compound).

In vitro and in vivo Induction of p53-Dependent Apoptosis by Extract of Euryale ferox Salisb in A549 Human Caucasian Lung Carcinoma Cancer Cells Is Mediated Through Akt Signaling Pathway.

Lung cancer is one of the leading causes of death, and mortality rates have steadily been increasing. Recently, several studies were conducted to develop novel, physiologically active compounds from medicinal plant extracts. Several plant-derived extracts and molecules regulate and inhibit signaling molecules associated with the growth and proliferation of cancer cells. Euryale ferox salisb is a medicinal plant that is effective against different types of cancers. In this study, we investigated the apoptotic effects of E. ferox salisb extract (ESE) in A549 lung cancer cells, exerted by the inhibition of the Akt protein and activation of the p53 protein. Our results show that ESE induces apoptosis via the regulation of mitochondrial outer membrane potential and generation of reactive oxygen species (ROS). We demonstrate that apoptosis is induced in a p53-dependent manner when cells are treated with pifithrin-α (a p53 inhibitor) and LY294002 (an Akt inhibitor). The apoptotic effects from ESE were observed in vivo in Balb/c-nu mice bearing A549 xenografts. Altogether, these results suggest that E. ferox salisb extracts exert anti-cancer effects in a p53-dependent manner.

Bacillus/Trapa japonica Fruit Extract Ferment Filtrate enhances human hair follicle dermal papilla cell proliferation via the Akt/ERK/GSK-3ß signaling pathway.

BACKGROUND: Despite advances in medical treatments, the proportion of the population suffering from alopecia is increasing, creating a need for new treatments to control hair loss and prevent balding. Treatments based on plant-derived compounds could potentially prevent hair loss. Human hair follicle dermal papilla (HDP) cells, a type of specialized fibroblast in the hair bulb, play an essential role in controlling hair growth and in conditions such as androgenic alopecia. We examined the effect of Bacillus/Trapa japonica fruit ferment filtrate extracts (TJFs) on HDP cells to determine whether activation of the Akt/ERK/GSK-3ß signaling pathway improved HDP cell proliferation. METHODS: We prepared TJFs using various methods. The extract properties were analyzed using WST-1, Lowry, and cell migration assays as well as immunofluorescence staining. We also determined the cell cycle stage and performed western blotting and an in ovo chick chorioallantoic membrane assay. Last, we constructed an organotypic three-dimensional cell culture model for immunohistochemical use. RESULTS: Our study confirmed that the TJFs contained numerous peptides and five unknown fractions. The TJFs stimulated HDP cell proliferation and migration via the Akt/ERK/GSK-3ß signaling pathway. To verify that the Akt/ERK/GSK-3ß pathway affected HDP cell proliferation, we treated HDP cells with LY294002 (an Akt inhibitor), BIO (a GSK-3ß inhibitor), and PD98059 (an ERK inhibitor). The TJFs also induced cell cycle progression, inhibited type І 5α-reductase, decreased apoptosis, and enhanced angiogenesis (vascular expansion). In addition to these signaling pathways, proteins including insulin-like growth factor-1 and keratinocyte growth factor, stimulating hair growth, were detected in the three-dimensional cell culture model. CONCLUSIONS: Our results confirmed that TJFs enhance HDP cell proliferation via the Akt/ERK/GSK-3ß signaling pathway, suggesting a potential treatment for alopecia.