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Daniel Berner, Matthias Zenkel, Francesca Pasutto, Ursula Hoja, Panah Liravi, Gabriele C Gusek-Schneider, Friedrich E Kruse, Johannes Schödel, Andre Reis, Ursula Schlötzer-Schrehardt
Purpose: Alternative mRNA splicing coupled to nonsense-mediated decay (NMD) is a common mRNA surveillance pathway also known to dynamically modulate gene expression in response to cellular stress. Here, we investigated the involvement of this pathway in the regulation of lysyl oxidase-like 1 (LOXL1) expression in response to pseudoexfoliation (PEX)-associated pathophysiologic factors. Methods: Transcript levels of LOXL1 isoforms were determined in ocular tissues obtained from donor eyes without and with PEX syndrome...
November 1, 2017: Investigative Ophthalmology & Visual Science
Meryem Bouchemi, Christophe Roubeix, Karima Kessal, Luisa Riancho, Anne-Laure Raveu, Hayet Soualmia, Christophe Baudouin, Françoise Brignole-Baudouin
PURPOSE: To validate a new culture model of primary human trabecular meshwork cells (p-hTMCs) using Matrigel®, in order to mimic in vitro 3D-TM organization, and to investigate the proinflammatory effect of benzalkonium chloride (BAK) in 3D p-hTMC cultures. METHODS: p-hTMCs, seeded onto Matrigel®-coated inserts were stimulated with BAK (10(-4)%), dexamethasone (DEX) (10(-6)M) or transforming growth factor-beta 2 (TGF-β2) (5ng/ml) for 48h and observed with confocal microscopy...
February 16, 2017: Toxicology in Vitro: An International Journal Published in Association with BIBRA
Wan Wendy Wang, Kelsey A Watson, S Jeffrey Dixon, Hong Liu, Amin S Rizkalla, Cindy Ml Hutnik
BACKGROUND: Microinvasive glaucoma surgery (MIGS) is a relatively new addition to the glaucoma treatment paradigm. Small metallic stents are inserted into the trabecular meshwork in order to increase aqueous humour drainage. MIGS procedures are rapidly being adopted owing to a more favourable side effect profile when compared with traditional surgery. Remarkably, this rapid rate of utilization has occurred without any published studies on the effect of metal alloys used in these stents on human trabecular meshwork cells (HTMCs)...
July 2017: Clinical & Experimental Ophthalmology
Josiane Semaan, Aline Pinon, Benjamin Rioux, Lama Hassan, Youness Limami, Christelle Pouget, Catherine Fagnère, Vincent Sol, Mona Diab-Assaf, Alain Simon, Bertrand Liagre
Increasing incidence and mortality of colorectal cancer brings the necessity to uncover new possibilities in its prevention and treatment. Chalcones have been identified as interesting compounds having chemopreventive and antitumor properties. In this study, we investigated the effects of the synthetic chalcone derivative 3-hydroxy-3',4,4',5'-tetra-methoxy-chalcone (3-HTMC) on proliferation, cell cycle distribution, apoptosis, and its mechanism of action in human colorectal HT-29 (COX-2 sufficient) and HCT116 (COX-2 deficient) cancer cells...
December 2016: Journal of Cellular Biochemistry
Xiao-Ling Luo, Si-Yu Liu, Li-Jun Wang, Qiu-Yan Zhang, Peng Xu, Li-Long Pan, Jin-Feng Hu
Neuroinflammation underlies the pathogenesis and progression of neurodegenerative diseases. 2׳-hydroxy-4,3׳,4׳,6׳-tetramethoxychalcone (HTMC) is a known chalcone derivative isolated from Chloranthus henryi with anti-inflammatory activities in BV2 macrophages. However, its pharmacological effects on microglial cells have not been demonstrated. To this end, we examined the effects of HTMC on lipopolysaccharide (LPS)-induced inflammatory responses in BV2 microglial cells. HTMC concentration-dependently inhibited LPS-induced expression of inflammatory enzymes including inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), nitric oxide (NO) production, and the secretion of inflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6...
March 5, 2016: European Journal of Pharmacology
Megan E Cavet, Thomas R Vollmer, Karen L Harrington, Karl VanDerMeid, Mary E Richardson
PURPOSE: Previous in vivo studies demonstrated that latanoprostene bunod (LBN), a nitric oxide (NO)-donating prostaglandin F2α receptor agonist, results in greater intraocular pressure (IOP) lowering than latanoprost. The present series of investigations compared the effects of LBN and latanoprost on primary human trabecular meshwork cell (HTMC) contractility and underlying signaling pathways to determine whether LBN might mediate this additional IOP lowering via the conventional outflow pathway...
June 2015: Investigative Ophthalmology & Visual Science
Caitlin Chang, Angela Q Zhang, Dov B Kagan, Hong Liu, Cindy M L Hutnik
BACKGROUND: Benzalkonium chloride (BAK) is a controversial ophthalmic preservative because of its prominent side-effect profile. In this study, we examined the mechanism of BAK toxicity in human trabecular meshwork cells (HTMC) and compared the effects of BAK with tafluprost free acid, which is an active form of tafluprost commercially available in a preservative-free formulation. METHODS: Primary HTMC were treated with different BAK concentrations over various exposure times...
March 2015: Clinical & Experimental Ophthalmology
Tim C Lei, Omid Masihzadeh, Malik Y Kahook, David A Ammar
PURPOSE: The aim of this study was to nondestructively monitor morphological changes to the lipid membranes of primary cultures of living human trabecular meshwork cells (hTMC) without the application of exogenous label. METHODS: Live hTMC were imaged using two nonlinear optical techniques: coherent anti-Stokes Raman scattering (CARS) and two-photon autofluorescence (TPAF). The hTMC were treated with a commercial formulation of latanoprost (0.5 μg/mL) for 24 hours before imaging...
September 2013: Investigative Ophthalmology & Visual Science
David A Ammar, Malik Y Kahook
PURPOSE: Photodynamic therapy (PDT) laser light in conjunction with the benzoporphyrin derivative verteporfin is a current clinical treatment for choroidal vascular diseases such as age-related macular degeneration. The aim of this study was to examine the effects of PDT laser-activated and inactive verteporfin on various cultured ocular cells. METHODS: Primary human scleral fibroblasts (hFibro), primary human trabecular meshwork (TM) cells (hTMC), primary porcine TM cells (pTMC), and a human retinal pigment epithelial cell line (ARPE-19 cells) were treated with verteporfin with and without activation by PDT laser...
2013: Molecular Vision
R B Barbieri, N E Bufalo, L L Cunha, L V M Assumpção, R M B Maciel, J M Cerutti, L S Ward
CONTEXT: Different inherited profiles of genes involved in cellular mechanisms of activation and detoxification of carcinogenic products can provide specific protection or determine the risk for cancer. Low-penetrance polymorphic genes related to the biotransformation of environmental toxins have been associated with susceptibility to and the phenotype of, human tumours. OBJECTIVE: To investigate the role of germline inheritance of polymorphisms in CYP1A2*F, CYP1A1 m1, GSTP1, NAT2 and TP53 genes in hereditary medullary thyroid carcinoma (HMTC) patients...
August 2013: Clinical Endocrinology
David A Ammar, Naresh Mandava, Malik Y Kahook
PURPOSE: To investigate the effects of the vascular endothelial growth factor-neutralizing agent aflibercept on primary cultures of human trabecular meshwork cells (hTMC), human scleral fibroblasts (hFibro), and a retinal pigment epithelial cell line (ARPE-19). METHODS: Various concentrations of aflibercept were incubated with confluent cell cultures for 24 hours. Ranibizumab was used as an active control for comparison. Assays of cellular metabolism (MTT assay) and cell viability (calcein dye uptake) were performed...
May 2013: Retina
John Cuppoletti, Danuta H Malinowska, Kirti P Tewari, Jayati Chakrabarti, Ryuji Ueno
PURPOSE: Effects of cis-unoprostone isopropyl, its primary metabolite M1, trans-unoprostone isopropyl, latanoprost free acid, and fluprostenol were studied on Ca(2+)-activated K(+) (BK) channels, plasma membrane potential, [cAMP](i), [cGMP](i), and steady state [Ca(2+)](i), and protection against endothelin-1 (ET-1)-induced steady state [Ca(2+)](i) increases in human cortical neuronal (HCN-1A), trabecular meshwork (HTMC), and pulmonary artery smooth muscle (PASMC) cells. Effects on recombinant human prostaglandin (PG) receptors were determined...
August 7, 2012: Investigative Ophthalmology & Visual Science
Cheul Ho Park, Jae Woo Kim
PURPOSE: To investigate the effect of advanced glycation end products (AGE) on oxidative stress and cellular senescence in cultured human trabecular meshwork cells (HTMC). METHODS: Primarily cultured HTMC were exposed to 0, 10, 50, 100, 200 µg/mL of glycated bovine serum albumin (G-BSA) for 5 days. Also co-exposed were L-arginine, sepiapterin, and antioxidant N-acetylcysteine (NAC). Cellular survival and production of nitric oxide (NO), superoxide, and reactive oxygen species were assessed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide assay, Griess assay, cytochrome c assay, and dichlorofluorescin diacetate assay, respectively...
April 2012: Korean Journal of Ophthalmology: KJO
M Foresta, G Frosina, S C Sacca, C Cartiglia, M Longobardi, A Izzotti
Oxidative damage plays a pathogenic role in various chronic degenerative diseases. Oxidative damage targeting trabecular meshwork (TM) cells as a consequence of mitochondrial damage is a pathogenic mechanism for glaucoma, the most common cause of irreversible blindness worldwide. Consequences of oxidative damage are attenuated by endocellular activities involved in scavenging reactive oxidative species and DNA repair. Selected bacterial genes are highly efficient at protecting cells from oxidative DNA damage...
July 2011: Free Radical Research
Yerra Koteswara Rao, Te-Yu Kao, Jiunn-Liang Ko, Yew-Min Tzeng
The present Letter identified 2'-hydroxy-2,3,4',6'-tetramethoxychalcone (HTMC) as a potent in vitro cytotoxic agent with selective activity against cell lines derived from human lung cancer. In A549 lung adenocarcinoma cells, HTMC caused G1 phase cell-cycle arrest. HTMC treatment also led to an inhibition of cell-cycle regulatory proteins phosphorylation of cdc2 (Tyr(15) and Tyr(161)) and Rb (Ser(795) and Ser(807/811)), which was accompanied by the accumulation of tumor suppressor genes p53 and p21. In addition, in vivo data demonstrated that HTMC act as a tumor growth suppressing agent...
November 15, 2010: Bioorganic & Medicinal Chemistry Letters
Yerra Koteswara Rao, Te-Yu Kao, Ming-Fang Wu, Jiunn-Liang Ko, Yew-Min Tzeng
High telomerase activity (TA) is detected in most cancer cells; and thus, TA inhibition by drug or dietary food components is a new strategy for cancer prevention. In this report, we examined the effects of fourteen natural or synthetic compounds on TA in human lung adenocarcinoma A549 cells. The results demonstrated that some of the tested compounds inhibited TA, being 2'-hydroxy-2,3,4',6'-tetramethoxychalcone (HTMC) was the most potent among tested. In A549 cells, HTMC also inhibited the cell proliferation, decreased the expression of human telomerase reverse transcriptase (hTERT) and sequentially reduced the hTERT promoter...
October 1, 2010: Bioorganic & Medicinal Chemistry
Reza Ghassemifar, Sharon Redmond, Zainuddin, Traian V Chirila
Human tympanic membrane cells (hTMCs), harvested from tympanic membrane (TM) explants, were grown in culture and then seeded on membranes prepared from silkworm (Bombyx mori) silk fibroin (BMSF) and on tissue-culture plastic membranes (PET). Fibroin was isolated from silk cast into membranes with a thickness of 10-15 microm. The hTMCs were cultured on both materials for 15 days in a serum-containing culture medium. The cells grown on both substrata were subjected to nuclear staining (DAPI) and counted. Further, the cultures were immunostained for a number of protein markers related to the epithelial/keratinocyte phenotype and cell adhesion complexes...
March 2010: Journal of Biomaterials Applications
Richard Bartlome, Julien M Rey, Markus W Sigrist
Numerous gas-sensing devices are based on infrared laser spectroscopy. In this paper, the technique is further developed and, for the first time, applied to forensic urinalysis. For this purpose, a difference frequency generation laser was coupled to an in-house-built, high-temperature multipass cell (HTMC). The continuous tuning range of the laser was extended to 329 cm(-1) in the fingerprint C-H stretching region between 3 and 4 microm. The HTMC is a long-path absorption cell designed to withstand organic samples in the vapor phase (Bartlome, R...
July 15, 2008: Analytical Chemistry
Y Gu, H Lee, J R Kirchhoff, L Manzey, R A Hudson
The synthesis of 4- and 5-hydroxy-3-[(trimethylammonio)methyl]catechol (4- and 5-HTMC) was carried out to examine their proposed involvement as intermediates in the spontaneous redox-dependent half-of-sites inactivation of neurotoxin binding sites in the nicotinic acetylcholine receptor (nAcChR) mediated by the parent compound 3-[(trimethylammonio)methyl]catechol (TMC) [Nickoloff et al. (1985) Biochemistry 24, 999-1007]. Oxidation of 4- and 5-HTMC occurred with sodium periodate with facile conversion to the corresponding p-quinones which were intercepted with thiols and cyclopentadiene...
July 19, 1994: Biochemistry
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