Identification of the Thyroid Transcription Factor-1 as a Target for Rat MST2 Kinase

Abstract Thyroid transcription factor-1 (TTF-1) is a homeodomain-containing transcription factor that is required for thyroid-specific expression of the thyroglobulin and thyroperoxidase genes as well as for lung-specific expression of the surfactant protein A, B, and C and the CC10 and the HNF-3α genes. TTF-1 is a phosphoprotein, and the phosphorylation of TTF-1 has been studied already. However, the kinase(s) that could be responsible for this phosphorylation have not been known. In this paper we report the identification by in-gel kinase assay of a 56-kDa serine/threonine kinase that is able to phosphorylate TTF-1 in thyroid cells. The cloning of this kinase revealed that we had identified the rat homolog of the human MST2 kinase. The pathway in which human MST2 functions is not known; however, it does not appear to involve either mitogen-activated protein kinases such as Erk1 and Erk2 nor the stress-activated protein kinases such as JNK and p38. We show that the activity responsible for TTF-1 phosphorylation is constitutive in thyroid cells. Furthermore, we demonstrate that TTF-1 is phosphorylatedin vivo by rMST2 at the same residues that had been identified previously as the major phosphorylation sites. Thus, TTF-1 represents the first identified target of this class of protein kinases

The CRE-Like Element Inside the 5′-Upstream Region of the Rat Sodium/Iodide Symporter Gene Interacts with Diverse Classes of b-Zip Molecules that Regulate Transcriptional Activities through Strong Synergy with Pax-8

AbstractWe previously demonstrated that transcription of the rat sodium/iodide symporter (NIS) gene is regulated by NUE, an upstream enhancer located between nucleotides −2264 and −2495 of the 5′-flanking region. To elucidate the mechanism of TSH/cAMP-mediated regulation of NIS gene expression, we have characterized the putative cAMP response element (CRE)/activator protein (AP)-1 site (termed NUC) that is closely located between the two Pax-8 (paired box domain transcription factor-8) binding sites within NUE. In two different approaches using either gel supershift analyses or dominant-negative inhibitors of b-Zip molecules, we have shown that NUC can be recognized by several members of the AP-1 and CREB family transcription factors that modulate the transcriptional activity of NUE. Using tethered dimers of b-Zip molecules, we have also demonstrated that specific homo- or heterodimers of AP-1 can synergistically stimulate NUE activity in concert with Pax-8. To demonstrate further that NUC is a bona fide CRE, we made an artificial promoter with the five-time tandem repeat of this sequence (5xNUC). In comparison to the canonical CRE (5xCRE), 5xNUC manifested greater transcriptional activity and broader response to cAMP signaling. Hence, we postulate that the significance of this evolutionally conserved CRE-like site may lie in its broader cell type specificity

The IkB kinase inhibitor nuclear factor-kB essential modulator–binding domain peptide for inhibition of balloon injury-induced neointimal formation

Objective—The activation of nuclear factor-kB (NF-kB) is a crucial step in the arterial wall’s response to injury. The identification and characterization of the NF-kB essential modulator– binding domain (NBD) peptide, which can block the activation of the IkB kinase complex, have provided an opportunity to selectively abrogate the inflammation-induced activation of NF-kB. The aim of the present study was to evaluate the effect of the NBD peptide on neointimal formation.<br></br> Methods and Results—In the rat carotid artery balloon angioplasty model, local treatment with the NBD peptide (300 microg/site) significantly reduced the number of proliferating cells at day 7 (by 40%; P<0.01) and reduced injury-induced neointimal formation (by 50%; P<0.001) at day 14. These effects were associated with a significant reduction of NF-kB activation and monocyte chemotactic protein-1 expression in the carotid arteries of rats treated with the peptide. In addition, the NBD peptide (0.01 to 1 micromol/L) reduced rat smooth muscle cell proliferation, migration, and invasion in vitro. Similar results were observed in apolipoprotein E-/-, mice in which the NBD peptide (150 microg/site) reduced wire-induced neointimal formation at day 28 (by 47%; P<0.01).<br></br> Conclusion—The NBD peptide reduces neointimal formation and smooth muscle cell proliferation/migration, both effects associated with the inhibition of NF-kB activation

Effect of a moderately hypoenergetic Mediterranean diet and exercise program on body cell mass and cardiovascular risk factors in obese women

Objective: To assess the effects of a moderately hypoenergetic Mediterranean diet (MHMD) and exercise program on body cell mass (BCM) and cardiovascular disease risk factors in obese women. Subjects/Methods: Forty-seven obese women, 39.7 +/- 13.2 years of age, with a body mass index (BMI) 30.7 +/- 6.0 kg/m(2), completed the study. The following were measured at baseline, 2 and 4 months: BCM, BCM index (BCMI), body weight, BMI, fat-free mass (FFM), fat mass (FM), total body water (TBW), extracellular water (ECW) and intracellular water (ICW) using bioelectrical impedance analysis; fasting blood glucose (FBG), serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and triglyceride (TG) concentrations; systolic (SBP) and diastolic (DBP) blood pressure. Results: Body weight, BMI, FM, TC and TG significantly decreased (P<0.001; P<0.002 (TG)) at 2 and 4 months. FFM, TBW, ECW, FBG and DBP significantly decreased at 2 months (P<0.05 (FFM); P<0.001). LDL-C significantly decreased (P<0.001), while HDL-C significantly increased (P<0.002) at 4 months. BCM, BCMI, ICW and SBP remained stable over time. Conclusion: BCM was preserved and cardiovascular disease risk factors improved in obese women placed on a MHMD and exercise program for 4 months

Redundant domains contribute to the transcriptional activity of the thyroid transcription factor 1.

The thyroid transcription factor 1 (TTF-1) is a homeodomain-containing protein implicated in the activation of thyroid-specific gene expression. Here we report that TTF-1 is capable of activating transcription from thyroglobulin and, to a lesser extent, thyroperoxidase gene promoters in nonthyroid cells. Full transcriptional activation of the thyroglobulin promoter by TTF-1 requires the presence of at least two TTF-1 binding sites. TTF-1 activates transcription via two functionally redundant transcriptional activation domains that as suggested by competition experiments, could use a common intermediary factor

MicroRNA deregulation in thyroid cancer

In cancer microRNAs are often dysregulated with their expression patterns being correlated with clinically relevant tumor characteristics. Recently, microRNAs were shown to be directly involved in cancer initiation and progression. Despite the large amount of data showing strong correlations between cancer phenotype and microRNAs aberrant expression, very little is known about the molecular mechanisms inducing such deregulation. Thyroid carcinomas comprise a heterogeneous group of neoplasms with distinctive clinical and pathological characteristics. Activating mutations in Ras genes are frequently found in poorly differentiated and in anaplastic thyroid carcinomas. We have recently shown that oncogenic activation of Ras is able to change the expression of several microRNAs in thyroid epithelial cells. One of the top aberrantly expressed ones is miR-21, a microRNA prevoiusly reported overexpressed in a wide variety of cancers and causally linked to cellular proliferation, survival and migration. By using an inducible Ras oncogene we demonstrated that constitutively active Ras induce overexpression of miR-21 at very early times after its activation, and that such overexpression is maintained at later times as well as in chronically Ras-transformed cells. Analysis of a panel of thyroid tumors with different hystotypes revealed that miR-21 is overexpressed mainly in anaplastic carcinomas, thus correlating with the most aggressive phenotype. Interestingly, this induction seems to be cell-type specific, since the inducible Ras oncogene is unable to increase miR-21 levels in cultured fibroblasts. Moreover, our data show that at least two different Ras downstream pathways are necessary to induce miR-21 expression. We then asked if the ability of Ras in inducing miR-21 overexpression is verified in vivo. To answer this question we analyzed the expression of this microRNA in a mouse model of Ras-induced lung tumorigenesis, showing that Ras constitutive activation is able to increase miR-21 levels in normal lung and that the Ras-initiated lung cancer progression is accompained by a further increase in miR-21 expression. Taken together, our data strongly suggest that the oncogenic activation of Ras could be responsible for the increased expression of miR-21 frequently observed in human cancers

Il PON Reti e Mobilità e gli obiettivi di Sostenibilità: il ruolo del Piano di Monitoraggio Ambientale

The National Operative Program (PON) “Reti e mobilità” has been approved by European Commission on the 7th of December 2007. The strategic approach of the PON, exclusively devoted to the socalled “convergence regions” of Italy (Campania, Calabria, Sicilia, Puglia), aims to: 1. improve the modal balance by an economic, social and environmental perspective; 2. develop the inter-modality in order to move towards the integration of convergence area into the network of the European transport system; 3. improve the mobility and the accessibility, also to reduce the traffic congestion; 4. increase the efficiency related to security standards, to management techniques and to the quality of transportation services in the sector of freight; 5. guarantee the reduction of environmental impacts through a global improvement of the efficiency of the transport systems. In such a way it is clear that the PON “Reti e mobilità” takes greatly into account the Sustainability principles recognized at European scale as attested by the budget, namely over the 70% of the entire fund, associated with low-impacts infrastructures (railways and harbors). The PON has been submitted to the SEA procedure, following what the 2001/42/CE Directive establishes. The Environmental Report of PON devotes great attention to the monitoring activity as shown by the reported recommendation about the need for adequate measures for the environmental monitoring, also in order to apply corrective measures during the implementation of the program. By this point of view, a first and important step has been the elaboration of the “Environmental Monitoring Plan” (EMP) that represents the main methodological document for the following implementation of the monitoring activity. This paper, after a brief presentation of the program and of the objectives of Sustainability that the Programs aims to pursue through the realization of specific projects, is devoted to introduce the Environmental Monitoring Plan of the PON that has been approved by the Ministry of the Infrastructures and Transport -in charge as Management Authority of the PON- in February 2011. In detail, the PMA represents the tool through which, the Management Authority, that has specific responsibilities and functions in terms of monitoring and environmental assessment of the program, controls the significant impacts on the environment caused by the implementation of the PON and verify the level of achievement of the established objectives of environmental sustainability. The structure of EMP is based on three main aspects: 1) the adoption of the results of other interesting experiences carried out by experts institutions on the topic; 2) an approach favoring the creation of an “integrated” monitoring system with the others Operative Programs activated at regional scale; 3) the implementation of a cooperation and shared process with all the directly-involved actors

Prevalence of obesity and obesity-associated muscle wasting in patients on peritoneal dialysis

Background and aims: A progressive decrease in muscle mass until full-blown sarcopenia may occur in patients on peritoneal dialysis (PD) and worsen their life quality and expectancy. Here we investigate the prevalence of obesity and obesity-associated muscle wasting in PD patients. Patients and methods: The study design was observational, cross sectional. Body composition was assessed with BIA and BIVA in 88 PD patients (53.4 ± 13.1 years; 67% male). Patients with obesity and/or with reduced muscle mass were identified using FMI and SM/BW cutoff values, respectively. Inflammatory status was assessed by measuring CRP and fibrinogen blood levels. Results: A total of 44.3% of the patients showed a reduced muscle mass (37.5% moderate and 6.8% severe). The prevalence of obesity was 6.1%, 81.8%, and 100% in patients with normal, moderately, and severely reduced muscle mass, respectively (p < 0.05). Of the total, 15.2% of the patients with normal muscle mass, 18.4% of those with moderately reduced muscle mass, and 66.7% of those with severely reduced muscle mass had diabetes. The prevalence of severe muscle mass loss was higher in those with diabetes than in those without diabetes (22.2% vs. 2.8%, p < 0.05). Patients with obesity-associated muscle wasting showed higher fibrinogen (613.9 ± 155.1 vs. 512.9 ± 159.5 mg/dL, p < 0.05) and CPR (1.4 ± 1.3 vs. 0.6 ± 0.8 mg/dL, p < 0.05) blood concentrations than those with normal body composition. Conclusion: Obesity and diabetes were strongly associated with muscle mass loss in our PD patients. It remains to be established whether prevention of obesity with nutritional interventions can halt the occurrence of muscle mass loss in patients on PD

Thermal Conductivity Enhancement of Al2O3 Nanofluid in Ethylene Glycol and Water Mixture

AbstractThe ability of nanofluids that exhibits enhanced thermal performance is acknowledged by researchers through studies since decades ago. However, the observation of thermal properties for nanofluids in water and ethylene glycol based is not fully explored yet. Hence, this paper presents the thermal conductivity of water and ethylene glycol (EG) based Al2O3 nanofluid. The 13 nm sized Al2O3 nanoparticles were dispersed into three different volume ratio of water: EG such as 40:60, 50:50 and 60:40 using a two-step method. The measurement of thermal conductivity was performed using KD2 Pro Thermal Properties Analyzer at working temperatures of 30 to 70 ̊C for volume concentration of 0.5 to 2.0%. The results indicate that the thermal conductivity increases with the increase of nanofluid concentration and temperature. While the percentage of ethylene glycol increase, the range of thermal conductivity decreases due to ethylene glycol properties. The measurement data of the nanofluids give maximum enhancement of thermal conductivity at condition 2.0% volume concentration, temperature of 70 ̊C and for all base fluid