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Genistein, an isoflavone found in soybeans, exhibits antioxidant, anti-inflammatory, and anticancer properties. This study explored the molecular mechanisms behind genistein’s anticancer effects in prostate cancer DU145 cells. In this study, genistein decreased cell viability, increased annexin V-PE(+) cells, and enhanced the sub-G0/G1 peak by flow cytometric analysis. Increased reactive oxygen species increased mitochondrial depolarization indicating mitochondrial dysfunction and inhibition of ATP formation were also observed in genistein-treated DU145 cells. Genistein upregulated p53 at the mRNA and protein levels and increased caspase-3/7 activity along with the cleavage of Bax, procaspase-3, and PARP. With the increasing genistein concentrations, the percentage of cells in the sub-G0/G1 peak and G2/M phase increased, which was inhibited by treatment with the pan-caspase inhibitor Z-VAD together with 100 μM genistein, which had little toxicity to normal prostate epithelial HPrEC cells. Genistein treatment simultaneously inhibited the activation of STAT3 and other closely related oncogenic kinases such as AKT and ERK and p38 and decreased VEGF expression. Taken together, these results suggest that genistein inhibits the growth of DU145 cells and induces apoptosis by inhibiting STAT3, AKT, ERK, and p38 which provides a molecular basis for the anticancer activity of genistein and suggests its potential as a valuable therapeutic candidate for prostate cancer.

In high radiation fields, gamma cameras suffer from pulse pile-up, resulting in poor energy resolution, count losses, and image distortion. To overcome this problem, various methods have been introduced to reduce the size of the aperture or pixel, reject the pile-up events, and correct the pile-up events, but these technologies have limitations in terms of mechanical design and real-time processing. The purpose of this study is to develop a real-time gamma camera to evaluate the radioactive contamination in high radiation fields. The gamma camera is composed of a pinhole collimator, NaI(Tl) scintillator, position sensitive photomultiplier (PSPMT), signal processing board, and data acquisition (DAQ). The pulse pile-up is corrected in real-time with a field programmable gate array (FPGA) using the start time correction (STC) method. The STC method corrects the amplitude of the pile-up event by correcting the time at the start point of the pile-up event. The performance of the gamma camera was evaluated using a high dose rate 137Cs source. For pulse pile-up ratios (PPRs) of 0.45 and 0.30, the energy resolution improved by 61.5 and 20.3%, respectively. In addition, the image artifacts in the 137Cs radioisotope image due to pile-up were reduced.

This study aimed to optimize the ultrasonic-assisted extraction of polyphenolic compounds from red water lily (Nymphaea rubra) leaves (NRL) with heightened antioxidant activities and perform untargeted metabolite identification. Response surface methodology, employing Box-Behnken design, determined optimal conditions: 53.0 % ethanol, 32.0 min extraction time, and 53.0 °C temperature. The resulting extraction demonstrated significant total phenolic content of 45.12 ± 1.12 mgGAE/g and total flavonoid content of 44.05 ± 1.65 mgCAE/g. High-resolution mass spectrometry identified 150 secondary metabolites, encompassing phenols, flavonoids, tannins, terpenoids, and fatty acids. The optimized NRL extract exhibited notably increased radical scavenging abilities (19.21 ± 1.71 μM ASCE/g for DPPH and 24.59 ± 1.35 μM ASCE/g for ABTS) along with elevated reducing power potential (63.35 ± 4.28 μM ASCE/g for CUPRAC and 61.32 ± 2.56 μM ASCE/g for FRAP) in comparison to traditional extracts, which ranged from (7.23 ± 1.58 to 15.13 ± 1.79 μM ASCE/g, 13.29 ± 1.32 to 19.29 ± 1.69 μM ASCE/g, 22.74 ± 1.68 to 54.37 ± 3.38 μM ASCE/g, 24.14 ± 2.00 to 52.77 ± 2.33 μM ASCE/g for DPPH, ABTS, CUPRAC and FRAP, respectively). This suggests its promising potential for incorporation into antioxidant-rich functional foods. These findings underscore NRL's promise for commercial applications in the nutraceutical, food, and pharmaceutical industries.

Sargassum horneri (SH) is widely consumed as a healthy seaweed food in the Asia–Pacific region. However, the bioactive components contributing to its biological activity remain unknown. Herein, we optimized multifrequency ultrasonic-assisted extraction conditions to achieve higher antioxidant activity using a response surface methodology and an artificial neural network. High-resolution mass spectrometry (HRMS; negative mode) was used to tentatively identify the secondary metabolites in the optimized SH extract, which were further tested against oxidative stress in RAW264.7 cells. Additionally, the identified compounds were analyzed in silico to determine their binding energies with the Keap1 protein (4L7B). We identified 89 compounds using HRMS, among which 19 metabolites (8 polyphenolics, 2 flavonoids, 2 lignans, 2 terpenes, 2 tannins, 2 sulfolipids, and 1 phospholipid) were putatively reported for the first time in SH. The in vitro results revealed that optimized SH extract inhibited oxidative stress via the Nrf2/MAPKs/HO-1 pathway in a dose-dependent manner. This result was validated by performing in silico simulation, indicating that sargaquinoic acid and glycitein-7-O-glucuronide had the highest binding energies (−9.20 and −9.52 Kcal/mol, respectively) toward Keap1 (4L7B). This study offers a unique approach for the scientific community to identify potential bioactive compounds by optimizing the multivariant extraction processing conditions, which could be used to develop functional and nutraceutical foods.

This study sought to optimize the ultrasonic-assisted extraction of polyphenolic compounds from unmature Ajwa date seeds (UMS), conduct untargeted metabolite identification and assess antioxidant and depigmenting activities. Response surface methodology (RSM) utilizing the Box–Behnken design (BBD) and artificial neural network (ANN) modeling was applied to optimize extraction conditions, including the ethanol concentration, extraction temperature and time. The determined optimal conditions comprised the ethanol concentration (62.00%), extraction time (29.00 min), and extraction temperature (50 °C). Under these conditions, UMS exhibited total phenolic content (TPC) and total flavonoid content (TFC) values of 77.52 ± 1.55 mgGAE/g and 58.85 ± 1.12 mgCE/g, respectively, with low relative standard deviation (RSD%) and relative standard error (RSE%). High-resolution mass spectrometry analysis unveiled the presence of 104 secondary metabolites in UMS, encompassing phenols, flavonoids, sesquiterpenoids, lignans and fatty acids. Furthermore, UMS demonstrated robust antioxidant activities in various cell-free antioxidant assays, implicating engagement in both hydrogen atom transfer and single electron transfer mechanisms. Additionally, UMS effectively mitigated tert-butyl hydroperoxide (t-BHP)-induced cellular reactive oxygen species (ROS) generation in a concentration-dependent manner. Crucially, UMS showcased the ability to activate mitogen-activated protein kinases (MAPKs) and suppress key proteins including tyrosinase (Tyr), tyrosinase-related protein-1 and -2 (Trp-1 and -2) and microphthalmia-associated transcription factor (MITF), which associated melanin production in MNT-1 cell. In summary, this study not only optimized the extraction process for polyphenolic compounds from UMS but also elucidated its diverse secondary metabolite profile. The observed antioxidant and depigmenting activities underscore the promising applications of UMS in skincare formulations and pharmaceutical developments.

The well-known medicinal plant Portulaca oleracea L. (PO) is used as a traditional medicine and culinary herb to treat various diseases. Fatty acids, essential oils, and flavonoids were extracted from PO seeds and leaves using ultrasonic, microwave, and supercritical fluid extraction with RSM techniques. However, investigations on the secondary metabolites and antioxidant capabilities of the aerial part of PO (APO) are scarce. In order to extract polyphenols and antioxidants from APO as effectively as possible, this study used heat reflux extraction (HRE), response surface methodology (RSM), and artificial neural network (ANN) modeling. It also used high-resolution mass spectrometry to identify the APO secondary metabolite. A central-composite design (CCD) was used to establish the ideal ethanol content, extraction time, and extraction temperature to extract the highest polyphenolic compounds and antioxidant activity from APO. According to RSM, the highest amount of TPC (8.23 ± 1.06 mgGAE/g), TFC (43.12 ± 1.15 mgCAE/g), DPPH-scavenging activity (43.01 ± 1.25 % of inhibition) and FRAP (35.98 ± 0.19 µM ascorbic acid equivalent) were obtained at 60.0 % ethanol, 90.2 % time, and 50 °C. Statistical metrics such as the coefficient of determination (R2), root-mean-square error (RMSE), absolute average deviation (AAD), and standard error of prediction (SEP) revealed the ANN's superiority. Ninety-one (91) secondary metabolites, including phenolic, flavonoids, alkaloids, fatty acids, and terpenoids, were discovered using high-resolution mass spectrometry. In addition, 21 new phytoconstituents were identified for the first time in this plant. The results revealed a significant concentration of phytoconstituents, making it an excellent contender for the pharmaceutical and food industries.

We used in silico methods to predict the physiochemical and pharmacological characteristics, toxicity, and biological activities of the screened compounds. All compounds showed positive results while calculating their physiochemical and pharmacokinetic descriptors. Using the Prediction of Activity Spectra for Substances (PASS) software on compounds form non-oilseed legumes, we identified compounds (mainly 4 polyphenol compounds) with anti-infective, anti-eczematic, antimutagenic, muco-membranous protector, fibrinolytic, anticarcinogenic, hepato-protectant, cardio-protectant, antioxidant, and astringent effect. PASS predicted HO-1 expression enhancing and free radical scavenging properties for gallic acid, coumaric acid, catechin, and epicatechin. Data about validation protocols for molecular docking of ligand IVV to Keap1 was performed by root mean square deviation (RMSD) value is used to validate docking protocol and representation mainly for analyzing stability of protein and predicting conformational changes of protein. Molecular docking is a powerful technique for studies of receptor-ligand interaction and has led to the discovery of Keap1-Nrf2 small molecule inhibitors. Keap1 inhibits the degradation of Nrf2. Our results suggest that screened compounds from non-oilseed legumes can effectively interact with the Keap1 binding site and dissociate Keap1 and Nrf2. The screened compounds from non-oilseed legumes that displayed high binding affinities with Keap1 are promising new Nrf2 activators. We performed molecular docking to identify the molecular interactions of gallic acid, catechin, and epicatechin with Keap1. Non-oilseed legumes plant is a natural source of potent antioxidant that may prevent diseases and could be potentially used as functional food, nutraceuticals, and drugs.

In this study, expanded polystyrene (EPS) was photodegraded in vitro, and its water-soluble degradation products were extracted using solid-phase extraction. An in vivo test using the MTT assay showed that EPS exhibited increased toxicity upon photodegradation. This clearly demonstrates the necessity of investigating the degradation products at a molecular level, and thus, we conducted analyses using high-resolution mass spectrometry coupled with liquid chromatography. Overall, 68 compounds were detected, of which 13 were identified after comparing the retention times and tandem mass spectra of the extracted compounds with those of standards. The toxicity of the degraded EPS was studied by examination of the detected compounds combined with published reports. In addition, the quantification of the most abundant compounds showed that discarded EPS can generate a significant amount of chemicals by UV degradation. The data in this study show that degraded EPS can pose a significant environmental threat as a source of potentially hazardous chemicals. This study provides a fundamental basis for the evaluation of the fate and toxicity of photodegraded EPS.

Abstract Among the three isoforms encoded by Rtn4, Nogo-A has been intensely investigated as a central nervous system inhibitor. Although Nogo-A expression is increased in muscles of patients with amyotrophic lateral sclerosis, its role in muscle homeostasis and regeneration is not well elucidated. In this study, we discovered a significant increase in Nogo-A expression in various muscle-related pathological conditions. Nogo−/− mice displayed dystrophic muscle structure, dysregulated muscle regeneration following injury, and altered gene expression involving lipid storage and muscle cell differentiation. We hypothesized that increased Nogo-A levels might regulate muscle regeneration. Differentiating myoblasts exhibited Nogo-A upregulation and silencing Nogo-A abrogated myoblast differentiation. Nogo-A interacted with filamin-C, suggesting a role for Nogo-A in cytoskeletal arrangement during myogenesis. In conclusion, Nogo-A maintains muscle homeostasis and integrity, and pathologically altered Nogo-A expression mediates muscle regeneration, suggesting Nogo-A as a novel target for the treatment of myopathies in clinical settings.

Background The definitive pathologic diagnosis of cardiac sarcoidosis requires observation of a granuloma in the myocardial tissue. It is common, however, to receive a “negative” report for a clinically probable case. We would like to advise pathologists and clinicians on how to interpret “negative” biopsies. Methods Our study samples were 27 endomyocardial biopsies from 25 patients, three cardiac transplantation and an autopsied heart with suspected cardiac sarcoidosis. Pathologic, radiologic, and clinical features were compared. Results The presence of micro-granulomas or increased histiocytic infiltration was always (6/6 or 100%) associated with fatty infiltration and confluent fibrosis, and they showed radiological features of sarcoidosis. Three of five cases (60%) with fatty change and confluent fibrosis were probable for cardiac sarcoidosis on radiology. When either confluent fibrosis or fatty change was present, one-third (3/9) were radiologically probable for cardiac sarcoidosis. We interpreted cases with micro-granuloma as positive for cardiac sarcoidosis (five of 25, 20%). Cases with both confluent fibrosis and fatty change were interpreted as probable for cardiac sarcoidosis (seven of 25, 28%). Another 13 cases, including eight cases with either confluent fibrosis or fatty change, were interpreted as low probability based on endomyocardial biopsy. Conclusions The presence of micro-granuloma could be an evidence for positive diagnosis of cardiac sarcoidosis. Presence of both confluent fibrosis and fatty change is necessary for probable cardiac sarcoidosis in the absence of granuloma. Either of confluent fibrosis or fatty change may be an indirect pathological evidence but they are interpreted as nonspecific findings.

This study investigated the in vitro antioxidant properties (DPPH, ABTS, CUPRAC and FRAP), total phenolic content and flavonoid content of extracts from three non-oil seed legumes (Phaseolus lunatus red and white, and Canavalia ensiformis), local edible seeds from Indonesia, obtained using different solvent system (distilled water, 70% ethanol, and 100% ethanol). The variety of legume was a major source of variation in the phenolic contents, flavonoid content and antioxidant activity. HPLC analysis of the non-oil seed legume extracts identified gallic acid, epicatechin and coumaric acid. Among the varieties of non-oil seed legume extracts, the phenolic content varied from 15.21–38.60 mg gallic acid equivalents/g dry weight and the flavonoid content was 11.73–24.61 mg catechin equivalents/g dry weight. The antioxidant activity of the extracts suppressed the reactive oxygen species (ROS) generation and cellular damage induced by UV-B in HaCaT cells. These results showed that antioxidant activity (1.83–19.42% of inhibition DPPH; 2.99–37.29% of inhibition ABTS; 0.20–2.47 µM CUPRAC value; and 0.96–1.10 µM of FRAP value) of extracts possessed strong radical scavenging activity as well as inhibited ROS generation in a dose-dependent manner without showing any cytotoxicity. Collectively, the data presented that antioxidant of the extracts have potent antioxidant activity and decreasing ROS generation in HaCaT cells. It can be intimately used as alternative criterion for antioxidant and antiradical activities that can be utilized as a functional food and nutraceutical ingredients.

Each individual has a unique skin tone based on the types and quantities of melanin pigment, and oxidative stress is a key element in melanogenesis regulation. This research sought to understand the in vitro and in vivo antioxidant and depigmenting properties of betel leaves (Piper betle L.) extract (PBL) and the underlying mechanism. Ethyl acetate fractions of PBL (PBLA) demonstrated excellent phenolic content (342 ± 4.02 mgGAE/g) and strong DPPH, ABTS radicals, and nitric oxide (NO) scavenging activity with an IC50 value of 41.52 ± 1.02 μg/mL, 45.60 ± 0.56 μg/mL, and 51.42 ± 1.25 μg/mL, respectively. Contrarily, ethanolic extract of PBL (PBLE) showed potent mushroom, mice, and human tyrosinase inhibition activity (IC50 = 7.72 ± 0.98 μg/mL, 20.59 ± 0.83 μg/mL and 24.78 ± 0.56 μg/mL, respectively). According to gas chromatography–mass spectrometry, PBL is abundant in caryophyllene, eugenol, O-eugenol, 3-Allyl-6-methoxyphenyl acetate, and chavicol. An in vitro and in vivo investigation showed that PBLE suppressed tyrosinase (Tyr), tyrosinase-related protein-1 and -2 (Trp-1 and Trp-2), and microphthalmia-associated transcription factors (MITF), decreasing the formation of melanin in contrast to the untreated control. PBLE reduced the cyclic adenosine monophosphate (cAMP) response to an element-binding protein (CREB) phosphorylation by preventing the synthesis of cAMP. Additionally, it activates c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinases (p38), destroying Tyr and MITF and avoiding melanin production. Higher levels of microtubule-associated protein-light chain 3 (LC3-II), autophagy-related protein 5 (Atg5), Beclin 1, and lower levels of p62 demonstrate that PBLE exhibits significant anti-melanogenic effects via an autophagy-induction mechanism, both in vitro and in vivo. Additionally, PBLE significantly reduced the amount of lipid peroxidation while increasing the activity of several antioxidant enzymes in vivo, such as catalase, glutathione, superoxide dismutase, and thioredoxin. PBLE can therefore be employed in topical formulations as a potent skin-whitening agent.

The date palm (Phoenix dactylifera L.) is a popular edible fruit consumed all over the world and thought to cure several chronic diseases and afflictions. The profiling of the secondary metabolites of optimized ripe Ajwa date pulp (RADP) extracts is scarce. The aim of this study was to optimize the heat extraction (HE) of ripe Ajwa date pulp using response surface methodology (RSM) and artificial neural network (ANN) modeling to increase its polyphenolic content and antioxidant activity. A central composite design was used to optimize HE to achieve the maximum polyphenolic compounds and antioxidant activity of target responses as a function of ethanol concentration, extraction time, and extraction temperature. From RSM estimates, 75.00% ethanol and 3.7 h (extraction time), and 67 °C (extraction temperature) were the optimum conditions for generating total phenolic content (4.49 ± 1.02 mgGAE/g), total flavonoid content (3.31 ± 0.65 mgCAE/g), 2,2-diphenyl-1-picrylhydrazyl (11.10 ± 0.78 % of inhibition), and cupric-reducing antioxidant capacity (1.43 µM ascorbic acid equivalent). The good performance of the ANN was validated using statistical metrics. Seventy-one secondary metabolites, including thirteen new bioactive chemicals (hebitol II, 1,2-di-(syringoyl)-hexoside, naringin dihydrochalcone, erythron-guaiacylglycerol-β-syringaresinol ether hexoside, erythron-1-(4′-O-hexoside-3,5-dimethoxyphenyl)-2-syrngaresinoxyl-propane-1,3-diol, 2-deoxy-2,3-dehydro-N-acetyl-neuraminic acid, linustatin and 1-deoxynojirimycin galactoside), were detected using high-resolution mass spectroscopy. The results revealed a significant concentration of phytoconstituents, making it an excellent contender for the pharmaceutical and food industries.

Objective: To investigate the effect of Opuntia humifusa aqueous extract on gastric ulcers. Methods: An ethanol-induced model was used to examine the protective effect of Opuntia humifusa against gastric ulcers. The gastric ulcer index was evaluated via clinical observation and image analysis. Various inflammatory indicators were determined by RT-PCR and Western blotting assays. Results: The gastric ulcer index was reduced to 8% in the group treated with Opuntia humifusa aqueous extract compared with that in the control group. RT-PCR analysis revealed that MUC5AC expression was reduced to 39% in the control group compared with the non-treated group, whereas the omeprazole and Opuntia humifusa aqueous extract-treated groups increased the expression to 95% and 79%, respectively. Moreover, the expressions of various cytokines including TNF-α, IL-1β, and IL-6 were increased in the control group, while decreasing in Opuntia humifusa aqueous extract-treated group. Opuntia humifusa aqueous extract also suppressed the expressions of iNOS, COX-2, and its transcription factor NF-κB and increased mucus content considerably as compared to the control group. Conclusions: These results suggest that Opuntia humifusa aqueous extract is suitable as an alternative remedy for gastric ulcer treatment.

Sargassum fusiforme (SF) is a popular edible brown macroalga found in Korea, Japan, and China and is known for its health-promoting properties. In this study, we used two sophisticated models to obtain optimized conditions for high antioxidant activity and metabolite profiling using high-resolution mass spectrometry. A four-factor central composite design was used to optimize the microwave-assisted extraction and achieve the maximum antioxidant activities of DPPH (Y1: 28.01 % inhibition), ABTS (Y2: 36.07 % inhibition), TPC (Y3: 43.65 mg GAE/g), and TFC (Y4: 17.67 mg CAE/g), which were achieved under the optimized extraction conditions of X1: 47.67 %, X2: 2.96 min, X3: 139.54 °C, and X4: 600.00 W. Moreover, over 79 secondary metabolites were tentatively identified, of which 12 compounds were reported for the first time in SF, including five phenolic (isopropyl 3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate, 3,4-dihydroxyphenylglycol, scopoletin, caffeic acid 4-sulfate, and cinnamoyl glucose), two flavonoids (4’,7-dihydroxyisoflavone and naringenin), three phlorotannins (diphlorethohydroxycarmalol, dibenzodioxin-1,3,6,8-tetraol, and fucophlorethol), and two other compounds (dihydroxyphenylalanine and 5-hydroxybenzofuran-2(3H)-one) being identified for the first time in optimized SF extract. These compounds may also be involved in improving the antioxidant potential of the extract. Therefore, optimized models can provide better estimates and predictive capabilities that would assist in finding new bioactive compounds with improved biological activities that can be further applied at a commercial level.

Lactic acid bacteria are one of the potential natural remedies used worldwide, commonly known as probiotics. Here, the aim of this research investigation was to isolate a probiotic Lactobacilli strain, YLM015, from the popular Korean fermented vegetable “Kimchi” and to evaluate its anti-viral potential against influenza virus A (IFVA) H1N1 using the MDCK cell line in vitro, and in embryonated eggs in ovo. The YML015 strain was selected from among the 1200 Lactobacilli isolates for further studies based on its potent anti-viral efficacy. YML015 was identified and characterized as Lactoplantibacillus plantarum YML015 based on the 16S rRNA gene sequencing and biochemically with an API 50 CHL Kit. In ovo assay experienced with embryonated eggs and the hemagglutination inhibition method, as well as cytopathogenic reduction assay, was performed individually to observe anti-influenza viral activity of YML015 against influenza virus A H1N1. Additionally, YML015 was classified for its non-resistance nature as safe for humans and animals as confirmed by the antibiotic susceptibility (MIC) test, cell viability, and hemolysis assay. The heat stability test was also experienced by using different heat-treated cell-free supernatant (CFS) samples of YML015. As a result, YML015 showed highly potent anti-viral activity against influenza virus A H1N1 in vitro in the MDCK cell line. Overall findings suggest that anti-influenza viral activity of L. plantarum YML015 makes it a potential candidate of choice for use as an influential probiotic in pharmacological preparations to protect humans and animals from flu and viral infection.

Insulin resistance (IR) plays a key role in the pathogenesis and clinical outcome of patients with multiple diseases and diabetes. In this study, we examined the antidiabetic effects of a terpenoid-rich extract from Dillenia indica L. bark (TRDI) in palmitic acid-induced insulin resistance (PA-IR) in C2C12 myotube and a streptozotocin (STZ)-induced diabetic mice model and explored the possible underlying mechanism. TRDI showed potential DPPH- and ABTS-radical scavenging effects with a half-maximal inhibitory concentration (IC50) value of 9.76 ± 0.50 µg/mL and 17.47 ± 1.31 µg/mL, respectively. Furthermore, TRDI strongly mitigated α-glucosidase activity with an IC50 value of 3.03 ± 1.01 µg/mL, which was 92-fold higher than the positive control, acarbose (IC50 = 279.49 ± µg/mL). TRDI stimulated the insulin receptor substrarte-1 (INS-1), downregulated phosphoinositide-dependent kinase-1 (PDK1) and protein kinase B (Akt) in both normal and PA-IR C2C12 cells as well as in STZ-induced diabetic mice, enhanced glucose transporter 4 (GLUT4) translocation to the plasma membrane (PM), and increased glucose absorption. Furthermore, TRDI administration significantly reduced PA-induced reactive oxygen species (ROS) formation in C2C12 cells and increased the protein level of numerous antioxidant enzymes such as superoxide dismutase 1 (SOD1), catalase (CAT), glutathione peroxidase-1 (GPx-1) and thioredoxin reductase (TrxR) both in vitro and in vivo. Furthermore, TRDI facilitated nuclear factor erythroid 2 related factor 2 (Nrf2) nuclear translocation and increased HO-1 expression in PA-IR C2C12 cells and STZ-induced diabetic mice. However, for the inhibition of Nrf2, TRDI failed to resist the effects of IR. Thus, this study provides new evidence to support the use of TRDI for diabetes treatment.

Gastric ulcers are one of the most common gastrointestinal diseases. In this study, as an attempt to reduce the minimal error in clinical observations during the diagnosis of gastric ulcers, the applicability of improved ImageJ analysis (IA) was investigated by comparing the results of animal experiments and clinical data. As a result, IA exhibited a significantly improved potential for determining the ulcer index (UI) of clinical data sheets compared to those rated directly by conventional clinical observation (CCO). This indicated that IA enhanced the reproducibility of the measurement of gastric UI using a Bland–Altman plot, resulting in a reduced deviation of each UI value. In addition, it was confirmed that errors in gastric UI decisions can be reduced by adjusting RGB values in diagnostic clinical data (i.e., adjusting to 100 is relatively better than adjusting to 50 or 200). Together, these results suggest that the new enhanced IA could be compatible with novel applications for measuring and evaluating gastric ulcers in clinical settings, meaning that the developed method could be used not only as an auxiliary tool for CCO, but also as a pipeline for ulcer diagnosis.

Abstract Background Cancer is one of the most frequently occurring diseases and is the second leading cause of death worldwide. In this study, anthraquinone derivatives (Compounds 1–5) were evaluated for their anti-cancer potential against various skin and breast cancer cell lines to assess whether these anthraquinone derivatives may serve as a lead for the augmentation of anti-cancer drug. Methods Anthraquinone derivatives, 2-methyl-1,3,6-trihydroxy-9,10-anthraquinone-3-O-(6′-O-acetyl)-α-rhamnosyl(1 → 2)-β-glucoside (Comp 1), 2-methyl-1,3,6-trihydroxy-9,10-anthraquinone (Comp 2), and alizarin (Comp 3) were isolated from the dichloromethane fraction of the roots of Rubia philippinensis., whereas ethyl acetate fraction yielded xanthopurpurin (Comp 4) and lucidin-ω-methyl ether (Comp 5). Structures of all the isolated compounds were determined by spectral data analysis. All isolated compounds (Comp 1–5) were assessed for cytotoxicity by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay against four different cancer cell lines, i.e. human melanoma (SK-MEL-5), murine melanoma (B16F10), and human breast adenocarcinoma (MCF7 and MDA-MB-231). Results Significant activity of the compounds 4 and 5 was observed against the breast cancer cell line MDA-MB-231 with IC50 values of 14.65 ± 1.45 and 13.03 ± 0.33 μM, respectively. Encouragingly, IC50 values of 67.89 ± 1.02 and 79.01 ± 0.03 μM against normal kidney epithelial cells (MDCK) were also obtained for compounds 4 and 5, respectively, which indicated very low toxicity and favorable selectivity indices for compounds 4 and 5 in the range of 1.85 to 3.95 and 2.11 to 6.06 against skin cancer cell lines (SK-MEL-5, and B16F10), and breast cancer cell lines (MCF7 and MDA-MB-231), respectively. Conclusion Our results suggested that the compounds 4 (xanthopurpurin) and 5 (lucidin-ω-methyl ether) showed high selective toxicity towards breast cancer cells at lower concentrations without showing toxicity towards normal cells, thus could be of potential as new lead molecules in cancer treatment.

The anti-graying effect of the hexane fraction of Fuzhuan brick tea is investigated in Melan-A cells and C57BL/6 mice. As a result, it is found that reactive oxygen species-induced damage is associated with the reduction of melanogenesis in hair bulb melanocytes when reactive oxygen species generation in Melan-A cells occurred. The results revealed that the hexane fraction of Fuzhuan brick tea could remarkably reduce reactive oxygen species generation in Melan-A cells; meanwhile, it could increase the cellular tyrosinase and melanin content, as well as up-regulate the expression of tyrosinase, tyrosinase related protein-1, tyrosinase related protein-2, and microphthalmia-associated transcription factor, and activate the MAP-kinase pathway through activating the phosphorylation of p38 c-Jun N terminal kinase/extracellular signal-regulated kinase. Furthermore, high-pressure liquid chromatography analysis reveals that the tea’s major ingredients in hexane fraction include gallic acid, theaflavin, theobromine, caffeine, epicatechin, and quercetin. Together, the current results suggest that Fuzhuan brick tea proves to protect from the damage of hydroquinone, which induces hair pigment loss.

The secondary metabolites profiling of Nymphaea nouchali stem (NNSE) extract was carried out using a high-resolution mass spectroscopic technique. The antioxidant effects of NNSE, as well as the underlying mechanisms, were also investigated in tert-butyl hydroperoxide (t-BHP)-stimulated oxidative stress in RAW264.7 cells. Tandem mass spectroscopy with (−) negative mode tentatively revealed the presence of 54 secondary metabolites in NNSE. Among them, phenolic acids and flavonoids were predominant. Phenolic acids (brevifolincarboxylic acid, p-coumaroyltartaric acid, niazinin B, lalioside, 3-feruloylquinic acid, and gallic acid-O-rutinoside), flavonoids (elephantorrhizol, apigenin-6-C-galactoside 8-C-arabinoside, and vicenin-2), sialic acid (2-deoxy-2,3-dehydro-N-acetylneuraminic acid), and terpenoid (α-γ-onoceradienedione) were identified in NNSE for the first time. Unbridled reactive oxygen species/nitrogen species (ROS/RNS) and redox imbalances participate in the induction and development of many oxidative stress-linked diseases. The NNSE exhibited significant free radical scavenging capabilities and was also able to reduce t-BHP-induced cellular generation in RAW264.7 cells. The NNSE prevented oxidative stress by inducing the endogenous antioxidant system and the levels of heme oxygenase-1 (HO-1) by upregulating Nrf2 through the modulation of mitogen-activated protein kinases (MAPK), such as phosphorylated p38 and c-Jun N terminal kinase. Collectively, these results indicate that the NNSE exhibits potent effects in preventing oxidative stress-stimulated diseases and disorders through the modulation of the MAPK/Nrf2/HO-1 signaling pathway. Our findings provide new insights into the cytoprotective effects and mechanisms of Nymphaea nouchali stem extract against oxidative stress, which may be a useful remedy for oxidative stress-induced disorders.

Context: Aloe has been used for the prevention and cure of various diseases and symptoms including burns, injuries, oedema and pain. Objective: This study determines the specific inhibitory activity of matrix metalloproteinase (MMP)-9 induced by the low molecular-weight gel fraction of Aloe vera (L.) Burm.f. (lgfAv) on alcohol-induced acute gastric lesions. Materials and methods: We examined the protective effects of oral (p.o.) administration of lgfAv (molecular weight cutoff

The antioxidant effects of the ethyl acetate fraction of Dillenia indica bark (DIBEt) and the underlying mechanisms were investigated in tert-butyl hydroperoxide (t-BHP)-stimulated oxidative stress in RAW 264.7 cells. Paper spray ionization-mass spectroscopy with positive-ion mode tentatively revealed 27 secondary metabolites in D. indica bark extract; predominant among them were alkaloids, phenolic acids, and flavonoids. A new triterpenoid (nutriacholic acid) was confirmed in DIBEt for the first time. DIBEt had strong free radical-scavenging capabilities and was also able to reduce t-BHP-induced cellular reactive oxygen species (ROS) generation in RAW 264.7 cells. DIBEt was found to prevent oxidative stress by boosting the levels of heme oxygenase-1 (HO-1) through the up-regulation of nuclear factor erythroid 2-related factor 2 (Nrf2) via the regulation of extracellular signal-regulated kinase (ERK) phosphorylation in RAW 264.7 cells. These results support the potential of DIBEt for defense against oxidative stress-stimulated diseases.

As part of our continuous effort to find potential anti-inflammatory agents from endophytic fungi, a Fusarium solani strain, isolated from the plant Aponogeton undulatus Roxb., was investigated. Cerevisterol (CRVS) was identified from endophytic fungi, a Fusarium solani strain, and moreover exhibited anti-inflammatory activity. However, the underlying mode of action remains poorly understood. The aim of this study is to reveal the potential mechanisms of CRVS against inflammation on a molecular level in LPS-activated RAW 264.7 peritoneal macrophage cells. CRVS was isolated from F. solani and characterized based on spectral data analysis. The MTT assay was performed to measure cell viability in CRVS-treated macrophages. Anti-inflammatory activity was assessed by measurement of nitric oxide (NO) and prostaglandin E2 (PGE2) levels, as well as the production of various cytokines, such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and -6 (IL-6) in LPS-stimulated macrophages. RT-PCR and immunoblotting analyses were done to examine the expression of various inflammatory response genes. A reporter gene assay was conducted to measure the level of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and activator protein-1 (AP-1) transactivation. CRVS suppresses the LPS-induced production of NO and PGE2, which is a plausible mechanism for this effect is by reducing the expression of iNOS and COX-2. CRVS also decreases the expression of pro-inflammatory cytokines, such as TNF-α, IL-6, and IL-1β. CRVS halted the nuclear translocation of NF-κB by blocking the phosphorylation of inhibitory protein κBα (IκBα) and suppressing NF-κB transactivation. The mitogen-activated protein kinases (MAPK) signaling pathways are also suppressed. CRVS treatment also inhibited the transactivation of AP-1 and the phosphorylation of c-Fos. Furthermore, CRVS could induce the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) by down-regulating Kelch-like ECH-associated protein 1 (Keap-1) and up-regulating hemeoxygenases-1 (HO-1) expression. The results suggest that CRVS acts as a natural agent for treating inflammatory diseases by targeting an MAPK, NF-κB, AP-1, and Nrf2-mediated HO-1 signaling cascade.

Background Patients with diabetes mellitus often have a difficult life, suffering from foot ulceration or amputation. Diabetes is characterized by chronic inflammation, and one of the features of inflammation is hypoxia. Recently, it has been reported that KAI1 is a hypoxia target gene. There is no published research on hypoxia-related KAI1 protein levels in human diabetic skin. Therefore, we have investigated the expression of KAI1 protein in diabetic skin tissue in vivo. Methods The expression of KAI1 protein was evaluated by western blotting in 6 diabetic skin tissue samples and 6 normal skin samples. Immunohistochemical staining was carried out to identify KAI1 expression. Results The western blotting revealed significantly increased expression of the KAI1 protein in diabetic skin tissues as compared to normal skin tissues. Immunohistochemical examination demonstrated that KAI1 was expressed in all diabetic skin tissues with moderate-to-strong positivity and weakly expressed in normal skin tissues. Conclusions Our data suggest that a high expression of the KAI1 protein can be observed in diabetic skin tissue. To the best of our knowledge, this is the first report suggesting that KAI1 protein expression in diabetic skin tissues may be associated with chronic inflammatory states and hypoxia.

Recombinant human bone morphogenetic protein (rhBMP)-2 has been approved by the FDA for clinical application, but its use is limited due to high cost and a supra-physiological dose for therapeutic efficacy. Therefore, recent studies have focused on the generation of new therapeutic small molecules to induce bone formation or potentiate the osteogenic activity of BMP-2. Here, we show that [4-(7-chloroquinolin-4-yl) piperazino][1-phenyl-5-(trifluoromethyl)-1H-pyrazol-4-yl]methanone (KM11073) strongly enhances the BMP-2-stimulated induction of alkaline phosphatase (ALP), an early phase biomarker of osteoblast differentiation, in bi-potential mesenchymal progenitor C2C12 cells. The KM11073-mediated ALP induction was inhibited by the BMP antagonist noggin, suggesting that its osteogenic activity occurs via BMP signaling. In addition, a pharmacological inhibition study suggested the involvement of p38 activation in the osteogenic action of KM11073 accompanied by enhanced expression of BMP-2, -6, and -7 mRNA. Furthermore, the in vivo osteogenic activity of KM11073 was confirmed in zebrafish and mouse calvarial bone formation models, suggesting the possibility of its single use for bone formation. In conclusion, the combination of rhBMP-2 with osteogenic small molecules could reduce the use of expensive rhBMP-2, mitigating the undesirable side effects of its supra-physiological dose for therapeutic efficacy. Moreover, due to their inherent physical properties, small molecules could represent the next generation of regenerative medicine.

The increasing antimicrobial drug resistance of Salmonella typhimurium has been a major public health problem in many countries. The type III secretion system (TTSS) of S typhimurium is an attractive target for the development of new antibiotics against this bacterium. In this study, we determined whether berberrubine derived from Berberis vulgaris could inhibit TTSS expression in S typhimurium. Berberrubine inhibited TTSS expression at a concentration of 60.9 µM. Additionally, TTSS-related mRNA expression of sipA, sipB, sopA, and sopD was downregulated in the presence of berberrubine. Collectively, we anticipate that berberrubine can be a useful tool to prevent the risks of infections caused by pathogenic bacteria. [ABSTRACT FROM AUTHOR]

Sargassum fusiforme (SF) is a popular edible brown macroalga found in Korea, Japan, and China and is known for its health-promoting properties. In this study, we used two sophisticated models to obtain optimized conditions for high antioxidant activity and metabolite profiling using high-resolution mass spectrometry. A four-factor central composite design was used to optimize the microwave-assisted extraction and achieve the maximum antioxidant activities of DPPH (Y1 : 28.01 % inhibition), ABTS (Y2 : 36.07 % inhibition), TPC (Y3 : 43.65 mg GAE/g), and TFC (Y4 : 17.67 mg CAE/g), which were achieved under the optimized extraction conditions of X1 : 47.67 %, X2 : 2.96 min, X3 : 139.54 °C, and X4 : 600.00 W. Moreover, over 79 secondary metabolites were tentatively identified, of which 12 compounds were reported for the first time in SF, including five phenolic (isopropyl 3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate, 3,4-dihydroxyphenylglycol, scopoletin, caffeic acid 4-sulfate, and cinnamoyl glucose), two flavonoids (4’,7-dihydroxyisoflavone and naringenin), three phlorotannins (diphlorethohydroxycarmalol, dibenzodioxin-1,3,6,8-tetraol, and fucophlorethol), and two other compounds (dihydroxyphenylalanine and 5-hydroxybenzofuran2(3H)-one) being identified for the first time in optimized SF extract. These compounds may also be involved in improving the antioxidant potential of the extract. Therefore, optimized models can provide better estimates and predictive capabilities that would assist in finding new bioactive compounds with improved biological activities that can be further applied at a commercial level. [ABSTRACT FROM AUTHOR]