Globin synthesis in fibroblasts fused with erythroblasts. II. Human fibroblasts

1975 ◽  
Vol 1 (4) ◽  
pp. 323-333
Author(s):  
Elena Citkowitz ◽  
Michael G. Riggs ◽  
Vernon M. Ingram
Keyword(s):  
Human Fibroblasts ◽  
Globin Synthesis

Production of mouse globin in heterokaryons of mouse erythroleukaemia cells and human fibroblasts

1977 ◽  
Vol 26 (1) ◽  
pp. 347-357
Author(s):  
B.P. Alter ◽  
S.C. Goff ◽  
D.G. Hillman ◽  
A.B. Deisseroth ◽  
B.G. Forget
Keyword(s):  
Amino Acids ◽  
Human Fibroblasts ◽  
Fusion System ◽  
Globin Genes ◽  
Polyacrylamide Gels ◽  
Globin Mrna ◽  
Globin Chains ◽  
Cytoplasmic Rna ◽  
Friend Cells ◽  
Globin Synthesis

In an effort to activate the globin genes of non-erythroid cells, tetraploid murine erythroleukaemia cells (Friend cells) were fused with diploid human amniotic fibroblasts. When the Friend cells were pretreated with dimethylsulphoxide, an average of 27% heterokaryons was observed. These cells stained with benzidine, an indication that they contained haemoglobin. The cells incorporated radioactive amino acids into proteins. Electrophoresis of [3H]leucine-labelled lysates on SDS urea polyacrylamide gels indicated that up to 7% of the newly synthesized protein co-electrophoresed with globin. CM cellulose chromatography demonstrated the presence of mouse but not human globin chains. Hybridization analyses of cytoplasmic RNA also revealed only mouse globin mRNA in the heterokaryons. Although heterokaryons form readily between mouse erythroleukaemia cells and human fibroblasts, and globin synthesis does occur, only the erythroid partner in the fusion system employed here directs globin production.

The Growth of Herpesvirus mulatto in Human Fibroblast

1974 ◽  
Vol 32 ◽  
pp. 244-245
Author(s):  
Glennelle Washington ◽  
Philip P. McGrath ◽  
Peter R. Graze ◽  
Ivor Royston
Keyword(s):  
Rhesus Monkey ◽  
Microscopic Study ◽  
Electron Microscopic Study ◽  
Peripheral Blood ◽  
Human Fibroblast ◽  
Rhesus Monkeys ◽  
Human Fibroblasts ◽  
Electron Microscopic ◽  
Specific Antisera ◽  
Peripheral Blood Leucocytes

Herpes-like viruses were isolated from rhesus monkey peripheral blood leucocytes when co-cultivated with WI-38 cells. The virus was originally designated rhesus leucocyte-associated herpesvirus (LAHV) and subsequently called Herpesvirus mulatta (HVM). The original isolations were from juvenile rhesus monkeys shown to be free of antibody to rhesus cytomegalic virus. The virus could only be propagated in human or simian fibroblasts. Use of specific antisera developed from HVM showed no relationship between this virus and other herpesviruses. An electron microscopic study was undertaken to determine the morphology of Herpesvirus mulatta (HVM) in infected human fibroblasts.

Anti-Diabetic Activity of a Mineraloid Isolate, in vitro and in Genetically Diabetic Mice

2011 ◽  
Vol 81 (1) ◽  
pp. 34-42 ◽  
Author(s):  
Joel Deneau ◽  
Taufeeq Ahmed ◽  
Roger Blotsky ◽  
Krzysztof Bojanowski
Keyword(s):  
Dna Microarrays ◽  
Human Fibroblasts ◽  
Diabetic Animal ◽  
In Vitro Tests ◽  
Diabetic Mice ◽  
Fossil Material ◽  
Expression Of Genes ◽  
Enhanced Expression ◽  
Genetically Diabetic Mice

Type II diabetes is a metabolic disease mediated through multiple molecular pathways. Here, we report anti-diabetic effect of a standardized isolate from a fossil material - a mineraloid leonardite - in in vitro tests and in genetically diabetic mice. The mineraloid isolate stimulated mitochondrial metabolism in human fibroblasts and this stimulation correlated with enhanced expression of genes coding for mitochondrial proteins such as ATP synthases and ribosomal protein precursors, as measured by DNA microarrays. In the diabetic animal model, consumption of the Totala isolate resulted in decreased weight gain, blood glucose, and glycated hemoglobin. To our best knowledge, this is the first description ever of a fossil material having anti-diabetic activity in pre-clinical models.

Processing and Characterization of Recombinant von Willebrand Factor Expressed in Different Cell Types Using a Vaccinia Virus Vector

1992 ◽  
Vol 67 (01) ◽  
pp. 154-160 ◽  
Author(s):  
P Meulien ◽  
M Nishino ◽  
C Mazurier ◽  
K Dott ◽  
G Piétu ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Von Willebrand Factor ◽  
Human Fibroblasts ◽  
Active Form ◽  
Cell Types ◽  
Biologically Active ◽  
L Cells ◽  
Von Willebrand ◽  
Different Cell Types ◽  
Willebrand Factor

SummaryThe cloning of the cDNA encoding von Willebrand factor (vWF) has revealed that it is synthesized as a large precursor (pre-pro-vWF) molecule and it is now clear that the prosequence or vWAgll is responsible for the intracellular multimerization of vWF. We have cloned the complete vWF cDNA and expressed it using a recombinant vaccinia virus as vector. We have characterized the structure and function of the recombinant vWF (rvWF) secreted from five different cell types: baby hamster kidney (BHK), Chinese hamster ovary (CHO), human fibroblasts (143B), mouse fibroblasts (L) and primary embryonic chicken cells. Forty-eight hours after infection, the quantity of vWF antigen found in the cell supernatant varied from 3 to 12 U/dl depending on the cell type. By SDS-agarose gel electrophoresis, the percentage of high molecular weight forms of vWF varied from 39 to 49% relative to normal plasma for BHK, CHO, 143B and chicken cells but was less than 10% for L cells. In all cell types, the two anodic subbands of each multimer were missing. The two cathodic subbands were easily detected only in BHK and L cells. By SDS-PAGE of reduced samples, pro-vWF was present in similar quantity to the fully processed vWF subunit in L cells, present in moderate amounts in BHK and CHO and in very low amounts in 143B and chicken cells. rvWF from all cells bound to collagen and to platelets in the presence of ristocetin, the latter showing a high correlation between binding efficiency and degree of multimerization. rvWF from all cells was also shown to bind to purified FVIII and in this case binding appeared to be independent of the degree of multimerization. We conclude that whereas vWF is naturally synthesized only by endothelial cells and megakaryocytes, it can be expressed in a biologically active form from various other cell types.

Modeling of dynamic mosaicism on ring chromosomes in human fibroblasts and IPSCS

2020 ◽  
pp. 12-13
Author(s):  
Т.В. Никитина ◽  
А.А. Кашеварова ◽  
М.М. Гридина ◽  
А.А. Хабарова ◽  
А.Г. Мензоров ◽  
...  
Keyword(s):  
Genetic Structure ◽  
Human Fibroblasts ◽  
Cellular Heterogeneity ◽  
Mitotic Stability ◽  
Factors Affecting ◽  
Ring Chromosomes ◽  
Cell Clones ◽  
Mitotic Instability ◽  
Mitotic Behavior

Митотическая нестабильность кольцевых хромосом может приводить к появлению клеточных клонов с различной генетической структурой. В качестве модели нестабильности кольцевых хромосом в митозе мы использовали фибробласты от пациентов с r(8), r(13), r(18) и r(22) и полученные из них индуцированные плюрипотентные стволовые клетки (ИПСК). Линии ИПСК с r(22) имели относительно стабильный кариотип на протяжении десятков (до 60) пассажей и сохраняли неизменную структуру кольцевой хромосомы. Кариотип линий ИПСК с r(8) и r(18) на ранних пассажах стабильный, планируется его изучение на поздних пассажах. Наибольшее разнообразие кариотипа выявлено в линиях ИПСК с r(13), в которых наблюдали различные перестройки и выраженную клеточную гетерогенность. Определение факторов, влияющих на митотическую стабильность кольцевых хромосом, может иметь значение для консультирования пациентов. Mitotic instability of ring chromosomes can lead to the appearance of cell clones with different genetic structure. IPSCs from fibroblasts of patients with r(8), r(13), r(18), and r(22) were used as a model of ring chromosomes mitotic behavior. Karyotypes of iPSC lines with r(8) and r(18) have so far been evaluated only in the early passages, lines with r(22) have maintained a relatively stable karyotype up to 60 passages. The occurrence of rearrangements and cellular heterogeneity was found characteristic for r(13) iPSCs. The determination of factors affecting the ring chromosomes mitotic stability would be beneficial for the patient’s prognosis.

Erythrocyte and reticulocyte parameters during biological monitoring of lead exposure to the body

2020 ◽  
Vol 60 (11) ◽  
pp. 873-876
Author(s):  
Anastasiya G. Khotuleva ◽  
Mariya S. Kozyreva
Keyword(s):  
Biological Monitoring ◽  
Hematological Parameters ◽  
Lead Level ◽  
Statistical Processing ◽  
Absorption Spectrometry ◽  
The Body ◽  
Control Group ◽  
Processing Plant ◽  
Hematological Disorders ◽  
Globin Synthesis

Introduction. The most susceptible to lead is the hematopoietic system of hematopoietic organs due to lead inhibition of heme and globin synthesis and cytotoxic effect on the membrane of Mature red blood cells. The aim of study was to evaluate the informative value of the study of erythrocyte and reticulocyte parameters determined on modern hematological analyzers in patients working in contact with lead during medical and biological monitoring. Materials and methods. 45 employees of the lead battery processing plant and 30 persons of control group were examined. The level of lead in the blood was determined by atomic absorption spectrometry, δ-ALA in the urine-by the reaction of pyrol formation with acetylacetone in terms of gram of creatinine, the study of hematological parameters was performed on a Sysmex HT-2000i analyzer. Statistical processing of the results was performed using the program STATISTICA 10.0. Results. Significant changes in erythrocytic (RDW) and reticulocytic (RET, IRF, LFR, MFR, HFR, RET-He) parameters, erythropoietin in workers in contact with lead compared to the control group, changes in MCV, MCH, RDW, RET indicators in the group working in dynamics after 2 years were revealed. Associations of hematological parameters with biomarkers of exposure and effect (lead level in blood and ALA in urine) were revealed. Conclusions. Assessment of erythrocyte (MCV, MCH, RDW) and reticulocyte parameters (RET% and their distribution by maturity) in dynamics during periodic medical examinations of workers in contact with lead allows us to detect the development of hematological disorders at early stages.

Effect of Dinitrosyl Iron Complexes (NO Donors) on the Metabolic Processes of Human Fibroblasts

2018 ◽  
Vol 483 (4) ◽  
pp. 452-456
Author(s):  
A. Gizatullin ◽  
◽  
N. Akent'eva ◽  
N. Sanina ◽  
N. Shmatko ◽  
...  
Keyword(s):  
Human Fibroblasts ◽  
Iron Complexes ◽  
Dinitrosyl Iron Complexes ◽  
Metabolic Processes ◽  
No Donors ◽  
Dinitrosyl Iron
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