scholarly journals Quantitative Polymerase Chain Reaction Detection of Microchimerism in Female Transplant Renal Recipients

2020 ◽  
Vol 104 (11-12) ◽  
pp. 865-870
Author(s):  
Donata Villari ◽  
Francesca Salvianti ◽  
Maria Zanazzi ◽  
Alberto Martini ◽  
Pietro Spatafora ◽  
...  
Keyword(s):  
Renal Transplantation ◽  
Quantitative Polymerase Chain Reaction ◽  
Immunological Tolerance ◽  
Solid Organ ◽  
Kidney Graft ◽  
Polymerase Chain Reaction Detection ◽  
Male Donor ◽  
Polymerase Chain ◽  
Cadaveric Donors ◽  
The Relationship

<b><i>Introduction:</i></b> Microchimerism (MC) is the presence of a small amount of foreign cells or DNA within a person’s circulation or tissues. It has been identified also in recipients of solid organ transplants where it seems to be critical for the development and maintenance of immunological tolerance. Nevertheless, natural and/or iatrogenic MC can be acquired prior to transplantation, through pregnancy and/or blood transfusion. <b><i>Objective:</i></b> The aim of this study was to detect the presence of MC in women after renal transplantation from male cadaveric donors and its relationship with graft outcomes. <b><i>Methods:</i></b> We studied by qPCR the presence of the DYS14 gene sequence of the Y chromosome in 12 females who received a kidney graft from a male donor before transplantation (T0), after 15 days (T1) and 1 year of transplantation (T2). We found the sequence in all recipients after renal transplantation. <b><i>Results:</i></b> All the women were negative for this sequence prior to transplantation (T0). Mean (SD) Y-related DNA quantity was 0.80 (0.69) ng/mL plasma and 0.15 (0.26) ng/mL plasma at T1 and T2, respectively. No acute rejection was observed, and mean (SD) estimated Cr clearance was 68.8 (16.9) mL/min within 1 year from transplantation. <b><i>Conclusions:</i></b> Presence of MC was associated with good kidney graft outcomes after 1 year of transplantation, but further studies will be needed to investigate the relationship between clinical outcomes and the development of MC in renal transplant recipient.

A signaling mechanism for growth-related expression of fetal hemoglobin

Blood ◽  
2004 ◽  
Vol 103 (5) ◽  
pp. 1929-1933 ◽  
Author(s):  
Natarajan V. Bhanu ◽  
Tiffany A. Trice ◽  
Y. Terry Lee ◽  
Jeffery L. Miller
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Fetal Hemoglobin ◽  
Clinical Settings ◽  
Globin Mrna ◽  
Erythroid Progenitor ◽  
Signaling Mechanism ◽  
Erythroid Progenitor Cells ◽  
Polymerase Chain ◽  
New Treatment ◽  
The Relationship

Abstract Increases in fetal hemoglobin have been identified after birth in several clinical settings associated with stressed or malignant erythropoiesis. To better understand the relationship between the expression of this fetal protein and growth, donated human erythroid progenitor cells were cultured in the presence of erythropoietin (EPO) plus the growth-modifying cytokine stem cell factor (SCF), and several growth-related signaling pathways were interrogated. Only the MEK1/2 inhibitor (PD98059) demonstrated significant effects on fetal hemoglobin. In the absence of PD98059, levels of fetal hemoglobin averaged 27.4% ± 7.9% in EPO+SCF compared with 1.26% ± 1.7% in EPO alone (P = .02). A linear dose response in levels of fetal hemoglobin to PD98059 was detected (0.16 μM = 27.13%, 0.8 μM = 19.6%, 4 μM = 12.2%, 20 μM = 1.54%). Western blot analyses revealed that SCF was required for phosphorylation of MEK and p44MAPK in this setting, and quantitative polymerase chain reaction demonstrated a significant increase in γ-globin mRNA. Particular perturbations of growth-related signaling may also function to activate tissue-specific genes normally expressed during fetal development. This concept may be relevant for the development of new treatment rationales for beta hemoglobinopathies.

Two-Dimensional CoII Coordination Polymer for Photodegradation of Organic Dyes and Treatment of Temporary Osteoporosis of the Hip (TOH) by Regulating wnt/β-catenin/PPARγ Pathway

2020 ◽  
Vol 73 (1) ◽  
pp. 9
Author(s):  
Hui Chen ◽  
Xue-Jian Wu
Keyword(s):  
Coordination Polymer ◽  
Dna Cleavage ◽  
Organic Dyes ◽  
Quantitative Polymerase Chain Reaction ◽  
Organic Ligand ◽  
Annexin V ◽  
Two Dimensional ◽  
Polymerase Chain ◽  
The Relationship ◽  
Marrow Stem Cell

In this study, a new two-dimensional CoII-containing coordination polymer [Co(pda)(H2O)](DMF)2 (1, DMF=N,N-dimethylformamide) based on a light-harvesting triphenylamine type organic ligand has been successfully prepared by reaction of Co(NO3)2⋯6H2O and 4,4′-(phenylazanediyl)dibenzoic acid (H2pda) in a mixed solvent of water and DMF. Complex 1 has significant photocatalytic activity under visible light for the degradation of organic dyes such as rhodamine B (RhB), methylene blue (MB), as well as methyl orange (MO) assisted by H2O2. Compound 1 was investigated for anti-osteoporosis activity, the Annexin V-FITC/PI method was used to test the relationship between apoptosis and osteoporosis and the effect of 1 on bone marrow stem cell (BMSC) apoptosis. The expression levels of wnt/β, catenin, and PPARγ after treatment with 1 was determined by reverse transcription quantitative polymerase chain reaction. The DNA cleavage ability of 1 was further studied by molecular docking.

scholarly journals Quantification of Phytophthora capsici Oospores in Soil by Sieving-Centrifugation and Real-Time Polymerase Chain Reaction

Plant Disease ◽  
2008 ◽  
Vol 92 (1) ◽  
pp. 143-149 ◽  
Author(s):  
C. F. Pavón ◽  
M. Babadoost ◽  
K. N. Lambert
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Sucrose Solution ◽  
Quantitative Polymerase Chain Reaction ◽  
Phytophthora Capsici ◽  
Soil Samples ◽  
Infested Soil ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
The Relationship

A procedure was developed to quantify Phytophthora capsici oospores in soil by combining a sieving-centrifugation method and a real-time quantitative polymerase chain reaction (QPCR) assay. Five soil samples representing three different soil textures were infested with oospores of P. capsici to produce 101, 102, 103, 104, or 105 spores per 10 g of air-dried soil. Each 10-g sample of infested soil was suspended in 400 ml of water and then passed through 106-, 63-, and 38-μm metal sieves. The filtrate was then passed through a 20-μm mesh filter. Materials caught on the filter were washed with water into two 50-ml centrifuge tubes and spun for 4 min (900 × g). The pellet was suspended in 30 ml of 1.6 M sucrose solution and centrifuged for 45 s (190 × g). The supernatant was passed through the 20-μm mesh filter. The sucrose extraction process of oospores was repeated five times to maximize oospore extraction. Materials caught on the 20-μm mesh filter were washed with water into a 50-ml tube and spun for 4 min (900 × g). The pellet was suspended in 1 ml of water, and the number of oospores was determined with a haemocytometer. The relationship between number of oospores recovered from the soil and number of oospores incorporated into the soil was Ŷ = –0.95 + 1.31X – 0.03X2 (R2 = 0.98), in which Ŷ = log10 of number of oospores recovered from the soil and X = log10 of number of oospores incorporated into the soil. The oospores were germinated after treatment with 0.1% KMnO4 solution for 10 min to induce germination. On the basis of the detection of ribosomal DNA, a QPCR method for P. capsici oospores was developed. PCR inhibitors were eliminated by extracting oospores from the soil by sieving-centrifugation. DNA was extracted and quantified from P. capsici oospores with suspensions of 101, 101.5, 102, 102.5, 103, 103.5, 104, 104.5, and 105 oospores per ml of water. The relationship between the DNA quantities and number of P. capsici oospores was Ŷ = –3.57 – 0.54X + 0.30X2 (R2 = 0.93), in which Ŷ = log10 (nanogram of P. capsici DNA) and X = log10 (number of oospores). The relationship between the quantity of DNA of P. capsici oospores recovered from the soil and the number of oospores incorporated into the soil was determined by Ŷ = –3.53 – 0.73X + 0.32X2 (R2 = 0.955, P < 0.05), in which Ŷ = log10 (DNA quantity of P. capsici oospores recovered from the soil) and X = log10 (number of P. capsici oospores incorporated into the soil). Utilizing the sieving-centrifugation and QPCR methods, oospores of P. capsici were quantified in soil samples collected from commercial fields.

scholarly journals Stress reactivity elicits a tissue-specific reduction in telomere length in ageing zebrafish (Danio rerio)

2020 ◽  
Author(s):  
James R. Evans ◽  
Jose V. Torres-Pérez ◽  
Maria Elena Miletto Petrazzini ◽  
Riva Riley ◽  
Caroline H. Brennan
Keyword(s):  
Telomere Length ◽  
Stress Reactivity ◽  
Quantitative Polymerase Chain Reaction ◽  
Heart Tissue ◽  
Telomere Shortening ◽  
Tissue Specific ◽  
Age Related ◽  
Specific Manner ◽  
Polymerase Chain ◽  
The Relationship

ABSTRACTTelomere length reflects cellular ageing. Increased telomere shortening in leukocytes is associated with a range of neurodegenerative and cardiovascular diseases, the onset and progression of which may be mediated by behavioural traits such as anxiety and stress reactivity. However, the effects of the hypothalamus-pituitary-adrenal axis stress response are shown to be tissue specific. As such, leukocyte telomere length may not give an accurate measure of the relationship between stress-reactivity and telomere length in disease relevant tissues. To test the hypothesis that stress-reactivity contributes to age-related telomere shortening in a tissue specific manner, we examined the correlation between telomere length in heart and brain tissue and stress-reactivity in a population of young (6-9 month) and ageing (18 month) zebrafish. Stress-reactivity was assessed by tank diving, a zebrafish version of the rodent open-field test, and through gene expression. Telomere length was assessed using quantitative polymerase chain reaction. We show that ageing zebrafish have shorter telomeres in both heart and brain. Telomere length is inversely related to stress-reactivity in heart but not brain of ageing individuals. These data support the hypotheses that an anxious predisposition contributes to telomere shortening in heart tissue, and by extension age-related heart disease, and that stress-reactivity contributes to age-related telomere shortening in a tissue-specific manner.

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