Identification of marker compounds in fermented Benincasa hispida and validation of the method for its analysis.

Fermentation is a process that improves health functionality by inducing the production and increase of bioactive compounds. In this study, to standardize the fermentation process for Benincasa hispida, marker compounds that are increased or produced during fermentation were identified based on UPLC-QTOF-MS/MS. Analysis method verification and content analysis were conducted using HPLC-PDA. The marker compounds produced or increased in content were identified as 2-furoic acid, 2,3-dihydroxybenzoic acid, and rubinaphthin A by comparing their retention times, UV and MS spectra, and molecular formulas with those reported in previous studies. In addition, the increase in the content of the marker compounds by fermentation was confirmed, and the analytical method was validated by measuring its specificity, linearity, limit of detection and quantitation, precision, and accuracy. These results suggest that the developed fermentation process, marker compound identification, and verified analysis method can be applied to develop potential functional food ingredients from fermented B. hispida.

Autophagy inducers lead to transient accumulation of autophagosomes in Arabidopsis roots.

KEY MESSAGE: This study reveals that plant roots show a rapid termination of autophagy induction, offering a plant model for studying how excessive autophagy is deterred. In eukaryotes, autophagy is an intracellular mechanism that is important for recycling nutrients by degrading various macromolecules and organelles in vacuoles and lysosomes. Autophagy is induced when the nutrient supply to plant cells is limited. The protein kinase target of rapamycin (TOR) complex negatively regulates autophagy when nutrients are present in adequate amounts. The TOR inhibitor AZD8055 is an autophagy inducer that is useful for studying starvation-induced autophagy in plant cells. The mechanism by which AZD8055 increases the autophagic flux in plant cells has not been studied in detail. Here, we show that AZD8055-induced autophagy requires phosphatidylinositol 3-kinase activity and canonical AUTOPHAGY-RELATED (ATG) genes in Arabidopsis thaliana. Autophagic flux rapidly increased in seedlings treated with AZD8055. Unexpectedly, autophagy induction was transient in root cells and terminated earlier than in cotyledon cells. Transient induction is partly caused by a temporary effect of AZD8055 on phagophore initiation. These findings indicate a TOR-independent mechanism for terminating autophagy induction, thereby paving the way for elucidating how excess autophagy is prevented in plant roots.

Environmental obesogens (bisphenols, phthalates and parabens) and their impacts on adipogenic transcription factors in the absence of dexamethasone in 3T3-L1 cells.

Endocrine-disrupting chemicals (EDCs) are exogenous compounds that are capable of blocking or mimicking the action of bioidentical hormones. Obesogenic EDCs, commonly called obesogens, play an important role in adipogenesis. This study was carried out to determine the effects of select obesogens and their alternatives on adipogenesis in 3T3-L1 cells under dexamethasone (DEX)-free conditions. Preadipocytes were treated with a cocktail of 3-isobutyl-1-methylxanthine (IBMX) and insulin to which an obesogen (viz., bisphenol A (BPA) or its analogs BPS and BPF; dioctyl terephthalate; tris (2-ethylhexyl) trimellitate; or various parabens) had been added. A mixture containing IBMX, insulin, and DEX, which constitute the typical hormonal cocktail required for adipocyte differentiation, was used as the control against which the other groups were measured. The obesogens and the PBA analogs all had evident adipogenic effects under DEX-free conditions, as was determined by estimating the lipid accumulation levels in the cells using Oil Red O staining. Furthermore, the expression of adipogenic transcription factors (CCAAT/enhancer-binding protein-alpha, peroxisome proliferator-activated receptor-gamma, and adipocyte protein 2) was induced by 20 µM of BPA, BPS, or BPF at both the mRNA and protein levels, as determined through reverse transcription-polymerase chain reaction and western blot assays. Taken together, the results reveal that adipocyte differentiation can be induced by obesogens and their alternatives in the absence of DEX.

Method validation of 12 kinds of food dye in chewing gums and soft drinks, and evaluation of measurement uncertainty for soft drinks.

We developed and validated a method for the simultaneous quantitative analysis of food dyes in two food matrices, i.e., chewing gum and soft drinks. Furthermore, we evaluated the stability of food dyes in these matrices with respect to the pH and acid content. The optimized and validated method is based on high-performance liquid chromatography-photodiode array (HPLC-PDA) and liquid chromatography-tandem spectroscopy; the proposed method could identify and quantify 12 dyes in the two matrices. The recoveries of the food dyes identified by HPLC-PDA analysis ranged from 98.61% to 118.42%, with relative standard deviations of 0.06-4.90%. In addition, the expanded uncertainties of the measurements ranged from 0.57 to 3.12%. Finally, the food dyes were found to be stable in the matrices over 30 days. Thus, we believe that the proposed analytical method is suitable for the identification and quantification of food dyes in chewing gum and soft drink samples.

Physicochemical, nutritional and functional properties of Cucurbita moschata.

Cucurbita moschata is widely planted in most parts of the world, and is rich in carotenoids, vitamins, dietary fiber, minerals, and phenolic compounds. It also has important medicinal value. Some related research has proven that Cucurbita moschata has the potential ability to induce anti-obesity, anti-diabetic, antibacterial, and anticancer effects. At the same time, it has attracted more attention in the medical field. These nutrients and bioactive compounds in Cucurbita moschata have important effects on human health. In order to make better use of this crop, it still needs further study. Therefore, the purpose of this article is to summarize the physicochemical properties and nutritional components of Cucurbita moschata, and to provide a reference for further research on the benefits of on human health.

Antiobesity effect of Brassica juncea cultivated in Jeongseon with optimized sinigrin content using 3T3-L1 adipocytes.

In recent years, Brassica juncea has been selected as a special agricultural crop in Jeongseon, Gangwon-do, Korea, and is actively grown there. However, there have been no studies on B. juncea cultivated in Jeongseon (BJJ). Sinigrin, an index component of cruciferous vegetables, has been reported to have antiobesity effects. In this study, we developed a method for obtaining a BJJ extract with optimized sinigrin content, and investigate the antiobesity properties of the BJJ extract and sinigrin. The optimal extraction conditions for BJJ were found to be with 60% ethanol, at 70°C, for 3 hr. Lipid accumulation and ROS production were significantly suppressed in both the BJJ extract and sinigrin-treated groups. Furthermore, BJJ extract and sinigrin were effectively controlled the expression of proteins that regulate lipid accumulation, fatty acid oxidation, and energy metabolism. Thus, BJJ extract containing sinigrin may be used as a health functional food material. PRACTICAL APPLICATIONS: Brassica juncea has been reported to be rich in flavonoids, polyphenols, and glucosinolate, which are secondary vegetable metabolites. In this study, an extraction method to optimize the content of sinigrin in BJJ was established, and the antiobesity mechanism for the extract was confirmed. Lipid accumulation and ROS production were significantly suppressed in both the BJJ extract and sinigrin-treated groups in the study. It was confirmed that the expression of proteins that regulate lipid accumulation, lipid synthesis, fatty acid oxidation, heat generation, and energy metabolism was effectively controlled by the BJJ extract and sinigrin. Therefore, the ethanol extraction method of this study are considered to be useful for the preparation of extracts using cruciferous vegetables, and BJJ extract containing sinigrin have the potential to be used as a health functional food material for obesity.

Radical Scavenging-Linked Anti-Adipogenic Activity of Aster scaber Ethanolic Extract and Its Bioactive Compound.

Aster scaber is a wild vegetable cultivated in Korea and is known to contain phytochemicals with various biological activities. The potential antioxidant and anti-obesity effects of A. scaber and their mechanism are yet to be reported. We evaluated the total phenolic, flavonoid, and proanthocyanidin contents and oxygen radical absorbance capacity of A. scaber ethanolic extract (ASE), and analyzed the major phenolic compounds of ASE. Antioxidant activity was measured at the chemical level through 2,2-diphenyl-1-picrylhydrazyl (DPPH), reducing power assay, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), and fluorescence recovery after photobleaching (FRAP) assay. In addition, it was measured in vitro through inhibition of Reactive oxygen species (ROS) production in 3T3-L1 adipocyte, and inhibition of lipid accumulation was also evaluated. ASE reduced the expression of enzymes involved in the production of ROS and increased the expression of antioxidant enzymes that reduce increased ROS levels. They also reduced the expression of adipogenesis transcription factors that regulate adipocyte differentiation in relation to ROS production, inhibited the expression of lipogenesis-related genes related to fat accumulation through AMP-activated protein kinase (AMPK) activity, and increased expression of lipolysis-related genes. Thus, ASE containing CGA (chlorogenic acid) inhibits ROS production in 3T3-L1 adipocytes, owing to its strong antioxidant activity, and inhibits lipid accumulation caused by oxidative stress. The extract can be used as a potential functional food material for reducing oxidative stress and obesity.

Enhancement of Immune Activities of Mixtures with Sasa quelpaertensis Nakai and Ficus erecta var. sieboldii.

The objective of the present study was to develop a concoction of natural products that could dramatically improve immune function with minimal possible side effects. Sasa quelpaertensis Nakai and Ficus erecta var. sieboldii are plants that are native to Jeju Island, Korea and are known to be rich in physiologically active substances. We prepared a mixture of different proportions and extraction conditions using two natural plants and determined their optimum mixing ratio and extraction method by assessing immune function-related biomarkers in RAW264.7 macrophages. Optimal extract (HR02/04(8:2)-W) was selected from in vitro experiments and its immunity-enhancing efficacy was evaluated in mice. After oral administration of extract to BALB/c mice for 2 weeks, nitric oxide production in the peritoneal exudate cells, natural killer cell cytotoxicity, cytokine expression in splenocytes, and total cell number of immune tissues and phenotype analysis were evaluated. Our results demonstrated that HR02/04(8:2)-W significantly enhanced the immune system by increasing natural killer cell activity, cytokine expression, and total number of cells in immune tissues. In conclusion, our study validates the role of HR02/04(8:2)-W in enhancing immunity and its potential development as a functional food.

Areas of white matter hyperintensities and motor symptoms of Parkinson disease.

OBJECTIVE: To determine whether deep white matter and periventricular hyperintensities affect the motor symptoms of Parkinson disease (PD) differently, we analyzed MRI and dopamine transporter imaging. METHODS: We analyzed the medical records of patients with de novo PD who underwent dopamine transporter PET scanning and MRI at their first visit. Deep white matter and periventricular hyperintensities were scored with a visual rating scale, and motor symptoms were assessed by Unified Parkinson's Disease Rating Scale motor score and tremor, rigidity, bradykinesia, and axial symptom subscores. The influence of white matter hyperintensity on motor symptoms was explored using multivariable linear regression models. RESULTS: A total of 93 patients (mean age, 67.2 ± 9.9 years; 44 male) were included and the mean motor score was 25.0 ± 10.8. Subscores for bradykinesia and axial symptoms were correlated with both deep white matter and periventricular hyperintensities scores. Multivariable linear regression models revealed that deep white matter hyperintensities score was significantly associated with subscore for bradykinesia and periventricular hyperintensities score was associated with subscores for bradykinesia and axial symptoms after adjusting for putaminal dopamine transporter availability and clinical factors. CONCLUSIONS: These results demonstrate that deep white matter hyperintensities are associated with bradykinesia and periventricular hyperintensities are associated with bradykinesia and axial symptoms in patients with PD independently of the severity of dopaminergic depletion.

Prediction of ovarian aging using ovarian expression of BMP15, GDF9, and C-KIT.

IMPACT STATEMENT: Ovarian aging is becoming a more important issue in terms of fertility preservation and infertility treatment. Serum anti-Mullerian hormone (AMH) level and antral follicle count (AFC) are being practically used as markers of ovarian aging as well as ovarian reserve in human. However, these factors have some drawbacks in assessing ovarian aging and reserve. Therefore, the identification of ovarian expressions of BMP15, GDF9, and C-KIT according to female could be applied as a potent predictor of ovarian aging. This work provides new information on the development of diagnosis and treatment strategy of age-related fertility decline and premature ovarian insufficiency.

NGL-1/LRRC4C-Mutant Mice Display Hyperactivity and Anxiolytic-Like Behavior Associated With Widespread Suppression of Neuronal Activity.

Netrin-G ligand-1 (NGL-1), encoded by Lrrc4c, is a post-synaptic adhesion molecule implicated in various brain disorders, including bipolar disorder, autism spectrum disorder, and developmental delay. Although previous studies have explored the roles of NGL-1 in the regulation of synapse development and function, the importance of NGL-1 for specific behaviors and the nature of related neural circuits in mice remain unclear. Here, we report that mice lacking NGL-1 (Lrrc4c-/- ) show strong hyperactivity and anxiolytic-like behavior. They also display impaired spatial and working memory, but normal object-recognition memory and social interaction. c-Fos staining under baseline and anxiety-inducing conditions revealed suppressed baseline neuronal activity as well as limited neuronal activation in widespread brain regions, including the anterior cingulate cortex (ACC), motor cortex, endopiriform nucleus, bed nuclei of the stria terminalis, and dentate gyrus. Neurons in the ACC, motor cortex, and dentate gyrus exhibit distinct alterations in excitatory synaptic transmission and intrinsic neuronal excitability. These results suggest that NGL-1 is important for normal locomotor activity, anxiety-like behavior, and learning and memory, as well as synapse properties and excitability of neurons in widespread brain regions under baseline and anxiety-inducing conditions.

Inhibitory effect of Carnosol on UVB-induced inflammation via inhibition of STAT3.

Ultraviolet B (UVB) irradiation causes sunburn, inflammatory responses, dysregulation of immune function, oxidative stress, DNA damage and photocarcinogenesis on skin. Rosemary (Rosmarinus officinalis L.) has been reported to inhibit inflammation. Carnosol, a major component of Rosemary, has prominent anti-inflammatory effects. However, its protective effect on UVB-induced inflammatory skin responses has not yet been reported. Here, we investigated the effectiveness of carnosol on UVB-induced inflammation. We examined the anti-inflammation effect of topical application of carnosol (0.05 µg/cm2) on UVB (540 mJ/cm2, for 3 successive days)-induced skin inflammation in HR1 mice. Topical application of carnosol inhibited UVB-induced erythema, epidermal thickness, inflammatory responses in HR1 mice. Carnosol reduced the level of Immunoglobulin-E and IL-1ß in blood serum of UVB-induced mice. Carnosol also significantly inhibited the UVB-induced expression of inflammatory marker protein (iNOS and COX-2) in back skin of mice. In addition, carnosol treated skin decreased activation of STAT3, a transcriptional factor regulating inflammatory genes. Our study suggested that carnosol has protective effects on skin inflammatory skin damages by UVB.

Modulation of dendritic cell function by Trichomonas vaginalis-derived secretory products.

Trichomoniasis caused by the parasitic protozoan Trichomonas vaginalis is the most common sexually transmitted disease in the world. Dendritic cells are antigen presenting cells that initiate immune responses by directing the activation and differentiation of naïve T cells. In this study, we analyzed the effect of Trichomonas vaginalis-derived Secretory Products on the differentiation and function of dendritic cells. Differentiation of bone marrow-derived dendritic cells in the presence of T. vaginalis-derived Secretory Products resulted in inhibition of lipopolysaccharide-induced maturation of dendritic cells, down-regulation of IL-12, and up-regulation of IL-10. The protein components of T. vaginalis-derived Secretory Products were shown to be responsible for altered function of bone marrow- derived dendritic cells. Chromatin immunoprecipitation assay demonstrated that IL-12 expression was regulated at the chromatin level in T. vaginalis-derived Secretory Productstreated dendritic cells. Our results demonstrated that T. vaginalis- derived Secretory Products modulate the maturation and cytokine production of dendritic cells leading to immune tolerance.