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https://www.readbyqxmd.com/read/28930976/detection-of-residual-donor-erythroid-progenitor-cells-after-hematopoietic-stem-cell-transplantation-for-patients-with-hemoglobinopathies
#1
Roman Crazzolara, Gabriele Kropshofer, Michael Steurer, Sieghart Sopper, Wolfgang Schwinger
The presence of incomplete chimerism is noted in a large proportion of patients following bone marrow transplant for thalassemia major or sickle cell disease. This observation has tremendous implications, as subsequent therapeutic immunomodulation strategies can improve clinical outcome. Conventionally, polymerase chain reaction-based analysis of short tandem repeats is used to identify chimerism in donor-derived blood cells. However, this method is restricted to nucleated cells and cannot distinguish between dissociated single-cell lineages...
September 6, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28930975/streamlined-single-cell-tcr-isolation-and-generation-of-retroviral-vectors-for-in-vitro-and-in-vivo-expression-of-human-tcrs
#2
Maran L Sprouse, Gabriele Blahnik, Thomas Lee, Natalie Tully, Pinaki Benarjee, Eddie A James, Maria J Redondo, Matthew L Bettini, Maria Bettini
Although, several methods for sequencing of paired T cell receptor (TCR) alpha and beta chains from single T cells have been developed, none so far have been conducive to downstream in vivo functional analysis of TCR heterodimers. We have developed an improved protocol based on a two-step multiplex-nested PCR, which results in a PCR product that spans entire variable regions of a human TCR alpha and beta chains. By identifying unique restriction sites and incorporating them into the PCR primers, we have made the PCR product compatible with direct sub-cloning into the template retroviral vector...
September 10, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28930450/continuous-sorting-of-cells-based-on-differential-p-selectin-glycoprotein-ligand-expression-using-molecular-adhesion
#3
Bushra Tasadduq, Brynn Mcfarland, Muhymin Islam, Alexander Alexeev, Ali Fatih Sarioglu, Todd A Sulchek
Cell surface molecular adhesions govern many important physiological processes and are used to identify cells for analysis and purifications. But most effective cell adhesion separation technologies use labels or long-term attachments in their application. While label-free separation microsystems typically separate cells by size, stiffness, and shape, they often do not provide sufficient specificity to cell type that can be obtained from molecular expression. We demonstrate a label-free microfluidic approach capable of high throughput separation of cells based upon surface molecule adhesion...
September 20, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28930187/viruses-of-microbes
#4
EDITORIAL
Laurent Debarbieux, Matthias Fischer, Tessa E F Quax
Viruses of microbes encompass all viruses that infect archaea, bacteria, and single-celled eukaryotes, especially algae and protozoa [...].
September 20, 2017: Viruses
https://www.readbyqxmd.com/read/28929742/beta-hemolytic-bacteria-selectively-trigger-liposome-lysis-enabling-rapid-and-accurate-pathogen-detection
#5
Rongji Sum, Muthukaruppan Swaminathan, Sahil Kumar Rastogi, Obdulio Piloto, Ian Cheong
For more than a century, blood agar plates have been the only test for beta-hemolysis. Although blood agar cultures are highly predictive for bacterial pathogens, they are too slow to yield actionable information. Here, we show that beta-hemolytic pathogens are able to lyse and release fluorophores encapsulated in sterically stabilized liposomes whereas alpha and gamma-hemolytic bacteria have no effect. By analyzing fluorescence kinetics, beta-hemolytic colonies cultured on agar could be distinguished in real time with 100% accuracy within 6 h...
September 20, 2017: ACS Sensors
https://www.readbyqxmd.com/read/28929679/-identification-of-six-species-of-medicinal-diospyros-plants-based-on-leaf-macro-and-micro-morphology
#6
Ze Yi, Juan-Juan Qiao, Geng-Yu Lu, Gang Wu, Guo-Yong Xie, Min-Jian Qin
To establish a method for the identification of five species and one variety of medicinal plants from Diospyros, their leaf veins, epidermis, anatomic and powder characters were observed and compared with macro-morphological and microscopic methods. The results indicated the differences of secondary and tertiary veins among those Diospyros species. The single cell non-glandular hair and glandular hair exist in most species' epidermis while stone cells were only found in the leaf powders of two species. Through the study, the main differences of leaf macro- and micro-morphology of these species were obtained and practical keys were also established, which can provide scientific base not only for identification of these species during their vegetative stages, but also for accuracy authentication of the source of Kaki Folium...
November 2016: Zhongguo Zhong Yao za Zhi, Zhongguo Zhongyao Zazhi, China Journal of Chinese Materia Medica
https://www.readbyqxmd.com/read/28929569/evaluation-of-chemical-chaperones-based-on-the-monitoring-of-bip-promoter-activity-and-visualization-of-extracellular-vesicles-by-real-time-bioluminescence-imaging
#7
Tomohisa Horibe, Nanako Okushima, Aya Torisawa, Ryutaro Akiyoshi, Yoko Hatta-Ohashi, Hirobumi Suzuki, Koji Kawakami
It is known that endoplasmic reticulum (ER) stress in cells and extracellular vesicles (EVs) plays a significant role in cancer cells, therefore the evaluation of compounds that can regulate ER stress and EV secretion would be a suitable system for further screening and development of new drugs. In this study, we evaluated chemical chaperones derived from natural products based on monitoring Bip/GRP78 promoter activity during cancer cell growth, at the level of the single cell, by a bioluminescence microscopy system that had several advantages compared with fluorescence imaging...
September 20, 2017: Luminescence: the Journal of Biological and Chemical Luminescence
https://www.readbyqxmd.com/read/28928738/age-stratified-t-cell-responses-in-children-infected-with-mycobacterium-tuberculosis
#8
Alexandra Dreesman, Véronique Corbière, Violette Dirix, Kaat Smits, Sara Debulpaep, Iris De Schutter, Myriam Libin, Mahavir Singh, Anne Malfroot, Camille Locht, Françoise Mascart
Tuberculosis (TB) in young children differs from adult TB in that the risk of rapid progression to active TB (aTB) is higher in children than in adults. The reasons for this increased risk are not fully understood. Early differentiation remains difficult between children at risk to develop aTB from those who will remain healthy and develop a latent TB infection (LTBI). Biomarkers to differentiate aTB from LTBI in children, especially in very young children, are urgently needed. To identify M. tuberculosis-specific functional T cell subsets related to clinical manifestations in children, we enrolled 87 children exposed to M...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28928395/stochastic-priming-and-spatial-cues-orchestrate-heterogeneous-clonal-contribution-to-mouse-pancreas-organogenesis
#9
Hjalte List Larsen, Laura Martín-Coll, Alexander Valentin Nielsen, Christopher V E Wright, Ala Trusina, Yung Hae Kim, Anne Grapin-Botton
Spatiotemporal balancing of cellular proliferation and differentiation is crucial for postnatal tissue homoeostasis and organogenesis. During embryonic development, pancreatic progenitors simultaneously proliferate and differentiate into the endocrine, ductal and acinar lineages. Using in vivo clonal analysis in the founder population of the pancreas here we reveal highly heterogeneous contribution of single progenitors to organ formation. While some progenitors are bona fide multipotent and contribute progeny to all major pancreatic cell lineages, we also identify numerous unipotent endocrine and ducto-endocrine bipotent clones...
September 19, 2017: Nature Communications
https://www.readbyqxmd.com/read/28928248/carbonic-anhydrase-iii-protects-osteocytes-from-oxidative-stress
#10
Chao Shi, Yuhei Uda, Christopher Dedic, Ehab Azab, Ningyuan Sun, Amira I Hussein, Christopher A Petty, Keertik Fulzele, Maria C Mitterberger-Vogt, Werner Zwerschke, Renata Pereira, Kunzheng Wang, Paola Divieti Pajevic
Osteocytes are master orchestrators of bone remodeling; they control osteoblast and osteoclast activities both directly via cell-to-cell communication and indirectly via secreted factors, and they are the main postnatal source of sclerostin and RANKL (receptor activator of NF-kB ligand), two regulators of osteoblast and osteoclast function. Despite progress in understanding osteocyte biology and function, much remains to be elucidated. Recently developed osteocytic cell lines-together with new genome editing tools-has allowed a closer look at the biology and molecular makeup of these cells...
September 19, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/28927991/deciphering-cell-intrinsic-properties-a-key-issue-for-robust-organoid-production
#11
REVIEW
Nathalie Picollet-D'hahan, Monika E Dolega, Delphine Freida, Donald K Martin, Xavier Gidrol
We highlight the disposition of various cell types to self-organize into complex organ-like structures without necessarily the support of any stromal cells, provided they are placed into permissive 3D culture conditions. The goal of generating organoids reproducibly and efficiently has been hampered by poor understanding of the exact nature of the intrinsic cell properties at the origin of organoid generation, and of the signaling pathways governing their differentiation. Using microtechnologies like microfluidics to engineer organoids would create opportunities for single-cell genomics and high-throughput functional genomics to exhaustively characterize cell intrinsic properties...
September 16, 2017: Trends in Biotechnology
https://www.readbyqxmd.com/read/28927942/microfluidic-single-cell-technology-in-immunology-and-antibody-screening
#12
REVIEW
Yu Fen Samantha Seah, Hongxing Hu, Christoph A Merten
Single-cell technology has a major impact on the field of immunology. It enables the kinetics and logic of immune signaling and immune cell migration to be elucidated, facilitates antibody screening and allows massively parallelized analysis of B- and T-cell repertoires. Impressive progress has been made over the last decade, strongly boosted by microfluidic approaches. In this review, we summarize the most powerful microfluidic systems based on continuous-flow, nanowells, valves and droplets and we analyze their benefits for phenotypic characterization, drug discovery and next generation sequencing experiments...
September 16, 2017: Molecular Aspects of Medicine
https://www.readbyqxmd.com/read/28927434/sifit-inferring-tumor-trees-from-single-cell-sequencing-data-under-finite-sites-models
#13
Hamim Zafar, Anthony Tzen, Nicholas Navin, Ken Chen, Luay Nakhleh
Single-cell sequencing enables the inference of tumor phylogenies that provide insights on intra-tumor heterogeneity and evolutionary trajectories. Recently introduced methods perform this task under the infinite-sites assumption, violations of which, due to chromosomal deletions and loss of heterozygosity, necessitate the development of inference methods that utilize finite-sites models. We propose a statistical inference method for tumor phylogenies from noisy single-cell sequencing data under a finite-sites model...
September 19, 2017: Genome Biology
https://www.readbyqxmd.com/read/28926758/some-aspects-of-the-iodine-metabolism-of-the-giant-kelp-macrocystis-pyrifera-phaeophyceae
#14
Teresa M Tymon, Eric P Miller, Jennifer L Gonzales, Andrea Raab, Frithjof C Küpper, Carl J Carrano
Despite its paramount role in the functioning of coastal ecosystems, relatively little is known about halogen metabolism in giant kelp (Macrocystis pyrifera). This is an important shortcoming given the potential implications for marine and atmospheric chemical processes in the wide distribution range of Macrocystis. The work presented here constitutes the first in depth investigation of the uptake, efflux, and of the physiological function of iodide in this important kelp species. Iodide uptake and efflux rates were measured in adult sporophytes of Macrocystis under normal and stressed (exogenous hydrogen peroxide and an elicitor-triggered oxidative burst) conditions...
September 8, 2017: Journal of Inorganic Biochemistry
https://www.readbyqxmd.com/read/28926213/whole-genome-amplification-and-sequencing-of-a-daphnia-resting-egg
#15
Justin B Lack, Lawrence J Weider, Punidan D Jeyasingh
Resting eggs banks are unique windows that allow us to directly observe shifts in population genetics, and phenotypes over time as natural populations evolve. Although a variety of planktonic organisms also produce resting stages, the keystone freshwater consumer, Daphnia, is a well-known model for paleogenetics and resurrection ecology. Nevertheless, paleogenomic investigations are limited largely because resting eggs do not contain enough DNA for genomic sequencing. In fact, genomic studies even on extant populations include a laborious preparatory phase of batch culturing dozens of individuals to generate sufficient genomic DNA...
September 19, 2017: Molecular Ecology Resources
https://www.readbyqxmd.com/read/28924833/rho-inhibiting-c2in-c3-fusion-toxin-inhibits-chemotactic-recruitment-of-human-monocytes-ex-vivo-and-in-mice-in-vivo
#16
Tobias Martin, Amelie Möglich, Ina Felix, Christina Förtsch, Anne Rittlinger, Annette Palmer, Stephanie Denk, Julian Schneider, Lena Notbohm, Mona Vogel, Hartmut Geiger, Stephan Paschke, Markus Huber-Lang, Holger Barth
Bacterial protein toxins became valuable molecular tools for the targeted modulation of cell functions in experimental pharmacology and attractive therapeutics because of their potent and specific mode of action in human cells. C2IN-C3lim, a recombinant fusion toxin (~50 kDa) of the Rho-inhibiting C3lim from Clostridium (C.) limosum and a non-toxic portion of the C. botulinum C2 toxin (C2IN), is selectively internalized into the cytosol of monocytic cells where C3lim specifically ADP-ribosylates Rho A and -B, thereby inhibiting Rho-mediated signaling...
September 18, 2017: Archives of Toxicology
https://www.readbyqxmd.com/read/28924545/x-autosome-and-x-y-translocations-in-female-carriers-x-chromosome-inactivation-easily-detectable-by-5-ethynyl-2-deoxyuridine-edu
#17
M Donat, A Louis, K Kreskowski, M Ziegler, A Weise, I Schreyer, T Liehr
Here we report one new case each of an X-autosome translocation (maternally derived), and an X-Y-chromosome translocation. Besides characterizing the involved breakpoints and/or imbalances in detail by molecular cyto-genetics, also skewed X-chromosome inactivation was determined on single cell level using 5-ethynyl-2-deoxyuridine (EdU). Thus, we confirmed that the recently suggested EdU approach can be simply adapted for routine diagnostic use. The latter is important, as only by knowing the real pattern of the skewed X-chromosome inactivation, correct interpretation of obtained results and subsequent reliable genetic counseling, can be done...
June 30, 2017: Balkan Journal of Medical Genetics: BJMG
https://www.readbyqxmd.com/read/28924377/current-progresses-of-single-cell-dna-sequencing-in-breast-cancer-research
#18
REVIEW
Jianlin Liu, Ragini Adhav, Xiaoling Xu
Breast cancers display striking genetic and phenotypic diversities. To date, several hypotheses are raised to explain and understand the heterogeneity, including theories for cancer stem cell (CSC) and clonal evolution. According to the CSC theory, the most tumorigenic cells, while maintaining themselves through symmetric division, divide asymmetrically to generate non-CSCs with less tumorigenic and metastatic potential, although they can also dedifferentiate back to CSCs. Clonal evolution theory recapitulates that a tumor initially arises from a single cell, which then undergoes clonal expansion to a population of cancer cells...
2017: International Journal of Biological Sciences
https://www.readbyqxmd.com/read/28923977/prepatterning-by-rhogefs-governs-rho-gtpase-spatiotemporal-dynamics-during-wound-repair
#19
Mitsutoshi Nakamura, Jeffrey M Verboon, Susan M Parkhurst
Like tissues, single cells are subjected to continual stresses and damage. As such, cells have a robust wound repair mechanism comprised of dynamic membrane resealing and cortical cytoskeletal remodeling. One group of proteins, the Rho family of small guanosine triphosphatases (GTPases), is critical for this actin and myosin cytoskeletal response in which they form distinct dynamic spatial and temporal patterns/arrays surrounding the wound. A key mechanistic question, then, is how these GTPase arrays are formed...
September 18, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28923782/single-cell-epigenome-sequencing-technologies
#20
REVIEW
Lu Wen, Fuchou Tang
No abstract text is available yet for this article.
September 16, 2017: Molecular Aspects of Medicine
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