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https://www.readbyqxmd.com/read/29352609/induction-of-mitophagy-in-the-hei-oc1-auditory-cell-line-and-activation-of-the-atg12-lc3-pathway-in-the-organ-of-corti
#1
Cristian Setz, Anne-Sophie Benischke, Anna Catharina Pinho Ferreira Bento, Yves Brand, Soledad Levano, Franziska Paech, Katharina Leitmeyer, Daniel Bodmer
Autophagy is a highly evolutionary conserved quality control defense mechanism within cells, which has also been implicated in cell death processes. In the mammalian inner ear, autophagy has been shown to play a role during early morphogenesis as well as in adult cochlear hair cells exposed to ototoxic insults. Mitophagy, a selective autophagic cell process targeting mitochondria, hasn't been studied in the inner ear so far. On this work, we searched for molecular indicators of mitophagy within House Ear Institute-Organ of Corti-1 (HEI-OC1) cells as well as in the organ of Corti (OC)...
January 11, 2018: Hearing Research
https://www.readbyqxmd.com/read/29352284/sensitive-detection-of-fluorescence-in-western-blotting-by-merging-images
#2
Yukari Kondo, Shinichiro Higa, Takeshi Iwasaki, Tomoya Matsumoto, Kazumitsu Maehara, Akihito Harada, Yoshihiro Baba, Masatoshi Fujita, Yasuyuki Ohkawa
The western blotting technique is widely used to analyze protein expression levels and protein molecular weight. The chemiluminescence method is mainly used for detection due to its high sensitivity and ease of manipulation, but it is unsuitable for detailed analyses because it cannot be used to detect multiple proteins simultaneously. Recently, more attention has been paid to the fluorescence detection method because it is more quantitative and is suitable for the detection of multiple proteins simultaneously...
2018: PloS One
https://www.readbyqxmd.com/read/29352283/using-a-model-comparison-approach-to-describe-the-assembly-pathway-for-histone-h1
#3
Carlos Contreras, Minaya Villasana, Michael J Hendzel, Gustavo Carrero
Histones H1 or linker histones are highly dynamic proteins that diffuse throughout the cell nucleus and associate with chromatin (DNA and associated proteins). This binding interaction of histone H1 with the chromatin is thought to regulate chromatin organization and DNA accessibility to transcription factors and has been proven to involve a kinetic process characterized by a population that associates weakly with chromatin and rapidly dissociates and another population that resides at a binding site for up to several minutes before dissociating...
2018: PloS One
https://www.readbyqxmd.com/read/29352262/microbial-regulation-of-the-l-cell-transcriptome
#4
Tulika Arora, Rozita Akrami, Ramona Pais, Linda Bergqvist, Bengt R Johansson, Thue W Schwartz, Frank Reimann, Fiona M Gribble, Fredrik Bäckhed
L cells are an important class of enteroendocrine cells secreting hormones such as glucagon like peptide-1 and peptide YY that have several metabolic and physiological effects. The gut is home to trillions of bacteria affecting host physiology, but there has been limited understanding about how the microbiota affects gene expression in L cells. Thus, we rederived the reporter mouse strain, GLU-Venus expressing yellow fluorescent protein under the control of the proglucagon gene, as germ-free (GF). Lpos cells from ileum and colon of GF and conventionally raised (CONV-R) GLU-Venus mice were isolated and subjected to transcriptomic profiling...
January 19, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29352213/cucumber-mosaic-virus-coat-protein-induces-the-development-of-chlorotic-symptoms-through-interacting-with-the-chloroplast-ferredoxin-i-protein
#5
Yanhong Qiu, Yongjiang Zhang, Chaonan Wang, Rong Lei, Yupin Wu, Xinshi Li, Shuifang Zhu
Cucumber mosaic virus (CMV) infection could induce mosaic symptoms on a wide-range of host plants. However, there is still limited information regarding the molecular mechanism underlying the development of the symptoms. In this study, the coat protein (CP) was confirmed as the symptom determinant by exchanging the CP between a chlorosis inducing CMV-M strain and a green-mosaic inducing CMV-Q strain. A yeast two-hybrid analysis and bimolecular fluorescence complementation revealed that the chloroplast ferredoxin I (Fd I) protein interacted with the CP of CMV-M both in vitro and in vivo, but not with the CP of CMV-Q...
January 19, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29352081/expression-of-the-pseudomonas-aeruginosa-biofilm-antibiotic-resistance-gene-ndvb-requires-the-rpos-stationary-phase-sigma-factor
#6
Clayton W Hall, Aaron J Hinz, Luke B-P Gagnon, Li Zhang, Jean-Paul Nadeau, Sarah Copeland, Bratati Saha, Thien-Fah Mah
Chronic, biofilm-based bacterial infections are exceptionally difficult to eradicate due to the high degree of antibiotic recalcitrance exhibited by cells in biofilm communities. In the opportunistic pathogen Pseudomonas aeruginosa, biofilm recalcitrance is multifactorial and arises in part from the preferential expression of resistance genes in biofilms compared to exponential phase planktonic cells. One such mechanism involves ndvB, which we have previously shown to be expressed specifically in biofilms. In this study, we investigated the regulatory basis of this lifestyle-specific expression by developing an unstable GFP transcriptional reporter to observe the expression pattern of ndvB We found that, in addition to its expression in biofilms, ndvB was also upregulated in planktonic cells as they enter stationary phase...
January 19, 2018: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/29351997/spatiotemporal-organization-of-exocytosis-emerges-during-neuronal-shape-change
#7
Fabio L Urbina, Shawn M Gomez, Stephanie L Gupton
Neurite elongation and branching in developing neurons requires plasmalemma expansion, hypothesized to occur primarily via exocytosis. We posited that exocytosis in developing neurons and nonneuronal cells would exhibit distinct spatiotemporal organization. We exploited total internal reflection fluorescence microscopy to image vesicle-associated membrane protein (VAMP)-pHluorin-mediated exocytosis in mouse embryonic cortical neurons and interphase melanoma cells, and developed computer-vision software and statistical tools to uncover spatiotemporal aspects of exocytosis...
January 19, 2018: Journal of Cell Biology
https://www.readbyqxmd.com/read/29351860/microtubule-dynamics-moving-toward-a-multi-scale-approach
#8
REVIEW
Mahya Hemmat, Brian T Castle, David J Odde
Microtubule self-assembly dynamics serve to facilitate many vital cellular functions, such as chromosome segregation during mitosis and synaptic plasticity. However, the detailed atomistic basis of assembly dynamics has remained an unresolved puzzle. A key challenge is connecting together the vast range of relevant length-time scales, events happening at time scales ranging from nanoseconds, such as tubulin molecular interactions (Å-nm), to minutes-hours, such as the cellular response to microtubule dynamics during mitotic progression (μm)...
January 16, 2018: Current Opinion in Cell Biology
https://www.readbyqxmd.com/read/29351859/structural-and-immunological-characterization-of-hydroxyl-radical-modified-human-igg-clinical-correlation-in-rheumatoid-arthritis
#9
Sidra Islam, Abdul Rouf Mir, Mir Yasir Arfat, Farzana Khan, Masihuz Zaman, Asif Ali, Moinuddin
Structural alterations in proteins under oxidative stress have been widely implicated in the immuno-pathology of various disorders. This study has evaluated the extent of damage in the conformational characteristics of IgG by hydroxyl radical (OH) and studied its implications in the immuno-pathology of rheumatoid arthritis (RA). Using various biophysical and biochemical techniques, changes in aromatic microenvironment of the IgG and the protein aggregation became evident after treatment with OH. The SDS-PAGE study confirmed the protein aggregation while far ultraviolet circular dichroism spectroscopy (Far-UV CD) and fourier transform infrared spectroscopy (FTIR) inferred towards the alterations in secondary structure of IgG under OH stress...
January 10, 2018: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
https://www.readbyqxmd.com/read/29351850/systematic-gene-to-phenotype-arrays-a-high-throughput-technique-for-molecular-phenotyping
#10
Philipp A Jaeger, Lilia Ornelas, Cameron McElfresh, Lily R Wong, Randolph Y Hampton, Trey Ideker
We have developed a highly parallel strategy, systematic gene-to-phenotype arrays (SGPAs), to comprehensively map the genetic landscape driving molecular phenotypes of interest. By this approach, a complete yeast genetic mutant array is crossed with fluorescent reporters and imaged on membranes at high density and contrast. Importantly, SGPA enables quantification of phenotypes that are not readily detectable in ordinary genetic analysis of cell fitness. We benchmark SGPA by examining two fundamental biological phenotypes: first, we explore glucose repression, in which SGPA identifies a requirement for the Mediator complex and a role for the CDK8/kinase module in regulating transcription...
January 18, 2018: Molecular Cell
https://www.readbyqxmd.com/read/29351836/interrogating-interactions-and-modifications-of-histones-in-live-cells
#11
Xiao-Meng Li, Xiang David Li
In this issue of Cell Chemical Biology, new methods are reported to interrogate histone interactions and modifications. Kleiner et al. (2018) develop a chemical proteomics platform for profiling of direct, context-dependent histone-protein interactions in living cells, and Delachat et al. (2018) engineer fluorescent sensors for coexisting histone modifications in live stem cells.
January 18, 2018: Cell Chemical Biology
https://www.readbyqxmd.com/read/29351414/the-murine-choroid-plexus-epithelium-expresses-the-2cl-h-exchanger-clc-7-and-na-h-exchanger-nhe6-in-the-luminal-membrane-domain
#12
Helle H Damkier, Henriette L Christensen, Inga B Christensen, Qi Wu, Robert A Fenton, Jeppe Praetorius
The choroid plexus epithelium within the brain ventricles secretes the majority of the cerebrospinal fluid (CSF). The luminal Na+,K+-ATPase acts in concert with a host of other transport proteins to mediate efficient fluid secretion across the epithelium. The CSF contains little protein buffer, but the pH value seems nonetheless maintained within narrow limits, even when faced with acid/base challenges. The involvement of choroid plexus acid/base transporters in CSF pH regulation is highlighted by the expression of several acid/base transporters in the epithelium...
December 20, 2017: American Journal of Physiology. Cell Physiology
https://www.readbyqxmd.com/read/29351408/choroid-plexus-epithelial-cells-express-the-adhesion-protein-p-cadherin-at-cell-cell-contacts-and-syntaxin-4-in-the-luminal-membrane-domain
#13
Inga Baasch Christensen, Esben Nees Mogensen, Helle Hasager Damkier, Jeppe Praetorius
The choroid plexus epithelial cells (CPECs) belong to a small group of polarized cells, where the Na+,K+-ATPase is expressed in the luminal membrane. The basic polarity of the cells is, therefore, still debated. We investigated the subcellular distribution of an array of proteins known to play fundamental roles in either in establishing and maintaining basic cell polarity or the polarized delivery and recycling of plasma membrane proteins. Immunofluorescence histochemical analysis was applied to determine the subcellular localization of apical and basolateral membrane determinants...
January 10, 2018: American Journal of Physiology. Cell Physiology
https://www.readbyqxmd.com/read/29351321/methylglyoxal-produces-more-changes-in-biochemical-and-biophysical-properties-of-human-igg-under-high-glucose-compared-to-normal-glucose-level
#14
Mohd Adnan Khan, Zarina Arif, Mohd Asad Khan, Moinuddin, Khursheed Alam
Hyperglycaemia triggers increased production of methylglyoxal which can cause gross modification in proteins' structure vis-a-vis function though advanced glycation end products (AGEs). The AGEs may initiate vascular and nonvascular pathologies. In this study, we have examined the biochemical and biophysical changes in human IgG under normal and high glucose after introducing methylglyoxal into the assay mixture. This non-enzymatic reaction mainly engaged lysine residues as indicated by TNBS results. The UV results showed hyperchromicity in modified-IgG samples while fluorescence data supported AGEs formation during the course of reaction...
2018: PloS One
https://www.readbyqxmd.com/read/29351213/cyclin-b3-deficiency-impairs-germline-stem-cell-maintenance-and-its-overexpression-delays-cystoblast-differentiation-in-drosophila-ovary
#15
Dongsheng Chen, Lijuan Zhou, Fuling Sun, Mingzhong Sun, Xiaoqian Tao
It is well known that cyclinB3 (cycB3) plays a key role in the control of cell cycle progression. However, whether cycB3 is involved in stem cell fate determination remains unknown. The Drosophila ovary provides an exclusive model for studying the intrinsic and extrinsic factors that modulate the fate of germline stem cells (GSCs). Here, using this model, we show that DrosophilacycB3 plays a new role in controlling the fate of germline stem cells (GSC). Results from cycB3 genetic analyses demonstrate that cycB3 is intrinsically required for GSC maintenance...
January 19, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29351211/wide-field-fluorescence-microscopy-of-real-time-bioconjugation-sensing
#16
Marcin Szalkowski, Karolina Sulowska, Justyna Grzelak, Joanna Niedziółka-Jönsson, Ewa Roźniecka, Dorota Kowalska, Sebastian Maćkowski
We apply wide-field fluorescence microscopy to measure real-time attachment of photosynthetic proteins to plasmonically active silver nanowires. The observation of this effect is enabled, on the one hand, by sensitive detection of fluorescence and, on the other hand, by plasmonic enhancement of protein fluorescence. We examined two sample configurations with substrates being a bare glass coverslip and a coverslip functionalized with a monolayer of streptavidin. The different preparation of the substrate changes the observed behavior as far as attachment of the protein is concerned as well as its subsequent photobleaching...
January 19, 2018: Sensors
https://www.readbyqxmd.com/read/29351078/asymmetric-peptidomimetics-containing-l-tartaric-acid-core-inhibit-the-aspartyl-peptidase-activity-and-growth-of-leishmania-amazonensis-promastigotes
#17
André L S Santos, Filipe P Matteoli, Leandro S Sangenito, Marta H Branquinha, Bruno A Cotrim, Gabriel O Resende
Aspartyl-type peptidases are promising chemotherapeutic targets in protozoan parasites. In the present work, we identified an aspartyl peptidase activity from the soluble extract of Leishmania amazonensis promastigotes, which cleaved the fluorogenic peptide 7-methoxycoumarin-4-acetyl-Gly-Lys-Pro-Ile-Leu-Phe-Phe-Arg-Leu-Lys(DNP)-D-Arg-amide (cathepsin D substrate) under acidic pH conditions at 37°C, showing a KM of 0.58 μM and Vmax of 129.87 fluorescence arbitrary units/s mg protein. The leishmanial aspartyl peptidase activity was blocked by pepstatin A (IC50 = 6...
March 26, 2018: Acta Parasitologica
https://www.readbyqxmd.com/read/29350877/effects-of-human-rhinovirus-on-epithelial-barrier-integrity-and-function-in-children-with-asthma
#18
Kevin Looi, Alysia G Buckley, Paul J Rigby, Luke W Garratt, Thomas Iosifidis, Graeme R Zosky, Alexander N Larcombe, Francis J Lannigan, Kak-Ming Ling, Kelly M Martinovich, Elizabeth Kicic-Starcevich, Nicole C Shaw, Erika N Sutanto, Darryl A Knight, Anthony Kicic, Stephen M Stick
BACKGROUND: Bronchial epithelial tight junctions (TJ) have been extensively assessed in healthy airway epithelium. However, no studies have yet assessed the effect of human rhinovirus (HRV) infection on the expression and resultant barrier function in epithelial tight junctions (TJ) in childhood asthma. OBJECTIVES: To investigate the impact of HRV infection on airway epithelial TJ expression and barrier function in airway epithelial cells (AEC) of children with and without asthma...
January 19, 2018: Clinical and Experimental Allergy: Journal of the British Society for Allergy and Clinical Immunology
https://www.readbyqxmd.com/read/29350433/amino-group-of-salicylic-acid-exhibits-enhanced-inhibitory-potential-against-insulin-amyloid-fibrillation-with-protective-aptitude-toward-amyloid-induced-cytotoxicity
#19
Masihuz Zaman, Mohsin Vahid Khan, Syed Mohammad Zakariya, Saima Nusrat, Syed Mustapha Meeran, Parvez Alam, Mohammad Rehan Ajmal, Wahiduzzaman Wahiduzzaman, Yasser E Shahein, Amira M Abouelella, Rizwan Hasan Khan
Protein misfolding and aggregation lead to amyloid generation that in turn may induce cell membrane disruption and leads to cell apoptosis. In an effort to prevent or treat amyloidogenesis, large number of studies has been paying attention on breakthrough of amyloid inhibitors. In the present work, we aim to access the effect of two drugs, that is, acetylsalicylic acid and 5-amino salicylic acid on insulin amyloids by using various biophysical, imaging, cell viability assay, and computational approaches. We established that both drugs reduce the turbidity, light scattering and fluorescence intensity of amyloid indicator dye thioflavin T...
December 11, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/29350381/macrophage-migration-inhibitory-factor-knockout-attenuates-endotoxin-induced-cardiac-dysfunction-in-mice
#20
Jie Zhang, Xin Zhang, Yuqi Cui, Lianqun Cui, Peng Zhao
BACKGROUND: The accumulated evidence suggests that macrophage migration inhibitory factor (MIF) plays a key role not only in acute and chronic inflammatory diseases but also in cardiovascular disease. The cardiac dysfunction is related to lipopolysaccharide (LPS) in sepsis. AIM: This study was designed to examine whether MIF mediates LPS-induced cardiac dysfunction and address the mechanisms. METHODS: Echocardiography, immunohistochemical analysis, cell shortening/relengthening and intracellular Ca2+ fluorescence evaluation were performed in whole hearts and isolated cardiomyocytes from C57 and MIF knockout mice treated with or without LPS...
January 19, 2018: Kardiologia Polska
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