Transforming Growth Factor-beta Levels Research Articles

Endothelial Dysfunction and 24-Hour Ambulatory Blood Pressure Monitoring in Patients with Diabetes

Abstract Arterial hypertension in patients with diabetes plays a main role in the earlier development of diabetic kidney disease (DKD), and endothelial dysfunction is considered to be a process involved in the development of diabetic complications. It is important to study the markers which gives the opportunity to identify DKD in early stage. In this article is presented a study of 24-hour Ambulatory Blood Pressure Monitoring (24hABPM) data in patients with diabetes and its interconnection with estimated glomerular filtration rate and endothelial dysfunction. The endothelial function was determined using the levels of TGF-b1(transforming growth factor-beta1) and VCAM-1 (vascular cell adhesion molecule-1). In our study, we obtained convincing results that complex including endothelial dysfunction and results of 24hABPM can be considered as early signs of DKD progression in patients with diabetes.

Vildagliptin Recruits Regulatory T Cells in Patients Undergoing Primary Percutaneous Coronary Intervention

ABSTRACTRegulatory T cells (Treg) has been documented to be protective against myocardial ischemia–reperfusion injury (MIRI). The administration of drugs which recruit Treg cells may participate in the cardioprotection of MIRI. The purpose of the present study was to investigate whether the add-on vildagliptin (vild) to standard treatment of MIRI prior to reperfusion could increase Treg recruitment, anti-inflammatory, and antioxidant effects of the standard treatment or not. Sixty diabetic patients with ST-segment elevation myocardial infarction were randomly divided into two equal groups: control group was given the standard medical treatment and vild group was given the standard medical treatment plus vild. There were no statistical differences between the mean of percentage of changes in nitric oxide, ischemia modified albumin, highly sensitive C reactive protein, and interferon-gamma levels in the studied groups. While, the percentages of changes of myeloperoxidase level, CD4+CD25+ Treg cells count, and transforming growth factor-beta1 level were significantly higher in vild group compared with control group. We concluded that addition of vild to standard medical treatment of MIRI could increase its effectiveness through recruitment of CD4+CD25+ Treg cells.

Treg and NK cells related cytokines are associated with deep rectosigmoid endometriosis and clinical symptoms related to the disease

The aim of this study was to evaluate Treg and NK cells related cytokines in deep infiltrating endometriosis lesions and its relationship with clinical symptoms of the disease. mRNA expression of Transforming Growth Factor Beta (TGFB), Interleukin (IL)10, Interferon Gamma (IFNG), IL7, and IL15 was analyzed by Real-Time PCR in eutopic endometrium and rectosigmoid lesions from 11 women with deep infiltrating endometriosis and in eutopic endometrium from 11 healthy women. IL10, IFNG, and IL7 expression was significantly higher in endometriotic bowel lesions than in eutopic endometrium from women with endometriosis. IL10 and TGFB expression was significantly higher in endometriotic bowel lesions than in eutopic endometrium from healthy women. In addition, TGFB and IL15 levels correlated positively with deep dyspareunia and cyclic dyschezia, respectively, while IL7 levels correlated negatively with dysmenorrhea. Deep infiltrating rectosigmoid endometriosis displays alterations in Treg and NK cells related cytokine, and TGFB, IL7 and IL15 expression is related with dyspareunia, dysmenorrhea and cyclic dyschezia, respectively, in patients with the disease.

Transforming growth factor beta1 (TGF-ß1) levels in a rat model of induced pleural empyema.

To evaluate the concentration of transforming growth factor beta 1 (TGFB1) levels in a rat pleural effusion obtained by inoculation of intrapleural bacteria or turpentine through thoracentesis. Thirty-Nine Wistar rats were divided into three groups: Staphylococcus aureus (SA, n = 17); Streptococcus pneumoniae (SP, n = 12); and turpentine (control, n = 10). Pleural fluid was collected through ultrasound-guided thoracentesis 12 h, 24 h, and 36 h after instillation of bacteria or turpentine. Levels of TGFB1 were measured in pleural fluid. At 12 h, mean TGFB1concentrations were 5.3450 pg/mL in the SA group, 5.3449 pg/mL in the SP group, and 5.3450 pg/mL in controls. At 24 h, they were 4.6700 pg/mL in the SA group, 4.6700 pg/mL in the SP group, and 4.6700 pg/mL in controls. At 36 h, they were 4.6699 pg/mL in the SA group and in control. No difference was observed among the groups in mean TGFB1concentration (p = 0.12); however, a significant intragroup reduction in mean TGFB1 was observed between 12 and 24 h (p < 0.01). The transforming growth factor beta 1 concentrations were not useful as a diagnostic tool or an early marker of infected pleural effusion.

Natural and environmental oestrogens induce TGFB1 synthesis in oviduct cells.

Autocrine/paracrine factors generated in response to 17β-oestradiol (E2), within the oviduct, facilitate early embryo development for implantation. Since transforming growth factor beta 1 (TGFB1) plays a key role in embryo implantation, regulation of its synthesis by E2 may be of biological/pathophysiological relevance. Here, we investigated whether oviduct cells synthesize TGFB1 and whether E2 and environmental oestrogens (EOEs; xenoestrogens and phytoestrogens) modulate its synthesis. Under basal conditions, bovine oviduct cells (OCs; oviduct epithelial cells and oviduct fibroblasts; 1:1 ratio) synthesized TGFB1. E2 concentration-dependent induced TGFB1 levels in OCs and these effects were mimicked by some, but not all EOEs (genistein, biochanin A and 4-hydroxy-2',4',6'-trichlorobiphenyl, 4-hydroxy-2',4',6'-dichlorobiphenyl); moreover, EOEs enhanced (P < 0.05) the stimulatory effects of E2 on TGFB1 synthesis. The OCs expressed oestrogen receptors alpha and beta and aryl hydrocarbon; moreover, co-treatment with ER antagonist ICI182780 blocked the stimulatory effects of E2 and EOEs on TGFB1 synthesis. Treatment with non-permeable E2-BSA failed to induce TGFB1, thereby ruling out the involvement of membrane ERs. Cycloheximide (protein synthesis inhibitor) blocked E2-induced TGFB1 synthesis providing evidence for de novo synthesis. The stimulatory effects of E2 and EOEs, were inhibited (P < 0.05) by MAPK inhibitor (PD98059), whereas intracellular-Ca2+ chelator (BAPTA-AM) and adenylyl cyclase inhibitor (SQ22536) abrogated the effects of E2, but not EOEs, suggesting that post-ER effects of E2 and EOEs involve different pathways. Our results provide the first evidence that in OCs, E2 and EOEs stimulate TGFB1 synthesis via an ER-dependent pathway. Exposure of the oviduct to EOEs may result in continuous/sustained induction of TGFB1 levels in a non-cyclic fashion and may induce deleterious effects on reproduction.

Correlation of nuclear factor-κB, regulatory T cell and transforming growth factor β with rheumatoid arthritis.

Objective: Investigated the correlation of nuclear factor-κB, regulatory cells and transforming growth factor-β with rheumatoid arthritis. Methods: Included 65 cases of RA patients admitted in our hospital from June 2015 to December 2016 into case group, and included 50 healthy people into control group during the same period. Collected the peripheral detection of nuclear factor-κB, regulatory cells and transforming growth factor beta levels, and compared them between two groups. Results: The percentage of CD4+, CD25+ T cells in the case group was significantly lower than that in the control group (P < .05); There was no significant difference in the percentage of CD4+, CD25+ CD127low/−, T cells between groups (P > .05); The levels of TGF - beta and NF - kappa B in the case group were higher than those in the control group, and the difference between the two groups was statistically significant (P < .05); The levels of ESR, CRP and RF in the case group were higher than those in the control group (P < .05). There was a negative correlation between the expression of nuclear factor-κB, transforming growth factor-β and RF level in RA patients by pearson correlation analysis, r = −0.652, P < .05. Conclusion: The expression levels of CD4+, CD25+ T cells in patients with RA are significantly decrease, which has a negative correlation with RA activity index RF, and showed that the pathogenesis of RA is related to the regulation of immune system.

Are There Any Differences Between the Distribution of Placental Bed Leukocyte Subtypes and Plasma Cytokine Levels of Preeclamptic and Healthy Pregnants?

Objectives: Preeclampsia (PE) is associated with impaired decidual leukocyte and plasma cytokine balance compared with normal pregnancy. We aimed to investigate maternal plasma levels of Interferon-gamma (IFN-g), Tumor Necrosis Factor-alpha (TNF-a), Transforming Growth Factor-beta (TGF-b), Interleukin-4 (IL4), IL6, IL10, IL17, IL35, suppressor of Cytokine Signalling-3 (SOCS3) and placental bed leukocytes in preeclamptic and healthy pregnants.Materials & Methods: This study was conducted with 40 preeclamptic and 40 normotensive pregnants. Cytokine levels were studied with enzyme-linked immunosorbent assay. CD8, CD56 and CD163 antigens were analysed by immunohistochemical study on placental bed biopsies. Results: In preeclamptic women; IFN-g and TGF-b levels were significantly higher and IL-35 levels were significantly lower than those of controls. CD8, CD56 and CD163 positivity of preeclamptic group were not significantly higher than those of controls. CD8 staining showed negative correlation with plasma IL17 levels. CD163 staining showed negative correlation with TNF-a/IL4 ratio. TNF-a/IL4 ratio showed minimal influence on placental bed CD163 staining.Conclusion: Slightly increased placental bed CD8, CD56 and CD163 positive leukocytes and increased plasma IFN-g, TGF-b and decreased plasma IL35 levels of preeclamptic pregnants indicate an aberrant cell mediated immunity in PE. We could not say yet that this condition is whether result or reason. New studies are needed to discuss our results.

The Beneficial Effects of Oral Cotrimoxazole upon Likely Biomarkers of Oxidative Stress in Advanced Fibrotic Lung Disease

Introduction: In cases of idiopathic pulmonary fibrosis, we have observed an elevation in mean red cell volume, serum gamma glutamyl transferase and peripheral monocyte counts, initially in a pilot study but also in new incident cases. These changes could not be explained by drug therapy, vitamin deficiency or other diseases. Method: We compared the peripheral blood abnormalities in 149 patients with lung fibrosis to 448 age and sex matched controls. We also examined the effect of cotrimoxazole treatment for 12 weeks on these abnormalities. From the pilot study of cotrimoxazole in lung fibrosis patients, the relationship of the peripheral blood monocyte count and serum cytokine transforming growth factor beta-1 was examined. Epstein Barr viral status was examined in a selection of patients in case it explained our observations. Results: The findings confirm the elevation in mean red cell volume, gamma glutamyl transferase and peripheral monocyte counts in patients compared with matched controls. Oral cotrimoxazole ameliorated these 3 blood abnormalities. Serological evidence of Epstein Barr viral infection was present in tested patients but active viral replication was absent. The monocyte count had a linear relationship with the serum transforming growth factor beta-1 levels, which increased by 600 pg/ml for every of 0.1 × 109/l increase in the monocyte count. Conclusion: These observations may reflect oxidative stress which was reduced by cotrimoxazole. A related sulphonamide “dapsone” is known to reduce oxidative stress through direct effects on neutrophil and monocyte function; similar effects may explain these findings and require a formal study.

557P Association of TGFB1 exon 1 129 T/C polymorphism with reduced risk of sporadic breast cancer in older North Indian women population

Background Transforming growth factor beta 1 (TGFB1) is a cytokine secreted by immune cells that plays an important role in breast tumorigenesis. It shows dual characteristics:anti-inflammatoryactivityandgrowthinhibitionofearlytumor cells and growth promoting activityin advanced tumor. Here, we investigated the association between TGFB1 +29 polymorphism and breast cancer risk in North Indian population. Methods We analyzed 285 patients with sporadic breast tumor and 363 healthy controlsinanassociationstudyforTGFB1+29T/C,TRAIL-716C/T,TP53codon72, IFNG CA repeat polymorphism using DNA sequencing or SSLP. The expression levels of TGFB1,TRAIL,DR4,DR5,DCR1,DCR2,CYCS,BCL2,CASP8,CASP8L,FLIPL, FLIPS,CASP3 andCASP3sintheDDRandapoptoticpathwayweremeasuredbyRT- PCR. Results The study of TGFB1 +29polymorphismindicatedasignificantdifferencein the genotype distribution between patients and controls. The predominant TT and TC genotypes, when compared against CC genotype, provided 1.7-fold risk. This association was mainly attributable to higher age group (> 45 years) women. Stratificationofgenotypicdatarevealedthatinpresenceofprotectorgenotypesof TGFB1 +29 CC and TRAIL -716 CT and TT genotypes, a 2.5-fold protection was observed. Studies carried out on TGFB1 +29 and TP53 codon 72 interaction showed that in presence ofthe protector genotypes a 2-fold protection was observed. Combined influence of TGFB1 and IFNG polymorphisms on breast cancer risk revealed that in presenceofprotectorgenotypeofIFNGCArepeat,TGFB1 +29riskgenotypeprovided 2.5-foldprotection.Furthermore,thecomparisonofTGFB1 transcriptlevelsintumors revealed significant difference for TGFB1 expression with different +29 genotype background.Also,TGFB1 +29genotypeinfluencedapoptosisthatoccurredthrough TRAIL mediated extrinsic pathway; CC with increased apoptosis compared to TT+TC in breast tumor patients greater than 45 years in age. Conclusions:Thus,weconcludethatasapotentinhibitorofcellproliferationand apoptosis,TGFB1 +29CCgenotypemaybeoneofthefactorsresponsibleforless aggressive and slow growing tumors in Legal entity responsible for the study: Jawaharlal Nehru University, New Delhi, India Funding Council of Scientific and Industrial Research, University Grants Commission, Department of Biotechnology Disclosure All authors have declared no conflicts of interest.

Associations of Selected Cytokines Levels in Organ Transplant Recipients Without and With Malignant Skin Neoplasms.

Skin malignancies are the most prevalent neoplasms seen in organ transplant recipients (OTRs). Immunosuppressive treatment has been attributed to play a causative role in malignancy development. The aim of the study was to assess cytokine concentrations involved in cytotoxic and regulatory responses in patients after organ transplantation (Tx). We compared two OTR subgroups: those with malignant skin tumors and those without any known changes developed after Tx. We enrolled 102 patients, 63: 3-360 (median: min-max) months after Tx, aged 54.3 ± 9.9 (mean ± SD) years (38.2% females). Seventeen patients werediagnosed with malignant skin neoplasms. The most frequent treatment schemes werecyclosporine A - mycophenolate mofetil - glucocorticosteroids (GS) (37.4%), mycophenolate mofetil-tacrolimus - GS (15.2%), and azathioprine-cyclosporine A-GS (14.1%). A 5-mL sample of venous blood was obtained from participants of two subgroups: those with malignant skin tumors and those without any known changes. The blood was tested for interleukin 2 (IL-2), interferon gamma, IL-10, and transforming growth factor beta concentrations (Multicytokine Flex Set, ELISA). The Kruskal-Wallis test was used to compare variables; P< .05 was considered valid. Age, gender distribution, and time from transplantation did not differ across the two subgroups. We found significantly lower blood concentrations of IL-2 and IL-10 in patients with post-transplantation skin cancers versus patients without any known skin changes (0pgmL(-1) vs. 21.22 pgmL(-1), and 4.93 pgmL(-1) vs. 7.36 pgmL(-1), respectively). The differences between interferon gamma and transforming growth factor beta levels were insignificant across studied groups. Our findings suggest that immunosuppressive response assessed by cytokine IL-2 and IL-10 levels may be used in the risk stratification for the development of skin cancer in organ recipient patients.

Clinical and prognostic significance of serum transforming growth factor-beta1 levels in patients with pancreatic ductal adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) has a poor 5-year survival rate of 5%. Biomarkers for the early detection of pancreatic cancer are urgently needed. Transforming growth factor-beta1 (TGF-β1) is elevated in the tissues and plasma of patients with PDAC. However, no studies systemically report prognostic significance of plasma TGF-β1 levels in PDAC. In the present study, we assessed the prognostic significance of serum TGF-β levels in patients with PDAC. TGF-β levels were determined in serum from 146 PDAC patients, and 58 patients with benign pancreatic conditions. Regression models were used to correlate TGF-β levels to gender, age, stage, class, and metastasis. Survival analyses were performed using multivariate Cox models. Serum levels of TGF-β1 distinguished PDAC from benign pancreatic conditions (P<0.001) and healthy control subjects (P<0.001). Serum levels of TGF-β also distinguished tumor stage (P=0.002) and lymph node metastasis (P=0.001). High serum levels of TGF-β1 were significantly correlated with reduced patient survival. Multivariate analysis revealed that TGF-β1, lymph node metastasis and tumor stage were independent factors for PDAC survival. Our results indicate that serum TGF-β1 may be used as a potential prognostic marker for PDAC.

Maternal insulin-like growth factor-1 and transforming growth factor beta-1 levels during pregnancy in rats fed with low and high salt

Maternal insulin-like growth factor-1 and transforming growth factor beta-1 levels during pregnancy in rats fed with low and high salt

Effects of amlodipine on ischaemia/reperfusion injury in the rat testis.

The aim of this study was to examine the effects of amlodipine (AML) in rat testicular torsion/detorsion damage. In this study, rats were divided into eight groups: (i) sham; (ii) testicular ischaemia, 2 h of ischaemia; (iii) testicular ischaemia/reperfusion (I/R), 2 h of ischaemia followed by 2 h of reperfusion; (iv) ischaemia + AML (5 mg kg(-1)) administered 30 min before ischaemia; (v) ischaemia + AML (10 mg kg(-1)) administered 30 min before ischaemia; (vi) and (vii) I/R + AML (5 mg kg(-1)) and I/R + AML (10 mg kg(-1)) administered 1.5 h after the induction of ischaemia, respectively, and at the end of a 2-h ischaemia period and a 2-h reperfusion period applied; and (viii) sham + AML (10 mg kg(-1)). Significant decreases in levels of superoxide dismutase and glutathione were observed in ischaemia and reperfusion groups when compared with healthy controls. These antioxidant levels increased in AML groups while malondialdehyde levels significantly decreased. While increases in tumour necrosis factor-alpha and transforming growth factor-beta levels were found in the torsion and detorsion groups, significant decreases in the levels of these inflammatory cytokines were observed in the treatment groups. These results demonstrate that AML significantly produced protective effects on testis tissue damage that occurs in the torsion/detorsion model via biochemical, histopathological and molecular pathways.

Abstract 80: Analysis of the regulation of angiogenesis pathway by inhibiting MDM2 function in LNCaP-MST prostate cancer cells using PCR array

Abstract Interfering with angiogenesis is a well-known strategy for cancer treatment due the fact that this process is essential for promoting cancer growth and metastatic spread by providing adequate supply of oxygen and nutrients to the tumor cells. The process of angiogenesis is highly regulated by both activator and inhibitor genes through their corresponding proteins. One of the suspected promoters of angiogenesis is MDM2 (Murine Double Minute 2 gene). The overexpression of MDM2 has been found in many cancer types and also correlated well with poor prognosis of certain cancers. In addition to some of the newly emerging mechanistic abilities, MDM2 is well known to regulate p53 levels and its functions by dual mechanisms, which plays a major role in many cellular processes such as cell cycle control and apoptosis. However, the role for MDM2 in regulating angiogenesis is slowly evolving which can explain the potential metastatic status of the cancer where there is no p53 mutation. In this connection, to further understand its role in tumor angiogenesis, the status of MMP9 (Matrix metallopeptidase 9 also called gelatinase B), THSB1 (Thrombospondin 1), TGB1 (Transforming growth factor-beta 1) were studied, by inhibiting MDM2. The ultimate goal of our study was to determine how the angiogenesis pathway is regulated by the overexpression of MDM2. To explore the possible mechanisms, LNCaP (prostate cancer cells), and LNCaP-MST (MDM2 transfected prostate cancer cells) cells were treated with Nutlin-3 (20 uM for 24 hours). Indeed LNCaP-MST cells are known to show 10 times higher expression of MDM2 compared to LNCaP cells. Therefore, Nutlin-3 treatment was expected to inhibit the interaction between p53 and MDM2 in these cells and thereby impact the expression of angiogenesis pathway related genes. When we analyzed the gene expression profile using a PCR array that was designed to explore the above mentioned pathway we were able to confirm that MDM2 overexpression could positively regulate MMP9, THSB1, and TGB1 levels in a highly significant manner. Interestingly, the genes expression profile that was found to be elevated in LNCaP-MST cells was reversed after Nutlin-3 treatment confirming that some of these angiogenesis related genes are positively regulated by MDM2 overexpression. In Conclusion, we are proposing that targeting MDM2 can effectively control the pro-angiogenesis mechanisms that can be found activated in MDM2 positive cancer types. We are conducting further studies to fully unravel the role of MDM2 in promoting tumor angiogenesis and metastatic ability (The financial support from the Royal Dames of Cancer Research Inc., Ft. Lauderdale is gratefully acknowledged). Citation Format: Ali Alaseem, Thiagarajan Venkatesan, Khalid Alhazzani, Appu Rathinavelu. Analysis of the regulation of angiogenesis pathway by inhibiting MDM2 function in LNCaP-MST prostate cancer cells using PCR array. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 80. doi:10.1158/1538-7445.AM2015-80

Adiponectin inhibits oxidative stress and modulates TGF-b1 and COL-1 expression via the AMPK pathway in HSC-T6 cells

To investigate the anti-oxidative stress and anti-fibrotic mechanisms of adiponectin by examining effects on oxidative stress levels and expression of fibrosis-related signaling factors, including transforming growth factor-beta 1 (TGFb1), collagen I (COL-1), and the adenosine monophosphate-activated protein kinase (AMPK) pathway by using an in vitro HSC-T6 cultured cell system. Activated HSC-T6 cells were pre-treated with 1.0 mug/mL adiponectin for 0, 30, 60 and 120 min, or left untreated to serve as controls, and both groups were then exposed to 5 mumol/L H2O2; a portion of the adiponectin-treated oxidative stress-induced cells were treated with an AMPK inhibitor (Compound C). The effects on mRNA levels of TGFb1. and COL-1 were analyzed by real-time PCR, in the levels of secreted TGF-b1 and COL-1 were detected by enzyme-linked immunosorbent assay of supernatants, and in the phosphoAMPK and AMPK protein expressions were detected by Western blotting. Compared to the H2O2 group without adiponectin pre-treatment, the H2O2 group with adiponectin pre-treatment showed significantly increased activity of superoxide dismutase (SOD), decreased content of malondialdehyde (MDA), and decreased gene and protein expressions of TGF-b1 and COL-1 (P less than 0.05). Moreover, inhibition of the AMPK pathway inhibited these adiponectin-mediated effects. The H2O2 group with adiponectin pre-treatment also showed increased levels of phospho-AMPK protein expression, with the maximum effect detected after 120 min of the adiponectin pre-treatment (P less than 0.01). Inhibition of oxidative stress is one of the mechanisms of the anti-fibrotic effects of adiponectin. Adiponectin can attenuate oxidative stress levels, resulting in down-regulation of TGFb1 and COL-1 expression through activation of the AMPK pathway.

Nitric Oxide As a Mediator of Bone Marrow Fibrosis in Patients with Myelofibrosis

Bone marrow (BM) fibrosis is a key pathomorphologic feature of patients (pts) with primary myelofibrosis (PMF) and the fibrotic phases of essential thrombocythemia (post-ET MF) and polycythemia vera (post-PV MF). The degree of BM fibrosis appears to correlate with survival. Indeed worse survival has been associated with increased BM fibrosis. The BM stromal microenvironment is important in the pathogenesis of BM fibrosis. Cellular components (fibroblasts, macrophages, endothelial cells, adipocytes), structural fibrils (collagen, reticulin) and extracellular matrix components are all forming elements of the BM stroma. Increased stromal fibrosis has been linked to abnormalities in the number/ function of megakaryocytes and platelets in hematologic diseases. Several cytokines like Platelet Derived Growth Factor (PDGF) and Transforming Growth Factor-Beta (TGF-b) have been also linked to the pathophysiology of BM fibrosis. PDGF has been shown to increase fibroblast growth in megakaryocytes and platelets although increased PDGF did not correlate with increased production of either reticulin or collagenous fibrosis. Moreover, PMF pts have increased TGF-b levels in platelets, megakaryocytes, and monocytes. Nitric Oxide (NO) is a ubiquitous gas important in physiologic processes particularly vasodilatation. Dysregulation of NO levels has been implicated in pulmonary hypertension (PH), hemoglobinopathies, and cardiovascular diseases. In Peyronie’s disease, a localized fibrosis of the penile tunica albuginea, increased NO production by expression of iNOS decreases collagen deposition by neutralization of profibrotic reactive oxygen species and decreased myofibroblast formation. Aside from its role in maintaining normal vascular tone, NO also plays a role in fibroblast formation and collagen biosynthesis. We previously reported that ruxolitinib, a JAK1/2 inhibitor restores NO levels leading to improvement of PH in MF pts (Tabarroki et al., Leukemia 2014). We now hypothesize that plasma/serum NO level is a key regulator of BM fibrosis in MF and that ruxolitinib treatment (Tx) leads to improvement of BM fibrosis by NO modulation. Using a Sievers 280i NO analyzer we measured the plasma/serum NO level of a large cohort (n=75) of pts with myeloid and myeloproliferative neoplasms (MPN) [MDS, RARS/RCMD=8; MPN, ET=8, PV=8, MF=24, Mastocytosis=7; MDS/MPN, CMML=11, MDS/MPN-U, RARS-T=9]. Healthy subjects (n=10) were used as a control. MPN pts had low NO (nM) levels among the pts studied with the lowest level found in MF pts: MF=30.31±11.8, PV=39.0±16.1, ET=36±20.3, RARS=74.6±41.7 (P=.01), CMML=84.4±89.2 (P=.04), RCMD=163.4±103.8 (P<.001), RARS-T=131.1±99.8 (P<.001). In total, NO levels were lower in classic MPN (n=40, 35.3±16.6) compared to MDS (n=8, 119±62.8; P=.001) and MDS/MPN (n=20, 105±94.6; P=.008). When we looked at the correlation between NO levels and BM fibrosis grade we found that there is an inverse correlation between NO levels and worsening BM fibrosis grade from grade MF1 to MF3. NO levels in normal (n=10) vs MF1 (n=3) were 53.3 vs 39.1, P=.025; normal vs MF2 (n=7) were 53.3 vs 37, P=.021; normal vs MF3 (n=12) were 53.3 vs 34.4, P=.006. A total of 8 pts who were treated with ruxolitinib and had at least 1 pre and 1 post Tx (≥3 months from initiation of ruxolitinib) were tested for NO levels. Among the 8 pts, 4 pts who demonstrated improvement in BM scores had a trend towards improved NO levels after ruxolitinib Tx [NO pre vs post; pt #1: 6 vs 10.5; pt#2: 4.3 vs 6.4; pt#3 49.7 vs 52.1; pt#4 36 vs 41.3; P=.02] while 4 had worsening or had no change in BM fibrosis grade and had a minimal change or decline in the NO (pt#5: 18.4 vs 23, pt#6: 14.29 vs 12.1, pt#7: 32.7 vs 32.1, pt#8: 110.9 vs 40.4). One pt who had improvement in BM fibrosis grade after ruxolitinib Tx had increased iNOS expression by Western blotting (pt#1) while no iNOS expression (pt#5) was noted in the pt who did not have improvement in BM fibrosis. Of note, multi-analytic cytokines profile also showed an overall decrease in cytokines especially a 2.8 fold-decrease in IL8 levels post-Tx in the pt with improvement in BM fibrosis. In conclusion, NO is decreased in MPN particularly in MF and may be a key mediator of BM fibrosis in MF. Pharmacologic therapies such as JAK inhibitors may mediate improvement of BM fibrosis by modulation of NO levels in MF. DisclosuresTiu:Gilead: Membership on an entity’s Board of Directors or advisory committees; Novartis: Speakers Bureau; Incyte : Membership on an entity’s Board of Directors or advisory committees, Speakers Bureau.

Serum transforming growth factor-beta1 levels may have predictive and prognostic roles in patients with gastric cancer.

Transforming growth factor-beta1 (TGF-beta1) plays an important role in the pathogenesis of multiple malignancies, and also, its expression strongly affects the outcomes of cancer patients. The objective of this study was to determine the clinical significance of the serum levels of TGF-beta1 in gastric cancer patients. A total of 63 patients with a pathologically confirmed diagnosis of gastric cancer were enrolled into this study. Serum TGF-beta1 concentrations were determined by the solid-phase sandwich ELISA method. Thirty healthy age- and sex-matched controls were included in the analysis. The median age at diagnosis was 62 years, range 28 to 82 years. There was no significant difference in baseline serum TGF-beta1 levels between gastric cancer patients and the healthy control group (p = 0.08). The known clinical variables including age of patient, gender, site of lesion, histology, histological grade, stage of disease, and serum levels of lactate dehydrogenase (LDH), CEA, and carbohydrate antigen (CA) 19.9 were not found to be correlated with serum TGF-beta1 concentrations (p > 0.05). However, the chemotherapy-responsive patients had higher serum TGF-beta1 levels compared with chemotherapy-unresponsive ones (median values 330.50 v 49.54 pg/mL, respectively, p = 0.01). Moreover, patients with elevated serum TGF-beta1 concentrations had significantly favorable overall survival compared with those with lower levels (median 71.1 v 39.9 weeks, respectively, p = 0.04). In conclusion, serum levels of TGF-beta1 may have predictive and prognostic roles in patients with gastric cancer.

FOXP3 DNA Methylation Levels as a Potential Biomarker in the Development of Periapical Lesions

IntroductionEpigenetic mechanisms, such as DNA methylation, can modify gene expression patterns without changing the DNA sequence, comprising a tool that cells use to lock genes in the “off” position. Variations in the methylation profile have been correlated to a variety of human diseases. Here, we hypothesize that DNA methylation in immune response–related genes may contribute to the development of periapical lesions. MethodsThe DNA methylation patterns of 22 immune response–related gene promoters were evaluated in 137 human periapical granulomas, 8 apical cysts, and 31 healthy gingival tissues from 2 independent cohorts using a pathway-specific real-time polymerase chain reaction array (EpiTect Methyl II; Qiagen Inc, Valencia, CA). Messenger RNA expression analysis by qualitative polymerase chain reaction was also performed. SABiosciences's hierarchical clustering and methylation (Qiagen, Valencia, CA) and Prism6 software (GraphPad Software, Inc, La Jolla, CA) were used for data analysis. ResultsFOXP3 gene promoter showed the highest level of methylation in both periapical granulomas and apical cysts (P < .001), and methylation levels were inversely correlated with FOXP3 messenger RNA expression in the lesions. Furthermore, FOXP3 expression was prevalent in inactive lesions and was positively correlated with interleukin-10 and transforming growth factor beta levels. ConclusionsOur results suggest that FOXP3 acts as a master switch governing the development and function of T-regulatory cells, whose functions include the inhibition of immune responses and temper inflammation. The observed differential methylation patterns of FOXP3 in periapical lesions may be crucial in determining its suppressive activity and may be involved in periapical lesion development.

Inhibitory effects of maslinic acid upon human esophagus, stomach and pancreatic cancer cells

Apoptotic, anti-invasive and anti-migratory effects of maslinic acid (MA) at 4, 8, or 16 µM in human esophageal squamous cancer cell line, OE33; gastric cancer cell line, SGC-7901; and pancreatic cancer cell line, Panc-1, were examined. MA treatments inhibited OE33 and SGC-7901 cells growth at 21–66% and 32–75%, respectively; but lowered Panc-1 viability at 13–27% only. MA treatments increased cleaved caspase-3 and Bax expression, and raised caspase-3 and caspase-8 activities in OE33 and SGC-7901 cells. MA treatments also increased DNA fragmentation and decreased reactive oxygen species production in these two cell lines. MA treatments declined invasion and migration in OE33 and SGC-7901 cells, and lowered vascular endothelial growth factor and transforming growth factor-beta1 levels in these cells. MA suppressed hypoxia-inducible factor-1alpha, matrix metalloproteinase (MMP)-2 and MMP-9 expression in OE33 and SGC-7901 cells. These findings indicated that this triterpene was a potent agent against esophagus and stomach cancers.

Fenugreek seed powder mitigates cadmium-induced testicular damage and hepatotoxicity in male rats

Cadmium is a potential environmental and industrial pollutant affecting human tissues and organs including liver and testes. The protective role of fenugreek seed powder (FSP) was investigated in male rats subjected to cadmium-induced testicular injury and hepatic dysfunction. Testicular damage and hepatotoxicity were induced by oral administration of cadmium chloride (5mg/kg body weight, once a day) for 7 weeks. FSP was given at 5% w/w in chow diet for 8 weeks, starting 1 week before cadmium administration. FSP intake significantly increased serum testosterone level and testis weight that were reduced by cadmium. FSP also compensated deficits in hepatic and testicular antioxidant defense system, interleukin-4 and nitric oxide levels, reduced serum liver function enzyme activities and suppressed lipid peroxidation in hepatic and testicular tissues resulted from cadmium administration. Additionally, FSP attenuated the cadmium-induced elevations in hepatic and testicular tumor necrosis factor-α and transforming growth factor-beta1 levels as well as cadmium deposition and hydroxyproline content. The protective effect afforded by FSP was mainly due its antioxidant, antifibrotic and anti-inflammatory effects. In conclusion, the results of the present work indicated that FSP may represent a promising medicinal herb to protect hepatic and testicular tissues from the detrimental effects of cadmium.