Involvement of stress-related genes polB and PA14_46880 in biofilm formation of Pseudomonas aeruginosa.
AuthorsAlshalchi SA, Anderson GG,
Journal
PubMed ID25156741
Chronic infections of Pseudomonas aeruginosa are generally established through production of biofilm. During biofilm formation, production of an extracellular matrix and establishment of a distinct bacterial phenotype make these infections difficult to eradicate. However, biofilm studies have been hampered by the fact that most assays utilize nonliving surfaces as biofilm ... More
Human Primordial Germ Cells Are Specified from Lineage-Primed Progenitors.
AuthorsChen D, Sun N, Hou L, Kim R, Faith J, Aslanyan M, Tao Y, Zheng Y, Fu J, Liu W, Kellis M, Clark A
JournalCell Rep
PubMed ID31875561
'In vitro gametogenesis is the process of making germline cells from human pluripotent stem cells. The foundation of this model is the quality of the first progenitors called primordial germ cells (PGCs), which in vivo are specified during the peri-implantation window of human development. Here, we show that human PGC (hPGC) specification ... More
ePro-ClearSee: a simple immunohistochemical method that does not require sectioning of plant samples.
AuthorsNagaki K, Yamaji N, Murata M
JournalSci Rep
PubMed ID28176832
'Investigations into the epigenetic status of individual cells within tissues can produce both epigenetic data for different cell types and positional information of the cells. Thus, these investigations are important for understanding the intra- and inter-cellular control systems of developmental and environmental responses in plants. However, a simple method to ... More
Revealing hidden complexities of genomic rearrangements generated with Cas9.
AuthorsBoroviak K, Fu B, Yang F, Doe B, Bradley A
JournalSci Rep
PubMed ID28993641
'Modelling human diseases caused by large genomic rearrangements has become more accessible since the utilization of CRISPR/Cas9 in mammalian systems. In a previous study, we showed that genomic rearrangements of up to one million base pairs can be generated by direct injection of CRISPR/Cas9 reagents into mouse zygotes. Although these ... More
SosA inhibits cell division in Staphylococcus aureus in response to DNA damage.
'Inhibition of cell division is critical for viability under DNA-damaging conditions. DNA damage induces the SOS response that in bacteria inhibits cell division while repairs are being made. In coccoids, such as the human pathogen, Staphylococcus aureus, this process remains poorly studied. Here, we identify SosA as the staphylococcal SOS-induced ... More
Light needle microscopy with spatially transposed detection for axially resolved volumetric imaging.
AuthorsKozawa Y, Sato S
JournalSci Rep
PubMed ID31406209
'The demand for rapid three-dimensional volumetric imaging is increasing in various fields, including life science. Laser scanning fluorescence microscopy has been widely employed for this purpose; however, a volumetric image is constructed by two-dimensional image stacking with a varying observation plane, ultimately limiting the acquisition speed. Here we propose a ... More
Nanoscale Surveillance of the Brain by Microglia via cAMP-Regulated Filopodia.
AuthorsBernier LP, Bohlen CJ, York EM, Choi HB, Kamyabi A, Dissing-Olesen L, Hefendehl JK, Collins HY, Stevens B, Barres BA, MacVicar BA
JournalCell Rep
PubMed ID31167136
'Microglia, the brain''s immune cells, maintain homeostasis and sense pathological changes by continuously surveying the parenchyma with highly motile large processes. Here, we demonstrate that microglia also use thin actin-dependent filopodia that allow fast nanoscale sensing within discrete regions. Filopodia are distinct from large processes by their size, speed, and ... More
Homology-mediated end joining-based targeted integration using CRISPR/Cas9.
AuthorsYao X, Wang X, Hu X, Liu Z, Liu J, Zhou H, Shen X, Wei Y, Huang Z, Ying W, Wang Y, Nie YH, Zhang CC, Li S, Cheng L, Wang Q, Wu Y, Huang P, Sun Q, Shi L, Yang H
JournalCell Res
PubMed ID28524166
Targeted integration of transgenes can be achieved by strategies based on homologous recombination (HR), microhomology-mediated end joining (MMEJ) or non-homologous end joining (NHEJ). The more generally used HR is inefficient for achieving gene integration in animal embryos and tissues, because it occurs only during cell division, although MMEJ and NHEJ ... More
Characterization of cytoskeleton features and maturation status of cultured human iPSC-derived cardiomyocytes.
AuthorsZuppinger C, Gibbons G, Dutta-Passecker P, Segiser A, Most H, Suter TM
JournalEur J Histochem
PubMed ID28735524
Recent innovations in stem cell technologies and the availability of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have opened new possibilities for studies and drug testing on human cardiomyocytes in vitro. Still, there are concerns about the precise nature of such 'reprogrammed' cells. We have performed an investigation using immunocytochemistry ... More
CRISPR/Cas9 - Mediated Precise Targeted Integration In Vivo Using a Double Cut Donor with Short Homology Arms.
AuthorsYao X, Wang X, Liu J, Hu X, Shi L, Shen X, Ying W, Sun X, Wang X, Huang P, Yang H
JournalEBioMedicine
PubMed ID28527830
Precisely targeted genome editing is highly desired for clinical applications. However, the widely used homology-directed repair (HDR)-based genome editing strategies remain inefficient for certain in vivo applications. We here demonstrate a microhomology-mediated end-joining (MMEJ)-based strategy for precisely targeted gene integration in transfected neurons and hepatocytes in vivo with efficiencies up ... More
A role for the locus coeruleus in the analgesic efficacy of N-acetylaspartylglutamate peptidase (GCPII) inhibitors ZJ43 and 2-PMPA.
N-acetylaspartylglutamate (NAAG) is the third most prevalent and widely distributed neurotransmitter in the mammalian nervous system. NAAG activates a group II metabotropic glutamate receptor (mGluR3) and is inactivated by an extracellular enzyme, glutamate carboxypeptidase II (GCPII) in vivo. Inhibitors of this enzyme are analgesic in animal models of inflammatory, neuropathic and ... More
Shape-dependent antibacterial effects of non-cytotoxic gold nanoparticles.
AuthorsPenders J, Stolzoff M, Hickey DJ, Andersson M, Webster TJ
JournalInt J Nanomedicine
PubMed ID28408817
Gold nanoparticles (AuNPs) of various shapes (including spheres, stars and flowers), with similar dimensions, were synthesized and evaluated for their antibacterial effects toward Staphylococcus aureus, a bacterium responsible for numerous life-threatening infections worldwide. Optical growth curve measurements and Gompertz modeling showed significant AuNP shape- and concentration-dependent decreases in bacterial growth ... More
Cytotrophoblast, Not Syncytiotrophoblast, Dominates Glycolysis and Oxidative Phosphorylation in Human Term Placenta.
AuthorsKolahi KS, Valent AM, Thornburg KL
JournalSci Rep
PubMed ID28230167
The syncytiotrophoblast (SCT) at the maternal-fetal interface has been presumed to be the primary driver of placental metabolism, and the underlying progenitor cytotrophoblast cells (CTB) an insignificant contributor to placental metabolic activity. However, we now show that the metabolic rate of CTB is much greater than the SCT. The oxygen ... More
Myogenic Differentiation from MYOGENIN-Mutated Human iPS Cells by CRISPR/Cas9.
AuthorsHigashioka K, Koizumi N, Sakurai H, Sotozono C, Sato T
JournalStem Cells Int
PubMed ID28473859
It is well known that myogenic regulatory factors encoded by the Myod1 family of genes have pivotal roles in myogenesis, with partially overlapping functions, as demonstrated for the mouse embryo. Myogenin-mutant mice, however, exhibit severe myogenic defects without compensation by other myogenic factors. MYOGENIN might be expected to have an ... More
Spatial organization of the somatosensory cortex revealed by osmFISH.
AuthorsCodeluppi S, Borm LE, Zeisel A, La Manno G, van Lunteren JA, Svensson CI, Linnarsson S
JournalNat Methods
PubMed ID30377364
Global efforts to create a molecular census of the brain using single-cell transcriptomics are producing a large catalog of molecularly defined cell types. However, spatial information is lacking and new methods are needed to map a large number of cell type-specific markers simultaneously on large tissue areas. Here, we describe ... More
easySLM-STED: Stimulated emission depletion microscopy with aberration correction, extended field of view and multiple beam scanning.
AuthorsGörlitz F, Guldbrand S, Runcorn TH, Murray RT, Jaso-Tamame AL, Sinclair HG, Martinez-Perez E, Taylor JR, Neil MAA, Dunsby C, French PMW
JournalJ Biophotonics
PubMed ID29978591
We demonstrate a simplified set-up for STED microscopy with a straightforward alignment procedure that uses a single spatial light modulator (SLM) with collinear incident excitation and depletion beams to provide phase modulation of the beam profiles and correction of optical aberrations. We show that this approach can be used to ... More
Tunneling nanotubes, a novel mode of tumor cell-macrophage communication in tumor cell invasion.
AuthorsHanna SJ, McCoy-Simandle K, Leung E, Genna A, Condeelis J, Cox D
JournalJ Cell Sci
PubMed ID30659112
The interaction between tumor cells and macrophages is crucial in promoting tumor invasion and metastasis. In this study, we examined a novel mechanism of intercellular communication, namely membranous actin-based tunneling nanotubes (TNTs), that occurs between macrophages and tumor cells in the promotion of macrophage-dependent tumor cell invasion. The presence of ... More
Differential contributions of nonmuscle myosin IIA and IIB to cytokinesis in human immortalized fibroblasts.
AuthorsYamamoto K, Otomo K, Nemoto T, Ishihara S, Haga H, Nagasaki A, Murakami Y, Takahashi M
JournalExp Cell Res
PubMed ID30711568
Nonmuscle myosin II (NMII) plays an important role in cytokinesis by constricting a contractile ring. However, it is poorly understood how NMII isoforms contribute to cytokinesis in mammalian cells. Here, we investigated the roles of the two major NMII isoforms, NMIIA and NMIIB, in cytokinesis using a WI-38 VA13 cell ... More
Immunofluorescence and image analysis pipeline for
AuthorsBrown JR, Phongthachit C, Sulkowski MJ
JournalBiol Methods Protoc
PubMed ID31403085
The neuromuscular junction (NMJ) of larval
Cellular and molecular assessment of rose bengal photodynamic antimicrobial therapy on keratocytes, corneal endothelium and limbal stem cell niche.
AuthorsNaranjo A, Pelaez D, Arrieta E, Salero-Coca E, Martinez JD, Sabater AL, Amescua G, Parel JM
JournalExp Eye Res
PubMed ID31539544
Rose Bengal Photodynamic Antimicrobial Therapy (RB-PDAT) is a novel potential treatment for progressive infectious keratitis. The principle behind this therapy is using Rose Bengal as a photosensitizer that can be activated by green light and results in the production of oxygen free radicals which in turn eradicate the microorganism. Given ... More
AuthorsIshii H, Otomo K, Hung JH, Tsutsumi M, Yokoyama H, Nemoto T
JournalBiomed Opt Express
PubMed ID31467771
We developed two-photon excitation stimulated emission depletion (STED) nanoscopy using high-peak-power sub-nanosecond 655-nm pulses. The STED pulse exhibited ideal optical properties and sufficient pulse energy to realize a 70-nm spatial resolution in the compact setup with electrically controllable components. For biological applications, we screened suitable fluorescent dyes or proteins and ... More