Calmodulins (CaMs) regulate numerous Ca2+-mediated cellular processes in plants by interacting with their respective downstream effectors. Due to the limited number of CaMs, other calcium sensors ...modulate the regulation of Ca2+-mediated cellular processes that are not managed by CaMs. Of 50 CaM-like (CML) proteins identified in Arabidopsis thaliana, we characterized the function of CML10.
Yeast two-hybrid screening revealed phosphomannomutase (PMM) as a putative interaction partner of CML10. In vitro and in vivo interaction assays were performed to analyze the interaction mechanisms of CML10 and PMM. PMM activity and the phenotypes of cml10 knock-down mutants were studied to elucidate the role(s) of the CML10–PMM interaction.
PMM interacted specifically with CML10 in the presence of Ca2+ through its multiple interaction motifs. This interaction promoted the activity of PMM. The phenotypes of cml10 knock-down mutants were more sensitive to stress conditions than wild-type plants, corresponding with the fact that PMM is an enzyme which modulates the biosynthesis of ascorbic acid, an antioxidant.
The results of this research demonstrate that a calcium sensor, CML10, which is an evolutionary variant of CaM, modulates the stress responses in Arabidopsis by regulating ascorbic acid production.
Piperine is a bioactive alkaloid that possesses various health benefits and is responsible for the pungent aroma of pepper. Piperine content in whole and ground black pepper (n = 132) was analyzed by ...near‐infrared spectroscopy (NIRS) in the 950 to 1650 nm wavelength window. Chemometric modeling using partial least square regression was performed, and outliers were checked and removed during the preparation of the calibration curve by considering sample residual variance and sample leverage. Model accuracy was evaluated with a low root‐mean‐square error of cross‐validation (RMSECV) and a high ratio performance to deviation (RPD). The optimal model had a coefficient of determination (R2) of 0.726, RMSECV of 0.289 g/100 g, and RPD of 1.744 for the whole black pepper. The results of R2, RMSECV, and RPD for the ground black pepper were 0.850, 0.231 g/100 g, and 2.424, respectively. Therefore, based on the perspective of onsite process, the proposed NIRS method can be employed for selecting abnormal samples during the inspection of black pepper raw material and for quantifying piperine contents of final black pepper product.
Practical Application
Generally, the quality indicators of black pepper are graded solely based on their external appearance, quality, and size. This study discloses the development of a near‐infrared spectroscopy‐based fast and accurate nondestructive analytical method for the detection of piperine, a bioactive constituent of pepper, as a tool for the quality control of whole and ground black pepper.
Accumulated clinical and biomedical evidence indicates that the gut microbiota and their metabolites affect brain function and behavior in various central nervous system disorders. This study was ...performed to investigate the changes in brain metabolites and composition of the fecal microbial community following injection of amyloid β (Aβ) and donepezil treatment of Aβ-injected mice using metataxonomics and metabolomics. Aβ treatment caused cognitive dysfunction, while donepezil resulted in the successful recovery of memory impairment. The Aβ + donepezil group showed a significantly higher relative abundance of Verrucomicrobia than the Aβ group. The relative abundance of 12 taxa, including Blautia and Akkermansia, differed significantly between the groups. The Aβ + donepezil group had higher levels of oxalate, glycerol, xylose, and palmitoleate in feces and oxalate, pyroglutamic acid, hypoxanthine, and inosine in brain tissues than the Aβ group. The levels of pyroglutamic acid, glutamic acid, and phenylalanine showed similar changes in vivo and in vitro using HT-22 cells. The major metabolic pathways in the brain tissues and gut microbiota affected by Aβ or donepezil treatment of Aβ-injected mice were related to amino acid pathways and sugar metabolism, respectively. These findings suggest that alterations in the gut microbiota might influence the induction and amelioration of Aβ-induced cognitive dysfunction via the gut–brain axis. This study could provide basic data on the effects of Aβ and donepezil on gut microbiota and metabolites in an Aβ-induced cognitive impairment mouse model.
Obesity can be caused by microbes producing metabolites; it is thus important to determine the correlation between gut microbes and metabolites. This study aimed to identify gut ...microbiota-metabolomic signatures that change with a high-fat diet and understand the underlying mechanisms. To investigate the profiles of the gut microbiota and metabolites that changed after a 60% fat diet for 8 weeks, 16S rRNA gene amplicon sequencing and gas chromatography-mass spectrometry (GC-MS)-based metabolomic analyses were performed. Mice belonging to the HFD group showed a significant decrease in the relative abundance of Bacteroidetes but an increase in the relative abundance of Firmicutes compared to the control group. The relative abundance of Firmicutes, such as Lactococcus, Blautia, Lachnoclostridium, Oscillibacter, Ruminiclostridium, Harryflintia, Lactobacillus, Oscillospira, and Erysipelatoclostridium, was significantly higher in the HFD group than in the control group. The increased relative abundance of Firmicutes in the HFD group was positively correlated with fecal ribose, hypoxanthine, fructose, glycolic acid, ornithine, serum inositol, tyrosine, and glycine. Metabolic pathways affected by a high fat diet on serum were involved in aminoacyl-tRNA biosynthesis, glycine, serine and threonine metabolism, cysteine and methionine metabolism, glyoxylate and dicarboxylate metabolism, and phenylalanine, tyrosine, and trypto-phan biosynthesis. This study provides insight into the dysbiosis of gut microbiota and metabolites altered by HFD and may help to understand the mechanisms underlying obesity mediated by gut microbiota.
The purpose of this study is to investigate the effect of fungi on kimchi metabolites during fermentation. A gas chromatography-mass spectrometry (GC-MS) based metabolite profiling approach in ...combination with principal component analysis (PCA) is performed to differentiate metabolites produced by fungi or bacteria. To avoid bacterial growth, kimchi is treated with 100 μg/mL of ampicillin every three days from 30 to 50 days of fermentation. The relative content of the major fungi at 50 days of fermentation, between the control group and the ampicillin treatment group, was not significantly different. The administration of ampicillin changed the metabolites in kimchi by affecting the growth of kimchi bacteria. Based on the pattern of change of each metabolite, the changed metabolites are grouped into four categories: (1) metabolites produced or consumed by fungi, (2) metabolites involving both fungi and bacteria, (3) metabolites produced or consumed by bacteria, and (4) metabolites of undetermined origin. Alanine, thymine, galacturonic acid, and malonic acid can be regarded as the metabolites produced by fungi between 30 and 50 days of fermentation. In contrast, malic acid, oxaloacetic acid, galactitol, glucose, and mannitol are presumed to be the metabolites mainly consumed by fungi. This study is meaningful as the first study conducted by inhibiting growth of bacteria to identify the metabolites contributed by fungi or bacteria in the kimchi fermentation process. These results could be used to make customized kimchi that controls the production of desired metabolites by selectively controlling the formation of microbial communities in the kimchi industry.
Pectinase is a well‐known enzyme used in the food processing industry to produce fruit juice and concentrate. This study evaluated the anticancer and antiangiogenesis activities of pectinase‐treated ...Prunus mume fruit concentrate (PC) and its phenolic components. PC treatment (250 to 1,000 µg/mL) resulted in decreased proliferation of SW480 human colorectal cancer cells through S‐phase cell cycle arrest; however, equivalent concentrations of PC did not show toxicity toward CRL‐1539 colon normal cells. Furthermore, PC‐induced caspase‐dependent apoptosis in SW480 cells, which was characterized by accumulation of apoptotic cell population, cell shrinkage, formation of apoptotic bodies, upregulation of proapoptotic Bax, cleaved PARP, caspase‐3, caspase‐8, and caspase‐9, and downregulation of antiapoptotic Bcl‐2. Antiangiogenesis effects of PC were assessed using human umbilical vein endothelial cells (HUVECs). We found that PC did not inhibit HUVECs proliferation at concentrations of 500 to 1,500 µg/mL. In addition, treatment with PC at nontoxic concentrations (500 to 1,000 µg/mL) blocked vascular endothelial growth factor induced cell migration, invasion, capillary‐like tube formation, and angiogenesis from rat aortic rings. HPLC‐PDA analysis showed that there were at least four different phenolics including 5‐HMF, neochlorogenic acid, protocatechuic acid, and syringic acid. Taken together, these results indicated that PC could be used as a good source of phenolic compounds with selective anticancer and antiangiogenesis activities.
Practical Application
Pectinases are one of the well‐known enzyme used in the part of food processing. Treatment of pectinase is a useful strategy to reduce viscosity, turbidity, and pulp particles in the production of fruit juice, extract, and concentrate. In the present study, we found that pectinase‐treated P. mume fruit concentrate significantly suppresses colorectal cancer proliferation and angiogenesis of human umbilical vein endothelial cells. The significance of our findings is that pectinase‐treated P. mume concentrate may be used as a commercial functional food material to inhibit colorectal cancer and angiogenesis.
•Convolutional neural networks (CNN) were trained using RGB images of strawberries.•CNN can determine the quality of strawberries with a high accuracy of about 97 %.•The efficient discrimination ...model was developed at a low cost and in a short time.
Deterioration in the appearance of strawberries is attributed to rapid metabolic changes, cellular damage, and softening occurring during their distribution and storage. To quickly and non-destructively monitor the external quality of strawberries, recognition models based on 750 Red Green Blue (RGB) image classifications and using convolutional neural networks (CNNs) were developed. A model using eight configurations was used to compare the discrimination accuracies according to the following variables: training–test set distributions, number of learning images, and number of epochs. Strawberry samples were classified as fresh, bruised, or moldy. According to our validation results, training the model with 90 % of the image data ensured a high learning performance. Using our test dataset, we found that the accuracy, precision, specificity, and sensitivity of the model reached 97 %. In the feature map derived from convolutional layers, the bruised and moldy areas of the strawberry were also identified. Together, these results suggest that CNNs have potential use in the non-destructive monitoring of quality changes in the food industry.
The objective of this study was to evaluate anti-obesity effects of kimchi microbial community (KMC) on obesity and gut microbiota using a high fat diet-induced mouse model compared to effects of a ...single strain. Administration of KMC decreased body weight, adipose tissue, and liver weight gains. Relative content of
Muribaculaceae
in the gut of the KMC-treated group was higher than that in the high-fat diet (HFD) group whereas relative contents of
Akkermansiaceae, Coriobacteriaceae
, and
Erysipelotrichaceae
were lower in KMC-treated group. Metabolic profile of blood was found to change differently according to the administration of KMC and a single strain of
Lactobacillus plantarum
. Serum metabolites significantly increased in the HFD group but decreased in the KMC-treated group included arachidic acid, stearic acid, fumaric acid, and glucose, suggesting that the administration of KMC could influence energy metabolism. The main genus in KMC was not detected in guts of mice in KMC-treated group. Since the use of KMC has advantages in terms of safety, it has potential to improve gut microbial community for obese people.
Recent studies have demonstrated that natural agents targeting the accumulation of reactive oxygen species (ROS) that selectively kill, leaving normal cells undamaged, can suppress prostate cancer. ...Here, we show that auriculasin, derived from Flemingia philippinensis, induces significant cell death and apoptosis via ROS generation in prostate cancer cells. Auriculasin treatment resulted in selective apoptotic cell death in LNCaP prostate cancer cells, characterized by DNA fragmentation, accumulation of sub-G1 cell population, cleavage of poly (ADP-ribose) polymerase (PARP), regulation of Bax/Bcl-2 ratio, increase of cytosolic apoptosis-inducing factor (AIF) and endonuclease G (EndoG), in addition to inhibiting tumor growth in a xenograft mouse model. Interestingly, auriculasin-induced apoptosis did not result in caspase-3, -8, and -9 activations. We found that auriculasin treatment decreased phosphorylation of AKT/mTOR/p70s6k in a dose- and time-dependent manner. Further, cellular ROS levels increased in LNCaP cells treated with auriculasin and blocking ROS accumulation with ROS scavengers resulted in inhibition of auriculasin-induced PARP cleavage, AIF increase, upregulation of Bax/Bcl-2 ratio, and decrease in AKT/mTOR phosphorylation. Taken together, these data suggest that auriculasin targets ROS-mediated caspase-independent pathways and suppresses PI3K/AKT/mTOR signaling, which leads to apoptosis and decreased tumor growth.
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•Auriculasin sensitively inhibited growth of human prostate cancer cells compared with human prostate epithelial cells.•Auriculasin induces caspase-independent apoptosis in LNCaP human prostate cancer cells.•Auriculasin-induced apoptosis in LNCaP cells was regulated by ROS generation.•Auriculasin suppressed tumor growth in xenograft athymic mouse model.
Nowadays, new types of vinegar have been developed using various raw materials and biotechnological processes. The fruit of
has been extensively distributed in East Asia and used as a folk medication ...for fatigue. In this study, the
vinegar (PV) was produced by a two-step fermentation and was evaluated for its anti-fatigue activity by C2C12 myoblasts and high-intensity exercised rats. The administration of PV significantly improved running endurance and glycogen accumulation in the liver and muscle of PV supplemented rats compared to sedentary and exercised control groups. In addition, PV supplementation elicited lower fatigue-related serum biomarkers, for instance, ammonia, inorganic phosphate, and lactate. PV administered rats exhibited higher lactate dehydrogenase activity and glutathione peroxidase activity, and lower creatine kinase activity and malondialdehyde levels. Furthermore, phenolic compounds in PV were identified using HPLC analysis. The phenolic acids analyzed in PV were protocatechuic acid, syringic acid, chlorogenic acid, and its derivates. These results indicate that the administration of PV with antioxidative property contributes to the improvement of fatigue recovery in exhausted rats. The findings of this study suggest that the PV containing various bioactive constituents can be used as a functional material against fatigue caused by high-intensity exercise.
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