scholarly journals Is there an inflammatory stimulus to human term labour?

PLoS ONE ◽  
2021 ◽  
Vol 16 (8) ◽  
pp. e0256545
Author(s):  
Natasha Singh ◽  
Bronwen Herbert ◽  
Garvin Sooranna ◽  
Nishel M. Shah ◽  
Ananya Das ◽  
...  
Keyword(s):  
Statistical Significance ◽  
P Value ◽  
Mrna Levels ◽  
Inflammatory Stimulus ◽  
Rt Pcr ◽  
Protein Levels ◽  
Decidua Basalis ◽  
Early Labour ◽  
Transcription Factor Activation ◽  
Term Labour

Inflammation is thought to play a pivotal role in the onset of term and some forms of preterm labour. Although, we recently found that myometrial inflammation is a consequence rather than a cause of term labour, there are several other reproductive tissues, including amnion, choriodecidua parietalis and decidua basalis, where the inflammatory stimulus to labour may occur. To investigate this, we have obtained amnion, choriodecidual parietalis and decidua basalis samples from women at various stages of pregnancy and spontaneous labour. The inflammatory cytokine profile in each tissue was determine by Bio-Plex Pro® cytokine multiplex assays and quantitative RT-PCR. Active motif assay was used to study transcription activation in the choriodecidua parietalis. Quantitative RT-PCR was use to study the pro-labour genes (PGHS-2, PGDH, OTR and CX43) in all of the tissues at the onset of labour and oxytocin (OT) mRNA expression in the choriodecidual parietalis and decidua basalis. Statistical significance was ascribed to a P value <0.05. In the amnion and choriodecidua parietalis, the mRNA levels of various cytokines decreased from preterm no labour to term no labour samples, but the protein levels were unchanged. The choriodecidua parietalis showed increase in the protein levels of IL-1β and IL-6 in the term early labour samples. In the amnion and decidua basalis, the protein levels of several cytokines rose in term established labour. The multiples of the median derived from the 19-plex cytokine assay were greater in term early labour and term established labour samples from the choriodecidua parietalis, but only in term established labour for myometrium. These data suggest that the inflammatory stimulus to labour may begin in the choriodecidua parietalis, but the absence of any change in prolabour factor mRNA levels suggests that the cytokines may act on the myometrium where we observed changes in transcription factor activation and increases in prolabour gene expression in earlier studies.

Negative Correlation Between Serum Levels of Homocysteine and Apolipoprotein M

2019 ◽  
Vol 19 (2) ◽  
pp. 120-126
Author(s):  
J. Wei ◽  
Y. Yu ◽  
Y. Feng ◽  
J. Zhang ◽  
Q. Jiang ◽  
...  
Keyword(s):  
Negative Correlation ◽  
Hepg2 Cells ◽  
Serum Levels ◽  
Significant Negative Correlation ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Apolipoprotein M ◽  
Protein Mass ◽  
Protein Levels ◽  
Phosphoinositide 3 Kinase

Background: Homocysteine (Hcy) has been suggested as an independent risk factor for atherosclerosis. Apolipoprotein M (apoM) is a constituent of the HDL particles. The goal of this study was to examine the serum levels of homocysteine and apoM and to determine whether homocysteine influences apoM synthesis. Methods: Serum levels of apoM and Hcy in 17 hyperhomocysteinemia (HHcy) patients and 19 controls were measured and their correlations were analyzed. Different concentrations of homocysteine (Hcy) and LY294002, a specific phosphoinositide 3- kinase (PI3K) inhibitor, were used to treat HepG2 cells. The mRNA levels were determined by RT-PCR and the apoM protein mass was measured by western blot. Results: We found that decreased serum apoM levels corresponded with serum HDL levels in HHcy patients, while the serum apoM levels showed a statistically significant negative correlation with the serum Hcy levels. Moreover, apoM mRNA and protein levels were significantly decreased after the administration of Hcy in HepG2 cells, and this effect could be abolished by addition of LY294002. Conclusions: resent study demonstrates that Hcy downregulates the expression of apoM by mechanisms involving the PI3K signal pathway.

scholarly journals Expression of Tachykinins and Tachykinin Receptors and Interaction with Kisspeptin in Human Granulosa and Cumulus Cells1

2016 ◽  
Vol 94 (6) ◽  
Author(s):  
Jordán García-Ortega ◽  
Francisco M. Pinto ◽  
Nicolás Prados ◽  
Aixa R. Bello ◽  
Teresa A. Almeida ◽  
...  
Keyword(s):  
Cell Function ◽  
Oocyte Retrieval ◽  
Cumulus Cells ◽  
Mrna Levels ◽  
Human Ovary ◽  
Rt Pcr ◽  
Protein Levels ◽  
Preovulatory Follicles ◽  
Nk3 Receptor ◽  
Nk2 Receptor

Abstract The neurokinin B/NK3 receptor (NK3R) and kisspeptin/kisspeptin receptor (KISS1R), two systems which are essential for reproduction, are coexpressed in human mural granulosa (MGC) and cumulus cells (CCs). However, little is known about the presence of other members of the tachykinin family in the human ovary. In the present study, we analyzed the expression of substance P (SP), hemokinin-1 (HK-1), NK1 receptor (NK1R), and NK2 receptor (NK2R) in MGCs and CCs collected from preovulatory follicles of oocyte donors at the time of oocyte retrieval. RT-PCR, quantitative RT-PCR, immunocytochemistry, and Western blotting were used to investigate the patterns of expression of tachykinin and tachykinin receptor mRNAs and proteins and the possible interaction between the tachykinin family and kisspeptin. Intracellular free Ca2+ levels ([Ca2+]i) in MGCs after exposure to SP or kisspeptin in the presence of SP were also measured. We found that SP, HK-1, the truncated NK1R isoform NK1R-Tr, and NK2R were all expressed in MGCs and CCs. NK1R-Tr mRNA and NK2R mRNA and protein levels were higher in MGCs than in CCs from the same patients. Treatment of cells with kisspeptin modulated the expression of HK-1, NK3R, and KISS1R mRNAs, whereas treatment with SP regulated kisspeptin mRNA levels and reduced the [Ca2+]i response produced by kisspeptin. These data demonstrate that the whole tachykinin system is expressed and acts in coordination with kisspeptin to regulate granulosa cell function in the human ovary.

scholarly journals Amniotic IGF-I supplementation of growth-restricted fetal sheep alters IGF-I and IGF receptor type 1 mRNA and protein levels in placental and fetal tissues

2005 ◽  
Vol 186 (1) ◽  
pp. 145-155 ◽  
Author(s):  
S Shaikh ◽  
F H Bloomfield ◽  
M K Bauer ◽  
H H Phua ◽  
R S Gilmour ◽  
...  
Keyword(s):  
Late Gestation ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Igf Binding Proteins ◽  
Fetal Sheep ◽  
Hepatic Expression ◽  
Protein Levels ◽  
Igf I ◽  
Igf Receptor

We have previously reported that chronic intra-amniotic supplementation of the late gestation growth-restricted (IUGR) ovine fetus with IGF-I (20 μg/day) increased gut growth but reduced liver weight and circulating IGF-I concentrations. Here we report mRNA and protein levels of IGF-I, the type 1 IGF receptor (IGF-1R) and IGF-binding proteins (IGFBP)-1, -2 and -3 in fetal gut, liver, muscle and placenta from fetuses in that earlier study in an attempt to explain these contrasting results. mRNA and protein were extracted from tissues obtained at post mortem at 131 days of gestation (term, 145 days) from three groups of fetuses (control, IUGR+saline and IUGR+IGF-I, n=9 per group). Control fetuses were unembolised and untreated. In the IUGR groups, growth restriction was induced from 113 to 120 days by placental embolisation; from 120 to 130 days fetuses were treated with daily intra-amniotic injections of either saline or 20 μg IGF-I. mRNA was measured by RT-PCR or real-time RT-PCR, and protein by Western blot. In liver, muscle and placenta, IGF-I mRNA and protein levels were reduced by between 8 and 30% in IGF-I-treated fetuses compared with saline-treated fetuses and controls with no change in IGF-1R mRNA or protein levels. In contrast, in the gut, IGF-I mRNA and protein levels were not significantly altered with IGF-I treatment, but IGF-1R levels were increased, especially in the jejunum. Immunolocalisation demonstrated that IGF-1R expression was confined to the luminal aspect of the gut. mRNA levels of all three IGFBPs were reduced in the gut of IGF-I-treated fetuses, but hepatic expression was significantly increased. These data demonstrated tissue-specific regulation of IGF-I, IGF-1R and IGFBPs-1, -2 and -3 in response to intra-amniotic IGF-I supplementation, though the underlying mechanisms remain obscure.

scholarly journals Stimulation of iodide uptake by human chorionic gonadotropin in FRTL-5 cells: effects on sodium/iodide symporter gene and protein expression

2002 ◽  
pp. 655-661 ◽  
Author(s):  
F Arturi ◽  
I Presta ◽  
D Scarpelli ◽  
JM Bidart ◽  
M Schlumberger ◽  
...  
Keyword(s):  
Western Blot ◽  
Chorionic Gonadotropin ◽  
Human Chorionic Gonadotropin ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Iodide Uptake ◽  
Protein Levels

BACKGROUND: Various clinical and experimental findings support the concept that human chorionic gonadotropin (hCG) can stimulate iodide uptake in thyroid cells. DESIGN: We investigated the molecular mechanisms underlying the effects of hCG on iodide uptake, and particularly its action on the expression of Na+/I- symporter (NIS) mRNA and protein. METHODS: Iodide uptake was analyzed in FTRL-5 cells by measuring (125)I concentrations in cells after a 30-min exposure to 0.1 microCi carrier-free Na (125)I in the presence or absence of hCG or, for control purposes, TSH. Expression of NIS mRNA and NIS protein synthesis were evaluated, respectively, with a semiquantitative 'multiplex' RT-PCR method and Western blot analysis. RESULTS: Iodide uptake was increased by hCG in a dose- and time-dependent manner: maximal effects were observed after 72 h of stimulation. The effect was cAMP dependent and paralleled that of TSH, although it lacked the early cycloheximide-independent component seen with TSH, and its peak effect was lower. Semiquantitative multiplex RT-PCR revealed that hCG produced a significant increase in NIS mRNA levels that was detectable after 4 h and peaked after 48 h. In contrast, in TSH-stimulated FRTL-5 cells, maximum NIS mRNA expression was observed after 24 h of stimulation. Western blot analysis demonstrated that hCG also caused a 2.5-fold increase over basal values in NIS protein levels, which was similar to that observed after TSH stimulation although the peak effects of the latter hormone were less marked and occurred earlier. CONCLUSION: Our data demonstrated that hCG stimulates iodide uptake in FRTL-5 cells by increasing NIS mRNA and protein levels. Thus, the functional status of the thyroid may be influenced by hCG-dependent changes in NIS expression occurring during pregnancy.

scholarly journals 203.Studies of the activin pathway in dominant and subordinate bovine follicles before, during and after selection

2004 ◽  
Vol 16 (9) ◽  
pp. 203
Author(s):  
B. C. Sisco ◽  
A. N. Shelling ◽  
P. L. Pfeffer
Keyword(s):  
Feedback Loop ◽  
Mrna Levels ◽  
Dominant Follicle ◽  
Rt Pcr ◽  
Protein Levels ◽  
Follicular Wave ◽  
Follicular Size ◽  
Quantitative Changes ◽  
Follicle Selection ◽  
Inhibin Subunits

In monovulatory species such as cattle, one of a cohort of developing follicles assumes dominancy and continues to grow in each follicular wave. After dominant follicle selection, pituitary-derived FSH levels decrease through a negative feedback loop mediated by oestradiol and inhibin A produced by the dominant follicle. The dominant follicle itself only requires very low basal levels of FSH, thus escaping atresia which is the fate of the subordinate follicles. The mechanisms involved in dominant follicle (DF) selection remain unclear. Most studies have focused on the stages following selection. To investigate what roles activin and inhibin play in DF selection we looked at the quantitative changes in the expression of the genes coding for the activin/inhibin subunits (Inhibin α, βA and βB) as well as other genes in the activin pathway (SMAD2, ActRIIA/B, follistatin (FST), FSHR). We examined mRNA levels in follicular granulosa cells (GCs) before (d1.5), during (d2.5) and after (d3.5 and 7) DF selection using real-time RT-PCR. Prior to DF selection, highest levels of inhibin βA, FST and SMAD2 transcripts converged on the largest follicles. Inhibin α, ActRIIA/B and FSHR levels did not correlate with follicular size at this stage. At Day 2.5, highest levels of inhibin βA, inhibin α, FST and SMAD2 transcripts were seen in a single putative DF. ActRIIA/B and FSHR did not show any difference between follicles. By Days 3.5 and 7, a dramatic difference in expression levels of inhibin βA, inhibin α and FST were seen in DF compared to SF. Yet in absolute terms inhibin βA levels decreased after selection, whereas inhibin α levels increased. Inhibin βB expression was only detected in Day 7 GCs and was significantly higher in the DF. These results suggest a shift from an activin environment during the pre and peri-DF selection period, to an inhibin environment following DF selection. Inhibin/activin protein levels in the follicular fluid using western ligand blotting confirmed this. We postulate that the higher activin activity within DF influences the selection mechanism as activin and inhibin have been shown to play a role in gonadotropin regulation in the ovary around the time of selection.

scholarly journals PKA and AKIP1 interact to mediate cAMP-driven COX-2 expression: A potentially pivotal interaction in preterm and term labour

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0252720
Author(s):  
Angela Yulia ◽  
Natasha Singh ◽  
Alice J. Varley ◽  
Kaiyu Lei ◽  
Danijela Markovic ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Human Myometrium ◽  
Interacting Protein ◽  
Protein Levels ◽  
Early Labour ◽  
Myometrial Contractility ◽  
Cox 2 ◽  
Nuclear Levels ◽  
Term Labour

Previously, we showed that cAMP increased COX-2 expression in myometrial cells via MAPK. Here, we have extended these observations, using primary myometrial cell cultures to show that the cAMP agonist, forskolin, enhances IL-1β-driven COX-2 expression. We then explored the role of A-kinase interacting protein (AKIP1), which modulates the effect of PKA on p65 activation. AKIP1 knockdown reversed the effect of forskolin, such that its addition inhibited IL-1β-induced COX-2 mRNA expression and reduced the IL-1β-induced increase in nuclear levels of p65 and c-jun. Forskolin alone and with IL-1β increased IκBα mRNA expression suggesting that in the context of inflammation and in the presence of AKIP1, cAMP enhances p65 activation. AKIP1 knockdown reversed these changes. Interestingly, AKIP1 knockdown had minimal effect on the ability of forskolin to repress either basal OTR expression or IL-1β-stimulated OTR mRNA expression. AKIP1 was up-regulated by IL-1β, but not stretch and was repressed by cAMP. The mRNA expression of AKIP1 increased in early labour in tandem with an increase in COX-2 mRNA and protein. AKIP1 protein levels were also increased with inflammation and stretch-induced preterm labour. Our results identify a second important cAMP effector-switch occurring at term in human myometrium and suggest that a hitherto unrecognized interaction may exist between AKIP1, NFκB and AP-1. These data add to the proposition that cAMP acts as a key regulator of human myometrial contractility.

255 GENE EXPRESSION OF Cox 5a, 5b, AND 6b1 AND THEIR ROLES IN PRE-IMPLANTATION OF MOUSE EMBRYOS

2006 ◽  
Vol 18 (2) ◽  
pp. 235
Author(s):  
S.-E. Lee ◽  
X.-Y. Li ◽  
X.-S. Cui ◽  
N.-H. Kim
Keyword(s):  
Rna Interference ◽  
Mrna Expression ◽  
Real Time ◽  
Oocyte Maturation ◽  
Blastocyst Stage ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Target Mrna ◽  
Protein Levels

Despite clear evidence of regulation of mitochondrial respiration by nuclear encoded genes, cytochrome oxidase (Cox), little information is available on their expression and functional roles during early embryonic development. To examine the role of Cox in oocyte maturation and embryogenesis, we first characterized mRNA and protein levels of nuclear encoded genes, Cox 5a, 5b, and 6b1, in mouse oocytes and during early embryogenesis, using real-time RT-PCR and immunocytochemistry. We then examined the possible role of these genes in oocyte maturation and pre-implantation development using RNA interference analysis. The relative abundances of Cox 5a, 5b, and 6b1 transcripts was measured by real time RT-PCR. After normalization by comparison to histone H2a mRNA levels, the mRNA expression of Cox 5a, 5b, and 6b1 were found to be considerable in mature oocytes and zygotes, but reduced slightly in 2-cell embryos. From the 2-cell to the blastocyst stage, mRNA expression is dependent on the number of blastomeres, as expression increases only gradually with development. Immunocytochemical studies revealed that Cox 5a, 5b, and 6b1 proteins were expressed in all blastomeres of the blastocyst. Injection of Cox 5a, 5b, or 6b1 siRNA into GV stage oocytes decreased expression of the target mRNA specifically, while not affecting the expression of mRNAs for the other subunits in mature oocytes. Similarly, each siRNA injection into zygotes specifically reduced target mRNA expression at the 2-cell, morula and blastocyst stages (P < 0.05). Silencing of mRNA expression by RNA interference (siRNA) did not inhibit oocyte maturation or developmental events up to the morula and blastocyst stages. The expression level of mtDNA9, as well as overall levels of mitochondrial mRNAs, was not different following injection of siRNA for Cox 5a, 5b, or 6b1. However, it is evident that the number of mitochondria in siRNA treated blastocysts was greatly reduced, and they appeared to be morphologically abnormal. Significantly higher apoptosis and lower cell numbers were observed in siRNA treated blastocysts. Real time RT PCR revealed that silencing of Cox 5a, 5b, and 6b1 decreased mRNA and protein levels of E-cadherin. These results suggest that the Cox subunits, Cox 5a, 5b, and 6b1, play an important role in mitochondrial function during pre-implantation development. This work was funded by a grant from the National Research Laboratory Program in Korea.

Evaluation of MMP-12 expression in chronic rhinosinusitis with nasal polyposis

2021 ◽  
Vol 0 (0) ◽  
pp. 0-0
Author(s):  
S. Lygeros ◽  
G. Danielides ◽  
G.C. Kyriakopoulos ◽  
K. Grafanaki ◽  
F. Tsapardoni ◽  
...  
Keyword(s):  
Chronic Rhinosinusitis ◽  
Gene Expression Regulation ◽  
Statistical Significance ◽  
Sinus Surgery ◽  
Mrna Levels ◽  
Tissue Samples ◽  
Potential Implication ◽  
Protein Levels ◽  
Healthy Mucosa ◽  
The Difference

Background: The purpose of this study was to evaluate the expression of MMP-12 in patients with chronic rhinosinusitis with polyps (CRSwNP). Methodology: Tissue samples from 37 patients with CRSwNP undergoing functional endoscopic sinus surgery and healthy mucosa specimens from 12 healthy controls were obtained intraoperatively. The mRNA and protein expression levels of MMP-12 were quantified by real-time polymerase chain reaction and Western blotting, respectively. Results: mRNA levels of MMP-12 were significantly elevated in the CRSwNP tissue samples compared to those in control ones. The protein levels of MMP-12 showed a trend of increasing but with no statistical significance. Conclusions: Elevation of MMP-12 in patients with CRSwNP suggests its potential implication in the pathogenesis of the disease. The difference in the expression profile observed between mRNA and protein levels could be due to post-translational gene expression regulation. Our findings provide evidence that MMP-12 along with other MMPs may serve as a biomarker and therapeutic target in the management of the disease.

Molecular profiles of gastric cancer: The UCSD experience.

2013 ◽  
Vol 31 (4_suppl) ◽  
pp. 29-29
Author(s):  
Devon Marcus McGee ◽  
John P. Shen ◽  
Paul Timothy Fanta ◽  
Andrew M. Lowy
Keyword(s):  
Gastric Cancer ◽  
Statistical Significance ◽  
Accurate Determination ◽  
Prognostic Significance ◽  
Mrna Levels ◽  
Her2 Expression ◽  
Rt Pcr ◽  
Expression Levels ◽  
Platinum Based Chemotherapy ◽  
Molecular Studies

29 Background: Gastric cancer is the 2nd leading cause of cancer mortality globally. A small number of studies reported that low TS and ERCC1 mRNA levels are associated with improved survival, and may be important as prognostic factors. Methods: Intratumoral gene expression levels were assessed using laser-captured micro-dissection and quantitative Real-Time PCR on formalin fixed paraffin embedded tumor samples from 22 gastric adenocarcinomas. A retrospective chart review was performed to measure clinical parameters. Primary objectives were to determine the range of expression of TS, ERCC1, and HER2, and to investigate if these biomarkers are predictive of survival. Results: TS, ERCC1, and HER2 median expression levels using RT-PCR were 3.56, 1.54, and 0.085, respectively. Median OS was 62 months. Subjects with low TS trended towards improved survival (92 months vs. 34 months, p: 0.17), as did subjects with low ERCC1 (92 months vs. 55 months, p: 0.44). There was no association between HER2 expression and OS. All subjects had HER2 levels below 0.55. With regard to treatment, 90.9% of patients received platinum-based chemotherapy and 4.5% received HER2-guided therapy. Conclusions: Our results are consistent with prior reports that associate low TS and low ERCC1 with improved OS, and may imply prognostic significance. Although these trends did not reach statistical significance, they are consistent with prior studies. Further molecular studies are needed to better assess the utility of these markers as prognostic indicators. There was no clear trend between HER2 levels and OS, most likely because all subjects had low HER2 levels. However, as accurate determination of HER2 expression is becoming increasingly important in the management of gastric cancer patients, further research into the degree of concordance between RT-PCR assessment and FISH assessment of HER2 gene amplication is warranted. Overall, more molecular studies are needed to better stratify this disease into subtypes, and identify new drug targets for chemo-resistant subtypes.

Vitamin D-inducible calcium transport and gene expression in three Caco-2 cell lines

2002 ◽  
Vol 283 (3) ◽  
pp. G618-G625 ◽  
Author(s):  
James C. Fleet ◽  
Faria Eksir ◽  
Kenneth W. Hance ◽  
Richard J. Wood
Keyword(s):  
Vitamin D ◽  
Calcium Absorption ◽  
Parental Cell ◽  
P Value ◽  
Parental Cell Line ◽  
Facilitated Diffusion ◽  
Mrna Levels ◽  
Transport Channel ◽  
Protein Levels ◽  
Calbindin D

The parental cell line (P) of Caco-2 cells and two clones, BBe and TC7, were studied at 11 days postconfluence to test the facilitated diffusion model of vitamin D-mediated intestinal calcium absorption (CaTx). Nuclear vitamin D receptor (nVDR) and calbindin D9k (CaBP) were measured by Western blot; 1,25-hydroxyvitamin D3 24-hydroxylase (CYP24), CaBP, plasma membrane Ca-ATPase (PMCA), and Ca transport channel (CaT1) mRNA levels were examined by RT-PCR; and net apical-to-basolateral CaTx was examined after treating cells with vehicle or 10 nM calcitriol for 8 (mRNA levels) or 48 h (protein, CaBP mRNA, CaTx). nVDR level was lowest in BBe (38% P value) and directly related to CYP24 induction (TC7 = P, which were 1.56 times greater than BBe). nVDR was inversely related to the vitamin D-induced levels of CaT1 mRNA, CaBP mRNA, PMCA mRNA, and net CaTx, with the highest induction seen in BBe. Basal CaBP mRNA (86 times greater than P) and protein levels were highest in TC7 cells and were not associated with higher net CaTx, suggesting CaBP may not be rate limiting for CaTx in these cells.

Sign in / Sign up

Export Citation Format

Share Document