Abundance of ADAM-8, -9, -10, -12, -15 and -17 and ADAMTS-1 in mouse uterus during the oestrous cycle

2005 ◽  
Vol 17 (5) ◽  
pp. 543 ◽  
Author(s):  
Jiyoung Kim ◽  
Haekwon Kim ◽  
Joon Yeong Lee ◽  
Young Min Choi ◽  
Su-Jae Lee ◽  
...  
Keyword(s):  
Protein Expression ◽  
Mrna Level ◽  
Oestrous Cycle ◽  
Mrna Levels ◽  
Uterine Tissue ◽  
Tissue Remodelling ◽  
Mouse Uterus ◽  
A Disintegrin And Metalloproteinase ◽  
Epithelial Layers ◽  
Immunohistochemical Studies

The aim of the present study was to determine whether a disintegrin and metalloproteinase (ADAM)-8, -9, -10, -12, -15 and -17 and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-1 are involved in the remodelling process of the mouse uterus during the oestrous cycle. The mRNA expression of ADAM was observed in all uterine tissues throughout the entire cycle. The levels of ADAM-8 mRNA were maximal at pro-oestrus, whereas the expression of ADAM-9 and ADAMTS-1 mRNA was maximal at oestrus. The minimum mRNA level of all ADAM genes always occurred at dioestrus. The mRNA levels of ADAM-10, -12, -15 and -17 did not vary significantly, regardless of the stage of the oestrous cycle. Immunoblot analyses demonstrated the presence of all ADAM proteins throughout the cycle. In terms of protein intensities, ADAM-8, -12 and -17 were maximal at pro-oestrus, whereas ADAM-10 and ADAMTS-1 were maximal at metoestrus and ADAM-9 was maximal at oestrus. Regardless of the ADAM species, minimal protein expression always occurred at dioestrus. Immunohistochemical studies showed ADAM protein expression in luminal and glandular epithelial layers, but not in the stromal layer. Moreover, ADAM proteins were found to be heterogeneously localised and their individual localisations depended on the stage of the oestrous cycle. From these observations, we suggest that the ADAM genes play an important role in mouse uterine tissue remodelling during the oestrous cycle.

Expression of ADAMTS-1, -4, -5 and TIMP-3 in normal and multiple sclerosis CNS white matter

2006 ◽  
Vol 12 (4) ◽  
pp. 386-396 ◽  
Author(s):  
G Haddock ◽  
A K Cross ◽  
J Plumb ◽  
J Surr ◽  
D J Buttle ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
White Matter ◽  
Normal Tissue ◽  
Chondroitin Sulphate ◽  
Mrna Level ◽  
Mrna Levels ◽  
Protein Levels ◽  
Chondroitin Sulphate Proteoglycans ◽  
A Disintegrin And Metalloproteinase ◽  
Immunohistochemical Studies

ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) -1, -4 and -5 proteases have been identified in the CNS at the mRNA level. These glutamyl endopeptidases, inhibited by tissue inhibitor of metalloproteinases (TIMP)-3, are key enzymes in the degradation of the aggregating chondroitin sulphate proteoglycans (CSPGs), and may therefore play a role in CNS extracellular matrix (ECM) changes in multiple sclerosis (MS). We have investigated ADAMTS and TIMP-3 expression in normal and MS CNS white matter by real-time RT-PCR, western blotting and immunohistochemistry. We report for the first time the presence of ADAMTS-1, -4 and -5 in normal and MS white matter. Levels of ADAMTS-1 and -5 mRNA were decreased in MS compared to normal tissue, with no significant change in ADAMTS-4 mRNA levels. Protein levels of ADAMTS-4 were significantly higher in MS tissue compared to normal tissue. Immunohistochemical studies demonstrated that ADAMTS-4 was associated predominantly with astrocytes with increased expression within MS lesions. TIMP-3 mRNA was significantly decreased in MS compared to controls. These studies suggest a role for ADAMTS-4 in the pathogenesis of MS. Further studies on the activity of ADAMTS-4 will enable a better understanding of its role in the turnover of the ECM of white matter in MS.

scholarly journals The effect of calnexin deletion on the expression level of PDI in Saccharomyces cerevisiae under heat stress conditions

2008 ◽  
Vol 13 (1) ◽  
Author(s):  
Huili Zhang ◽  
Jianwei He ◽  
Yanyan Ji ◽  
Akio Kato ◽  
Youtao Song
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Heat Stress ◽  
Protein Expression ◽  
Molecular Chaperone ◽  
Mrna Level ◽  
Stress Conditions ◽  
Mrna Levels ◽  
Wild Type ◽  
Wild Type Yeast

AbstractWe cultured calnexin-disrupted and wild-type Saccharomyces cerevisiae strains under conditions of heat stress. The growth rate of the calnexin-disrupted yeast was almost the same as that of the wild-type yeast under those conditions. However, the induced mRNA level of the molecular chaperone PDI in the ER was clearly higher in calnexin-disrupted S. cerevisiae relative to the wild type at 37°C, despite being almost the same in the two strains under normal conditions. The western blotting analysis for PDI protein expression in the ER yielded results that show a parallel in their mRNA levels in the two strains. We suggest that PDI may interact with calnexin under heat stress conditions, and that the induction of PDI in the ER can recover part of the function of calnexin in calnexin-disrupted yeast, and result in the same growth rate as in wild-type yeast.

scholarly journals The Role of CD40 in Allergic Rhinitis and Airway Remodelling

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Ke-Jia Cheng ◽  
Min-Li Zhou ◽  
Yong-Cai Liu ◽  
Chen Wang ◽  
Ying-Ying Xu
Keyword(s):  
Allergic Rhinitis ◽  
Mrna Level ◽  
Costimulatory Molecule ◽  
Airway Remodelling ◽  
Mrna Levels ◽  
Collagen Deposition ◽  
Tissue Remodelling ◽  
Ifn Γ ◽  
Sirna Therapy

Background. Allergic rhinitis (AR) affects millions of people and is lack of effective treatment. CD40 is an important costimulatory molecule in immunity. However, few studies have focused on the role of CD40 in AR. Methods. In this study, we built mouse model of chronic AR. The mice were divided into the AR, control, intravenous CD40 siRNA, and nasal CD40 siRNA groups ( n = 6 each). We detected OVA-sIgE, IL-4, IL-5, IL-13, IL-10, IFN-γ, and TGF-β levels in serum and supernatant by ELISA, CD40+ splenic DCs, and Foxp3+ Tregs by flow cytometry and CD40 mRNA by RT2-PCR. We also used PAS and MT stains to assess tissue remodelling. Results. (1) The OVA-sIgE, IL-4, IL-5, and IL-13 levels in the serum or supernatant of nasal septal membrane of AR mice were significantly higher than control. After treated with CD40 siRNA, those indicators were significantly decreased. The IFN-γ, IL-10, and TGF-β levels in AR mice were significantly lower than that in control and were increased by administration of CD40 siRNA. (2) AR mice had significantly fewer Foxp3+ Tregs in the spleen than control mice. After treated with CD40 siRNA, AR mice had significantly more Foxp3+ Tregs. (3) AR mice exhibited a significantly higher CD40 mRNA levels than control. Administration of CD40 siRNA significantly reduced the CD40 mRNA level. (4) The AR mice showed significantly greater collagen deposition than the control in MT staining. Applications of CD40 siRNA significantly reduced the collagen deposition in AR mice. Conclusion. CD40 siRNA therapy shows promise for chronic AR as it significantly attenuated allergic symptoms and Th2-related inflammation and upregulated Foxp3+ Tregs. CD40 plays a role in tissue remodelling in AR, which can be inhibited by CD40 siRNA application.

Alterations in the Expression of Amyloid Precursor Protein Cleaving Enzymes mRNA in Alzheimer Peripheral Blood

2018 ◽  
Vol 16 (1) ◽  
pp. 29-38 ◽  
Author(s):  
Prapimpun Wongchitrat ◽  
Nattaporn Pakpian ◽  
Kuntida Kitidee ◽  
Kamonrat Phopin ◽  
Pornpatr A. Dharmasaroja ◽  
...  
Keyword(s):  
Amyloid Precursor Protein ◽  
Cognitive Performance ◽  
Peripheral Blood ◽  
Mrna Level ◽  
Precursor Protein ◽  
Mrna Levels ◽  
App Processing ◽  
Chain Reactions ◽  
Expression Of Genes ◽  
A Disintegrin And Metalloproteinase

Background: Alzheimer’s disease (AD) is the most common cause of dementia in elderly populations. Changes in the expression of the Amyloid Precursor Protein (APP)-cleaving enzymes directly affect the formation of Amyloid Beta (Aβ) plaques, a neuropathological hallmark of AD. Objective: We used peripheral blood from AD patients to investigate the expression of genes related to APP-processing [(β-site APP-cleaving enzyme 1 (BACE1), presenilin1 (PSEN1), and a disintegrin and metalloproteinase family 10 (ADAM10) and 17 (ADAM17)] and the epigenetic genes sirtuin (SIRT)1-3, which regulate Aβ production. Method: Real-time polymerase chain reactions were performed to determine the specific mRNA levels in plasma. The mRNA levels in AD patients were compared to those in healthy persons and assessed in relation to the subjects’ cognitive performance. Results: BACE1 mRNA level in AD subjects was significantly higher than those of healthy controls, whereas ADAM10 level was significantly lower in the AD subjects. The SIRT1 level was significantly decreased, while that of SIRT2 was increased in AD subjects and elderly controls compared to levels in healthy young control. In addition, correlations were found between the expression levels of BACE1, ADAM10 and SIRT1 and cognitive performance scores. Total Aβ (Aβ40+Aβ42) levels and the Aβ40/Aβ42 ratio were significantly increased in the AD subjects, whereas decrease in plasma Aβ42 was found in AD subjects. There was a negative correlation between Aβ40 or total Aβ and Thai Mental State Examination (TMSE) while there was no correlation between Aβ40/Aβ42 ratio or Aβ42 and TMSE. Conclusion: The present findings provide evidence and support for the potential roles of these enzymes that drive Aβ synthesis and for epigenetic regulation in AD progression and development, which can possibly be considered peripheral markers of AD.

Preferential expression of long form prolactin receptor mRNA in the rat brain during the oestrous cycle, pregnancy and lactation: hormones involved in its gene expression

1994 ◽  
Vol 141 (2) ◽  
pp. 325-333 ◽  
Author(s):  
T Sugiyama ◽  
H Minoura ◽  
N Kawabe ◽  
M Tanaka ◽  
K Nakashima
Keyword(s):  
Rat Brain ◽  
Short Form ◽  
Prolactin Receptor ◽  
Mrna Level ◽  
Oestrous Cycle ◽  
Ovariectomized Rats ◽  
Mrna Levels ◽  
Long Form ◽  
Pregnancy And Lactation ◽  
The Brain

Abstract The mRNA species for prolactin receptor (PRL-R) isoforms, long and short form PRL-Rs, were estimated by the reverse transcription-polymerase chain reaction method in the rat brain (cerebrum) during the oestrous cycle, pregnancy and lactation. The levels of long form PRL-R mRNA increased at pro-oestrus and oestrus, at the same time as serum prolactin levels increased, whereas the mRNA level of short form PRL-R was relatively unchanged. Long form PRL-R mRNA expression was also markedly increased in the brain at mid- and late gestation, and this elevated mRNA level was maintained during the period of lactation. In contrast, basal levels of short form PRL-R mRNA were also maintained throughout these periods of gestation and lactation. Ovariectomy moderately reduced brain mRNA levels of both long and short form PRL-R from the levels of those in control dioestrous rats, and hypophysectomy further suppressed them to the lowest levels. Administration of oestradiol valerate (E2V) or 17α-hydroxyprogesterone caproate (17OHPC) to ovariectomized rats resulted in dramatic increases in long form PRL-R mRNA levels in the brain, whereas no significant increase in short form PRL-R mRNA was observed. In rats which were ovariectomized and hypophysectomized, the administration of 17OHPC, rat prolactin or rat GH partially restored the brain level of long form PRL-R mRNA but not short form PRL-R mRNA. E2V, on the other hand, had no effect on the expression of brain PRL-R mRNAs in these hypophysectomized rats, suggesting that the stimulatory effect of E2V on long form PRL-R mRNA expression in ovariectomized rats was mediated by an enhanced secretion of a pituitary hormone, prolactin. These results suggest that the expression of long form PRL-R mRNA in the rat brain is directly induced by progesterone, prolactin or GH during the oestrous cycle, pregnancy and lactation. Journal of Endocrinology (1994) 141, 325–333

scholarly journals Effects of 1950 MHz radiofrequency electromagnetic fields on Aβ processing in human neuroblastoma and mouse hippocampal neuronal cells

2017 ◽  
Vol 59 (1) ◽  
pp. 18-26 ◽  
Author(s):  
Jeongyeon Park ◽  
Jong Hwa Kwon ◽  
Nam Kim ◽  
Kiwon Song
Keyword(s):  
Protein Expression ◽  
Electromagnetic Fields ◽  
Mrna Level ◽  
Neuronal Cells ◽  
Neural Cells ◽  
Continuous Exposure ◽  
Mrna Levels ◽  
Human Neuroblastoma ◽  
Radiofrequency Electromagnetic Fields ◽  
The Brain

Abstract Alzheimer’s disease (AD) is a neurodegenerative disease leading to progressive loss of memory and other cognitive functions. One of the well-known pathological markers of AD is the accumulation of amyloid-beta protein (Aβ), and its plaques, in the brain. Recent studies using Tg-5XFAD mice as a model of AD have reported that exposure to radiofrequency electromagnetic fields (RF-EMF) from cellular phones reduced Aβ plaques in the brain and showed beneficial effects on AD. In this study, we examined whether exposure to 1950 MHz RF-EMF affects Aβ processing in neural cells. We exposed HT22 mouse hippocampal neuronal cells and SH-SY5Y human neuroblastoma cells to RF-EMF (SAR 6 W/kg) for 2 h per day for 3 days, and analyzed the mRNA and protein expression of the key genes related to Aβ processing. When exposed to RF-EMF, mRNA levels of APP, BACE1, ADAM10 and PSEN1 were decreased in HT22, but the mRNA level of APP was not changed in SH-SY5Y cells. The protein expression of APP and BACE1, as well as the secreted Aβ peptide, was not significantly different between RF-EMF–exposed 7w-PSML, HT22 and SH-SY5Y cells and the unexposed controls. These observations suggest that RF-EMF exposure may not have a significant physiological effect on Aβ processing of neural cells in the short term. However, considering that we only exposed HT22 and SH-SY5Y cells to RF-EMF for 2 h per day for 3 days, we cannot exclude the possibility that 1950 MHz RF-EMF induces physiological change in Aβ processing with long-term and continuous exposure.

scholarly journals The inhibitory effects of rosmarinic acid on catabolism induced by IL-1β in rat chondrocyte

2018 ◽  
Author(s):  
Zhong-Nan Hu ◽  
Li-Juan Huang ◽  
Wei-Ping Chen
Keyword(s):  
Gene Expression ◽  
Extracellular Matrix ◽  
Protein Expression ◽  
Rosmarinic Acid ◽  
Mrna Level ◽  
Inhibitory Effects ◽  
Gene And Protein Expression ◽  
Ecm Degradation ◽  
Interleukin 1Beta ◽  
A Disintegrin And Metalloproteinase

The effects of rosmarinic acid (RosA) on osteoarthritis (OA) was investigated in rat chondrocytes. Chondrocytes were isolated from rat cartilage, incubated with RosA in the presence of interleukin-1beta (IL-1β) (10 ng/ml). The production of IL-6, as well as the mRNA level of aggrecan (ACAN) and collagen 2 (COL2), were assessed. The gene and protein expression of A disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4, ADAMTS-5 were also measured. RosA inhibited the production of IL-6 as well as the gene and protein expression of ADAMTS-4 and ADAMTS-5, in addition, RosA abolished IL-1β-induced inhibition of ACAN and COL2 gene expression. Our results suggest that RosA can inhibit extracellular matrix (ECM) degradation in OA, thus, RosA may be a possible agent in the treatment of OA.

Cisplatin Nano-Liposomes Promoting Apoptosis of Retinoblastoma Cells Both In Vivo and In Vitro

2020 ◽  
Vol 12 (4) ◽  
pp. 536-542
Author(s):  
Lijuan Zhao ◽  
Fei Wang ◽  
Wei Fan
Keyword(s):  
Protein Expression ◽  
Nude Mice ◽  
Caspase 3 ◽  
Mrna Level ◽  
Control Group ◽  
Mrna Levels ◽  
Bax Protein ◽  
Experimental Group

This study was established to investigate the effects of cisplatin nano-liposomes on the apoptosis of the human retinoblastoma (RB) cell line Y79 in vitro and in vivo. Y79 cells were cultured and then exposed to Annexin V/PI to test their apoptosis, tested with the Caspase-3 activity detection kit to examine the change in activity of Caspase-3, and subjected to western blotting to test Bcl-2 and Bax protein expression. Y79-cell-transplanted tumor model in nude mice was also established and divided into three groups, with five nude mice in each. Cisplatin nano-liposomes were applied to the experimental group, cisplatin was injected into the control group, while saline was administered to the blank group, after which the nude mice were killed and the tumor was removed. Tumor volumes and weights in the three groups were compared. Nucleic acid extraction from magnetic beads was adopted to extract DNA, RT-PCR was employed to test Bcl-2 and Bax mRNA levels in tumor tissues, and in situ cell death assay kit was applied to test apoptotic cells. In comparison to the cisplatin solution and DMSO groups, the cisplatin liposome group showed higher Y79 apoptotic rate, Caspase-3 activity, and Bax protein expression, and lower Bcl-2 protein expression (all P < 0 05). In comparison with the control and blank groups, the experimental group showed lower tumor volume, weight, and Bcl-2 mRNA level of nude mice. In addition, in comparison with the control group, the experimental group showed higher cellular apoptotic rate and Bax mRNA level. In terms of the clinical effects of cisplatin nano-liposomes on a tumor transplant in nude mice with cervical cancer, they were shown to promote tumor apoptosis.

Relationship of thymidylate synthase levels to outcome of malignant pleural mesothelioma patients treated with pemetrexed-based chemotherapy

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. 7508-7508
Author(s):  
G. Selvaggi ◽  
L. Righi ◽  
P. Ceppi ◽  
E. Bacillo ◽  
A. Billè ◽  
...  
Keyword(s):  
Multivariate Analysis ◽  
Protein Expression ◽  
Malignant Pleural Mesothelioma ◽  
Thymidylate Synthase ◽  
Mrna Level ◽  
Single Agent ◽  
Outcome Data ◽  
Rank Test ◽  
Pleural Mesothelioma ◽  
Mrna Levels

7508 Background: Pemetrexed has shown activity in malignant pleural mesothelioma (MPM) but scanty data are available on the expression of thymidylate synthase (TS), its most important molecular target. Methods: From a database of 75 non-surgical, chemotherapy-naive MPM patients from our Institution in the period 2004–2008, 50 (male/female: 37/13, median age: 65 years) met the selection criteria i.e. epithelial type, availability of thoracoscopic tissue and outcome data. Pemetrexed was administered as single agent (14/50) or in combination with cisplatin or carboplatin (36/50). Retrospectively TS protein expression levels were evaluated by immunohistochemistry and quantified with H-score method. In addition, mRNA extraction was performed in 23 micro-dissected tissues and TS relative levels quantified by RT-PCR. Survival probability was assessed by Kaplan-Meier method and results compared by log-rank test. Cox multivariate analysis for survival was performed adjusting for clinical-pathological variables. Results: Thirty-two patients had progressive disease and 24 had died at the time of the analysis. Median time to progression (TTP) and median survival time (MST) were 11.6 and 20.9 months, respectively. Median TS H-score value was 90 (5–240). No correlation were found with sex, age, PS, stage and chemotherapy regimen. Patients with high TS H-score (4th quartile) had a significantly shorter MST (13.3 vs 21.1 months, p<0.01) and showed a trend for shorter TTP (8.3 vs 11.9 months, p=0.07). Median TS mRNA level was 1.88 (1–3.7 unit-less ratio) and a significant correlation between mRNA and protein expression (RS=0.67, p<0.0001) was found. Patients with high TS mRNA levels (4th quartile) had significantly shorter TTP (8.7 vs 14.7 months, p=0.019) and MST (11.7 vs 24.7, p=0.018). Multivariate analysis for survival indicated that TS protein levels were an independent prognostic factor (HR=2.17; CI 1.04–4.54; p=0.038). Conclusions: TS (protein and mRNA) levels predict outcome of epithelial MPM patients treated with pemetrexed-based chemotherapy. TS quantification, if confirmed in larger prospective studies, could be used to select those patients more likely to respond to chemotherapy. [Table: see text]

Cloning and expression of progesterone receptor isoforms A and B in bovine corpus luteum

2015 ◽  
Vol 27 (7) ◽  
pp. 1029 ◽  
Author(s):  
Robert Rekawiecki ◽  
Magdalena Karolina Kowalik ◽  
Jan Kotwica
Keyword(s):  
Mrna Level ◽  
First Trimester ◽  
Oestrous Cycle ◽  
Cloning And Expression ◽  
Corpora Lutea ◽  
Mrna Levels ◽  
Variable Expression ◽  
Progesterone Receptor Isoforms ◽  
Protein Levels ◽  
Receptor Isoforms

Progesterone (P4) affects a cell through its nuclear receptor (PGR), which has two main isoforms: A (PGRA) and B (PGRB). A partial section of previously unknown PGRB cDNA from cattle was cloned. Next, mRNA and protein levels for these two isoforms in corpora lutea (CL) collected during different stages of the oestrous cycle and pregnancy were determined. The PGRB mRNA level was highest on Days 2–5 of the oestrous cycle, decreased over the next few days (P < 0.01) and increased again slightly on Days 17–20 (P < 0.05). During pregnancy, PGRB mRNA was at its lowest level during Weeks 3–5 (P < 0.01) and highest during Weeks 6–12 (P < 0.01). The profile of PGRA mRNA levels was similar to that of PGRB throughout the oestrous cycle. The PGRA protein level was highest on Days 2–10 of the oestrous cycle, decreased continuously to its lowest concentration on Days 17–20 (P < 0.01) and during Weeks 3–5 of pregnancy (P > 0.05) and increased during Weeks 6–12 (P < 0.05). PGRB protein concentration followed a similar pattern but at a markedly lower level. Both PGRA and PGRB isoforms are involved in the regulation of P4 action, especially in the newly formed CL and developed CL in the first trimester of pregnancy. These data suggest that the variable expression of these isoforms during the oestrous cycle may depend on the influence of P4.

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