scholarly journals 344. En Route to Non-Viral Genetic Engineering: Kinetics of DNA Uptake and Transgene Expression Following Repeated Transfection with Multiple Episomal Plasmids in Human Primary Fibroblast

2016 ◽  
Vol 24 ◽  
pp. S138
Author(s):  
Charlie Y. Hsu ◽  
Derrick E. Rancourt
Keyword(s):  
Genetic Engineering ◽  
Transgene Expression ◽  
Dna Uptake ◽  
Primary Fibroblast ◽  
Human Primary Fibroblast ◽  
Kinetics Of

Expression of the familial Mediterranean fever gene and activity of the C5a inhibitor in human primary fibroblast cultures

Blood ◽  
2000 ◽  
Vol 96 (2) ◽  
pp. 727-731 ◽  
Author(s):  
Yaacov Matzner ◽  
Suzan Abedat ◽  
Eli Shapiro ◽  
Shlomit Eisenberg ◽  
Ariela Bar-Gil-Shitrit ◽  
...  
Keyword(s):  
Familial Mediterranean Fever ◽  
Human Fibroblasts ◽  
Skin Fibroblasts ◽  
Sterile Inflammation ◽  
Inhibitor Activity ◽  
Primary Fibroblast ◽  
Fibroblast Cultures ◽  
M694v Mutation ◽  
Human Primary Fibroblast ◽  
Mediterranean Fever

Abstract Familial Mediterranean fever (FMF) is an inherited disease whose manifestations are acute but reversible attacks of sterile inflammation affecting synovial and serosal spaces. The FMF gene (MEFV) was recently cloned, and it codes for a protein (pyrin/marenostrin) homologous to known nuclear factors. We previously reported the deficient activity of a C5a/interleukin (IL)–8 inhibitor, a physiologic regulator of inflammatory processes, in FMF serosal and synovial fluids. We now describe the concomitant expression ofMEFV and C5a/IL-8–inhibitor activity in primary cultures of human fibroblasts. Fibroblasts grown from synovial and peritoneal tissues displayed C5a/IL-8–inhibitor activity that could be further induced with phorbol myristate acetate (PMA) and IL-1β. Very low levels of chemotactic inhibitor were evident in skin fibroblast cultures or in peritoneal and skin fibroblasts obtained from FMF patients. MEFV was expressed in peritoneal and skin fibroblasts at a lower level than in neutrophils and could be further induced by PMA and IL-1β. In the FMF cultures, the MEFV transcript carried the M694V mutation, consistent with the genetic defect found in patients with this disease. MEFV was also expressed in other cell lines that do not produce C5a/IL-8 inhibitor. These findings suggest that human primary fibroblast cultures express MEFV and produce C5a/IL-8–inhibitor activity. The interrelationship between pyrin, the MEFV product, and the C5a/IL-8 inhibitor requires further investigation.

scholarly journals Genetic engineering for in vivo optical interrogation of neuronal responses to cell type-specific silencing

2021 ◽  
Author(s):  
Firat Terzi ◽  
Johannes Knabbe ◽  
Sidney B. Cambridge
Keyword(s):  
High Resolution ◽  
Genetic Engineering ◽  
Transgene Expression ◽  
Neuronal Morphology ◽  
Dependent Manner ◽  
Cell Type ◽  
Fluorescent Marker ◽  
Genetic Perturbations ◽  
Cell Type Specific

SummaryGenetic engineering of quintuple transgenic brain tissue was used to establish a low background, Cre-dependent version of the inducible Tet-On system for fast, cell type-specific transgene expression in vivo. Co-expression of a constitutive, Cre-dependent fluorescent marker selectively allowed single cell analyses before and after inducible, tet-dependent transgene expression. Here, we used this method for acute, high-resolution manipulation of neuronal activity in the living brain. Single induction of the potassium channel Kir2.1 produced cell type-specific silencing within hours that lasted for at least three days. Longitudinal in vivo imaging of spontaneous calcium transients and neuronal morphology demonstrated that prolonged silencing did not alter spine densities or synaptic input strength. Furthermore, selective induction of Kir2.1 in parvalbumin interneurons increased the activity of surrounding neurons in a distance-dependent manner. This high-resolution, inducible interference and interval imaging of individual cells (high I5, ‘HighFive’) method thus allows visualizing temporally precise, genetic perturbations of defined cells.

scholarly journals Bioprocess and genetic engineering aspects of ascomycin production: a review

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Krishika Sambyal ◽  
Rahul Vikram Singh
Keyword(s):  
Genetic Engineering ◽  
Biosynthetic Pathway ◽  
Genetic Manipulation ◽  
Screening Method ◽  
Structural Complexity ◽  
Biological Properties ◽  
Therapeutic Effects ◽  
Main Body ◽  
Kinetics Of ◽  
Biosynthetic Machinery

Abstract Background Ascomycin is a highly valuable multifunctional drug which exhibits numerous biological properties. Being an immunosuppressant, it is known to prevent graft rejection in humans and has potential to treat varying skin ailments. Its derivatives represent a novel class of anti-inflammatory macrolactams. But the biosynthetic machinery of ascomycin is still unclear. Main body of the abstract Due to the structural complexity, there occurs difficulty in its chemical synthesis; therefore, microbial production has been preferred by using Streptomyces hygroscopicus subsp. ascomyceticus. Through several genetic manipulation and mutagenesis techniques, the yield can be increased by several folds without any difficulties. Genetic engineering has played a significant role in understanding the biosynthetic pathway of ascomycin. Short conclusion Recently, many efforts have been made to utilize the therapeutic effects of ascomycin and its derivatives. This article covers concepts related to the production kinetics of ascomycin including an update of the ongoing yield improvement techniques as well as screening method of novel strains for ascomycin production.

scholarly journals A dynamical systems model for the measurement of cellular senescence

2019 ◽  
Vol 16 (159) ◽  
pp. 20190311 ◽  
Author(s):  
Daniel Galvis ◽  
Darren Walsh ◽  
Lorna W. Harries ◽  
Eva Latorre ◽  
James Rankin
Keyword(s):  
Dynamical Systems ◽  
Replicative Senescence ◽  
Population Doubling ◽  
High Temporal Resolution ◽  
Model Parameters ◽  
Ki 67 ◽  
Primary Fibroblast ◽  
Systems Model ◽  
Human Primary Fibroblast

Senescent cells provide a good in vitro model to study ageing. However, cultures of ‘senescent’ cells consist of a mix of cell subtypes (proliferative, senescent, growth-arrested and apoptotic). Determining the proportion of senescent cells is crucial for studying ageing and developing new anti-degenerative therapies. Commonly used markers such as doubling population, senescence-associated β-galactosidase, Ki-67, γH2AX and TUNEL assays capture diverse and overlapping cellular populations and are not purely specific to senescence. A newly developed dynamical systems model follows the transition of an initial culture to senescence tracking population doubling, and the proportion of cells in proliferating, growth-arrested, apoptotic and senescent states. Our model provides a parsimonious description of transitions between these states accruing towards a predominantly senescent population. Using a genetic algorithm, these model parameters are well constrained by an in vitro human primary fibroblast dataset recording five markers at 16 time points. The computational model accurately fits to the data and translates these joint markers into the first complete description of the proportion of cells in different states over the lifetime. The high temporal resolution of the dataset demonstrates the efficacy of strategies for reconstructing the trajectory towards replicative senescence with a minimal number of experimental recordings.

Sugarcane genetic engineering research in South Africa: From gene discovery to transgene expression

Sugar Tech ◽  
2010 ◽  
Vol 12 (2) ◽  
pp. 85-90 ◽  
Author(s):  
D. A. Watt ◽  
D. L. Sweby ◽  
B. A. M. Potier ◽  
S. J. Snyman
Keyword(s):  
South Africa ◽  
Genetic Engineering ◽  
Transgene Expression ◽  
Gene Discovery ◽  
Engineering Research

Expression of the familial Mediterranean fever gene and activity of the C5a inhibitor in human primary fibroblast cultures

Blood ◽  
2000 ◽  
Vol 96 (2) ◽  
pp. 727-731 ◽  
Author(s):  
Yaacov Matzner ◽  
Suzan Abedat ◽  
Eli Shapiro ◽  
Shlomit Eisenberg ◽  
Ariela Bar-Gil-Shitrit ◽  
...  
Keyword(s):  
Familial Mediterranean Fever ◽  
Human Fibroblasts ◽  
Skin Fibroblasts ◽  
Sterile Inflammation ◽  
Inhibitor Activity ◽  
Primary Fibroblast ◽  
Fibroblast Cultures ◽  
M694v Mutation ◽  
Human Primary Fibroblast ◽  
Mediterranean Fever

Familial Mediterranean fever (FMF) is an inherited disease whose manifestations are acute but reversible attacks of sterile inflammation affecting synovial and serosal spaces. The FMF gene (MEFV) was recently cloned, and it codes for a protein (pyrin/marenostrin) homologous to known nuclear factors. We previously reported the deficient activity of a C5a/interleukin (IL)–8 inhibitor, a physiologic regulator of inflammatory processes, in FMF serosal and synovial fluids. We now describe the concomitant expression ofMEFV and C5a/IL-8–inhibitor activity in primary cultures of human fibroblasts. Fibroblasts grown from synovial and peritoneal tissues displayed C5a/IL-8–inhibitor activity that could be further induced with phorbol myristate acetate (PMA) and IL-1β. Very low levels of chemotactic inhibitor were evident in skin fibroblast cultures or in peritoneal and skin fibroblasts obtained from FMF patients. MEFV was expressed in peritoneal and skin fibroblasts at a lower level than in neutrophils and could be further induced by PMA and IL-1β. In the FMF cultures, the MEFV transcript carried the M694V mutation, consistent with the genetic defect found in patients with this disease. MEFV was also expressed in other cell lines that do not produce C5a/IL-8 inhibitor. These findings suggest that human primary fibroblast cultures express MEFV and produce C5a/IL-8–inhibitor activity. The interrelationship between pyrin, the MEFV product, and the C5a/IL-8 inhibitor requires further investigation.

Hepatic injury and the kinetics of bone marrow-derived hepatocyte transgene expression

2008 ◽  
Vol 43 (8) ◽  
pp. 1511-1519 ◽  
Author(s):  
Scott V. Perryman ◽  
D. Denny Jenkins ◽  
Konrad L. Streetz ◽  
Michael T. Longaker ◽  
Karl G. Sylvester
Keyword(s):  
Bone Marrow ◽  
Transgene Expression ◽  
Hepatic Injury ◽  
Kinetics Of

scholarly journals Biased Allelic Expression in Human Primary Fibroblast Single Cells

2015 ◽  
Vol 96 (1) ◽  
pp. 70-80 ◽  
Author(s):  
Christelle Borel ◽  
Pedro G. Ferreira ◽  
Federico Santoni ◽  
Olivier Delaneau ◽  
Alexandre Fort ◽  
...  
Keyword(s):  
Single Cells ◽  
Allelic Expression ◽  
Primary Fibroblast ◽  
Human Primary Fibroblast

scholarly journals Grammatophyllum speciosum Ethanolic Extract Promotes Wound Healing in Human Primary Fibroblast Cells

2018 ◽  
Vol 2018 ◽  
pp. 1-6
Author(s):  
Saraporn Harikarnpakdee ◽  
Verisa Chowjarean
Keyword(s):  
Wound Healing ◽  
Ethanolic Extract ◽  
Skin Wound ◽  
Total Phenolic ◽  
Fibroblast Cells ◽  
Skin Wound Healing ◽  
Primary Fibroblast ◽  
Human Primary Fibroblast ◽  
Phytochemical Compounds

Grammatophyllum speciosum is a plant in Orchidaceae family which contains a variety of phytochemical compounds that might be beneficial for medicinal use. This study aimed to evaluate the activity of pseudobulb of G. speciosum extract (GSE) in wound healing processes in human primary fibroblast cells along with in vitro antioxidant activity and total phenolic content of GSE. Scratch wound healing assay indicated that GSE was capable of increasing migration rate after 6 and 9 hours of treatment. Besides, the extract was able to scavenge DPPH, ABTS, and superoxide anion radicals indicating the antioxidative property of GSE. This study suggested a novel role of the of pseudobulb extract of G. speciosum as a wound healing enhancer. The results from this study might be beneficial for the development of further novel active compounds for skin wound healing.

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