scholarly journals The effect of isoferritins on granulopoiesis

Blood ◽  
1986 ◽  
Vol 67 (2) ◽  
pp. 436-443
Author(s):  
G Sala ◽  
M Worwood ◽  
A Jacobs
Keyword(s):  
Monoclonal Antibody ◽  
Colony Formation ◽  
Colony Growth ◽  
Conditioned Media ◽  
Regulatory Role ◽  
Consistent Effect ◽  
Cell Extracts ◽  
In Vitro Inhibition

The evidence for a regulatory role of acidic isoferritins on hemopoiesis is not entirely consistent with our knowledge of ferritin biochemistry, and no clear picture of this phenomeonon has emerged. In the present study, we have been unable to confirm a consistent effect of purified heart (acidic), spleen (basic), or serum (glycosylated) isoferritins on CFU-GM colony formation in vitro. Inhibition of colony formation by cell extracts or conditioned media does not relate to the presence of acidic isoferritins, nor is this effect neutralized by a monoclonal antibody to acidic isoferritins. The composition of ferritin preparations previously described as inhibitory to CFU-GM colony growth could not be confirmed, and they were not found to be predominantly acidic in nature. Our data do not support a role for acidic isoferritins as inhibitors of granulopoiesis.

scholarly journals The effect of isoferritins on granulopoiesis

Blood ◽  
1986 ◽  
Vol 67 (2) ◽  
pp. 436-443 ◽  
Author(s):  
G Sala ◽  
M Worwood ◽  
A Jacobs
Keyword(s):  
Monoclonal Antibody ◽  
Colony Formation ◽  
Colony Growth ◽  
Conditioned Media ◽  
Regulatory Role ◽  
Consistent Effect ◽  
Cell Extracts ◽  
In Vitro Inhibition

Abstract The evidence for a regulatory role of acidic isoferritins on hemopoiesis is not entirely consistent with our knowledge of ferritin biochemistry, and no clear picture of this phenomeonon has emerged. In the present study, we have been unable to confirm a consistent effect of purified heart (acidic), spleen (basic), or serum (glycosylated) isoferritins on CFU-GM colony formation in vitro. Inhibition of colony formation by cell extracts or conditioned media does not relate to the presence of acidic isoferritins, nor is this effect neutralized by a monoclonal antibody to acidic isoferritins. The composition of ferritin preparations previously described as inhibitory to CFU-GM colony growth could not be confirmed, and they were not found to be predominantly acidic in nature. Our data do not support a role for acidic isoferritins as inhibitors of granulopoiesis.

scholarly journals The regulatory role of interleukin 2-responsive T lymphocytes on early and mature erythroid progenitor proliferation

Blood ◽  
1986 ◽  
Vol 67 (6) ◽  
pp. 1607-1610
Author(s):  
Z Estrov ◽  
C Roifman ◽  
YP Wang ◽  
T Grunberger ◽  
EW Gelfand ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Colony Formation ◽  
Interleukin 2 ◽  
Erythroid Differentiation ◽  
Colony Growth ◽  
Dependent Manner ◽  
Erythroid Progenitor ◽  
Recombinant Interleukin 2

To analyze the role of T lymphocytes in human erythropoiesis, we evaluated the effect of recombinant interleukin 2 (IL 2) on marrow CFU- E and BFU-E colony formation in vitro. IL 2 resulted in an increase in CFU-E and BFU-E colony numbers in a dose-dependent manner. This increase could be prevented by anti-Tac, a monoclonal antibody to the IL 2 receptor. Moreover, anti-Tac on its own resulted in an overall decrease in colony numbers. Depletion of marrow adherent cells did not alter the effect of either IL 2 or anti-Tac on colony growth. Following the removal of marrow T lymphocytes, CFU-E and BFU-E colony formation proceeded normally; however, the effects of IL 2 and anti-Tac were markedly diminished. Readdition of T lymphocytes to the cultures restored the IL 2 effect. Although T lymphocytes were not themselves essential for in vitro erythropoiesis, our studies suggest that IL 2 and IL 2-responsive T cells can regulate both early and mature stages of erythroid differentiation.

scholarly journals The regulatory role of interleukin 2-responsive T lymphocytes on early and mature erythroid progenitor proliferation

Blood ◽  
1986 ◽  
Vol 67 (6) ◽  
pp. 1607-1610 ◽  
Author(s):  
Z Estrov ◽  
C Roifman ◽  
YP Wang ◽  
T Grunberger ◽  
EW Gelfand ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Colony Formation ◽  
Interleukin 2 ◽  
Erythroid Differentiation ◽  
Colony Growth ◽  
Dependent Manner ◽  
Erythroid Progenitor ◽  
Recombinant Interleukin 2

Abstract To analyze the role of T lymphocytes in human erythropoiesis, we evaluated the effect of recombinant interleukin 2 (IL 2) on marrow CFU- E and BFU-E colony formation in vitro. IL 2 resulted in an increase in CFU-E and BFU-E colony numbers in a dose-dependent manner. This increase could be prevented by anti-Tac, a monoclonal antibody to the IL 2 receptor. Moreover, anti-Tac on its own resulted in an overall decrease in colony numbers. Depletion of marrow adherent cells did not alter the effect of either IL 2 or anti-Tac on colony growth. Following the removal of marrow T lymphocytes, CFU-E and BFU-E colony formation proceeded normally; however, the effects of IL 2 and anti-Tac were markedly diminished. Readdition of T lymphocytes to the cultures restored the IL 2 effect. Although T lymphocytes were not themselves essential for in vitro erythropoiesis, our studies suggest that IL 2 and IL 2-responsive T cells can regulate both early and mature stages of erythroid differentiation.

scholarly journals Nerve Growth Factor (NGF) modulates in vitro induced myofibroblasts by highlighting a differential protein signature

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Graziana Esposito ◽  
Bijorn Omar Balzamino ◽  
Egidio Stigliano ◽  
Filippo Biamonte ◽  
Andrea Urbani ◽  
...  
Keyword(s):  
Nuclear Translocation ◽  
Cell Signalling ◽  
Specific Protein ◽  
Conditioned Media ◽  
Cell Extracts ◽  
Dose Dependent Decrease ◽  
Significant Expression ◽  
Biomolecular Analysis ◽  
Protein Signature

AbstractWe previously described the profibrogenic effect of NGF on conjunctival Fibroblasts (FBs) and its ability to trigger apoptosis in TGFβ1-induced myofibroblasts (myoFBs). Herein, cell apoptosis/signalling, cytokines’ signature in conditioned media and inflammatory as well as angiogenic pathway were investigated. Experimental myoFBs were exposed to NGF (0.1–100 ng/mL), at defined time-point for confocal and biomolecular analysis. Cells were analysed for apoptotic and cell signalling activation in cell extracts and for some inflammatory and proinflammatory/angiogenic factors’ activations. NGF triggered cJun overexpression and phospho-p65-NFkB nuclear translocation. A decreased Bcl2:Bax ratio and a significant expression of smad7 were confirmed in early AnnexinV-positive myoFBs. A specific protein signature characterised the conditioned media: a dose dependent decrease occurred for IL8, IL6 while a selective increase was observed for VEGF and cyr61 (protein/mRNA). TIMP1 levels were unaffected. Herein, NGF modulation of smad7, the specific IL8 and IL6 as well as VEGF and cyr61 modulation deserve more attention as opening to alternative approaches to counteract fibrosis.

scholarly journals Influence of Gelatin-Thrombin Matrix Tissue Sealant on Bacterial Colony Formation and Risk of Pelvic Infection

2016 ◽  
Vol 2016 ◽  
pp. 1-6
Author(s):  
Michael J. Jarrett ◽  
Andres Vázquez-Torres ◽  
Daniel N. Frank ◽  
Bruce D. McCollister ◽  
Patrick K. Henthorn ◽  
...  
Keyword(s):  
Colony Formation ◽  
Rrna Gene ◽  
Contributing Factors ◽  
Vaginal Cuff ◽  
Pelvic Infection ◽  
Bacterial Colony ◽  
E Coli ◽  
Tissue Sealant

Objective. Gelatin-thrombin matrix (GTM) tissue sealant use was previously identified as an independent predictor of pelvic infection following hysterectomies. We aim to elucidate contributing factors by assessing influence of GTM on bacterial colony formation and characterizing bacteria present at the vaginal cuff.Methods.Escherichia coliwas incubated in phosphate-buffered saline (PBS) and pelvic washings with and without GTM to assess influence on colony formation. Pelvic washings of the vaginal cuff were collected from hysterectomies occurring from June through October 2015.In vitrotechniques, 16S rRNA gene qPCR, and 16S amplicon sequencing were performed with washings to characterize bacteria at the vaginal cuff.Results. Mean bacterial colony formation in PBS was greater forE. coliincubated in the presence of GTM (1.48 × 107 CFU/mL) versus without (9.95 × 105 CFU/mL) following 20-hour incubation (p=0.001). Out of 61 pelvic washings samples, 3 were culture positive (≥5000 CFU/mL) withEnterococcus faecalis.Conclusion.In vitroexperiments support a facilitating role of GTM on colony formation ofE. coliin PBS. However, given the negative results of surgical site washings following adequate disinfection, the role of GTM in promoting posthysterectomy pelvic infections may be limited. Analysis of pelvic washings revealed presence ofE. faecalis, but results were inconclusive. Further studies are recommended.

scholarly journals Postrecruitment Function of Yeast Med6 Protein during the Transcriptional Activation by Mediator Complex

2018 ◽  
Vol 2018 ◽  
pp. 1-9
Author(s):  
Gwang Sik Kim ◽  
Young Chul Lee
Keyword(s):  
Gene Expression ◽  
Reporter Gene ◽  
Transcriptional Activation ◽  
Reporter Gene Expression ◽  
Temperature Sensitive ◽  
Internal Deletion ◽  
Cell Extracts ◽  
Evolutionarily Conserved

Med6 protein (Med6p) is a hallmark component of evolutionarily conserved Mediator complexes, and the genuine role of Med6p in Mediator functions remains elusive. For the functional analysis ofSaccharomyces cerevisiaeMed6p (scMed6p), we generated a series of scMed6p mutants harboring a small internal deletion. Genetic analysis of these mutants revealed that three regions (amino acids 33–42 (Δ2), 125–134 (Δ5), and 157–166 (Δ6)) of scMed6p are required for cell viability and are located at highly conserved regions of Med6 homologs. Notably, the Med6p-Δ2 mutant was barely detectable in whole-cell extracts and purified Mediator, suggesting a loss of Mediator association and concurrent rapid degradation. Consistent with this, the recombinant forms of Med6p having these mutations partially (Δ2) restore or fail (Δ5 and Δ6) to restore in vitro transcriptional defects caused by temperature-sensitivemed6mutation. In an artificial recruitment assay, Mediator containing a LexA-fused wild-type Med6p or Med6p-Δ5 was recruited to thelexAoperator region with TBP and activated reporter gene expression. However, the recruitment of Mediator containing LexA-Med6p-Δ6 tolexAoperator region resulted in neither TBP recruitment nor reporter gene expression. This result demonstrates a pivotal role of Med6p in the postrecruitment function of Mediator, which is essential for transcriptional activation by Mediator.

scholarly journals Investigations on the Role of the MicroRNA-338-5p/Wnt Family Member 2B (WNT2B) Axis in Regulating the Pathogenesis of Nasopharyngeal Carcinoma (NPC)

2021 ◽  
Vol 11 ◽  
Author(s):  
Suzhen Wang ◽  
Tianning Yang ◽  
Zhengxiang He
Keyword(s):  
Family Member ◽  
Colony Formation ◽  
Epithelial Mesenchymal Transition ◽  
Colony Formation Assay ◽  
Transwell Assay ◽  
Mesenchymal Transition ◽  
Downstream Target

BackgroundThe involvement of microRNA-338-5p in modulating NPC pathogenesis is still largely unknown, and this study aimed to investigate this issue.MethodsThe expressions of cancer associated genes were determined by Real-Time qPCR and Western Blot, and cell apoptosis was determined by flow cytometer (FCM). CCK-8 assay and colony formation assay were respectively used to determine cell proliferation and colony formation abilities. Transwell assay was used to evaluate cell migration. The expression levels of Ki67 protein in mice tissues were measured by Immunohistochemistry (IHC) assay.ResultsThe present study found that microRNA-338-5p suppressed NPC progression by degrading its downstream target, Wnt family member 2B (WNT2B). Specifically, microRNA-338-5p tended to be low-expressed in NPC tissues and cell lines, compared to the non-tumor nasopharyngeal mucosa tissues and normal nasopharyngeal cell line (NP69). Upregulation of microRNA-338-5p inhibited proliferation, mobility, and epithelial-mesenchymal transition (EMT) in NPC cells in vitro, while silencing of microRNA-338-5p had opposite effects. Consistently, microRNA-338-5p suppressed tumorigenesis of NPC cells in vivo. In addition, microRNA-338-5p targeted WNT2B for degradation and inhibition, and the inhibiting effects of microRNA-338-5p overexpression on NPC development were reversed by upregulating WNT2B.ConclusionsTaken together, we concluded that microRNA-338-5p targeted WNT2B to hinder NPC development.

Sign in / Sign up

Export Citation Format

Share Document