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Hao-Hsiang Hsu, John-Kevin Kracht, Laura Elisabeth Harder, Kerstin Rudnik, Gerd Lindner, Katharina Schimek, Uwe Marx, Ralf Pörtner
In vitro cultivated skin models have become increasingly relevant for pharmaceutical and cosmetic applications, and are also used in drug development as well as substance testing. These models are mostly cultivated in membrane-insert systems, their permeability toward different substances being an essential factor. Typically, applied methods for determination of these parameters usually require large sample sizes (e.g., Franz diffusion cell) or laborious equipment (e.g., fluorescence recovery after photobleaching (FRAP))...
February 23, 2018: Journal of Visualized Experiments: JoVE
Chenju Yi, Jérémy Theillon, Annette Koulakoff, Hugues Berry, Christian Giaume
Intercellular communication through gap junction channels plays a key role in cellular homeostasis and in synchronizing physiological functions, a feature that is modified in number of pathological situations. In the brain, astrocytes are the cell population that expresses the highest amount of gap junction proteins, named connexins. Several techniques have been used to assess the level of gap junctional communication in astrocytes, but so far they remain very difficult to apply in adult brain tissue. Here, using specific loading of astrocytes with sulforhodamine 101, we adapted the gap-FRAP (Fluorescence Recovery After Photobleaching) to acute hippocampal slices from 9 month-old adult mice...
March 15, 2018: Journal of Neuroscience Methods
Mariana Maciel de Oliveira, Regina Gomes Daré, Érica Oliveira Barizão, Jesuí Vergílio Visentainer, Mariza Barion Romagnolo, Celso Vataru Nakamura, Maria da Conceição Torrado Truiti
The total phenolic content (TP) and antioxidant activity of the ethanolic extract and fractions that were obtained from the leaves of Nectandra hihua were assessed using different methods. The ethanolic extract (EE) and ethyl acetate fraction (EAF) had the best antioxidant capacity, which was comparable to butylated hydroxytoluene and quercetin (ABTS+ 2.55 ± 0.06, 3.54 ± 0.03 mmol TE/g; DPPH IC50 10.27 ± 0.05, 9.88 ± 0.02 μg/mL; FRAP 2.17 ± 0.08, 2.38 ± 0.04 mmol TE/g; ORAC 5...
March 9, 2018: Journal of Photochemistry and Photobiology. B, Biology
Yara Rafaella Ribeiro Santos Rezende, Juliete Pedreira Nogueira, Narendra Narain
The acerola pulp and residue extracts were microencapsulated by spray and freeze-drying, using gum arabic and maltodextrin as encapsulating agents. Total anthocyanins, carotenoids (CA), ascorbic acid (AA), phenolic compounds (PC), total flavonoids, antioxidant activity, color, moisture, water activity, solubility, hygroscopicity and microstructure of the powders were analyzed. The acerola residue extract had higher concentrations of bioactive compounds (except for AA) and lower antioxidant activity. The microencapsulated powders by spray and freeze-drying showed good physico-chemical properties...
July 15, 2018: Food Chemistry
Rong Liang, Sheng Cheng, Xiuying Wang
We used a pulsed electric field (PEF) to treat four pentapeptides with similar amino acid sequences (KCHKP, KCHQP, QCHKP, and QCHQP). We then evaluated antioxidant activity of the pentapeptides according to ferric reducing antioxidant power (FRAP) and hydroxyl radical scavenging capacity. Structures of the peptides were determined by high performance liquid chromatography (HPLC), mid-infrared (MIR), circular dichroism (CD) spectroscopy, and nuclear magnetic resonance (NMR). The results indicated that antioxidant activities of KCHQP and QCHKP were increased by PEF, whereas those of KCHKP and QCHQP were reduced...
July 15, 2018: Food Chemistry
Siyabulela S B N Ntuli, Wentzel C A Gelderblom, David R Katerere
BACKGROUND: Sutherlandia frutescens (L.) R. Br is endemic to Southern Africa where it has been traditionally used for cancer and diabetes. In recent times it has been marketed for its reputed (but not proven) anticancer, antidiabetic and anti-HIV properties. Little is known about the mutagenic and antimutagenic potential of extracts and common marker compounds of Sutherlandia frutescens. Therefore this study aimed to investigate the putative efficacy and possible long-term adverse effects of using this herb...
March 15, 2018: BMC Complementary and Alternative Medicine
Samia Hadj Ahmed, Wafa Kharroubi, Nadia Kaoubaa, Amira Zarrouk, Fathi Batbout, Habib Gamra, Mohamed Fadhel Najjar, Gérard Lizard, Isabelle Hininger-Favier, Mohamed Hammami
BACKGROUND: Nutritional choices, which include the source of dietary fatty acids (FA), have an important significant impact on coronary artery disease (CAD). We aimed to determine on patients with CAD the relationships between Trans fatty acids (Trans FA) and different CAD associated parameters such as inflammatory and oxidative stress parameters in addition to Gensini score as a vascular severity index. METHODS: Fatty acid profiles were established by gas chromatography from 111 CAD patients compared to 120 age-matched control group...
March 15, 2018: Lipids in Health and Disease
Anna Baldisserotto, Piergiacomo Buso, Matteo Radice, Valeria Dissette, Ilaria Lampronti, Roberto Gambari, Stefano Manfredini, Silvia Vertuani
Moringa oleifera has gained increasing popularity as a food supplement but not in the pharmaceutical and cosmetic area. The aim of this study was the preparation, characterization, and evaluation of extracts from the leaves of Moringa oleifera as a herbal sun care phytocomplex. Three different extracts of Moringa oleifera leaves, from Senegal, have been prepared and chemically characterized in the phenolic fraction by HPLC-DAD and Folin-Ciocalteu test. To explore photoprotective properties, an extensive evaluation of UV filtering, antioxidant (DPPH, FRAP, ORAC, PCL), and anti-hyperproliferative (human melanoma Colo38 cells) capacities have been conducted...
March 15, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
Ren-You Gan, Kin-Weng Kong, Hua-Bin Li, Kao Wu, Ying-Ying Ge, Chak-Lun Chan, Xian-Ming Shi, Harold Corke
The red sword bean ( Canavalia gladiata ) is an underutilized edible bean cultivated in China. It was previously found to have the highest content of antioxidant polyphenols among 42 edible beans, mainly gallic acid, and gallotannins in its red bean coat, an apparently unique characteristic among edible beans. In this study, the main phenolic compounds in red sword bean coats were further separated by Sephadex LH-20 column chromatography, and identified by LC-MS/MS. Furthermore, the FRAP and ABTS antioxidant activities and antibacterial activity (diameter of inhibition zone, DIZ) of main gallotannin-rich fractions were tested...
2018: Frontiers in Chemistry
James L Kingsley, Jeffrey P Bibeau, S Iman Mousavi, Cem Unsal, Zhilu Chen, Xinming Huang, Luis Vidali, Erkan Tüzel
Fluorescence recovery after photobleaching (FRAP) is an important tool used by cell biologists to study the diffusion and binding kinetics of vesicles, proteins, and other molecules in the cytoplasm, nucleus, or cell membrane. Although many FRAP models have been developed over the past decades, the influence of the complex boundaries of 3D cellular geometries on the recovery curves, in conjunction with regions of interest and optical effects (imaging, photobleaching, photoswitching, and scanning), has not been well studied...
March 13, 2018: Biophysical Journal
Agnete B Madsen, Jonas R Knudsen, Carlos Henriquez-Olguin, Yeliz Angin, Kristien J Zaal, Lykke Sylow, Peter Schjerling, Evelyn Ralston, Thomas E Jensen
Studies in skeletal muscle cell cultures suggest that the cortical actin cytoskeleton is a major requirement for insulin-stimulated glucose transport, implicating the β-actin isoform which, in many cell types, is the main actin isoform. However, it is not clear that β-actin plays such a role in mature skeletal muscle. Neither dependency of glucose transport on β-actin nor actin reorganization upon glucose transport have been tested in mature muscle. To investigate the role of β-actin in fully differentiated muscle, we have performed a detailed characterization of wildtype and muscle-specific β-actin knockout (KO) mice...
March 13, 2018: American Journal of Physiology. Endocrinology and Metabolism
Cynthia P Hsu, Behzad Moghadaszadeh, John H Hartwig, Alan H Beggs
The α-actinin proteins are a highly conserved family of actin crosslinkers that mediate interactions between several cytoskeletal and sarcomeric proteins. Non-sarcomeric α-actinin-1 and α-actinin-4 crosslink actin filaments in the cytoskeleton, while sarcomeric α-actinin-2 and α-actinin-3 serve a crucial role in anchoring actin filaments to the muscle Z-line. To assess the difference in turnover dynamics and structure/function properties between the α-actinin isoforms at the sarcomeric Z-line, we used Fluorescence Recovery After Photobleaching (FRAP) in primary myofiber cultures...
March 8, 2018: Cytoskeleton
Maria F Quintero-Soto, Ana G Saracho-Peña, Jeanett Chavez-Ontiveros, Jose A Garzon-Tiznado, Karen V Pineda-Hidalgo, Francisco Delgado-Vargas, Jose A Lopez-Valenzuela
Chickpea (Cicer arietinum L.) genotypes, nine kabuli from Mexico and 9 desi from other countries, were investigated for their phenolic profiles and antioxidant activity (AA). Phenolics in methanol extracts (ME) were analyzed by ultra-performance liquid chromatography coupled to diode array detection and mass spectrometry (UPLC-DAD-MS), whereas the AA was measured as Trolox equivalents (TE) by ABTS, DPPH and FRAP methods. Twenty phenolic compounds were identified in the ME and their levels showed a great variability among the chickpea genotypes...
March 7, 2018: Plant Foods for Human Nutrition
Małgorzata Dżugan, Monika Wesołowska, Grzegorz Zaguła, Czesław Puchalski
BACKGROUND: The pomegranate (Punica granatum L.) has gained a reputation as a fruit with many health- promoting properties. It is considered to help prevent or treat various disease risk factors including high blood pressure, high cholesterol, oxidative stress, hyperglycemia and inflammatory activities. It has been demonstrated that certain components of pomegranates, such as polyphenols, have potential antioxidant, anti-inflammatory and anticarcinogenic effects. METHODS: Five commercially available (CA) and three homemade (HM) pomegranate (Puni- ca granatum L...
January 2018: Acta Scientiarum Polonorum. Technologia Alimentaria
Xuegui Liu, Yixing Gao, Danqi Li, Changfeng Liu, Mei Jin, Jun Bian, Mengchao Lv, Yuqiu Sun, Lixin Zhang, Pinyi Gao
Rosa laevigata fruit has been known as a functional foodstuff for a long time. Recently, increasing attention has been given to polysaccharides from R. laevigata fruit due to their numerous medicinal and nutritional properties. In this study, a rapid and effective approach for the extraction and separation of polysaccharides from the title fruit was developed using microwave-assisted aqueous two-phase extraction (MA-ATPE) with a PEG/ammonium sulfate system. After analysis of the response surface methodology (RSM) data based on a Box-Behnken design (BBD), a model was proposed and was found to predict an optimum yield value of 258...
March 7, 2018: Food & Function
Balaje Vijayaraghavan, Ricardo A Figueroa, Cecilia Bergqvist, Amit J Gupta, Paulo Sousa, Einar Hallberg
Samp1, spindle associated membrane protein 1, is a type II integral membrane protein localized in the inner nuclear membrane. Recent studies have shown that the inner nuclear membrane protein, Emerin and the small monomeric GTPase, Ran are direct binding partners of Samp1. Here we addressed the question whether Ran could regulate the interaction between Samp1 and Emerin in the inner nuclear membrane. To investigate the interaction between Samp1 and Emerin in live cells, we performed FRAP experiments in cells overexpressing YFP-Emerin...
March 3, 2018: Biochimica et Biophysica Acta
Mingliang Zhang, Zhen Zhang, Kangmin He, Jimin Wu, Nan Li, Rong Zhao, Jinghe Yuan, Han Xiao, Youyi Zhang, Xiaohong Fang
The dynamics and stoichiometry of receptors newly delivered on plasma membrane play a vital role in cell signal transduction, yet the knowledge on this process is limited due to the lack of suitable methods. Here we developed a new strategy, which combines single-molecule imaging (SMI) and fluorescence recovery after photobleaching (FRAP), named FRAP-SMI, to monitor and quantify the newly delivered/inserted single transmembrane receptor on the plasma membrane of living cells. Transforming growth factor-β type II receptor (TβRII), a typical serine/threonine kinase receptor, was studied with this method...
March 6, 2018: Analytical Chemistry
Li-Ching Lin, Chung-Yi Chen, Chia-Hung Kuo, Yun-Sheng Lin, Byeong Hee Hwang, Tina Kaiting Wang, Yueh-Hsiung Kuo, Hui-Min David Wang
N -Hydroxycinnamoylphenalkylamides (36H) exhibited both antioxidation and antityrosinase abilities. The compound was studied for its antioxidative properties, using a 1,1-diphenyl-2-picrylhydrazul- (DPPH-) scavenging test, a ferric ion-reducing antioxidant power assay (FRAP) assessment, and a metal-chelating power assay. The results showed that 36H had antioxidative capabilities in the DPPH-scavenging and ferric-reducing power examinations but the chelating power assay did not demonstrate antioxidative capability...
2018: Oxidative Medicine and Cellular Longevity
Lucas Olivo Martins Pereira, Wagner Vilegas, Marcelo Marucci Pereira Tangerina, Karuppusamy Arunachalam, Sikiru Olaitan Balogun, Paulo Eduardo Orlandi Matos, Domingos Tabajara de Oliveira Martins
ETHNOPHARMACOLOGICAL IMPORTANCE: Lafoensia pacari A. St.-Hil., (Lythraceae) is a native tree of Brazilian Cerrado and commonly known in Brazil as "mangava-brava". Its leaves are used in Brazilian folk medicine in wound healing, cutaneous mycoses, and in the treatment of gastritis and ulcers. AIM OF THE STUDY: The present study was designed to evaluate the wound healing activity and mechanism of action of the hydroethanolic extract of Lafoensia pacari A...
February 28, 2018: Journal of Ethnopharmacology
A Fatih Aydın, Gamze Kondakçı, Songül Hatipoğlu, Semra Doğru-Abbasoğlu, Müjdat Uysal
High levels of homocysteine (Hcy) have neurotoxic effects. Homocysteine thiolactone (HcyT), a thioester of Hcy, plays a role in Hcy-induced toxicity. In this study, effects of HcyT treatment (500 mg/kg body weight/day in drinking water) for 6 weeks on serum Hcy levels and brain prooxidant-antioxidant balance were investigated in rats. The effect of N-acetylcysteine (NAC) treatment (1 g/kg body weight/day in chow) for 6 weeks on HcyT-induced neurotoxicity was evaluated. Reactive oxygen species (ROS), thiobarbituric acid reactive substances (TBARS), diene conjugate (DC), protein carbonyl (PC) and advanced oxidation protein products (AOPP) were determined in the brain tissue...
February 21, 2018: Pathophysiology: the Official Journal of the International Society for Pathophysiology
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