scholarly journals NF-κB memory coordinates transcriptional responses to dynamic inflammatory stimuli

2022 ◽  
Author(s):  
Andrew G Wang ◽  
Minjun Son ◽  
Nicholas Thom ◽  
Savas Tay
Keyword(s):  
Single Cells ◽  
Cell Analysis ◽  
Complex Dynamic ◽  
Transcriptional Responses ◽  
Inflammatory Conditions ◽  
Small Molecule Inhibition ◽  
Signal Transduction Networks ◽  
Inflammatory Stimuli ◽  
History Of ◽  
Full Activation

Many scenarios in cellular communication requires cells to interpret multiple dynamic signals. It is unclear how exposure to immune stimuli alters transcriptional responses to subsequent stimulus under inflammatory conditions. Using high-throughput microfluidic live cell analysis, we systematically profiled the NF-κB response to different signal sequences in single cells. We found that NF-κB dynamics stores the history of signals received by cells: depending on the dose and type of prior pathogenic and cytokine signal, the NF-κB response to subsequent stimuli varied widely, from no response to full activation. Using information theory, we revealed that these stimulus-dependent changes in the NF-κB response encode and reflect information about the identity and dose of the prior stimulus. Small-molecule inhibition, computational modeling, and gene expression profiling show that this encoding is driven by stimulus-dependent engagement of negative feedback modules. These results provide a model for how signal transduction networks process sequences of inflammatory stimuli to coordinate cellular responses in complex dynamic environments.

scholarly journals Oral findings during follow-up of nasopharyngeal squamous cell carcinoma treatment: A case report

2021 ◽  
Vol 9 ◽  
pp. 2050313X2110330
Author(s):  
Atsushi Musha ◽  
Nobuteru Kubo ◽  
Naoko Okano ◽  
Hidemasa Kawamura ◽  
Yuhei Miyasaka ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Squamous Cell ◽  
Concurrent Chemoradiotherapy ◽  
Nasopharyngeal Cancer ◽  
Inflammatory Conditions ◽  
Positron Emission ◽  
History Of ◽  
Oral Area

A 50-year-old woman with a long history of nasopharyngeal cancer (T2N2M0, squamous cell carcinoma) underwent chemoradiotherapy and surgery. In the past, to prevent tumor recurrence or metastasis, she underwent concurrent chemoradiotherapy or neck dissection. However, during a follow-up 10 years after the surgery, intense F-18 fluorodeoxyglucose uptake was detected in the oral area (SUVmax 6.0). A biopsy of the area with F-18 fluorodeoxyglucose uptake revealed pathological inflammation. Radiography showed the presence of a wisdom tooth, located at the F-18 fluorodeoxyglucose accumulation site, and pericoronitis of this tooth was detected. Our findings indicate the importance of considering the effect of inflammatory conditions, such as periodontal disease, in using F-18 fluorodeoxyglucose positron emission tomography/computed tomography during follow-up after head and neck cancer treatment.

scholarly journals Microfluidic platform accelerates tissue processing into single cells for molecular analysis and primary culture models

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jeremy A. Lombardo ◽  
Marzieh Aliaghaei ◽  
Quy H. Nguyen ◽  
Kai Kessenbrock ◽  
Jered B. Haun
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Cell Analysis ◽  
Processing Technologies ◽  
Microfluidic Platform ◽  
Tissue Processing ◽  
Single Cell Rna Sequencing ◽  
Culture Models ◽  
Tissue Digestion

AbstractTissues are complex mixtures of different cell subtypes, and this diversity is increasingly characterized using high-throughput single cell analysis methods. However, these efforts are hindered, as tissues must first be dissociated into single cell suspensions using methods that are often inefficient, labor-intensive, highly variable, and potentially biased towards certain cell subtypes. Here, we present a microfluidic platform consisting of three tissue processing technologies that combine tissue digestion, disaggregation, and filtration. The platform is evaluated using a diverse array of tissues. For kidney and mammary tumor, microfluidic processing produces 2.5-fold more single cells. Single cell RNA sequencing further reveals that endothelial cells, fibroblasts, and basal epithelium are enriched without affecting stress response. For liver and heart, processing time is dramatically reduced. We also demonstrate that recovery of cells from the system at periodic intervals during processing increases hepatocyte and cardiomyocyte numbers, as well as increases reproducibility from batch-to-batch for all tissues.

Dermal Discoloration Secondary to Triamcinolone Acetonide Injection Observed in Patients with Autoimmune Disorders

2021 ◽  
Vol 111 (4) ◽  
Author(s):  
James A. Wright ◽  
Jessica A. Wenz ◽  
Gabrielle Jackson Madrigal
Keyword(s):  
Psoriatic Arthritis ◽  
Side Effects ◽  
Autoimmune Disease ◽  
Triamcinolone Acetonide ◽  
Current Literature ◽  
Side Effect ◽  
Inflammatory Conditions ◽  
Rare Side Effect ◽  
History Of ◽  
Synthetic Glucocorticoid

Triamcinolone acetonide is a synthetic glucocorticoid used to treat numerous acute and chronic inflammatory conditions. The various side effects of this drug from parenteral administration are well documented in the literature. In this study, three patients present with a rare side effect of violaceous dermal pigmentation. To the best of the authors' knowledge, this finding is rarely presented in the current literature. The purpose of this study is to provide awareness of a less-documented, delayed side effect from triamcinolone acetonide administration. Although all patients presenting in this study had a known history of autoimmune disease (eg, lupus, psoriatic arthritis) further research is needed to suggest a possible association between dermal violaceous change and the use of triamcinolone.

scholarly journals More than efficacy revealed by single-cell analysis of antiviral therapeutics

2019 ◽  
Author(s):  
Wu Liu ◽  
Mehmet U. Caglar ◽  
Zhangming Mao ◽  
Andrew Woodman ◽  
Jamie J. Arnold ◽  
...  
Keyword(s):  
Single Cell ◽  
Drug Targets ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Principal Component ◽  
Viral Population ◽  
Cell Analysis ◽  
Toxic Dose ◽  
Time Required

SUMMARYDevelopment of antiviral therapeutics emphasizes minimization of the effective dose and maximization of the toxic dose, first in cell culture and later in animal models. Long-term success of an antiviral therapeutic is determined not only by its efficacy but also by the duration of time required for drug-resistance to evolve. We have developed a microfluidic device comprised of ~6000 wells, with each well containing a microstructure to capture single cells. We have used this device to characterize enterovirus inhibitors with distinct mechanisms of action. In contrast to population methods, single-cell analysis reveals that each class of inhibitor interferes with the viral infection cycle in a manner that can be distinguished by principal component analysis. Single-cell analysis of antiviral candidates reveals not only efficacy but also properties of the members of the viral population most sensitive to the drug, the stage of the lifecycle most affected by the drug, and perhaps even if the drug targets an interaction of the virus with its host.

scholarly journals μCB-seq: Microfluidic cell barcoding and sequencing for high-resolution imaging and sequencing of single cells

2020 ◽  
Author(s):  
Tyler N. Chen ◽  
Anushka Gupta ◽  
Mansi Zalavadia ◽  
Aaron M. Streets
Keyword(s):  
High Resolution ◽  
Single Cell ◽  
Single Cell Analysis ◽  
Protein Localization ◽  
Single Cells ◽  
Optical Measurements ◽  
Detection Sensitivity ◽  
High Resolution Imaging ◽  
Cell Analysis ◽  
Resolution Imaging

AbstractSingle-cell RNA sequencing (scRNA-seq) enables the investigation of complex biological processes in multicellular organisms with high resolution. However, many phenotypic features that are critical to understanding the functional role of cells in a heterogeneous tissue or organ are not directly encoded in the genome and therefore cannot be profiled with scRNA-seq. Quantitative optical microscopy has long been a powerful approach for characterizing diverse cellular phenotypes including cell morphology, protein localization, and chemical composition. Combining scRNA-seq with optical imaging has the potential to provide comprehensive single-cell analysis, allowing for functional integration of gene expression profiling and cell-state characterization. However, it is difficult to track single cells through both measurements; therefore, coupling current scRNA-seq protocols with optical measurements remains a challenge. Here, we report Microfluidic Cell Barcoding and Sequencing (μCB-seq), a microfluidic platform that combines high-resolution imaging and sequencing of single cells. μCB-seq is enabled by a novel fabrication method that preloads primers with known barcode sequences inside addressable reaction chambers of a microfluidic device. In addition to enabling multi-modal single-cell analysis, μCB-seq improves gene detection sensitivity, providing a scalable and accurate method for information-rich characterization of single cells.

scholarly journals Enhancer priming enables fast and sustained transcriptional responses to Notch signaling

2018 ◽  
Author(s):  
Julia Falo-Sanjuan ◽  
Nicholas C Lammers ◽  
Hernan G Garcia ◽  
Sarah Bray
Keyword(s):  
Transcription Factors ◽  
Dynamic Response ◽  
Notch Signaling ◽  
Real Time ◽  
Single Cells ◽  
Transcriptional Responses ◽  
Sustained Activity ◽  
Developmental Signaling ◽  
Cellular Context ◽  
Nascent Transcripts

SummaryInformation from developmental signaling pathways must be accurately decoded to generate transcriptional outcomes. In the case of Notch, the intracellular domain (NICD) transduces the signal directly to the nucleus. How enhancers decipher NICD in the real time of developmental decisions is not known. Using the MS2/MCP system to visualize nascent transcripts in single cells in Drosophila embryos we reveal how two target enhancers read Notch activity to produce synchronized and sustained profiles of transcription. By manipulating the levels of NICD and altering specific motifs within the enhancers we uncover two key principles. First, increased NICD levels alter transcription by increasing duration rather than frequency of transcriptional bursts. Second, priming of enhancers by tissue-specific transcription factors is required for NICD to confer synchronized and sustained activity; in their absence, transcription is stochastic and bursty. The dynamic response of an individual enhancer to NICD thus differs depending on the cellular context.

Ruthenium red: a highly efficient and versatile cell staining agent for single-cell analysis using inductively coupled plasma time-of-flight mass spectrometry

The Analyst ◽  
2021 ◽  
Author(s):  
Wen Qin ◽  
Hans-Joachim Stärk ◽  
Thorsten Reemtsma
Keyword(s):  
Inductively Coupled Plasma ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Ruthenium Red ◽  
Cell Analysis ◽  
Inductively Coupled ◽  
Flight Mass Spectrometry ◽  
Cell Staining ◽  
A Cell ◽  
Tof Ms

A new method for determining the concentration of elements in single cells by the SC-ICP-TOF-MS method based on a metal-containing stain as a cell volume proxy has been developed and validated.

scholarly journals Methods and sensors for functional genomic studies of cell-cycle transitions in single cells

2020 ◽  
Vol 52 (10) ◽  
pp. 468-477
Author(s):  
Alexander C. Zambon ◽  
Tom Hsu ◽  
Seunghee Erin Kim ◽  
Miranda Klinck ◽  
Jennifer Stowe ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Single Cell ◽  
Single Cell Analysis ◽  
Imaging System ◽  
Single Cells ◽  
Cell Types ◽  
Cell Analysis ◽  
Mcf7 Cells ◽  
Genomic Studies ◽  
Cell Subpopulations

Much of our understanding of the regulatory mechanisms governing the cell cycle in mammals has relied heavily on methods that measure the aggregate state of a population of cells. While instrumental in shaping our current understanding of cell proliferation, these approaches mask the genetic signatures of rare subpopulations such as quiescent (G0) and very slowly dividing (SD) cells. Results described in this study and those of others using single-cell analysis reveal that even in clonally derived immortalized cancer cells, ∼1–5% of cells can exhibit G0 and SD phenotypes. Therefore to enable the study of these rare cell phenotypes we established an integrated molecular, computational, and imaging approach to track, isolate, and genetically perturb single cells as they proliferate. A genetically encoded cell-cycle reporter (K67p-FUCCI) was used to track single cells as they traversed the cell cycle. A set of R-scripts were written to quantify K67p-FUCCI over time. To enable the further study G0 and SD phenotypes, we retrofitted a live cell imaging system with a micromanipulator to enable single-cell targeting for functional validation studies. Single-cell analysis revealed HT1080 and MCF7 cells had a doubling time of ∼24 and ∼48 h, respectively, with high duration variability in G1 and G2 phases. Direct single-cell microinjection of mRNA encoding (GFP) achieves detectable GFP fluorescence within ∼5 h in both cell types. These findings coupled with the possibility of targeting several hundreds of single cells improves throughput and sensitivity over conventional methods to study rare cell subpopulations.

scholarly journals Synchrotron macro ATR-FTIR microspectroscopy for high-resolution chemical mapping of single cells

The Analyst ◽  
2019 ◽  
Vol 144 (10) ◽  
pp. 3226-3238 ◽  
Author(s):  
Jitraporn Vongsvivut ◽  
David Pérez-Guaita ◽  
Bayden R. Wood ◽  
Philip Heraud ◽  
Karina Khambatta ◽  
...  
Keyword(s):  
High Resolution ◽  
Single Cell ◽  
Spatial Resolution ◽  
Environmental Science ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Cell Analysis ◽  
Chemical Mapping ◽  
Ftir Microspectroscopy ◽  
Resolution Single

Coupling synchrotron IR beam to an ATR element enhances spatial resolution suited for high-resolution single cell analysis in biology, medicine and environmental science.

Differential Processing of Noxious Colonic Input by Thoracolumbar and Lumbosacral Dorsal Horn Neurons in the Rat

2005 ◽  
Vol 94 (6) ◽  
pp. 3788-3794 ◽  
Author(s):  
Gexin Wang ◽  
Bin Tang ◽  
Richard J. Traub
Keyword(s):  
Dorsal Horn ◽  
Single Cells ◽  
Male Rats ◽  
Colonic Inflammation ◽  
Dorsal Horn Neurons ◽  
Inflammatory Stimuli ◽  
Before And After ◽  
Spinal Segments ◽  
Colorectal Distention ◽  
Multiple Cells

Previous studies suggest the lumbosacral (LS) spinal cord processes acute colorectal stimuli whereas the thoracolumbar (TL) and LS spinal segments process inflammatory stimuli. In this study, the effects of colorectal distention (CRD) on TL and LS dorsal horn neuronal activity were recorded in Nembutal-anesthetized male rats both with and without colonic inflammation. Both single cells (before and after inflammation) and populations (multiple cells from noninflamed or inflamed rats) were studied. CRD-responsive neurons had excitatory Abrupt (on–off with stimulus) or Sustained (prolonged after discharge) responses or were Inhibited by CRD. In noninflamed rats, a significantly greater percentage of LS neurons (63% Abrupt, 27% Sustained) were excited by CRD than TL neurons (61% Abrupt, 3% Sustained). The remaining cells were Inhibited (10% LS, 36% TL). LS Abrupt neurons had lower thresholds and greater response magnitudes to CRD compared with TL Abrupt neurons. After colonic inflammation, TL neurons became more excitable: the percentage of Inhibited neurons decreased, the response magnitude of Abrupt neurons increased, and the threshold decreased. In contrast, in single-cell recordings, the response of LS Sustained neurons increased, whereas LS Abrupt neurons decreased. These data suggest that in noninflamed rats, the net response to CRD of TL visceroceptive spinal sensory neurons is less than that of LS neurons. Colonic inflammation increases the net response of TL neurons and differentially modulates the response of LS neurons. These differences may contribute to the functional dichotomy between the TL and LS spinal segments in processing acute and inflammatory colorectal pain.

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