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R-L Luan, P-C Wang, M-X Yan, J Chen
OBJECTIVE: Multidrug resistance and toxicity significantly compromise the therapeutic efficacy for sarcomas. We aimed at evaluating the effect of lutein-doxorubicin (DOX) combinatorial therapy on inhibiting S180 (Sarcoma 180) cell proliferation and tumor growth. MATERIALS AND METHODS: S180 cells in logarithmic growth phase were treated with lutein, DOX, or lutein-DOX combinatorial therapy for 48 h. The cell survival rate was determined by MTT assay. Apoptosis was detected by flow cytometry...
March 2018: European Review for Medical and Pharmacological Sciences
Yipeng Wang, Liping Guo, Lin Feng, Wen Zhang, Ting Xiao, Xuebing Di, Guoji Chen, Kaitai Zhang
Circulating tumour cell (CTC) behaviours are distinct from those of bulk tissues. Thus, treatments to eliminate CTCs differ from the regimens followed to reduce the primary tumour and its metastases. Accordingly, comprehensively deciphering the single nucleotide variant (SNV) profiles in CTCs, which partially determine CTC behaviours, is a priority. Using viable CTCs isolated with the oHSV1‑hTERT‑GFP virus coupled with fluorescence‑activated cell sorting (FACS), the whole genome was amplified using the multiple annealing and looping‑based amplification cycle (MALBAC) method...
May 2018: Oncology Reports
Octavia Ramayanti, Mitch Brinkkemper, Sandra A W M Verkuijlen, Leni Ritmaleni, Mei Lin Go, Jaap M Middeldorp
Epstein-Barr virus (EBV) persists in nasopharyngeal (NPC) and gastric carcinomas (EBVaGC) in a tightly latent form. Cytolytic virus activation (CLVA) therapy employs gemcitabine and valproic acid (GCb+VPA) to reactivate latent EBV into the lytic phase and antiviral valganciclovir to enhance cell death and prevent virus production. CLVA treatment has proven safe in phase-I/II trials with promising clinical responses in patients with recurrent NPC. However, a major challenge is to maximize EBV lytic reactivation by CLVA...
March 22, 2018: Cancers
Pei-Tseng Lee, Jonathan Zirin, Oguz Kanca, Wen-Wen Lin, Karen L Schulze, David Li-Kroeger, Rong Tao, Colby Devereaux, Yanhui Hu, Verena Chung, Ying Fang, Yuchun He, Hongling Pan, Ming Ge, Zhongyuan Zuo, Benjamin E Housden, Stephanie E Mohr, Shinya Yamamoto, Robert W Levis, Allan C Spradling, Norbert Perrimon, Hugo J Bellen
We generated a library of ~1,000 Drosophila stocks in which we inserted a construct in the intron of genes allowing expression of GAL4 under control of endogenous promoters while arresting transcription with a polyadenylation signal 3' of the GAL4. This allows numerous applications. First, ~90% of insertions in essential genes cause a severe loss-of-function phenotype, an effective way to mutagenize genes. Interestingly, 12/14 chromosomes engineered through CRISPR do not carry second-site lethal mutations. Second, 26/36(70%) of lethal insertions tested are rescued with a single UAS- cDNA construct...
March 22, 2018: ELife
Elena Guerini-Rocco, Antonio Passaro, Chiara Casadio, Valeria Midolo De Luca, Juliana Guarize, Filippo de Marinis, Davide Vacirca, Massimo Barberis
CONTEXT: - Molecular testing is essential for the diagnostic workup of patients with advanced non-small cell lung cancers. Cytology specimens from minimally invasive procedures, such as endobronchial ultrasound-guided transbronchial needle aspiration, are often the only available samples for these patients. The implementation of molecular diagnostic testing, and in particular next-generation sequencing-based testing, on these cytologic specimens is currently an evolving field for lung cytopathology...
April 2018: Archives of Pathology & Laboratory Medicine
Daniel Bonhenry, François Dehez, Mounir Tarek
The low bioavailability of most therapeutic compounds is often counterbalanced by association with molecular vectors capable of crossing cell membranes. Previous studies demonstrated that for vectors bearing titratable chemical groups, the translocation process might be accompanied by a change in the protonation state. For simple compounds e.g. a lysine analog, free energy calculations, using a single collective variable, namely the insertion depth, suggest that such a transition could only take place if the amino acid diffuses deep enough into the hydrophobic core of the membrane, a situation thermodynamically unfavorable...
March 22, 2018: Physical Chemistry Chemical Physics: PCCP
Jeannine Steinmeyer, Heidi-Kristin Walter, Mathilde A Bichelberger, Violetta Schneider, Tomáš Kubař, Franziska Rönicke, Bettina Olshausen, Karin Nienhaus, Gerd Ulrich Nienhaus, Ute Schepers, Marcus Elstner, Hans-Achim Wagenknecht
Two fluorescent dyes covalently attached in diagonal interstrand orientation to siRNA undergo energy transfer and thereby enable a dual color fluorescence readout (red/green) for hybridization. Three different structural variations were carried out and compared by their optical properties, including (i) the base surrogate approach with an acyclic linker as a substitute of the 2-deoxyriboside between the phosphodiester bridges, (ii) the 2'-modification of conventional ribofuranosides and (iii) the arabino-configured 2'-modification...
March 22, 2018: Organic & Biomolecular Chemistry
Claire M Ouimet, Mohamed Dawod, James Grinias, Victoria A Assimon, Jean Lodge, Anna K Mapp, Jason E Gestwicki, Robert T Kennedy
Tools for measuring affinities and stoichiometries of protein-protein complexes are valuable for elucidating the role of protein-protein interactions (PPIs) in governing cell functions and screening for PPI modulators. Such measurements can be challenging because PPIs can span a wide range of affinities and include stoichiometries from dimers to high order oligomers. Also, most techniques require large amounts of protein which can hamper research for difficult to obtain proteins. Protein cross-linking capillary electrophoresis (PXCE) has the potential to directly measure PPIs and even resolve multiple PPIs while consuming attomole quantities...
March 22, 2018: Analyst
José Luis Martínez-Rodríguez, Rosalinda Gutiérrez-Hernández, Claudia Araceli Reyes-Estrada, Angelica Judith Granados-López, Tatiana Arcos-Ortega, Jesús Adrián López
The metabolism of aromatic hydrocarbons by the organism forms products that cause cell death depending on the type of exposure. Benzene exposure has been linked to oxidative stress, hepatic damage, aplastic anemia and hematopoietic cancer as lymphoid and myeloid leukemia. However, there are not fast methods to evaluate chronically benzene exposure in human blood. The objective of this work was the evaluation of the correlation between oxidative damage with benzene exposure and the level of Cellular Plasma Membrane Stability (CPMS) in erythrocytes to use it as a future indicator to determine the grade of benzene intoxications...
March 22, 2018: Toxicology Mechanisms and Methods
Matthaeus Schwarz-Schilling, Aurore Dupin, Fabio Chizzolini, Swati Krishnan, Sheref Mansy, Friedrich C Simmel
Molecular complexes composed of RNA molecules and proteins are promising multifunctional nanostructures for a wide variety of applications in biological cells or in artificial cellular systems. In this study, we systematically address some of the challenges associated with the expression and assembly of such hybrid structures using cell-free gene expression systems. As a model structure, we investigated a pRNA-derived RNA scaffold functionalized with four distinct aptamers, three of which bind to proteins - streptavidin and two fluorescent proteins - while one binds the small molecule dye malachite green (MG)...
March 22, 2018: Nano Letters
Cayetana Vázquez-Diez, Greg FitzHarris
Chromothripsis is a phenomenon observed in cancer cells, wherein a single or few chromosome(s) exhibit vast genomic rearrangements. Recent studies elucidated a striking series of events in which defective segregation of chromosomes causes their incorporation into micronuclei, where they are subject to extensive DNA damage prior to re-joining the main mass of chromosomes in a subsequent cell cycle, which provide an appealing mechanism for the etiology of chromothripsis. Micronuclei are well known to be common in human preimplantation embryos...
2018: Methods in Molecular Biology
Veronica Ortega, Christina Mendiola, Gopalrao V N Velagaleti
One of the well-known hallmarks of cancer is genomic instability. Although gradualism is a well-established process of cancer evolution, recent studies have shown that chromothripsis or chromoanasynthesis can result in complex genomic rearrangements by a single catastrophic event rather than several incremental steps. These two novel phenomena suggest an evolutionary modality for cancer cells to circumvent individual mutational events with one simultaneous shattering of chromosomes or chromosome regions resulting in the random reassembling of shattered genetic material to form complex derivative chromosomes...
2018: Methods in Molecular Biology
Ruth N MacKinnon
Fluorescence in situ hybridization (FISH) to metaphase chromosomes, in conjunction with SNP array, array CGH, or whole genome sequencing, can help determine the organization of abnormal genomes after chromothripsis and other types of complex genome rearrangement. DNA microarrays can identify the changes in copy number, but they do not give information on the organization of the abnormal chromosomes, balanced rearrangements, or abnormalities of the centromeres and other regions comprised of highly repetitive DNA...
2018: Methods in Molecular Biology
Chase B Hellmer, Tomomi Ichinose
Retinal first-order neurons, photoreceptors, receive visual inputs and convert them to neural signals. The second-order neurons, bipolar cells then sort out the visual signals and encode them through multiple neural streams. Approximately 15 morphological types of bipolar cells have been identified, which are thought to encode different aspects of visual signals such as motion and color (Ichinose et al. J Neurosci 34(26):8761-8771, 2014; Euler et al. Nat Rev Neurosci 15(8):507-519, 2014). To investigate functional aspects of OFF bipolar cells, single cell recordings are preferred; however, bipolar cells in the mouse retina are small and hard to distinguish from other types of cells...
2018: Methods in Molecular Biology
Naoyuki Tanimoto, Mathias W Seeliger
Electroretinography (ERG) is important for functional diagnostics of the retina. Types of information about retinal function obtainable by ERG differ depending on recording conditions, e.g., a combination of light stimulus and adaptation. In terms of stimulation, single-flash and flicker stimuli are frequently used because response properties have been well investigated, allowing an assessment of fundamental retinal functionality; for example, how photoreceptors and bipolar cells, including signal transmission between them, are affected under pathological conditions...
2018: Methods in Molecular Biology
Clint L Makino, Teresa Duda, Alexandre Pertzev, Rameshwar K Sharma
Our ability to see is based on the activity of retinal rod and cone photoreceptors. Rods function when there is very little light, while cones operate at higher light levels. Photon absorption by rhodopsin activates a biochemical cascade that converts photic energy into a change in the membrane potential of the cell by decreasing the levels of a second messenger, cGMP, that control the gating of cation channels. But just as important as the activation of the cascade are the shut-off and recovery processes. The timing of shutoff and recovery ultimately affects sensitivity, temporal resolution and even the capacity for counting single photons...
2018: Methods in Molecular Biology
Bhubanananda Sahu, Akiko Maeda
The retinal pigmented epithelium (RPE) is a single layer of polarized epithelial cells which plays many important roles for visual function. One of such roles is production of visual chromophore, 11-cis-retinal through the visual cycle. The visual cycle consists of biochemical processes for regenerating chromophore by a collective action of the RPE and photoreceptor. Photoreceptors harbor the G protein-coupled receptors, opsin which enables to receive light when it bounds to 11-cis-retinal. With absorption of a photon of light, 11-cis-retinal photoisomerizes to all-trans-retinal...
2018: Methods in Molecular Biology
Abdalla Ahmed, Paul Delgado-Olguin
Isolation of highly purified populations of embryonic cardiomyocytes enables the study of congenital cardiac phenotypes at the cellular level. Fluorescent-activated cell sorting (FACS) is normally used to isolate fluorescently tagged cells. Here we describe the isolation of differentiating mouse embryonic cardiac progenitors and cardiomyocytes at embryonic day (E) 9.5 and E13.5, respectively by FACS. Over 50,000 differentiating cardiac progenitors and 200,000 cardiomyocytes can be obtained in a single prep using the methods described...
2018: Methods in Molecular Biology
Ruirui Li, Pengfei Gu, Xiangyu Fan, Junyu Shen, Yulian Wu, Lixuan Huang, Qiang Li
A polyhydroxyalkanoate (PHA)-producing strain was isolated from propylene oxide (PO) saponification wastewater activated sludge and was identified as Brevundimonas vesicularis UJN1 through 16S rDNA sequencing and Biolog microbiological identification. Single-factor and response surface methodology experiments were used to optimize the culture medium and conditions. The optimal C/N ratio was 100/1.04, and the optimal carbon and nitrogen sources were sucrose (10 g/L) and NH4 Cl (0.104 g/L) respectively. The optimal culture conditions consisted of initial pH of 6...
March 21, 2018: Applied Biochemistry and Biotechnology
Letícia Pereira Úngari, André Luiz Quagliatto Santos, Lucia Helena O'Dwyer, Maria Regina Lucas da Silva, Natália Nasser de Melo Fava, Guilherme Carrara Moreira Paiva, Rogério de Melo Costa Pinto, Márcia Cristina Cury
Based on morphological, morphometric, and molecular data, we describe a new hemoparasite of the genus Haemogregarina Danilewsky 1885, isolated from the Brazilian aquatic turtle Podocnemis unifilis (Testudines: Podocnemididae). The new species, Haemogregarina podocnemis sp. nov. (Apicomplexa: Haemogregarinidae), is characterized by small trophozoites with a single cytoplasmic vacuole on one side; pre-meronts with nuclear chromatin dispersed in the cytoplasm, with or without cytoplasmic vacuoles; meronts that are usually broad and slightly curved (kidney-shaped), with an average of eight small rectangular nuclei; immature gamonts (bean-shaped) with two morphological types: one with nuclear chromatin dispersed in the cytoplasm and the other with nuclei in the middle of the cell; mature gamonts of two morphological types: one with a length equal to or greater than that of the erythrocyte and the width of the nuclei similar to that of the hemoparasite and the other smaller than the erythrocyte with the width of the nuclei less than that of the hemoparasite...
March 22, 2018: Parasitology Research
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