Specification of skeletal muscle differentiation by repressor element-1 silencing transcription factor (REST)-regulated Kv7.4 potassium channels

Changes in the expression of potassium (K(+)) channels is a pivotal event during skeletal muscle differentiation. In mouse C(2)C(12) cells, similarly to human skeletal muscle cells, myotube formation increased the expression of K(v)7.1, K(v)7.3, and K(v)7.4, the last showing the highest degree of regulation. In C(2)C(12) cells, K(v)7.4 silencing by RNA interference reduced the expression levels of differentiation markers (myogenin, myosin heavy chain, troponinT-1, and Pax3) and impaired myotube formation and multinucleation. In K(v)7.4-silenced cells, the differentiation-promoting effect of the K(v)7 activator N-(2-amino-4-(4-fluorobenzylamino)-phenyl)-carbamic acid ethyl ester (retigabine) was abrogated. Expression levels for the repressor element-1 silencing transcription factor (REST) declined during myotube formation. Transcript levels for K(v)7.4, as well as for myogenin, troponinT-1, and Pax3, were reduced by REST overexpression and enhanced upon REST suppression by RNA interference. Four regions containing potential REST-binding sites in the 5′ untranslated region and in the first intron of the K(v)7.4 gene were identified by bioinformatic analysis. Chromatin immunoprecipitation assays showed that REST binds to these regions, exhibiting a higher efficiency in myoblasts than in myotubes. These data suggest that K(v)7.4 plays a permissive role in skeletal muscle differentiation and highlight REST as a crucial transcriptional regulator for this K(+) channel subunit

Resveratrol via sirtuin-1 downregulates RE1-silencing transcription factor (REST) expression preventing PCB-95-induced neuronal cell death

Resveratrol (3,5,4'-trihydroxystilbene) (RSV), a polyphenol widely present in plants, exerts a neuroprotective function in several neurological conditions; it is an activator of class III histone deacetylase sirtuin1 (SIRT1), a crucial regulator in the pathophysiology of neurodegenerative diseases. By contrast, the RE1-silencing transcription factor (REST) is involved in the neurotoxic effects following exposure to polychlorinated biphenyl (PCB) mixture A1254. The present study investigated the effects of RSV-induced activation of SIRT1 on REST expression in SH-SY5Y cells. Further, we investigated the possible relationship between the non-dioxin-like (NDL) PCB-95 and REST through SIRT1 to regulate neuronal death in rat cortical neurons. Our results revealed that RSV significantly decreased REST gene and protein levels in a dose- and time-dependent manner. Interestingly, overexpression of SIRT1 reduced REST expression, whereas EX-527, an inhibitor of SIRT1, increased REST expression and blocked RSV-induced REST downregulation. These results suggest that RSV downregulates REST through SIRT1. In addition, RSV enhanced activator protein 1 (AP-1) transcription factor c-Jun expression and its binding to the REST promoter gene. Indeed, c-Jun knockdown reverted RSV-induced REST downregulation. Intriguingly, in SH-SY5Y cells and rat cortical neurons the NDL PCB-95 induced necrotic cell death in a concentration-dependent manner by increasing REST mRNA and protein expression. In addition, SIRT1 knockdown blocked RSV-induced neuroprotection in rat cortical neurons treated with PCB-95. Collectively, these results indicate that RSV via SIRT1 activates c-Jun, thereby reducing REST expression in SH-SY5Y cells under physiological conditions and blocks PCB-95-induced neuronal cell death by activating the same SIRT1/c-Jun/REST pathway

BioEnterics Intragastric Balloon (BIB) versus Spatz Adjustable BalloonSystem (ABS): Our experience in the elderly

The BioEnterics Intragastric Balloon (BIB) and the Spatz Adjustable Balloon System (ABS) are in fact recommended for weight reduction as a bridge to bariatric surgery. We retrospected studied patients with body mass index (BMI) and age ranges of 37e46 and 70e80 years, respectively, who had undergone BIB from January 2010 to July 2012 and prospected studied patients who had undergone Spatz balloon from July 2012 to August 2014. The aim of this study is to compare BIB and Spatz in terms of weight loss, complications, and maintenance of weight after removal. For both procedures, the median weight loss was 20 ± 3 kg, median BMI at the end of the therapy was 32 ± 2, and no severe complication occurred

Improved Porosity of Insect Proof Screens Enhances Quality Aspects of Zucchini Squash without Compromising the Yield

none7siIn a global climate change environment, assuring optimal growing conditions is a difficult challenge, compromising the food supply for a rapidly rising population. The climatic conditions in the protected environment lead to high temperatures and fast insect development, impacting productivity and vegetables qualitative attributes. Consumers’ interest in healthy food requires sustainable tools to manage biotic and abiotic factors and, from this perspective, anti-insect nets represent an excellent “green” solution. For this purpose, our goal was to compare two different anti-insect nets on microclimate, production, and qualitative traits of Cucurbita pepo L. fresh fruits. The experiment was conducted in three separate polyethylene high tunnels, with 50 mesh anti-insect nets of different porosities being installed on the openings of two tunnels, while the third tunnel was a control without nets. Microclimate measurements, as well as yield, physiological, and phytochemicals variables, were assessed. The 50 mesh net led to a decrease in marketable yield (22.5%), fruit number (18.0%), CO2 net assimilation rate (6.0%), and transpiration rate (29.5%). Total soluble solids, antioxidant activities and total ascorbic acid concentration had an opposite trend. The 50 mesh AirPlus net improved quality aspects of zucchini fruits by increasing total ascorbic acid, total phenols, and antioxidant compounds, with no negative impact on yield.openFormisano, Luigi; Pannico, Antonio; El-Nakhel, Christophe; Starace, Giuseppe; Poledica, Milena; Pascale, Stefania De; Rouphael, YoussefFormisano, Luigi; Pannico, Antonio; El-Nakhel, Christophe; Starace, Giuseppe; Poledica, Milena; Pascale, Stefania De; Rouphael, Yousse

Assessment of the design criteria for concentric V-braced steel structures according to Italian and European codes

The critical review of design methodologies provided by the NTC2008, in agreement with the European seismic code (Eurocode 8) for steel Concentrically Braced Frames with chevron (or inverted V) diagonals (CBF-V), carried out by deepening the seismic behaviour of such typical steel seismic resistant structures, aims to provide more efficient design criteria able to ensure adequate safety levels under seism. As reference case studies, common structural configurations of CBF-V are designed according to the NTC2008 provisions. Each case study is designed through both the Linear Static (LS) and Dynamic (LD) analysis. For braces either Circular Hollow Sections (CHS) or HE profiles are used. General critical issues have been evidenced in the design process. The seismic performance of investigated structures is evaluated by non-linear static analyses, in order to appraise the most relevant behavioural issues, like the behaviour factor, the failure modes and the effectiveness of the capacity design criteria. A discussion on the obtained results has allowed to point out the pros and cons of the current design approach

Role of Laryngopharyngeal Reflux in Eustachian Tube Dysfunction in Adults

We have here studied the relationship between Eustachian tube dysfunction and laryngopharyngeal reflux, evaluating also the results of medical therapy in patients with such problems. Based on clinical, endoscopic and cytological investigations, we found that acid laryngopharyngeal reflux was the basis of audiological symptoms and chronic dysfunction of the Eustachian tube

Methylmercury upregulates RE-1 silencing transcription factor (REST) in SH-SY5Y cells and mouse cerebellum

Methylmercury (MeHg) is a highly neurotoxic compound that, in adequate doses, can cause damage to the brain, including developmental defects and in severe cases cell death. The RE-1-silencing transcription factor (REST) has been found to be involved in the neurotoxic effects of environmental pollutants such as polychlorinated biphenyls (PCBs). In this study, we investigated the effects of MeHg treatment on REST expression and its role in MeHg-induced neurotoxicity in neuroblastoma SH-SY5Y cells. We found that MeHg exposure caused a dose- and time- dependent apoptotic cell death, as evidenced by the appearance of apoptotic hallmarks including caspase-3 processing and annexin V uptake. Moreover, MeHg increased REST gene and gene product expression. MeHg-induced apoptotic cell death was completely abolished by REST knockdown. Interestingly, MeHg (1. μM/24. h) increased the expression of REST Corepressor (Co-REST) and its binding with REST whereas the other REST cofactor mammalian SIN3 homolog A transcription regulator (mSin3A) was not modified. In addition, we demonstrated that the acetylation of histone protein H4 was reduced after MeHg treatment and was critical for MeHg-induced apoptosis. Accordingly, the pan-histone deacetylase inhibitor trichostatin-A (TSA) prevented MeHg-induced histone protein H4 deacetylation, thereby reverting MeHg-induced neurotoxic effect. Male mice subcutaneously injected with 10 mg/kg of MeHg for 10 days showed an increase in REST expression in the granule cell layer of the cerebellum together with a decrease in histone H4 acetylation. Collectively, we demonstrated that methylmercury exposure can cause neurotoxicity by activating REST gene expression and H4 deacetylation

Histone deacetylase 4 promotes ubiquitin-dependent proteasomal degradation of Sp3 in SH-SY5Y cells treated with di(2-ethylhexyl)phthalate (DEHP), determining neuronal death.

Phthalates, phthalic acid esters, are widely used as plasticizers to produce polymeric materials in industrial production of plastics and daily consumable products. Animal studies have shown that di(2-ethylhexyl)phthalate (DEHP) may cause toxic effects in the rat brain. In the present study, chronic exposure to DEHP (0.1–100 μM) caused dose-dependent cell death via the activation of caspase-3 in neuroblastoma cells. Intriguingly, this harmful effect was prevented by the pan-histone deacetylase (HDAC) inhibitor trichostatin A, by the class II HDAC inhibitor MC-1568, but not by the class I HDAC inhibitor MS-275. Furthermore, DEHP reduced specificity protein 3 (Sp3) gene expression, but not Sp3 mRNA, after 24 and 48 h exposures. However, Sp3 protein reduction was prevented by pre-treatment with MC-1568, suggesting the involvement of class II HDACs in causing this effect. Then, we investigated the possible relationship between DEHP-induced neuronal death and the post-translational mechanisms responsible for the down-regulation of Sp3. Interestingly, DEHP-induced Sp3 reduction was associated to its deacetylation and polyubiquitination. Co-immunoprecipitation studies showed that Sp3 physically interacted with HDAC4 after DEHP exposure, while HDAC4 inhibition by antisense oligodeoxynucleotide reverted the DEHP-induced degradation of Sp3. Notably, Sp3 overexpression was able to counteract the detrimental effect induced by DEHP. Taken together, these results suggest that DEHP exerts its toxic effect by inducing deacetylation of Sp3 via HDAC4, and afterwards, Sp3-polyubiquitination

HMGA1 overexpression is associated with a particular subset of human breast carcinomas

Breast cancer represents the second leading cause of cancer mortality among American women and accounts for more than 40 000 deaths annually. High-mobility group A1 (HMGA1) expression has been implicated in the pathogenesis and progression of human malignant tumours, including breast carcinomas. The aim of this study was to evaluate HMGA1 detection as an indicator for the diagnosis and prognosis of human breast carcinoma

Association of FGFR1 with ER-alpha maintains ligand-independent ER transcription and mediates resistance to estrogen deprivation in ER+ breast cancer

In a cohort of patients with ER+ breast cancer, tumors with FGFR1 amplification retained high proliferation upon estrogen deprivation with the aromatase inhibitor letrozole. Estrogen deprivation increased total and nuclear FGFR1 and FGF ligands in ER+/FGFR1-amplified primary tumors and breast cancer cells. In estrogen-free conditions, FGFR1 associated with ER-alpha in tumor cell nuclei and regulated the transcription of ER-dependent genes. This interaction and transcriptional output were inducible by FGF ligands and blocked by a kinase-dead FGFR1 mutant or FGFR kinase inhibitors. ChIP-seq of FGFR1 amplified cells treated with FGF3 showed binding of FGFR1 and ERα to DNA. Treatment with the ER downregulator fulvestrant and/or the FGFR inhibitor lucitanib reduced binding of ERα or FGFR1 to DNA. RNA-seq data from FGFR1-amplified patients’ tumors treated with letrozole showed enrichment of estrogen response and E2F target genes. Finally, growth of ER+/FGFR1-amplified breast cancer cells and patient-derived xenografts was more potently inhibited by fulvestrant and lucitanib combined than each drug alone, suggesting a causal association of aberrant FGFR signaling with endocrine resistance. These data suggest the ER-alpha pathway is still active in estrogen-deprived ER+/FGFR1-amplified breast cancers and, therefore, these tumors should be considered for treatment with a combination of ER and FGFR antagonists