scholarly journals Human Synovia Contains Trefoil Factor Family (TFF) Peptides 1–3 Although Synovial Membrane Only Produces TFF3: Implications in Osteoarthritis and Rheumatoid Arthritis

2019 ◽  
Vol 20 (23) ◽  
pp. 6105
Author(s):  
Popp ◽  
Schicht ◽  
Garreis ◽  
Klinger ◽  
Gelse ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Real Time ◽  
Real Time Pcr ◽  
Synovial Membrane ◽  
Mrna Level ◽  
Trefoil Factor ◽  
Healthy Donors ◽  
Trefoil Factor Family ◽  
Secretion Product ◽  
Tff Peptides

Objective: Trefoil factor family peptide 3 (TFF3) has been shown to support catabolic functions in cases of osteoarthritis (OA). As in joint physiology and diseases such as OA, the synovial membrane (SM) of the joint capsule also plays a central role. We analyze the ability of SM to produce TFF compare healthy SM and its secretion product synovial fluid (SF) with SM and SF from patients suffering from OA or rheumatoid arthritis (RA). Methods: Real-time PCR and ELISA were used to measure the expression of TFFs in healthy SM and SM from patients suffering from OA or RA. For tissue localization, we investigated TFF1-3 in differently aged human SM of healthy donors by means of immunohistochemistry, real-time PCR and Western blot. Results: Only TFF3 but not TFF1 and -2 was expressed in SM from healthy donors as well as cases of OA or RA on protein and mRNA level. In contrast, all three TFFs were detected in all samples of SF on the protein level. No significant changes were observed for TFF1 at all. TFF2 was significantly upregulated in RA samples in comparison to OA samples. TFF3 protein was significantly downregulated in OA samples in comparison to healthy samples and cases of RA significantly upregulated compared to OA. In contrast, in SM TFF3 protein was not significantly regulated. Conclusion: The data demonstrate the production of TFF3 in SM. Unexpectedly, SF contains all three known TFF peptides. As neither articular cartilage nor SM produce TFF1 and TFF2, we speculate that these originate with high probability from blood serum.

scholarly journals Trefoil Factor Family (TFF) Peptides and Their Diverse Molecular Functions in Mucus Barrier Protection and More: Changing the Paradigm

2020 ◽  
Vol 21 (12) ◽  
pp. 4535 ◽  
Author(s):  
Werner Hoffmann
Keyword(s):  
Thiol Group ◽  
Suppressor Gene ◽  
Immune Defense ◽  
Trefoil Factor ◽  
Gastric Mucus ◽  
Free Thiol Group ◽  
Trefoil Factor Family ◽  
Molecular Functions ◽  
Mucus Barrier ◽  
Tff Peptides

Trefoil factor family peptides (TFF1, TFF2, TFF3) are typically co-secreted together with mucins. Tff1 represents a gastric tumor suppressor gene in mice. TFFs are also synthesized in minute amounts in the immune and central nervous systems. In mucous epithelia, they support rapid repair by enhancing cell migration (“restitution”) via their weak chemotactic and anti-apoptotic effects. For a long time, as a paradigm, this was considered as their major biological function. Within recent years, the formation of disulfide-linked heterodimers was documented for TFF1 and TFF3, e.g., with gastrokine-2 and IgG Fc binding protein (FCGBP). Furthermore, lectin activities were recognized as enabling binding to a lipopolysaccharide of Helicobacter pylori (TFF1, TFF3) or to a carbohydrate moiety of the mucin MUC6 (TFF2). Only recently, gastric TFF1 was demonstrated to occur predominantly in monomeric forms with an unusual free thiol group. Thus, a new picture emerged, pointing to diverse molecular functions for TFFs. Monomeric TFF1 might protect the gastric mucosa as a scavenger for extracellular reactive oxygen/nitrogen species. Whereas, the TFF2/MUC6 complex stabilizes the inner layer of the gastric mucus. In contrast, the TFF3–FCGBP heterodimer (and also TFF1–FCGBP) are likely part of the innate immune defense of mucous epithelia, preventing the infiltration of microorganisms.

scholarly journals Discrepancy between Jun/Fos Proto-Oncogene mRNA and Protein Expression in the Rheumatoid Arthritis Synovial Membrane

J ◽  
2020 ◽  
Vol 3 (2) ◽  
pp. 181-194 ◽  
Author(s):  
René Huber ◽  
Bruno Stuhlmüller ◽  
Elke Kunisch ◽  
Raimund W. Kinne
Keyword(s):  
Rheumatoid Arthritis ◽  
Protein Expression ◽  
Synovial Membrane ◽  
Rna Binding ◽  
Mrna Level ◽  
Joint Disease ◽  
Protein Levels ◽  
Modifying Factors ◽  
Mrna And Protein Expression ◽  
Joint Trauma

Rheumatoid arthritis (RA) is a chronic inflammatory and destructive joint disease characterized by overexpression of pro-inflammatory/pro-destructive mediators, whose regulation has been the focus of our previous studies. Since the expression of these proteins commonly depends on AP-1, the expression of the AP-1-forming subunits cJun, JunB, JunD, and cFos was assessed in synovial membrane (SM) samples of RA, osteoarthritis (OA), joint trauma (JT), and normal controls (NC) using ELISA and qRT-PCR. With respect to an observed discrepancy between mRNA and protein levels, the expression of the mRNA stability-modifying factors AU-rich element RNA-binding protein (AUF)-1, tristetraprolin (TTP), and human antigen R (HuR) was measured. JunB and JunD protein expression was significantly higher in RA-SM compared to OA and/or NC. By contrast, jun/fos mRNA expression was significantly (cjun) or numerically decreased (junB, junD, cfos) in RA and OA compared to JT and/or NC. Remarkably, TTP and HuR were also affected by discrepancies between their mRNA and protein levels, since they were significantly decreased at the mRNA level in RA versus NC, but significantly or numerically increased at the protein level when compared to JT and NC. Discrepancies between the mRNA and protein expression for Jun/Fos and TTP/HuR suggest broad alterations of post-transcriptional processes in the RA-SM. In this context, increased levels of mRNA-destabilizing TTP may contribute to the low levels of jun/fos and ttp/hur mRNA, whereas abundant mRNA-stabilizing HuR may augment translation of the remaining mRNA into protein with potential consequences for the composition of the resulting AP-1 complexes and the expression of AP-1-dependent genes in RA.

Trefoil factor family peptides – friends or foes?

2015 ◽  
Vol 6 (5-6) ◽  
pp. 343-359 ◽  
Author(s):  
Maike Busch ◽  
Nicole Dünker
Keyword(s):  
Cancer Research ◽  
Tumor Suppressors ◽  
Current Knowledge ◽  
Tumor Biology ◽  
Epigenetic Mechanisms ◽  
Trefoil Factor ◽  
Protective Function ◽  
Trefoil Factor Family ◽  
Tff Peptides

AbstractTrefoil factor family (TFF) peptides are a group of molecules bearing a characteristic three-loop trefoil domain. They are mainly secreted in mucous epithelia together with mucins but are also synthesized in the nervous system. For many years, TFF peptides were only known for their wound healing and protective function, e.g. in epithelial protection and restitution. However, experimental evidence has emerged supporting a pivotal role of TFF peptides in oncogenic transformation, tumorigenesis and metastasis. Deregulated expression of TFF peptides at the gene and protein level is obviously implicated in numerous cancers, and opposing functions as oncogenes and tumor suppressors have been described. With regard to the regulation of TFF expression, epigenetic mechanisms as well as the involvement of various miRNAs are new, promising aspects in the field of cancer research. This review will summarize current knowledge about the expression and regulation of TFF peptides and the involvement of TFF peptides in tumor biology and cancerogenesis.

scholarly journals Role of protein kinase Cβ in rhythmic contractions of human pregnant myometrium

Reproduction ◽  
2007 ◽  
Vol 133 (4) ◽  
pp. 797-806 ◽  
Author(s):  
Katsuhiko Yasuda ◽  
Tsuyoshi Nakamoto ◽  
Masahiro Yasuhara ◽  
Hidetaka Okada ◽  
Tatsuya Nakajima ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Real Time ◽  
Real Time Pcr ◽  
Contractile Activity ◽  
Mrna Level ◽  
Human Myometrium ◽  
Uterine Contractions ◽  
Activity Frequency ◽  
Rhythmic Contractions

To assess the role of protein kinase Cβ (PKCβ) in human myometrial contractions during pregnancy, we evaluated the effect of a PKCβ inhibitor (LY333531) on the pregnant and nonpregnant myometrial contractions and compared the level of PKCβ in the pregnant myometrium with that in the nonpregnant myometrium. The effects of LY333531 on the myometrial contractions were examined by measuring contractile activity (frequency and amplitude). PKCβ in human myometrium was assessed at mRNA level using real-time PCR method. The characteristics of contractile activity were different between the pregnant and the nonpregnant myometrium. The amplitude of rhythmic contractions in the preterm and term myometrium was increased 2- to 2.5-fold when compared with that in the nonpregnant myometrium, but the frequency of rhythmic contractions was decreased by about half. LY333531 (10−6M) reduced the increased amplitude in the preterm and term myometrium by about 50%, and the inhibitory effects of LY333531 in the pregnant myometrium were significantly greater than that in the nonpregnant myometrium (about 50 vs 25%). However, the frequency in the pregnant and nonpregnant myometrium was not influenced by LY333531. Real-time PCR revealed a significant, five- to sevenfold increase in the expression of PKCβ mRNA in the preterm and term myometrium when compared with the nonpregnant myometrium. These findings suggest that the increased amplitude of human myometrial contractions during pregnancy is related to the increased level of PKCβ. A PKCβ inhibitor may reduce preterm uterine contractions and prevent preterm delivery.

P–341 Epigenetic role of the H2BK5ac histone modification on the expression of ALX1 and PDHX genes in the endometriotic tissues and normal endometrium

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
S Sarshomar ◽  
S Sarshomar ◽  
F Chitsazian ◽  
F Ghaffari ◽  
M Shahhoseini ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Chromatin Immunoprecipitation ◽  
Mrna Level ◽  
Rna Extraction ◽  
Migration And Invasion ◽  
Epigenetic Mark ◽  
Promoter Regions ◽  
Normal Endometrium

Abstract Study question Evolution of the ALX1 and PDHX genes expression and incorporation of H2BK5ac mark on the promoter of these genes in endometriotic tissues versus normal endometrium Summary answer Lower incorporation of H2BK5ac mark in ALX1 and PDHX promoters can because to downregulation of these genes in endometriotic tissues compared to normal endometrium. What is known already Endometriosis is considered as multifactorial disease affected by genetic, hormonal, and environmental factors. Recent evidences suggest the role of epigenetic mechanisms in this disease. Aristaless-like homeobox1 (ALX1) and Pyruvate Dehydrogenase Protein X (PDHX) genes are considered in this study. Studies show that upregulation of the ALX1 gene cause cell proliferation, migration, and invasion in cancer cells. PDHX is involved in cellular metabolism and acts as a tumor suppressor gene while maintaining normal homeostasis. It is Hypothesized that H2BK5ac which is known as a dynamic marker in promoter regions of active genes, may be involved in regulation of these gene expression. Study design, size, duration Ten eutopic and ectopic endometrium tissue, as well as ten normal endometrium, were collected. Ectopic biopsies were obtained using diagnostic laparoscopy, while the endometrial control samples and eutopic samples were collected via pipelle. Participants/materials, setting, methods RNA extraction and cDNA synthesis were done then the expression of ALX1 and PDHX genes evaluated by quantitative real-time PCR. Promoter regions of mentioned genes investigated for the incorporation of the epigenetic mark of H2BK5ac using Chromatin immunoprecipitation (ChIP) followed by real-time PCR. Data analysis performed using One-way ANOVA analysis (SPSS software) considered the significant level of P < 0.05. Main results and the role of chance Results showed that the expression of ALX1 was significantly decreased in eutopic endometrial samples compared to normal endometrium (p = 0.007). Also, there was a significant reduction in PDHX mRNA level in the eutopic and ectopic samples vs. normal endometrium (p = .017 and p =.021, respectively). The chromatin immunoprecipitation real-time PCR (ChIP PCR) analyses showed significantly lower incorporation of H2BK5ac epigenetic mark in ALX1 promoter in eutopic endometrial samples compared to normal endometrium (p = 0.007). Also, reduced incorporation of H2BK5ac at the PDHX promoter region was observed in both eutopic and ectopic endometrial samples compared to normal endometrium (p = 0.004 and p = 0.003, respectively). Limitations, reasons for caution The main limitation of our study is the low number of samples. Wider implications of the findings: Our results suggest that the marked lower levels of H2BK5ac in regulatory regions of ALX1and PDHX might lead to deregulation of these genes in tissue endometriotic samples. Trial registration number ‘not applicable’ for non-clinical trials

Synthesis and localization of trefoil factor family (TFF) peptides in the human urinary tract and TFF2 excretion into the urine

2010 ◽  
Vol 339 (3) ◽  
pp. 639-647 ◽  
Author(s):  
Margarita Rinnert ◽  
Margitta Hinz ◽  
Peter Buhtz ◽  
Frank Reiher ◽  
Wolfgang Lessel ◽  
...  
Keyword(s):  
Urinary Tract ◽  
Trefoil Factor ◽  
Trefoil Factor Family ◽  
Tff Peptides

The importance for standardization and validation of real-time PCR for the detection of mature miR-21 in the blood of patients

2020 ◽  
pp. 66-67
Author(s):  
Л.В. Шуленина ◽  
Д.В. Салеева
Keyword(s):  
Real Time ◽  
Correlation Coefficient ◽  
Real Time Pcr ◽  
Coefficient Of Variation ◽  
Healthy Donors ◽  
Mature Micrornas ◽  
Pcr Method ◽  
Detection And Quantification

Процессы обнаружения и количественного определения методом ПЦР в реальном времени в крови пациентов зрелых микроРНК, в том числе и miR-21, должны быть стандартизированы и валидированы. Проведенные нами исследования, показывают, что специфичность метода ПЦР для miR-21 в крови здоровых доноров может быть демонстрирована образованием продукта-ампликона размером 67 нуклеотидов, прецизионность в условиях сходимости и воспроизводимости, выраженная в виде коэффициента вариации, составляет менее 2% и 3 % соответственно, а линейность метода подтверждается коэффицентом корреляции r≥0,99. The processes of detection and quantification by real-time PCR in blood of mature microRNAs, including miR-21, should be standardized and validated. Our studies show that the specificity of PCR method for miR-21 in the blood of healthy donors can be demonstrated by the formation of a 67 bp amplicon product, precision under conditions of convergence and reproducibility, expressed using the coefficient of variation, is less than 2% and 3%, respectively, and the linearity of PCR method for miR-21 is confirmed by the correlation coefficient r≥0.99.

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