Excessive polyamine (PA) consumption, including putrescine (Put), spermidine (Spd), and spermine (Spm), accelerates cancer cell proliferation and migration in cancer patients. In this study, 4-dimethylamino-1-naphthyl isothiocyanate was employed as a derivatization reagent, and microwave-assisted derivatization (300 W, 5 min) was applied to label PA with good chromophores. Following derivatization, a proton donor (citric acid) and an effervescent salt (KHCO3) were added. The resulting neutralization reaction was conducted in an ice bath to generate CO2 under ice-bath conditions for simultaneous salting-out and effervescence. The use of low temperature improved the salting-out effect and effervescence. This improved the extraction efficiency without the need for auxiliary equipment, allowing the extraction process to be completed within three minutes. The quantitative linear range was 1-200 mu M, with detection limits of 0.3, 0.1, and 0.1 mu M for Put, Spd, and Spm, respectively. This research successfully quantified the PA contents in 16 food samples and six cancer cell lines, achieving 400-700-fold greater sensitivity compared to methods using non-derivatized PAs. This improvement offers significant potential for improved clinical and dietary monitoring.

Preservatives and antioxidants are widely used in consumer products to inhibit microbial growth and oxidative degradation. However, their excessive use may pose potential health risks, highlighting the need for regulatory control and reliable analytical methods to ensure safety and compliance. This study aimed to develop a simple, environmentally friendly, and unified sample preparation strategy based on a pH-responsive deep eutectic solvent (pH-DES) coupled with homogeneous liquid-liquid microextraction (HLLME). The method was applied to extract six preservatives and one antioxidant from aqueous and oily matrices, followed by analysis using narrow-bore high-performance liquid chromatography with UV detection. The pH-DES was generated in situ in the sample using 4-methylcyclohexanol as the hydrogen bond acceptor and decanoic acid as the hydrogen bond donor at a 1:1molar ratio. As the pH-responsive component, decanoic acid, together with 4-methylcyclohexanol, facilitates the selective capture of target analytes through hydrogen bonding and hydrophobic interactions. The developed method exhibited satisfactory linearity in both aqueous and oily matrices and was successfully applied to 15 commercial food, pharmaceutical, and cosmetic products, yielding good recoveries (90.6-104.6%). In addition, AGREEprep evaluation confirmed the green performance of the proposed method. Overall, the in situ pH-DES-HLLME approach provides a simple and environmentally friendly strategy for the extraction of preservatives and an antioxidant from complex real-world samples.

Oral squamous cell carcinoma (OSCC), one of the most common cancers in Taiwan, needs new therapeutic agents and treatments. The aim of this study was to investigate the anti-proliferative activity of {N-[3-chloro-4-[5-[3-[[[4-[(cyclopropylcarbonyl)-amino]3-(trifluoromethyl)phenylamino]carbonyl]amino]phenyl]-1,2,4-oxadiazol-3-yl]phenyl]-3-pyridine-carboxamide} (COC), a synthetic molecule, in OSCC cells. COC exhibits potent tumor-suppressive efficacy with IC50 values of 195 nM and 204 nM toward SCC2095 and SCC4 OSCC cells, respectively. Our data revealed that COC caused caspase-dependent apoptosis and downregulated the MAPK signaling pathway. In addition, COC modulated the levels of E-cadherin and β-catenin and inhibited migration. COC also decreased p-STAT3 levels, and the overexpression of STAT3 partially attenuated COC-induced cytotoxicity. Therefore, our findings suggest the use of COC as a new approach to oral cancer treatment.

Spermidine (Spd) and spermine (Spm) are biomarkers for various cancers. In the present study, we used 4-dimethylamino-1-naphthyl isothiocyanate as a derivatizing reagent to achieve microwave-assisted nucleophilic addition to Spd and Spm. We designed a strategy comprising an ice bath combined with salting out to increase the efficiency of Spd and Spm derivative extractions. Ice-bath-effervescence-assisted salting-out extraction (IEASOE) was conducted using a proton donor (citric acid) and an effervescent salt (KHCO3), which simultaneously provided salting-out and effervescence effects. The ice bath stopped the derivatization and improved the extraction efficiency of the IEA-SOE. Moreover, under optimized derivatization and extraction conditions, the total time required was only 10 min. Finally, the extraction layer was collected and analyzed using a narrow-bore liquid chromatograph equipped with a UV detector. The quantitative linear range was 1-200 mu M, and the detection limit was 0.1 mu M. The intra- and inter-batch relative standard deviations were in the range of 3.96-7.74 %, with relative errors in the range of -10.55-4.70 %. The proposed analytical method was successfully applied to determine Spd and Spm concentrations in whole-blood samples. Our method will serve as a useful tool for the accurate assessment of essential biomarkers in blood and subsequently, disease diagnosis.

Natural marine products possess pharmacological effects and have been a source of novel drugs for centuries. In this study, we explored the anti-tumor activity and underlying mechanism of action of ilimaquinone, a sesquiterpene quinone from marine sponges, in gastric cancer. Three gastric cancer cell lines (KATO III, SCM-1, and AZ521) were cultured. Cell viability, cell cycle, and apoptosis were determined using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay and flow cytometry. The levels of apoptotic proteins, proteins related to autophagy, and signal transducer and activator of transcription 3 (STAT3) were detected using western blotting. Transient transfection experiments were conducted to assess the expression of STAT3 in the gastric cancer cells. Ilimaquinone inhibited cell growth, caused cell cycle arrest in the S phase, and induced apoptosis. In addition, ilimaquinone modulates autophagy, and the viability of gastric cancer cells decreases upon exposure to a combination of the autophagic inhibitor, chloroquine. Notably, ilimaquinone downregulated the expression of p-STAT3 in a concentration- and time-dependent manner in both gastric cancer cell lines. Moreover, the compound-mediated inhibition of gastric cancer cell proliferation was restored by ectopic STAT3 expression. Collectively, these findings demonstrate the value of ilimaquinone in the treatment of gastric cancer.

Alzheimer's disease is a progressive neurodegenerative condition that causes brain cell death and is the leading cause of dementia. Most patients with Alzheimer's disease are diagnosed with late-onset Alzheimer's disease (LOAD), with apolipoprotein E (APOE) genotypes being highly associated with the frequency of LOAD risk. A fluorescence detection system coupled with oligonucleotide ligation and magnetic separation was developed to identify two single-nucleotide polymorphisms (SNPs) for the APOE gene and recognize APOE alleles for LOAD. The system utilized a fluorescence probe with one base-discriminating nucleoside for SNP (F probe) and a perfectly complementary biotin-modified sequence against the target DNA (P probe). When the F and P probes matched the target DNA sequences, DNA ligation occurred, and ligation products were produced. Streptavidin magnetic beads were subsequently employed to remove the ligation products, and a decrease in fluorescence intensity was observed in the supernatant compared to when there was no target DNA. This system detected two SNPs of APOE alleles, namely rs429358 and rs7412. The results indicated that the R-values ((F0 - F1)/F0) for rs429358 were 0.92 ± 0.002 for the T/T target, 0.47 ± 0.004 for the T/C target and 0.11 ± 0.004 for the C/C target, respectively. The R-values for rs7412 were 0.73 ± 0.009 for the C/C target, 0.42 ± 0.001 for the C/T target and 0.16 ± 0.007 for the T/T target, respectively. F0 and F1 represent the fluorescence intensity of the F probe without and with target DNA, respectively. Based on fluorescence intensity, the fluorescence detection system was able to identify the genotypes of the APOE gene accurately to evaluate the risk of Alzheimer's disease.

Histamine causes allergic reactions and can serve as an indicator for assessing food quality. This study designed and developed a dispersive micro solid-phase extraction (D-μSPE) method that combined the advantages of dispersive liquid–liquid extraction and solid-phase extraction (SPE). Molecularly imprinted polymers (MIPs) were employed as the solid phase in the D-μSPE method to extract histamine in wine samples. We used microwave energy to significantly reduce the synthesis time, achieving an 11.1-fold shorter synthesis time compared to the conventional MIP synthetic method. Under optimized D-μSPE conditions, our results showed that the dispersive solvent could effectively increase the adsorption performance of MIPs in wine samples by 97.7

The enzyme pyruvate kinase M2 (PKM2) is involved in glycolysis, which plays an important role in the regulation of tumor progression. In this study, we investigated the anti-tumor activity of N-(4-(3-(3-(methylamino)-3-oxopropyl)-5-(4'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)-1H-pyrazol-1-yl)phenyl)propiolamide (MTP), a PKM2 inhibitor, in oral squamous cell carcinoma (OSCC) cells. Our results showed that MTP inhibited cell growth with IC50 values of 0.59 μM and 0.78 μM in SCC2095 and HSC-3 OSCC cells, respectively. MTP induced caspase-dependent apoptosis, which was associated with the modulation of PKM2 and oncogenic biomarkers epidermal growth factor receptor and β-catenin. In addition, MTP increased the generation of reactive oxygen species (ROS) and modulated the expression of autophagic gene products, including LC3B-II and p62. Western blotting showed that MTP inhibited Janus kinase 2 (JAK2) signaling, and JAK2 overexpression partially reversed MTP-mediated cytotoxicity. Taken together, these data indicate the potential use of MTP as a therapeutic agent for OSCC.

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is a rapid and low-solvent-consumption technique. However, almost every mass in the low mass-to-charge-ratio region of the mass spectrum appears as strongly fluctuating matrix background signals. Thus, it is difficult to identify small molecules using this technique. In this study, we used methanol to methylate valsartan, an angiotensin II receptor blocker that is commonly used to treat high blood pressure and heart failure. The methylation derivatization of valsartan enhanced the detection sensitivity and transformed the detection m/z ratio. The liquid-phase microextraction of valsartan in human plasma (20 μL) was achieved by acidifying valsartan with HCl aqueous solution and extracting it with toluene. An acetyl chloride/anhydrous methanol mixture was added for methylation derivatization, which was completed within 30 min at 30 °C. Finally, the residue was re-dissolved in irbesartan methanolic solution, which together with the matrix 2-mercaptobenzothiazole was spotted on an AnchorChip target plate for MALDI-TOF MS analysis. Liquid-phase microextraction was performed and the methylation-derivatization parameters were investigated. The valsartan calibration range was 0.2-10 μg mL-1 with good linearity in human plasma. In the within- and between-run analyses, the relative standard deviation and relative error were both <11.32%. This method was successfully applied to determine the valsartan concentration in the plasma of 10 patients with hypertension.

In this study, the anti-proliferative effect of ilimaquinone, a sesquiterpene derivative from the marine sponge, in breast cancer cells was investigated. Ilimaquinone inhibited the proliferation of MCF-7 and MDA-MB-231 breast cancer cells with IC50 values of 10.6 μM and 13.5 μM, respectively. Non-tumorigenic human breast epithelial cells were less sensitive to ilimaquinone than breast cancer cells. Flow cytometric and Western blot analysis showed that ilimaquinone induced S-phase arrest by modulating the expression of p-CDC-2 and p21. Ilimaquinone induces apoptosis, which is accompanied by multiple biological biomarkers, including the downregulation of Akt, ERK, and Bax, upregulation of p38, loss of mitochondrial membrane potential, increased reactive oxygen species generation, and induced autophagy. Collectively, these findings suggest that ilimaquinone causes cell cycle arrest as well as induces apoptosis and autophagy in breast cancer cells.

Biogenic amines are quality control criteria for foods that are potentially toxic to humans. In this study, amidation derivatization for biogenic amines and liquid-solid phase transition microextraction were carried out simultaneously for food sample pretreatment. The derivatization reaction was executed in one pot with coumarin-3-carboxylic acid as the derivatizing reagent and (1-cyano-2ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate as the coupling agent. Liquid-solid phase transition microextraction was achieved by the salting-out effect, using a phase change salt (1 M disodium hydrogen phosphate) solution. The combined derivatization and microextraction process was completed within 3 min at 30 degrees C, and the liquid top phase was easily obtained by placing the tube in an ice bath. Finally, a narrowbore liquid chromatograph coupled with a UV detector was used to determine the levels of six biogenic amines. The coupling agent-assisted derivatization and liquid-solid phase transition microextraction parameters were also investigated. The quantitative linear ranges were 3-400 mu M for histamine, putrescine, spermidine, cadaverine, and tyramine and 5-400 mu M for spermine, and the detection limit was 1 mu M. The relative standard deviations of the intra- and interbatches were <5.3% and 8.4%, respectively, while the relative error was <4.5% for both. We successfully applied this simultaneous derivatization-microextraction method to determine the biogenic amines in fermented foods. (C) 2021 Elsevier B.V. All rights reserved.

Secondary metabolites in marine organisms exhibit various pharmacological activities against diseases, such as cancer. In this study, the anti-proliferative effect of JBIR-100, a macrolide isolated from Streptomyces sp., was investigated in breast cancer cells. Cell growth was inhibited in response to JBIR-100 treatment concentration- and time-dependently in both MCF-7 and MDA-MB-231 breast cancer cells. JBIR-100 caused apoptosis, as verified by caspase activation and the cleavage of PARP. Western blotting revealed that JBIR-100 modulated the expression of Akt/NF-κB signaling components and Bcl-2 family members. Overexpression of Mcl-1 partially rescued MCF-7 cells from JBIR-100-induced cytotoxicity. In addition, transmission electron microscopy analyses, confocal analysis, and western blot assay indicated that JBIR-100 inhibited autophagy in MCF-7 cells. Exposure to the autophagy inhibitor did not synergize JBIR-100-induced apoptosis. In summary, our results suggested that JBIR-100 may be potentially used for breast cancer therapy.

We recently isolated a cardiac glycoside (CG), αldiginoside, from an indigenous plant in Taiwan, which exhibits potent tumor-suppressive efficacy in oral squamous cell carcinoma (OSCC) cell lines (SCC2095 and SCC4, IC50 < 0.2 µM; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays). Here, we report that αldiginoside caused Sphase arrest and apoptosis, through the inhibition of a series of signaling pathways, including those mediated by cyclin E, phospho-CDC25C (p-CDC25C), and janus kinase/signal transducer and activator of transcription (JAK/STAT)3. αldiginoside induced apoptosis, as indicated by caspase activation and poly (ADP-ribose) polymerase (PARP) cleavage. Equally important, αldiginoside reduced Mcl-1 expression through protein degradation, and overexpression of Mcl-1 partially protected SCC2095 cells from αldiginoside's cytotoxicity. Taken together, these data suggest the translational potential of αldiginoside to foster new therapeutic strategies for OSCC treatment.

Global climate change has led to a significant increase in temperature over the last century and has been associated with significant increases in the severity and frequency of heat injury (HI). The consequences of HI included dehydration and rhabdomyolysis, leading to acute kidney injury, which is now recognized as a clear risk factor for chronic kidney disease (CKD). We aimed to investigate the effects of HI on the risk of CKD. This nationwide longitudinal population-based retrospective cohort study utilized the Taiwan National Health Insurance Research Database (NHIRD) data. We enrolled patients with HI who were followed in NHIRD system between 2000 and 2013.We excluded patients diagnosed with CKD or genital-urinary system-related disease before the date of the new HI diagnosis. The control cohort consisted of individuals without HI history. The patients and control cohort were selected by 1:4 matching according to the following baseline variables: sex, age, index year, and comorbidities. The outcome measure was CKD diagnosis. In total, 815 patients diagnosed with HI were identified. During the 13 year observation period, we identified 72 CKD events (8.83%) in the heat stroke group and 143 (4.38%) CKD events in the control group. Patients with heat stroke had an increased risk of CKD than the control patients (adjusted HR = 4.346, P < 0.001) during the follow-up period. The risk of end-stage renal disease was also significantly increased in the heat stroke group than in the control group (adjusted hazards ratio: 9.078, p < 0.001). HI-related CKD may represent one of the first epidemics due to global warming. When compared to those without HI, patients with HI have an increased CKD risk.

This study explores the amounts of common chemical ultraviolet (UV) filters (i.e., avobenzone, bemotrizinol, ethylhexyl triazone, octocrylene, and octyl methoxycinnamate) in cosmetics and the human stratum corneum. An ultrasound–vortex-assisted dispersive liquid–liquid microextraction (US–VA–DLLME) method with a high-performance liquid chromatography–diode array detector was used to analyze UV filters. A bio-derived solvent (i.e., anisole) was used as the extractant in the US–VA–DLLME procedure, along with methanol as the dispersant, a vortexing time of 4 min, and ultrasonication for 3 min. The mass-transfer rate of the extraction process was enhanced due to vortex-ultrasound combination. Various C18 end-capped columns were used to investigate the separation characteristics of the UV filters, with XBridge BEH or CORTECS selected as the separation column. Calibration curves were constructed in the 0.05–5 μg/mL (all filters except octocrylene) and 0.1–10 μg/mL (octocrylene) ranges, and excellent analytical linearities with coefficients of determination (r2) above 0.998. The developed method was successfully used to analyze sunscreen. Moreover, experiments were designed to simulate the sunscreen-usage habits of consumers, and the cup method was used to extract UV filters from the human stratum corneum. The results suggest that a makeup remover should be employed to remove water-in-oil sunscreens from skin.

Oral squamous cell carcinoma (OSCC) is the fifth common cause of cancer mortality in Taiwan with high incidence and recurrence and needs new therapeutic strategies. In this study, ursolic acid (UA), a triterpenoid, was examined the antitumor potency in OSCC cells. Our results showed that UA inhibited the proliferation of OSCC cells in a dose- and time-dependent manner in both Ca922 and SCC2095 oral cancer cells. UA induced caspase-dependent apoptosis accompanied with the modulation of various biological biomarkers including downregulating Akt/mTOR/NF-κB signaling, ERK, and p38. In addition, UA inhibited angiogenesis as evidenced by abrogation of migration/invasion and blocking MMP-2 secretion in Ca922 cells. Interestingly, UA induced autophagy in OSCC cells, as manifested by LC3B-II conversion and increased p62 expression and accumulation of autophagosomes. Inhibition by autophagy inhibitor enhanced UA-mediated apoptosis in Ca922 cells. The experiment provides a rationale for using triterpenoid in the treatment of OSCC.

In this study, an ecofriendly analytical method was developed for determining glutathione (GSH) levels in biomatrix samples. 9-(bromomethyl)acridine was used for the first time as a derivatization reagent in GSH analysis. Microwave-assisted derivatization reduced the reaction time to 1 min. After derivatization, coacervative extraction was employed to extract GSH derivative from the complex biomatrix and to increase sensitivity. Because the negatively charged group of the GSH derivative was neutralized by the extracting agent Aliquat 336, aggregates formed without any coacervating agents. Furthermore, capillary liquid chromatography coupled with ultraviolet detection was applied to decrease waste generation and increase selectivity. This method successfully quantified GSH levels in various biomatrices, including erythrocytes, HaCaT cells, BALB/3T3 cells, and 313-L1 fibroblasts. This method only required a low sample volume (< 10 A standard addition method was utilized to spike the biomatrix samples with 0-4.8 nmol GSH to construct calibration curves. The proposed method performed well, with a determination coefficient of 0.999 and relative standard deviations of less than 6.59% for the slope and the intercept, as determined by linear regression analysis. The limit of detection of GSH in the standard solution was 800 nM or 0.4 pmol. Compared to non-derivatized GSH, the proposed method for detecting derivatized GSH provides 750-fold greater sensitivity.

A simple cyclodextrin-mediated capillary zone electrophoresis method equipped with a laser-induced fluorescence detector was developed for chiral analysis of the excitatory amino acids aspartate (Asp) and glutamate (Glu). Plasma and cerebrospinal fluid (CSF) samples were pretreated with centrifugal filter devices before analysis to remove high-molecular-weight proteins (molecular weight cut-off: 3000) and then derivatized using 10 mM 6-carboxyfluorescein N-hydroxysuccinimide ester in DMSO under sonication at 25 degrees C for 2 h. After the derivatization reaction, reacted samples were diluted 100-fold with 5([4,6-dichlorotriazin-2-yljamino)fluorescein hydrochloride (DTAF) solution and then hydrodynamically subjected to capillary electrophoresis (0.5 psi for 5 s, injection volume 8.27 nL). The separation buffer consisted of 50 mM borate buffer (pH 9.0) with 6 mM gamma-CD and 0.1% polyvinylpyrrolidone, and the separation voltage was set at 20 kV. In the linearity calculations for the determination of D/L-Asp and n/L-Glu in plasma and CSF, a standard addition method was utilized to spike solutions with 0-20.0 mu g mL(-1) L-Glu and 0-2.0 mu g mL(-1) D-Glu and D/L-Asp to construct calibration curves. Correlation coefficients were above 0.998 for every analyte. The limits of detection (S/N=3) for OIL-Asp and D/L-Glu standard solutions were 0.85-0.96 mu g mL(-1). The proposed method was applied successfully to determine D/L-Asp and D/L-Glu concentrations in the plasma and CSF samples of 26 patients with Alzheimer's disease (AD), and the association between these concentrations and disease severity was investigated. Statistical analysis showed a moderately negative correlation (r = -0.158) between plasma L-Asp concentration and AD severity. (C) 2018 Published by Elsevier B.V.

The peroxisome proliferator-activated receptor γ (PPARγ) is a nuclear receptor that plays a key role in regulating cellular metabolism, and is a therapeutic target for cancer therapy. To search for potential PPARγ activators, a compound library comprising 11 marine compounds was examined. Among them, a sterol, 3β,11-dihydroxy-9,11-secogorgost-5-en-9-one (compound 1), showed the highest PPARγ activity with an IC50 value of 8.3 μM for inhibiting human breast adenocarcinoma cell (MCF-7) growth. Western blotting experiments showed that compound 1 induces caspase activation and PARP cleavage. In addition, compound 1 modulated the expression of various PPARγ-regulated downstream biomarkers including cyclin D1, cyclin-dependent kinase (CDK)6, B-cell lymphoma 2 (Bcl-2), p38, and extracellular-signal-regulated kinase (ERK). Moreover, compound 1 increased reactive oxygen species (ROS) generation, upregulated the phosphorylation and expression of H2AX, and induced autophagy. Interestingly, pre-treatment with the autophagy inhibitor 3-methyladenine rescued cells from compound 1-induced growth inhibition, which indicates that the cytotoxic effect of compound 1 is, in part, attributable to its ability to induce autophagy. In conclusion, these findings suggest the translational potential of compound 1 in breast cancer therapy.

Parabens are common preservatives and environmental hormones. As such, possible detrimental health effects could be amplified through their widespread use in foods, cosmetics, and pharmaceutical products. Thus, the determination of parabens in such products is of particular importance. This study explored vortex-assisted dispersive liquid-liquid microextraction techniques based on the solidification of a floating organic drop (VA-DLLME-SFO) and salt-assisted cloud point extraction (SA-CPE) for paraben extraction. Microanalysis was performed using a capillary liquid chromatography-ultraviolet detection system. These techniques were modified successfully to determine four parabens in 19 commercial products. The regression equations of these parabens exhibited good linearity (r2=0.998, 0.1-10μg/mL), good precision (RSD<5%) and accuracy (RE<5%), reduced reagent consumption and reaction times (<6min), and excellent sample versatility. VA-DLLME-SFO was also particularly convenient due to the use of a solidified extract. Thus, the VA-DLLME-SFO technique was better suited to the extraction of parabens from complex matrices.