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Linas Tamošaitis, C Mark Smales
Transcriptomics has been extensively applied to the investigation of the CHO cell platform for the production of recombinant biotherapeutic proteins to identify transcripts whose expression is regulated and correlated to (non)desirable CHO cell attributes. However, there have been few attempts to analyse the findings across these studies to identify conserved changes and generic targets for CHO cell platform engineering. Here we have undertaken a meta-analysis of CHO cell transcriptomic data and report on those genes most frequently identified as differentially expressed with regard to cell growth (µ) and productivity (Qp)...
June 19, 2018: Biotechnology Journal
Laura Pellegrini, David N Hauser, Yan Li, Adamantios Mamais, Alexandra Beilina, Ravindran Kumaran, Andrea Wetzel, George Heaton, Iakov Rudenko, Mor Alkaslasi, Natalie Ivanina, Heather L Melrose, Mark R Cookson, Kirsten Harvey
Mutations in leucine-rich repeat kinase 2 (LRRK2) segregate with familial Parkinson's disease (PD) and genetic variation around LRRK2 contributes to risk of sporadic disease. Although knockout of LRRK2 or knock-in of pathogenic mutations into the mouse germline does not result in a PD phenotype, several defects have been reported in the kidneys of LRRK2 knockout mice. To understand LRRK2 function in vivo, we used an unbiased approach to determine which protein pathways are affected in LRRK2 knockout kidneys...
June 18, 2018: Human Molecular Genetics
Parisa Lotfi, Dennis Y Tse, Alberto di Ronza, Michelle L Seymour, Giuseppe Martano, Jonathan D Cooper, Fred A Pereira, Maria Passafaro, Samuel M Wu, Marco Sardiello
The accumulation of undegraded molecular material leads to progressive neurodegeneration in a number of lysosomal storage disorders (LSDs) that are caused by functional deficiencies of lysosomal hydrolases. To determine whether inducing macroautophagy/autophagy via small-molecule therapy would be effective for neuropathic LSDs due to enzyme deficiency, we treated a mouse model of mucopolysaccharidosis IIIB (MPS IIIB), a storage disorder caused by deficiency of the enzyme NAGLU (alpha-N-acetylglucosaminidase [Sanfilippo disease IIIB]), with the autophagy-inducing compound trehalose...
June 19, 2018: Autophagy
Elias Flockerzi, Loay Daas, Ursula Schlötzer-Schrehardt, Annette Zimpfer, Rainer Bohle, Berthold Seitz
PURPOSE: Cystinosis is an autosomal recessive inherited lysosomal storage disease with an incidence of 1:100.000 up to 1:200.000 caused by a gene mutation of a lysosomal transport protein resulting in deposition of cystine in lysosomes in all cells and tissues. In the cornea, crystalline, gold-dust deposition of cystine leads to visual impairment, recurrent erosions, photophobia, epiphora and blepharospasmus. Standard therapy is topical and systemic application of cysteamine which may resolve the accumulated cystine crystals...
June 18, 2018: International Ophthalmology
Joana P Guedes, Cátia S Pereira, Lígia R Rodrigues, Manuela Côrte-Real
Prostate cancer and osteosarcoma are the second most common type of cancer affecting men and the fifth most common malignancy among adolescents, respectively. The use of non-toxic natural or natural-derived products has been one of the current strategies for cancer therapy, owing to the reduced risks of induced-chemoresistance development and the absence of secondary effects. In this perspective, lactoferrin (Lf), a natural protein derived from milk, emerges as a promising anticancer agent due to its well-recognized cytotoxicity and anti-metastatic activity...
2018: Frontiers in Oncology
Chengmin He, Zhi Yang, Ying Jin, Xiaoyang Qi, Jin Chu, Xiaoyuan Deng
Adult mammalian skin has a defective regenerative capacity following full-thickness cutaneous injury; this defect overshadows the complete physiological functions of the skin. Immune-mediated skin reconstruction driven by biological scaffolds is a recently developed innovative repair strategy to support regenerative wound healing. However, to date, little is known about how biological scaffolds orchestrate the immune response to promote regeneration. Here, using acellular dermal matrix (ADM) scaffolds, we discovered that the default pro-inflammatory response was altered in response to a pro-regenerative response characterized by specific M2 polarization...
2018: Frontiers in Physiology
Wenjing Qi, Yufei Zhang, Jue Wang, Guoqing Tao, Libin Wu, Zdravko Kochovski, Hongjian Gao, Guosong Chen, Ming Jiang
We proposed the deprotection-induced block copolymer self-assembly (DISA), that is, the deprotection of hydroxyl groups resulted in in situ self-assembly of glycopolymers. In the previous studies, block copolymers soluble in common organic solvents were employed as the starting material. In this paper, by using the protected glyco-block containing pre-assembled glyco-vesicles in water as the starting material, we moved forward and made two exceeding achievements. Firstly, we have observed a deprotection-induced morphology transition trigged by alkali in water...
June 18, 2018: Journal of the American Chemical Society
Domenico Mattoscio, Alessandro Medda, Susanna Chiocca
Human papilloma viruses (HPVs) are a group of double-stranded DNA viruses known to be the primary cause of cervical cancer. In addition, evidence has now established their role in non-melanoma skin cancers, head and neck cancer (HNC), and the development of other anogenital malignancies. The prevalence of HPV-related HNC, in particular oropharyngeal cancers, is rapidly increasing, foreseeing that HPV-positive oropharyngeal cancers will outnumber uterine cervical cancers in the next 15⁻20 years. Therefore, despite the successful advent of vaccines originally licensed for cervical cancer prevention, HPV burden is still very high, and a better understanding of HPV biology is urgently needed...
June 15, 2018: International Journal of Molecular Sciences
Rihab Khlifi, Zaineb Dhaouefi, Mouna Maatouk, Aicha Sassi, Najet Boudhiba, Irina Ioannou, Kamel Ghedira, Leila Chekir-Ghedira, Soumaya Kilani-Jaziri
Plants and natural molecules are generally consumed not in raw state but after different processing conditions (heating, mechanical agitation or cooking). The understanding of the chemistry and biological outcome of thermal treatment is still scarce. In the current study, Eriodictyol, a natural flavanone, has undergone heat treatment, generating hence three different products ((3-(3,4-dihydroxyphenyl)-3-hydroxypropanoic acid, (3-(3,4-dihydroxyphenyl) propanal) and an unidentified component). The consequences of aforementioned treatment on the immunomodulatory behavior of resulted molecules were evaluated...
June 15, 2018: International Immunopharmacology
Sofia Raniolo, Giulia Vindigni, Valeria Unida, Alessio Ottaviani, Elena Romano, Alessandro Desideri, Silvia Biocca
DNA has been used to build nanostructures with potential biomedical applications. However, their use is limited by the lack of information on the mechanism of entry, intracellular fate and degradation rate of nanostructures inside cells. We generated octahedral DNA nanocages functionalized with folic acid and investigated the cellular uptake mediated by two distinctive internalization pathways, using two cellular systems expressing the oxidized low-density lipoprotein receptor-1 (LOX-1) and the α isoform of the folate receptor (αFR), respectively...
June 18, 2018: Nanoscale
Juliette Morlon-Guyot, Hiba El Hajj, Kevin Martin, Adrien Fois, Amandine Carrillo, Laurence Berry, Richard Burchmore, Markus Meissner, Maryse Lebrun, Wassim Daher
Apicomplexans use the endolysosomal system for the biogenesis of their secretory organelles, namely micronemes, rhoptries and dense granules. In Toxoplasma gondii, our previous in silico search identified the HOPS tethering but not the CORVET complex and demonstrated a role of Vps11 (a common component for both complexes) in its secretory organelles biogenesis. Herein, we performed Vps11-GFP-Trap pull-down assays and identified by proteomic analysis, not only the CORVET specific subunit Vps8, but also a BEACH domain-containing protein (BDCP) conserved in eukaryotes...
June 17, 2018: Cellular Microbiology
S S Murray, A W Wong, J Yang, Y Li, U Putz, S-S Tan, J Howitt
The regulation of Trk receptors is critical for orchestrating multiple signalling pathways required for developing and maintaining neuronal networks. Activation of Trk receptors results in signalling, internalisation and subsequent degradation of the protein. Although ubiquitination of TrkA by Nedd4-2 has been identified as an important degradation pathway, much less is known about the pathways regulating the degradation of TrkB and TrkC. Critical to the interaction between TrkA and Nedd4-2 is a PPxY motif present within TrkA but absent in TrkB and TrkC...
June 18, 2018: Molecular Neurobiology
Du-Juan Huang, Zhi-Mei Huang, Hu-Yan Xiao, Zhen-Kun Wu, Li-Juan Tang, Jian-Hui Jiang
Efficient intracellular delivery of nucleic acids to achieve sensitive detection and gene regulation is essential for chemistry and biology. Here we developed a novel protein scaffolded DNA tetrad, a four-arm DNA nanostructure constructed using streptavidin (SA) protein and four biotinylated hairpin DNA probes for efficient nucleic acid delivery and ultrasensitive miRNA imaging through crosslinking hybridization chain reaction (cHCR). DNA tetrads were easy to prepare and allowed precise control of the structure of the probes...
June 7, 2018: Chemical Science
Yiting Wang, Huanbin Wang, Han Yao, Chushu Li, Jing-Yuan Fang, Jie Xu
Immune checkpoint blockade therapies (ICBTs) targeting programmed cell death 1 (PD-1) and its ligand programmed death ligand-1 (PD-L1/B7-H1/CD274) have exhibited momentous clinical benefits and durable responses in multiple tumor types. However, primary resistance is found in considerable number of cancer patients, and most responders eventually develop acquired resistance to ICBT. To tackle these challenges, it is essential to understand how PD-L1 is controlled by cancer cells to evade immune surveillance...
2018: Frontiers in Pharmacology
Paul K Ziegler, Julia Bollrath, Charles K Pallangyo, Takaji Matsutani, Özge Canli, Tiago De Oliveira, Michaela A Diamanti, Nina Müller, Jaba Gamrekelashvili, Tracy Putoczki, David Horst, Arun K Mankan, Meryem G Öner, Susanna Müller, Josef Müller-Höcker, Thomas Kirchner, Julia Slotta-Huspenina, M Mark Taketo, Thomas Reinheckel, Stefan Dröse, Andrew C Larner, Winfried S Wels, Matthias Ernst, Tim F Greten, Melek C Arkan, Thomas Korn, Dagmar Wirth, Florian R Greten
In colorectal cancer patients, a high density of cytotoxic CD8+ T cells in tumors is associated with better prognosis. Using a Stat3 loss-of-function approach in two wnt/β-catenin-dependent autochthonous models of sporadic intestinal tumorigenesis, we unravel a complex intracellular process in intestinal epithelial cells (IECs) that controls the induction of a CD8+ T cell based adaptive immune response. Elevated mitophagy in IECs causes iron(II)-accumulation in epithelial lysosomes, in turn, triggering lysosomal membrane permeabilization...
June 11, 2018: Cell
Guang Lin, Pei-Tseng Lee, Kuchuan Chen, Dongxue Mao, Kai Li Tan, Zhongyuan Zuo, Wen-Wen Lin, Liping Wang, Hugo J Bellen
Mutations in PLA2G6 (PARK14) cause neurodegenerative disorders in humans, including autosomal recessive neuroaxonal dystrophy and early-onset parkinsonism. We show that loss of iPLA2-VIA, the fly homolog of PLA2G6, reduces lifespan, impairs synaptic transmission, and causes neurodegeneration. Phospholipases typically hydrolyze glycerol phospholipids, but loss of iPLA2-VIA does not affect the phospholipid composition of brain tissue but rather causes an elevation in ceramides. Reducing ceramides with drugs, including myriocin or desipramine, alleviates lysosomal stress and suppresses neurodegeneration...
June 7, 2018: Cell Metabolism
Wei Liu, Xiuying Duan, Xuefei Fang, Weina Shang, Chao Tong
Neurodegeneration is characterized by protein aggregate deposits and mitochondrial malfunction. Reduction in Tom40 (translocase of outer membrane 40) expression, a key subunit of the translocase of the outer mitochondrial membrane complex, led to accumulation of ubiquitin (Ub)-positive protein aggregates engulfed by Atg8a-positive membranes. Other macroautophagy markers were also abnormally accumulated. Autophagy was induced but the majority of autophagosomes failed to fuse with lysosomes when Tom40 was downregulated...
June 18, 2018: Autophagy
M M Akhtar, P M Elliott
Anderson-Fabry disease is an X-linked lysosomal storage disorder caused by mutations in the GLA gene that result in deficiency of the enzyme alpha-galactosidase A. The worldwide incidence of Fabry's disease is reported to be in the range of 1 in 40,000-117,000, although this value may be a significant underestimate given under recognition of symptoms and delayed or missed diagnosis. Deficiency in alpha-galactosidase A causes an accumulation of neutral glycosphingolipids such as globotriaosylceramide (Gb3) in lysosomes within various tissues including the vascular endothelium, kidneys, heart, eyes, skin and nervous system...
June 16, 2018: Biophysical Reviews
Yasuhiko Kizuka, Naoyuki Taniguchi
Bisecting GlcNAc, a branch structure in N-glycan, has unique functions and is involved in several diseases including Alzheimer's disease (AD). In this review, we provide an overview of the biosynthesis of bisecting GlcNAc and its physiological and pathological functions, particularly in the nervous system where bisecting GlcNAc is most highly expressed. The biosynthetic enzyme of bisecting GlcNAc is N-acetylglucosaminyltransferase-III (GnT-III). Overexpression, knockdown, and knockout of GnT-III have so far revealed various functions of bisecting GlcNAc, which are mediated by regulating the functions of key carrier proteins...
June 16, 2018: Glycoconjugate Journal
Robert Fuchs, Anika Stracke, Viktoria Holzmann, Gerfried Luschin-Ebengreuth, Nathalie Meier-Allard, Nadine Ebner, Teresa Maria Lassacher, Markus Absenger-Novak, Eleonore Fröhlich, Matthias Schittmayer, Sara Cano Crespo, Manuel Palacin, Beate Rinner, Ruth Birner-Gruenberger
The quinazoline based drug prazosin (PRZ) is a potent inducer of apoptosis in human cancer cells. We recently reported that PRZ enters cells via endocytosis and induces tubulation of the endolysosomal system. In a proteomics approach aimed at identifying potential membrane proteins with binding affinity to quinazolines, we detected the oncoprotein CD98hc. We confirmed shuttling of CD98hc towards lysosomes and upregulation of CD98hc expression in PRZ treated cells. Gene knockout (KO) experiments revealed that endocytosis of PRZ still occurs in the absence of CD98hc - suggesting that PRZ does not enter the cell via CD98hc but misroutes the protein towards tubular lysosomes...
June 14, 2018: Biochimica et Biophysica Acta
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