scholarly journals Evaluating the Expression of Wnt Pathway Related Genes in Mouse Vitrified Preantral Follicles: An Experimental Study

2021 ◽  
Author(s):  
Shahla Babaki ◽  
Saeed Zavareh ◽  
Parisa Farrokh ◽  
Meysam Nasiri
Keyword(s):  
Wnt Pathway ◽  
Ovarian Follicle ◽  
Critical Role ◽  
Wnt Signaling Pathway ◽  
Culture Period ◽  
Control Group ◽  
P Value ◽  
Mrna Levels ◽  
Preantral Follicles ◽  
Expression Levels

Background: Wnt signaling pathway plays critical role in ovarian follicle development. Therefore, the aim of this study was to evaluate the effects of vitrification on the expression of Wnt pathway related genes in preantral follicles (PFs). Methods: Isolated PFs (n=982) of 14-16 day old female mice (n=45: 15 for each group) were divided into fresh (n=265), toxicity (n=272), and vitrified (n=265). The mRNA levels of Wnt2, Wnt4, Lrp5 and Fzd3 were evaluated by real-time PCR on the 2nd and 6th days of culture period. One-way ANOVA was conducted to analyze the data. Post hoc Tukey's HSD was used for multiple comparisons and p-value less than 0.05 was considered statistically significant. Results: The developmental parameters of fresh PFs were significantly higher than those of vitrified (p<0.001). There were no differences between fresh and vitrified PFs on the 2nd day of culture (p<0.001). Wnt4 expression levels decreased significantly in vitrified groups compared with fresh ones (p<0.001). Fzd3 and Lrp expression levels increased significantly in vitrified groups compared with those in the fresh group on the 2nd day (p<0.001). On the 6th day of culture period, the expression levels of Wnt2 and Fzd3 increased significantly in vitrified group compared to those of fresh group (p<0.001). Moreover, the expression levels of Wnt4 and Lrp increased significantly in toxicity groups compared to those of the control group (p<0.001). Conclusion: Vitrification increase the expression levels of Wnt2, Lrp and Fzd3 genes of PFs during in vitro culture

Overexpression of the LEF-1 and TCF4 Transcription Factors in B-CLL: Further Evidence for a Role of the Wnt Signaling Pathway in B-CLL Biology and Leukemogenesis

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 544-544 ◽  
Author(s):  
Albert Gutierrez ◽  
Renee Tschumper ◽  
Jeanette Eckel-Passow ◽  
Neil E. Kay ◽  
Diane F. Jelinek
Keyword(s):  
B Cells ◽  
Wnt Signaling ◽  
B Cell ◽  
Wnt Pathway ◽  
Perfect Match ◽  
Wnt Signaling Pathway ◽  
Healthy Adults ◽  
P Value ◽  
Expression Levels

Abstract Background: B-chronic lymphocytic leukemia (B-CLL) is the most common leukemia in the Western world but its pathogenesis remains largely unknown. Our strategy was to employ gene expression profiling (GEP) to discover genetic differences between CLL B cells and blood B cells from healthy adults. Previously, we and others identified the transcription factor (TF) lymphocyte enhancer factor-1 (LEF-1) as one of several genes significantly over expressed in CLL B cells suggesting a role for the oncogenic Wnt signaling pathway. LEF-1 acts as a central mediator of Wnt signaling and is crucial for the proliferation and survival of pro-B cells during development. Recently, deregulated LEF-1 activation has been directly linked with leukemogenesis. Methodology: The goals of our study were to: validate LEF-1 over expression in CLL B cells in a new cohort of B-CLL patients and normal controls; study CLL B cell LEF-1 protein expression; and investigate CLL B cell expression levels of other genes linked to Wnt signaling. To accomplish these goals, Affymetrix U133 GEP was performed on B cells from 41 B-CLL patients who were enrolled on a trial of combined pentostatin, cyclophosphamide, and rituximab and 11 healthy adults over age 60. Only the perfect match data were utilized for all analyses; the non-background corrected perfect match data were normalized using an intensity-dependent procedure and analyses were done using the base-2 logarithm transformed normalized values. Results: GEP analysis confirmed CLL B cells expressed the LEF-1 gene at ~28-fold higher levels than normal B cells (p&lt;.0001). These results were validated by quantitative PCR and all CLL B cells studied expressed LEF-1 mRNA. By contrast, LEF-1 mRNA expression was undetectable in blood B cells from healthy adults both before and after in vitro mitogenic stimulation suggesting aberrant LEF-1 expression in B-CLL. Moreover, LEF-1 protein was readily detected by flow cytometry in CLL B cell samples and all clonal B cells expressed a uniform level of LEF-1. Full length LEF-1 is more oncogenic and predominates in certain cancers, while a dominant negative short isoform is found at greater levels in normal lymphocytes. Of interest, western blot analysis revealed that CLL B cells predominantly expressed full length oncogenic LEF-1. Ongoing studies are focused on silencing LEF-1 expression to determine LEF-1 target genes. In this regard, IGFBP4 (IGF binding protein 4) mRNA expression levels are 19-fold higher in CLL vs control B cells (p&lt;0.0001) and the IGFBP4 protein was recently shown to antagonize Wnt signaling. A scan of the IGFBP4 promoter element identified 3 possible LEF/TCF consensus sites. Thus, there is a possible negative feedback loop in CLL, with activation of the Wnt pathway leading to expression of a negative Wnt regulator. Finally, we queried the GEP data for evidence of differential expression of 63 other Wnt-related genes. Surprisingly, in addition to LEF1, only five genes met our selection threshold of ≥1.5 fold change in expression between CLL and control B cells and p value &lt;.001. These genes were CSNK1D (casein kinase, delta 1; 1.9-fold higher in control vs CLL; p&lt;0.0001); CCND2 (cyclin D2; 2.4-fold higher in CLL vs control; p&lt;0.0001); JUN (3.2-fold higher in control vs CLL); WNT3 (6.4-fold higher in CLL vs control; p&lt;0.0001); and TCF4 (transcription factor 4, alias ITF-2; 4-fold higher in CLL vs control; p&lt;0.0001). To our knowledge, we are the first to report that the TCF4 (ITF-2) gene is over expressed in B-CLL. This TF is of great interest because it is a known downstream target of the Wnt/TCF pathway, is activated in human cancers with b-catenin defects, and it promotes neoplastic transformation. Using Spearman correlation analysis to explore the relationship between LEF1, TCF4, and WNT3 expression levels, we observed a marginal correlation between WNT3 expression and LEF1 (ρ=0.31; p value=0.05) and TCF4 (ρ=0.29; p value=0.07). Summary: These studies add new support for a critical role of the Wnt pathway in CLL. The WNT3 results are consistent with the literature; however, it is notable that we failed to corroborate other reports demonstrating that other Wnts and Fzd genes are differentially expressed in CLL B cells vs normal B cells. It is striking that CLL B cells over express two TFs in this pathway that have been linked with neoplastic transformation. Ongoing studies are aimed at further elucidating the roles of these genes in B-CLL.

Toll like Receptors (TLRs) in B Cell Chronic Lymphocytic Leukemia (B-CLL).

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4980-4980
Author(s):  
Maria S. Manoussaka ◽  
Azka Memon ◽  
Amit Nathwani ◽  
Nino Porakishvili ◽  
Edward Clark ◽  
...  
Keyword(s):  
B Cells ◽  
Adaptor Protein ◽  
Lymphocytic Leukemia ◽  
Control Group ◽  
Specific Gene ◽  
Mrna Levels ◽  
Cycle Number ◽  
Expression Levels ◽  
Significant Difference ◽  
Gene Status

Abstract Introduction: B-CLL is characterised by the accumulation of long-lived CD5+ B cells. Mature B cells can be activated via TLRs to produce immunoglobulin and proliferate. We have previously reported differential surface expression of CD180, TLR-4 associated receptor, by B-CLL cells in relation to IgVh gene status (Porakishvili N et al., 2004; Blood (ASH annual meeting abstracts) 104: 2807). To further characterise subsets of leukemic cells, we assessed B-CLL cells for TLR mRNA expression. Method: B cells from untreated (n=24) and treated (CHOP n=3, Thalidomide n=1 and chlorambucil and prednisolone n=2) CLL patients, and healthy aged matched controls, (n=14) were isolated by CD19+ selection using iMACs columns (Miltenyi Biotec), total RNA extracted (Qiagen) and primers for TLR2, TLR4, TLR9, CD180 and the adaptor protein MYD88 were used for TaqMan PCR analysis (ABI). GAPDH was used for normalization. Data were expressed as mean delta cycle number (ct) ± standard deviation, giving a value inversely related to the level of expression of the specific gene. Patients were separated into groups according to IgVh gene mutation status or level of Zap 70 expression. Statistical analysis was performed using by Mann-Whitney U test and Spearman rank correlation or Pearson’s correlation coefficient. Results: Overall, there was a heterogeneous pattern of TLR expression in patients and controls with no significant differences between them. Treated patients were characterised by higher mRNA levels for the all TLRs measured (TLR9 p=0.018) and MYD88 (MYD88 p=0.026) compared with untreated patients, despite no significant difference of their GAPDH levels (p&gt;0.05). Significant correlations were seen between mRNA levels for the TLRs within the age matched control group (all p&lt;0.05). Groups of patients were further analysed according to mutated (mutated &gt;2%) or unmutated IgVh genes (unmutated &lt;2%). No significant correlation in the TLR expression levels was found within the mutated patient group (n=8) or Zap 70 low expression group (&lt;0.7 level) (n=11). Preliminary data from patients with an unmutated IgVh gene status (n=4) or according to levels of ZAP 70high expression (n=5) showed correlation of mRNA levels for TLRs and MYD88 expression (all p&lt;0.05). Conclusion: Our data indicates a direct relationship between MYD88 and TLRs transcription levels in healthy control individuals. Lack of significant correlation amongst TLR mRNA levels in CLL patients with mutated IgVh genes and Zap70low expression levels suggests aberrant regulation of TLRs in this group.

Expression of Ara-C Metabolizing Enzymes Mediates in Vitro Sensitivity to Ara-C in Primary AML Cells From Patients with Denovo AML,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3481-3481
Author(s):  
Ajay Abraham ◽  
Savitha Varatharajan ◽  
Ashok kumar Jayavelu ◽  
Shaji R Velayudhan ◽  
Rayaz Ahmed ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Candidate Genes ◽  
P Value ◽  
Mrna Levels ◽  
Wild Type ◽  
Mutation Status ◽  
Expression Levels ◽  
In Vitro Sensitivity ◽  
Mrna Expression Levels

Abstract Abstract 3481 Wide inter-individual variation in terms of treatment outcome and toxic side effects of treatment exist among patients with AML receiving chemotherapy with cytarabine (ara-C) and daunorubicin. The pre-requisite for the cytotoxic action of pro-drug Ara-C is the enzymatic conversion to its active tri-phosphorylated form ara-CTP. Many drug activating (Deoxycytidine kinase (dCK) and human Equilibrative Nucleoside Transporter 1 (hENT1) and deactivating (Cytidine deaminase (CDA), 5'nucleotidase (NT5C2) genes and ribonucleoside reductase (RRM1), which are involved in transport and biotransformation of cytarabine contribute to the variation in ara-C sensitivity in AML patients. FLT3-ITD and NPM1 mutations act as major poor and good prognostic markers respectively in cytogenetically normal AML. The effect of these mutations in ara-C metabolism remains to be elucidated. The present study aims to determine independent as well as the combined effect of ara-C metabolizing genes mRNA expression on in-vitro ara-C cytotoxicity and the role of FLT3-ITD and NPM mutations on mRNA expression of these genes. Diagnostic bone marrow sample (median blasts 65%; range 21 – 98%) from 98 adult patients with de novo AML (other than AML-M3) were included in this study. mRNA expression levels for each target gene relative to housekeeping gene GAPDH was analyzed using Taqman based gene expression assays. In vitro cytotoxicity was assessed using MTT cell viability assay and IC-50 was calculated. In vitro sensitivity or resistance was classified on the basis of the IC-50 values <6uM and >6uM ara-C respectively. FLT3 ITD and NPM mutation status at diagnosis were determined through PCR followed by Genescan analysis using genomic DNA samples. Type of NPM mutation was identified by sequencing. When ara-C IC-50 values were compared with the mRNA expression levels of these candidate genes, Ara-C sensitive samples (n= 30; IC-50 < 6uM) showed significantly higher mRNA expression of dCK and hENT1 compared to those with Ara-C resistance (n=51) IC50 >6uM (median 314 (61.56 – 1232) vs. 180 (31.87 – 749.2); p = 0.0004 and median 172.1 (44.12 – 657.6) vs. 96.19 (37.49 – 432.4), p= 0.0008 respectively. RRM1 and NT5C2 did not show any association with in vitro Ara-C cytotoxicity, while CDA showed a trend towards association with lower CDA expression in ara-C sensitive samples. Based on these findings we put forward Ara-C resistance index (RI). RI is calculated by the formula RI = ΔCT (dCK X ENT1)/ ΔCT CDA. (Smaller ΔCT value= higher mRNA expression). RI values were significantly higher in resistant (IC50 >6uM) compared to sensitive cells (median: 6.084; range 1.89–11.82) vs. 3.702 (1.89–9.80); p=<0.0001). This association should now be validated in an independent cohort. Effects of NPM and FLT3 mutation status on Ara-C metabolizing genes were then evaluated. No significant association was found between FLT3-ITD status and the mRNA expression of these candidate genes. Interestingly, dCK mRNA levels were significantly higher in samples with NPM mutation (n=39) compared to NPM wild type (n=59); median 272.3 (41.64–1232) vs. 188.6 (31.87–1030); p value= 0.01. When analysed separately, patients with NPM type A mutation (n=27) showed significantly higher dCK expression (median 347.4 (41.64–1232) vs. 188.6 (31.87–1030); p value= 0.003 compared to those with wild type NPM1. This first report showing an association between expression profiles of ara-C metabolizing genes and NPM mutation should form the basis for evaluating their clinical correlations. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Wnt Modulating Agents Inhibit Human Cytomegalovirus Replication

2013 ◽  
Vol 57 (6) ◽  
pp. 2761-2767 ◽  
Author(s):  
Arun Kapoor ◽  
Ran He ◽  
Rajkumar Venkatadri ◽  
Michael Forman ◽  
Ravit Arav-Boger
Keyword(s):  
Stem Cell ◽  
Cancer Stem Cell ◽  
Human Cytomegalovirus ◽  
Wnt Pathway ◽  
Critical Role ◽  
Wnt Signaling Pathway ◽  
Negative Regulator ◽  
Infected Cells ◽  
Hcmv Replication

ABSTRACTInfection with human cytomegalovirus (HCMV) continues to be a threat for pregnant women and immunocompromised hosts. Although limited anti-HCMV therapies are available, development of new agents is desired. The Wnt signaling pathway plays a critical role in embryonic and cancer stem cell development and is targeted by gammaherpesviruses, Epstein-Barr virus (EBV), and Kaposi's sarcoma-associated herpesvirus (KSHV). HCMV infects stem cells, including neural progenitor cells, during embryogenesis. To investigate the role of Wnt in HCMV replicationin vitro, we tested monensin, nigericin, and salinomycin, compounds that inhibit cancer stem cell growth by modulating the Wnt pathway. These compounds inhibited the replication of HCMV Towne and a clinical isolate. Inhibition occurred prior to DNA replication but persisted throughout the full replication cycle. There was a significant decrease in expression of IE2, UL44, and pp65 proteins. HCMV infection resulted in a significant and sustained decrease in expression of phosphorylated and total lipoprotein receptor-related protein 6 (pLRP6 and LRP6, respectively), Wnt 5a/b, and β-catenin and a modest decrease in Dvl2/3, while levels of the negative regulator axin 1 were increased. Nigericin decreased the expression of pLRP6, LRP6, axin 1, and Wnt 5a/b in noninfected and HCMV-infected cells. For all three compounds, a correlation was found between expression levels of Wnt 5a/b and axin 1 and HCMV inhibition. The decrease in Wnt 5a/b and axin 1 expression was more significant in HCMV-infected cells than noninfected cells. These data illustrate the complex effects of HCMV on the Wnt pathway and the fine balance between Wnt and HCMV, resulting in abrogation of HCMV replication. Additional studies are required to elucidate how HCMV targets Wnt for its benefit.

scholarly journals Effect of Concentrated growth factor (CGF) on human gingival fibroblast viability on titanium discs

2021 ◽  
Author(s):  
Masoumeh Faramarzi ◽  
Leila Roshangar ◽  
Adileh Shirmohammadi ◽  
Mehrnoosh Sadighi ◽  
Azadeh Madanipour ◽  
...  
Keyword(s):  
Growth Factor ◽  
Critical Role ◽  
Control Group ◽  
P Value ◽  
Human Gingival Fibroblast ◽  
Gingival Fibroblast ◽  
Gingival Fibroblasts ◽  
Term Survival ◽  
Accelerate Wound Healing

Abstract Objective Attachment of peri- implant mucosa to implant is crucial for long term survival of implant. Soft tissue healing around implants has been of great interest due to its important role in long-term maintenance of implant therapy. Considering the critical role of gingival fibroblasts in periodontal repair, the aim of this study was to evaluate the Concentrated Growth Factor (CGF) as an innovative approach to accelerate wound healing and increase the connective tissue seal around dental implants. Results 40% and 80% Concentrations of CGF significantly improved human gingival fibroblasts (HGF) viability compared to the control group (P value = 0.001). But the comparison of the other group with the control group was not statistically significant. The difference between 40% and 80% concentrations of CGF was not statistically significant (P value = 0.061). Results showed that the viability of HGF treated with CGF on titanium discs(test groups 2) significantly increased as compared to the test groups 1(without CGF) at 24 hours (P value = 0.001).Our results showed that 40% concentration of CGF at 24 hours significantly increased HGF viability.

Aligned Expression of IFI16 and STING Genes in RRMS Patients’ Blood

2020 ◽  
Vol 20 (6) ◽  
pp. 878-886
Author(s):  
Sobhan Helbi ◽  
Behnam Ravanbakhsh ◽  
Mohammad Karimi ◽  
Wesam Kooti ◽  
Nahid Jivad
Keyword(s):  
Mrna Level ◽  
Critical Role ◽  
Control Group ◽  
Case Group ◽  
Common Disease ◽  
Type I ◽  
Mrna Levels ◽  
Dna Sensing ◽  
Blood Samples ◽  
Significant Difference

Objective: Multiple sclerosis (MS) is a chronic neurodegenerative disease of the central nervous system. The most common disease phenotype is Relapsing-Remitting MS (RRMS). Beta interferons are the first line of RRMS patients’ treatment. Interferon-inducible protein 16 (IFI16) as a DNA sensing molecule and its downstream complex stimulator of interferon genes (STING) play a critical role in the activation of type I interferons. Hence we aimed to evaluate the expression rate of IFI16 and STING in RRMS patients’ blood under a different type of IFNβ treatment. Methods: In the present study, 99 individuals participated. The participants were divided into 4 groups: 28 control subjects, 25 new cases of RRMS patients, 25 RRMS patients treated with IFNβ-1a (B1a), 21 RRMS patients treated with IFNβ-1b (B1b). The EDTA-treated blood samples were taken and transferred at standard conditions to the Cellular and Molecular Research Center of Shahrekord University of Medical Sciences, RNA was extracted and converted into cDNA. To evaluate the expression of IFI16 and STING, the Real-Time PCR method using SYBR Green/ROX qPCR master mix was performed done. The level of genes expression was measured using 2–ΔΔCt method. The obtained data were analyzed using SPSS v22 software. Results: Comparison of the IFI and STING mRNA expression in blood samples in association with gender and age showed no significant differences (p>0.05). Also, the evaluation of IFI16 mRNA level revealed that the IFI16 genes’ expressions were remarkably higher in the new case group compared to the control group, however, STING expression did not show any significant difference. The mRNA levels of IFI16 and STING in IFNβ-treated groups were significantly lower than the new case group (p<0.001). Also, the genes’ expressions in both the IFNβ-treated groups were significantly lower compared to the control group (p<0.001). In the assessment of the correlation of IFI16 and STING expressions with age and sex in different research groups, no statistically significant differences were seen (p>0.05). Conclusion: Perhaps the IFNβ therapy decreases the IFI16 and STING expression in a STINGdependent pathway as a negative feedback mechanism for regulation of the immune system and suppression of pro-inflammatory cytokines production. The important role of DNA sensing molecules and STING-dependent pathway in MS gives a new insight into future treatment based on STING-direct therapies.

P–648 In-vitro Evaluation of SCF and StxBP1 in Polycystic Ovary Syndrome (PCOS)

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
S Erol ◽  
S Zırh ◽  
L Karako. Sokmensuer ◽  
G Bozdag ◽  
S F Muftuoglu
Keyword(s):  
Granulosa Cells ◽  
Polycystic Ovary ◽  
Critical Role ◽  
Vesicle Fusion ◽  
Snare Complex ◽  
Control Group ◽  
Signal Molecules ◽  
Expression Levels ◽  
Before And After

Abstract Study question Is the interaction between intrafollicular cells in PCOS, impaired by the change of vesicular fusion and/or exocytosis in granulosa cells (GCs)? Summary answer StxBP1 expression leves impared in GCs of PCOS. What is known already PCOS characterised as follicular arrest on antral follicles, cystic follicle formation, and follicular development failure. GCs secretes wide variety of factors via exocytosis, and plays critical role during folliculogenesis. Secretory vesicles are transported to cellular membrane. This process requires local concentrations of SNAREs consisting of tSNARE, vSNARE, and other vesicle fusion associated proteins. SNARE proteins are involved in vesicle fusion, exocytosis, and intracellular trafficking. GCs secretes KITL which provides follicular activation and growth. Syntaxins are one of the members of SNARE complex. StxBP1 is a protein which has a crucial role in secretory vesicle fusion that provides fusion of syntaxins. Study design, size, duration Granulosa cells (GCs) were collected for primary cell culture, since 2019 from both PCOS (n = 10) and healthy (male factor infertility) (n = 10) women undergoing ART. Each GCs from participant divided into two groups as in-vitro stimulated group and in-vitro nonstimulated group. Participants/materials, setting, methods GCs have been isolated from follicular fluid taken from patients during oocyte pick-up at Hacettepe University In-Vitro Fertilization Unit. nGCs were cultured at most secod subcultures after the isolation. The stimulated groups of both PCOS and control groups were stimulated by hCG(10IU/ml) ve FSH(0.5IU/ml) for 24 hours. Vesicle fusion proteins (Stx6, StxBP1, and SNAP25), KITL, and FSHr expressions were analyzed on granulosa cells from each group via immunofluorescent (IF) labeling and cyto-ELISA. Main results and the role of chance FSHr were compared in both control and PCOS before and after stimulation. There was no difference between FSHr expression levels in both groups. Indirect IF is widely considered for SNAP25, Stx6, StxBP1 proteins in all groups of GCs screening with/without stimulation. Expression of SNAP25, StxBP1 mainly scattered through all cytoplasmic area,s and membranous localization was observed. Stx6 expressions were particularly distinguished at perinuclear area of cytoplasm. However, stimulated cells of control appeared more peripherally Stx6 expression. This pattern caused by stimulation wasn’t observed in PCOS. Expressions of SNAP25, Stx6, StxBP1 were observed with less expression in PCOS. Also, the response to stimulation was lower than the control group. The differences in Stx6, SNAP25, StxBP1 and KITL levels before and after stimulation was evaluated for both control and PCOS in Cyto-ELISA. However, both SNAP25 and Stx6 expressions in GCs of both groups were similar in response to stimulation. The expression levels of StxBP1 in response to stimulation were significantly lesser than control at PCOS. KITL expressions were lower in the PCOS as expected furthermore similar to StxBP1 in response to stimulation. According to our findings, the highest response to stimulation in GCs occurred for StxBP1 and KITL in the control. Limitations, reasons for caution Since human cells were used in the study and the cells of each patient do not exhibit the same characteristics, the lowest number of patient samples identified in the statistical power analysis were included in the study. Wider implications of the findings: Our view to the disruption in the secretion of signal molecules in terms of vesicle dynamics will offer a new perspective in female infertility or cross-talks in folliclar cells of the ovary. Trial registration number TSA–2019–18196

Abstract 514: Jak2 Tyrosine Kinase Mediates Angiotensin II Renal Pathogenesis via its Pressor Dependent Actions

Hypertension ◽  
2014 ◽  
Vol 64 (suppl_1) ◽  
Author(s):  
Peter P Sayeski ◽  
Sung O Park ◽  
Annet Kirabo ◽  
Rebekah Baskin ◽  
Dale M Seth ◽  
...  
Keyword(s):  
Angiotensin Ii ◽  
Renal Injury ◽  
Interstitial Fibrosis ◽  
Critical Role ◽  
Glomerular Sclerosis ◽  
Control Group ◽  
Mrna Levels ◽  
Ang Ii ◽  
Urine Albumin ◽  
Independent Events

We previously found that Jak2 kinase, expressed within vascular smooth muscle cells (VSMC), plays a critical role in angiotensin II (Ang II)-mediated hypertension. Given that Jak2 mediates both pressor-dependent and pressor-independent events, we sought to determine the role of blood pressure (BP), per se, on the deleterious effects of Jak2 within the kidney. To investigate this, three groups of mice were examined; i) wild type mice (Controls) that received Ang II infusion, ii) mice lacking Jak2 expression within the VSMC (VSMC Jak2 Null) that also received Ang II, and iii) Control mice that received Ang II plus an anti-hypertensive triple therapy (3Rx). After baseline BP recordings, Ang II was infused (1000 ng/kg/min, SC) to all groups and the 3Rx regimen (80 mg/L hydralazine, 5 mg/L reserpine, 30 mg/L hydrochlorothiazide in the drinking water) was initiated two days later to the 3Rx group, in order to maintain BP at similar levels to the VSMC Jak2 Null group. After 28 days of Ang II, mice were euthanized and the kidneys were assessed via histological, molecular, and functional approaches. Chronic Ang II infusion significantly increased the levels of intrarenal Ang II in all three groups; Control = 1,262±283 fmol/g, VSMC Jak2 Null = 1,655±666 fmol/g, and 3Rx = 2,174±588. While Ang II infusion significantly increased the mean BP in the Control group (152 ± 2 mm Hg), it was significantly, and similarly, lower in both the VSMC Jak2 Null and 3Rx groups (125 ± 5 mm Hg and 131 ± 5 mm Hg, respectively). Glomerular sclerosis was absent and interstitial fibrosis ranged from absent- mild- moderate, and was similar in all groups. The increases in i) perivascular infiltration of CD3+ lymphocytes, ii) CTGF gene expression, iii) tubule casts and iv) albuminuria that were observed in the Control mice, were significantly reduced in both the VSMC Jak2 Null and 3Rx groups. [CTGF mRNA Levels: Control = 100%±17, VSMC Jak2 Null = 70%±12*, 3Rx= 56%±17*. Urine Albumin (ng/day): Control = 414 ± 262, VSMC Jak2 Null = 138 ± 172*, 3Rx= 101 ± 89* (*, p<0.05 vs. Control)]. Thus, the early renal injury due to chronic Ang II infusion correlates with increased BP and not with the expression of VSMC-derived Jak2, suggesting that Jak2 contributes to early Ang II-mediated renal injury via its pressor-dependent actions.

scholarly journals Evaluation of Plasma MMP-9 and VEGF-A mRNAs as Non-invasive Diagnostic Biomarkers in Women with Endometriosis

2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Sepideh Abdollahi ◽  
Pantea Izadi ◽  
Shahla Noori Ardebili ◽  
Samaneh Chegeni ◽  
Mir Saead Yekaninejad
Keyword(s):  
Plasma Level ◽  
Quantitative Polymerase Chain Reaction ◽  
Mrna Level ◽  
Control Group ◽  
P Value ◽  
Mrna Levels ◽  
Diagnostic Biomarker ◽  
Diagnostic Biomarkers ◽  
Non Invasive ◽  
Significant Difference

Background: Endometriosis is one of the common gynecological diseases and can lead to pelvic pain, dysmenorrhea, dyspareunia, and infertility in women. Thus, accurate and early diagnosis is a pivotal issue and an essential need for managing this disorder. At the present, the gold standard diagnostic method for endometriosis is laparoscopic surgery that is an invasive method and can lead to delay in diagnosis. Thus, there is an immediate necessity to search for non-invasive diagnostic biomarkers, such as blood-based ones. Objectives: Matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor-A (VEGF-A) have essential roles in the pathogenesis of endometriosis. Therefore, in this study, we evaluated the plasma mRNA levels of MMP-9 and VEGF-A, as potential non-invasive diagnostic biomarkers for endometriosis. Methods: This study included 48 women (24 cases and 24 controls) who underwent laparoscopy for suspected endometriosis. Preoperative plasma samples were collected, and after RNA extraction, the levels of MMP-9 and VEGF-A mRNAs were determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Results: Plasma MMP-9 mRNA level was statistically higher in endometriosis patients compared with the control group (P value = 0.01). However, plasma VEGF-A mRNA level did not show a significant difference between the two groups (P value =0.5). Conclusions: It seems that the plasma level of MMP-9 mRNA in endometriosis patients is significantly higher than in non-endometriosis women. This finding can provide new insights regarding this mRNA’s applicability as a non-invasive diagnostic biomarker for discovering new cases of endometriosis (newly diagnosed). According to our results, despite the suggested role of VEGF-A in endometriosis pathogenesis, it seems that the plasma level of VEGF-A mRNA does not have the potential to be used as a non-invasive diagnostic biomarker.

scholarly journals Overexpressions of RHOA, CSNK1A1, DVL2, FZD8, and LRP5 Genes Enhance Gastric Cancer Development in the Presence of Helicobacter Pylori

2021 ◽  
Author(s):  
Ufuk Demirci ◽  
Seda Orenay-Boyacioglu ◽  
Elmas Kasap ◽  
Emre Gerceker ◽  
Fahri Bilgic ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Helicobacter Pylori ◽  
Wnt Signaling ◽  
Critical Role ◽  
Wnt Signaling Pathway ◽  
Study Groups ◽  
Tumor Burden ◽  
Control Group ◽  
Pcr Array ◽  
Qrt Pcr

Abstract Introduction: Intestinal metaplasia (IM), and Helicobacter pylori (HP) infection can be shown as risk factors in the development of gastric cancer (GC). WNT signaling pathway plays a critical role in carcinogenesis. However, the literature studies are limited on the significance of this pathway for the transition from IM to GC.Methods: We aimed to investigate the importance of the genes of WNT signaling pathways diagnostic and prognostic markers in the presence and absence of HP in conversion from IM to GC. 104 patients, (GC group n=35, IM group n=45, control group n=25) were included in this case-control study. Expression of genes in WNT signalling were searched in study groups with qRT-PCR array and qRT-PCR method. Data were analysed using PCR array data analysis software.Results: Statistically significant overexpression of RHOA, CSNK1A1, DVL2, FZD8 and LRP5 genes was detected in the GC and IM groups compared to the control group (p <0.05). Statistically significant overexpression of RHOA, CSNK1A1, DVL2, FZD8 and LRP5 genes was observed in patients with metastatic GC compared to patients with GC without metastasis (p <0.05). It was found that the RHOA, CSNK1A1, DVL2, FZD8 and LRP5 genes were statistically significantly over-expressed in diffuse GC patients compared to non-diffuse GC patients (p <0.05). Statistically significant overexpression of RHOA, CSNK1A1, DVL2, FZD8 and LRP5 genes was detected in HP positive IM patients compared to HP negative IM patients (p <0.05).Conclusion: Overexpression of RHOA, CSNK1A1, DVL2, FZD8 and LRP5 genes in IM may suggest that these genes are important markers in the development of IM and inflammation with HP. In addition, these genes are linked to tumor burden in the GC group. Consequently, we can conclude that these genes are poor prognosis biomarkers for GC and have the potential to be used as markers for future treatment monitoring.

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