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Chronic exposure to airborne particulate matter (PM) causes respiratory damage in humans owing to oxidative stress and inflammation. Chrysanthemum zawadskii Herbich var. latilobum (Maxim.) Kitamura (CZL) has been used in traditional medicine to treat several inflammatory diseases; however, studies on inflammatory pulmonary diseases are scarce. This study investigated the protective effects of CZL extract against PM10-induced lung injury in BALB/c mice. Cell type specific signaling pathways were explored using A549 and RAW264.7 cell lines. CZL extract noticeably attenuated PM10-induced lung injury and inflammatory cell infiltration in a mouse model. Protein markers, such as p-AKT, p-ERK, and p-NF-κB for PM10 induced lung inflammation were effectively reduced in CZL extract-treated mice and cells. Furthermore, CZL extracts considerably reduced the generation of reactive oxygen species and nitric oxide in cells. Collectively, CZL extract effectively reduced PM10-induced lung injury by suppressing pulmonary inflammation, potentially due to its anti-inflammatory and antioxidant properties. [ABSTRACT FROM AUTHOR]

Methylation induces epigenetic silencing of tumor suppressor genes in human lung cancer. Inhibition of DNA methyltransferases by decitabine (DAC) can demethylate and activate epigenetically silenced tumor suppressor genes. Epigenetic therapy using DAC should be an attractive strategy for lung cancer therapy. FBW7 is a tumor suppressor that functions as an Mcl-1 E3 ligase to degrade Mcl-1 by ubiquitination. Here we discovered that treatment of various human lung cancer cells with DAC resulted in activation of FBW7 expression, decreased levels of Mcl-1 protein, and growth inhibition. DAC-activated FBW7 expression promoted Mcl-1 ubiquitination and degradation leading to a significant reduction in the half-life of Mcl-1 protein. Mechanistically, treatment of lung cancer cells or lung cancer xenografts with DAC induced the conversion of the FBW7 gene from a methylated form to an unmethylated form, which was associated with the increased expression of FBW7 and decreased expression of Mcl-1 in vitro and in vivo. DAC suppressed lung cancer growth in a dose-dependent manner in vivo. Combined treatment with DAC and a Bcl2 inhibitor, venetoclax, exhibited strong synergistic potency against lung cancer without normal tissue toxicity. These findings uncover a novel mechanism by which DAC suppresses tumor growth by targeting the FBW7/Mcl-1 signaling pathway. Combination of DAC with Bcl2 inhibitor venetoclax provides more effective epigenetic therapy for lung cancer. [ABSTRACT FROM AUTHOR]

The present study investigated the terminal differentiation capacity into adipocytes and subsequent growth inhibition in A549 cancer cells treated with pioglitazone (PGZ), a PPARγ activator. The rate of cell growth in A549 cells was significantly (P <.05) inhibited in concentrations above 10 μM PGZ while maintaining less cytotoxic effects in MRC-5 fibroblasts. Following 50 μM PGZ treatment, population doubling time (PDT) was significantly (P <.05) increased by inhibition of cell growth, as per increasing PGZ exposure time by up to 4 weeks. The adiposome-like vesicles were commonly observed in the PGZ-treated A549 cells, and the vesicles were highly stained with Oil-Red O solution. In addition, the cell size and expression of GLUT4 and PPARγ were significantly (P <.05) increased, as per increasing PGZ exposure time by up to 4 weeks. The significant (P <.05) down-regulation of telomerase activity and up-regulation of senescence-associated β-galactosidase (SA β-GAL) activity was displayed in the PGZ-treated A549 cells, as per increasing PGZ exposure time by up to 4 weeks. The G1 phase of the cell cycle was also significantly (P <.05) increased in the PGZ-treated A549 cells compared with untreated A549 cells. The present results have demonstrated that activation of PPARγ using PGZ induces cellular differentiation into adipocytes and inhibits cell growth in the A549 cancer cells. The terminal differentiation into adipocytes could offer potent chemotherapy in the cancer cells showing high glucose metabolism. [ABSTRACT FROM AUTHOR]

Upon bioprinting, cells are mixed with a biomaterial to fabricate a living tissue, thus emphasizing the importance of biomaterials. The biomaterial used in this study was a bio-ink prepared using skin decellularized extracellular matrix (dECM). Skin dECM was extracted by treating the dermis with chemicals and enzymes; the basic structural and functional proteins of the ECM, including collagen, glycosaminoglycans (GAGs), bioreactive materials and growth factors, were preserved, whereas the resident cells that might cause immune rejection or inflammatory responses were removed. The bio-ink based on dECM powder, together with human dermal fibroblasts (HDFs), was loaded into the nozzle of the 3D bioprinter to create the 3D construct. This construct underwent gelation with changing temperature while its shape was maintained for 7 days. The cells showed over 90% viability and proliferation. By analysing the gene expression pattern in the cells of the construct, the skin regenerative mechanism of the bio-ink was verified. Microarray results confirmed that the gene expression related to skin morphology and development had been enhanced because the bioreactive molecules and growth factors, in addition to residual ECM in dECM, provided an optimal condition for the HDFs. [ABSTRACT FROM AUTHOR]

The present study investigated the cellular properties in the dental tissue-derived mesenchymal stem cells (DSCs) exposed to nevirapine (NVP), an inhibitor of reverse transcriptase (RTase). After a prolonged exposure of DSCs for 2 weeks, the population doubling time (PDT) was significantly (P <.05) increased by delayed cell growth in the DSCs treated with 250 and 500 μM NVP, compared with untreated DSCs. Furthermore, the G1 phase of cell cycle with high activity of senescence-associated β-galactosidase was also significantly (P <.05) increased in the 250 μM NVP-treated DSCs, compared with untreated DSCs. The level of telomerase activity was unchanged between control and treatment. However, following the treatment of NVP, negative surface markers for mesenchymal stem cells (MSCs), such as CD34 and CD45, were significantly (P <.05) increased, while positive surface markers for MSCs, such as CD90 and CD105, were significantly (P <.05) decreased in the NVP-treated DSCs than those of untreated DSCs. Furthermore, the differentiation capacity into mesodermal lineage was gradually decreased, and a significant (P <.05) decrease of expression level of NANOG, OCT-4 and SOX-2 transcripts was observed in the DSCs treated with NVP, compared with untreated control DSCs. Taken together, the present results have revealed that inhibition of RTase by NVP induces delayed cell growth and loss of stemness. [ABSTRACT FROM AUTHOR]

The onset and progression of Alzheimer's disease (AD) is closely associated with amyloidβ(Aβ) peptide-induced cytotoxicity and abnormal protein transportation caused by breakdown of endoplasmic reticulum (ER)–Golgi apparatus trafficking network. Although the fragmentation of Golgi apparatus has been reported in AD human patients and various AD model animals, the molecular mechanisms causing the morpho-functional impairments of Golgi apparatus during AD progression remain poorly understood. Thus, we investigated the ultrastructure of Golgi apparatus and coat protein I (COPI) expression inβ-amyloid precursor protein/presenilin-1 double transgenic mouse and Aβ-stimulated BV-2 cell as an AD model using cryo-immunogold electron microscopy. In the neurons of the hippocampus of transgenic mouse and BV-2 cell, the cisternae of Golgi stacks were fragmented in difference with that of wild type mouse and cells. In addition, we further showed the COPI depletion in Golgi apparatus, which demonstrated the impairment of molecular integrities of Golgi apparatus. Taken together, our results provide insights into the Golgi apparatus-involved morphopathology of AD and we suggest that the Golgi fragmentation is caused by the depletion of COPI affecting the intra-Golgi transport through stimulation and accumulation of Aβduring AD development. [ABSTRACT FROM PUBLISHER]

A simple and sensitive high-performance liquid chromatography (HPLC) method was developed for the determination of 19 preservatives in cosmetic matrices. The composition of the mobile phase was optimized as a gradient to achieve a lower detection limit when compared to previously validated methods, and sample preparation conditions were investigated to optimize separation of the 19 preservatives. A C18 column was used with methanol, 0.05 mol/L ammonium acetate buffer, and water as the mobile phase under gradient elution conditions. Preservatives in cosmetics were extracted with 70% methanol using an ultrasonicator, after which they were analyzed with an HPLC-photodiode array detector. All preservatives were separated within 55 min. The recoveries ranged from 94.9% to 102.8%, with relative standard deviations of less than 3.2% and no correlation coefficients lower than 0.9986. Additionally, the developed method has a low detection limit, which makes it possible to analyze trace levels of compounds in various cosmetic and ingredient matrices. [ABSTRACT FROM AUTHOR]

The hepatic uptake of pitavastatin is mediated by carriers, especially OATP1B1, which is encoded by the SLCO1B1 gene. Because the liver is a target organ of pitavastatin, OATP1B1 is responsible for both the pharmacological effects and clearance of pitavastatin. The effects of the SLCO1B1*15 allele on the pharmacokinetics (PK) of pitavastatin were studied. Pitavastatin 2 mg was orally administered to 38 subjects with SLCO1B1*1a/*1b (n = 20), *1b/*15 (n = 13), or *15/*15 (n = 5). After pitavastatin administration, the plasma concentrations of pitavastatin and pitavastatin lactone were assayed for up to 48 h using liquid chromatography-tandem mass spectrometry. In comparison to the SLCO1B1*1a/*1b subjects, only a Cmax was slightly higher in the SLCO1B1*1b/*15 subjects. However, the SLCO1B1*15/*15 subjects had a 1.74-fold higher AUCinf (285.5 ± 14.5 vs. 164.6 ± 41.3 ng·h/mL; p < 0.001), a 2.21-fold higher Cmax (106.7 ± 15.1 vs. 48.3 ± 13.4 ng/mL; p < 0.001), and a 47.3% lower apparent oral clearance (13.1 ± 3.9 vs. 6.9 ± 0.4 L/h; p < 0.001) of pitavastatin. For pitavastatin lactone, there were no significant differences in AUCinf, Cmax, t1/2, and tmax among the three genotypes. Unlike previous studies, the disposition of pitavastatin exposure was not altered in subjects with the SLCO1B1*1b/*15 genotype, except Cmax. However, pitavastatin exposure was significantly increased in subjects with the SLCO1B1*15/*15 genotype due to reduced hepatic absorption. [ABSTRACT FROM AUTHOR]

We assessed blood interaction with different micrometer-scale topographies under flow conditions using a micro-fluidic array system. The channels of the micro-fluidic array chip were coated with azobenzene polymer films, which were then topographically structured using a one-step non-contact optical technique. A set of surfaces with different topographies was produced varying laser irradiation duration. These surfaces were then exposed to blood flow. The blood flow rate was measured with a micro-channel array flow analyzer. The measured blood flow rates decreased with time for all the samples, indicating formation of platelet clots which obstruct the channels during flow. This effect appeared enhanced on polymer surfaces having a sinusoidal profile with 200-nm-high ridges and 1.2-μm-grating spacing. The morphology of platelets that adhered on the polymer films was studied by scanning electron microscopy. Platelets adhered on azobenzene surfaces with flat topographies, typically exhibiting filopodia. Platelets adhered on optically structured surfaces also exhibited lamellipodia and appeared flattened on surfaces with the highest ridges. We conclude that surface topography influences blood behaviour on azobenzene polymer films. [ABSTRACT FROM AUTHOR]

Glipizide is an oral hypoglycemic agent widely used to treat type 2 diabetes mellitus. In this study, an efficient HPLC-UV assay method for determining the plasma glipizide level was developed, validated, and used to assess the pharmacokinetic profile of the glipizide in healthy Korean volunteers. After extraction with diethyl ether, the chromatographic separation of glipizide was carried out using a Bondclone C18 column (10 µm, 300 × 3.9 mm) with a mobile phase of 10 mM potassium phosphate monobasic and methanol (40:60 [vol/vol], pH 3.5) and UV detection at 225 nm. The flow rate of the mobile phase was 1.0 mL/min and the retention time of glipizide and internal standard (I.S.) was approximately 11.5 and 8.6 minutes, respectively. The quantification limit was 15 ng/mL and the linear range of the calibration curve ranged from 15 to 800 ng/mL in plasma with a correlation coefficient >0.9999. The mean accuracy was 86-101%. The coefficient of variation (precision) in the intra- and inter-day validation was 1.8-14.2 and 1.7-8.1%, respectively. The pharmacokinetics of oral glipizide was evaluated after administering 5 mg to each of 13 healthy Korean subjects. The AUCinf, Cmax, Tmax, and T1/2 were 3432 ± 886 ng·h/mL, 629.0 ± 94.2 ng/mL, 2.8 ± 1.8 h, and 3.9 ± 0.9 h, respectively. The results showed large inter-individual differences in the AUCinf, Cmax and T1/2. [ABSTRACT FROM AUTHOR]

Amyloid beta peptide (Aβ)-induced oxidative stress may be linked to neurodegenerative disease. Ethanol extracts of Rosa laevigata protected PC12 cells from hydrogen peroxide-induced oxidative stress. (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) reduction assays revealed a significant increase in cell viability when oxidatively stressed PC12 cells were treated with R. laevigata extract. The effect of R. laevigata on oxidative stress-induced cell death was further investigated by lactate dehydrogenase release assays and trypan blue exclusion assays. Administration of 1,2-benzenedicarboxylic acid dinonyl ester from R. laevigata extract to mice infused with Aβ significantly reversed learning and memory impairment in behavioural tests. After behavioural testing, the mice were sacrificed and brains were collected for the examination of lipid peroxidation, catalase activity and acetylcholinesterase (AchE) activity. These results suggest that 1,2-benzenedicarboxylic acid dinonyl ester from R. laevigata extract may be able to reduce Aβ-induced neurotoxicity, possibly by reducing oxidative stress. Therefore, R. laevigata extract may be useful for the prevention of oxidative stress-induced neurodegenerative disorders. [ABSTRACT FROM AUTHOR]

Objectives. CXCL13 is a potent chemokine, produced by mature and recently recruited macrophages to sites of inflammation, which has antimicrobial and anti-angiogenic properties. The purpose of this study was to: (1) determine whether CXCL13 is present in maternal serum, umbilical cord blood, and amniotic fluid (AF); (2) to determine if AF concentration changes with intra-amniotic infection/inflammation (IAI); and (3) to localize the production of CXCL13 in chorioamniotic membranes and umbilical cord. Study design. A cross-sectional study on maternal serum was performed including patients in the following groups: (1) non-pregnant women (n = 20), (2) normal pregnant women (n = 49), (3) patients at term not in labor (n = 30), and (4) patients in spontaneous labor at term (n = 29). Umbilical cord blood was collected from term neonates with (n = 30) and without labor (n = 28). Amniotic fluid was obtained from patients in the following groups: (1) midtrimester (n = 65); (2) term not in labor (n = 22); (3) term in labor (n = 47); (4) preterm labor (PTL) with intact membranes leading to term delivery (n = 70); and (5) PTL leading to preterm delivery with IAI (n = 79) and without IAI (n = 60). CXCL13 concentrations were determined by enzyme-linked immunosorbent assay. Chorioamniotic membranes and umbilical cords were examined with immunohistochemistry. Non-parametric statistics were used for analysis. Results. (1) CXCL13 was present in 100% of serum and cord blood samples, and 99% of AF samples (339/343). (2) Serum CXCL13 concentration was significantly higher in pregnant women when compared to non-pregnant women (median 313.3 pg/mL (interquartile range (IQR) 197.2-646.9) vs. 40.5 pg/mL (IQR 29.5-93.5), respectively; p < 0.001). (3) Serum CXCL13 concentration decreased with advancing gestational age (Spearman's Rho = -0.424; p < 0.001). (4) There were no significant differences in the median serum CXCL13 concentration between women at term with and without labor (371.6 pg/mL (IQR 194.3-614.3) vs. 235.1 pg/mL (IQR 182.8-354.7), respectively; p = 0.6). (5) The concentration of CXCL13 in AF did not change with gestational age (p = 0.1). (6) Patients with PTL and delivery with IAI had a significantly higher median concentration of CXCL13 than those without IAI (median 513.2 pg/mL (IQR 199.7-2505.5) vs. 137.3 pg/mL (IQR 96.7-209.6), respectively; p < 0.001) and those who delivered at term (133.7 pg/mL (IQR 97.8-174.8); p < 0.001). (7) Spontaneous labor did not result in a change in the median AF concentration of CXCL13 (labor: 86.9 pg/mL (IQR 55.6-152.0) vs. no labor: 77.8 pg/mL (IQR 68.0-98.0); p = 0.8). (8) CXCL13 was immunolocalized to macrophages in fetal membranes and umbilical vein. Conclusions. (1) We report for the first time the presence of CXCL13 in AF. (2) AF CXCL13 concentrations are dramatically increased in IAI. (3) Unlike other chemokines, AF and serum CXCL13 concentrations did not change with spontaneous parturition. [ABSTRACT FROM AUTHOR]

This study was aimed to validate a reliable analytical method for the pharmacokinetic study of amiloride in human plasma by a high performance liquid chromatography (HPLC) system with UV detection. Triamterene was used as an internal standard. After extraction with ethylacetate, the supernatant was evaporated. Then, the residue was reconstituted and an aliquot was injected onto the HPLC system. Separation was performed on a Capcell Pak C18 UG120 column (4.6 mm × 150 mm, 5 µm particles) with a mobile phase of 12% acetonitrile containing 0.4% glacial acetic acid and UV detection at a wavelength of 360 nm. The intra- and inter-day precision expressed as the relative standard deviation was less than 15%. The flow rate of mobile phase was 1 mL/min and the retention time of amiloride and internal standard, triamterene, was found to be 3.06 and 8.80 min, respectively. The lower limit of quantification (LOQ) was 0.2 ng/mL of amiloride using 1 mL of plasma. The calibration curve was linear in the concentration range of 0.2-50 ng/mL (r2 = 1.0000). The mean accuracy was 85.1-101.7%. The coefficient of variation (precision) in the intra- and inter-day validation was 2.1-12.5 and 2.4-13.7%, respectively. The pharmacokinetics of amiloride was evaluated after a single oral administration of 10 mg to healthy volunteers. The AUC0-72hr, Cmax, Tmax, and T1/2 were 267.7 ± 60.0 ng·hr/mL, 22.9 ± 5.4 ng/mL, 3.0 ± 0.8 hr, and 13.6 ± 2.4 hr, respectively. These results demonstrated that this method was highly feasible and reproducible for pharmacokinetic studies of amiloride in eight volunteers after oral adminis-tration (10 mg as amiloride HCl). [ABSTRACT FROM AUTHOR]

The amyloid beta (Aβ) peptide is known to increase free radical production in nerve cells, leading to cell death. To investigate the effect of Rosa laevigata against Aβ-induced oxidative damage, in vitro assays and in vivo behavioral tests were performed. R. laevigata showed cell protective effects against oxidative stress-induced cytotoxicity. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) reduction assay exhibited significant increase in cell viability when rat pheochromocytoma (PC 12) cells were treated with R. laevigata extracts. Administration of R. laevigata extracts to mice significantly reversed the Aβ-induced learning and memory impairment in in vivo behavioral tests. These results suggest that R. laevigata extracts can reduce the cytotoxicity of Aβ in PC 12 cells, possibly by the reduction of oxidative stress, and these extracts may be useful in the prevention of Alzheimer's disease. [ABSTRACT FROM AUTHOR]