A Novel Role of Cdk9/CyclinT2 complexes in skeletal muscle and Rhabdomyosarcoma cells

Cyclin dependent kinase 9 (Cdk9) is a member of the cyclin dependent kinase family. The regulatory units of Cdk9 are the T family Cyclins (T1, T2) and Cyclin K1. Cyclin T2 has two forms termed CycT2a and CycT2b that arise by an alternative splicing of the primary transcript. Previous studies underscored a crucial role for Cdk9 in association of Cyclin T2 during skeletal myogenesis. Upon induction of muscle differentiation, MyoD recruits Cdk9/CycT2 on musclespecific gene promoter sequences. This complex is able to phosphorylate the C-terminal domain of RNA polymerase II, enhancing Myod function and promoting myogenic differentiation. Rhabdomyosarcoma (RMS), one of the most common childhood solid tumor, arises from muscle precursor cells and fails to complete both the differentiation program both the irreversibly cell cycle exit, resulting in uncontrolled proliferation and incomplete myogenesis. In RMS, Cdk9 fails to phosphorylate MyoD and the ability of MyoD to arrest cell proliferation and to activate the myogenic program is repressed. The result of this study confirmed the involvement of Cdk9/ CyclinT2 complexes during the myogenesis. Both isoforms of Cyclin T2 are able to activate the myogenic program at different stages of differentiation but CycT2b have a predominant role of in particular during the latest stages. Moreover we demonstred that EZH2 is probably responsible to inhibition of Cdk9 in RMS cells and her overexpression contribuite to inhibition of myogenesis

Role of enhancer of zeste homolog 2 polycomb protein and its significance in tumor progression and cell differentiation

Epigenetics is a branch of genetics that focuses on the heritable changes of DNA or associated proteins, other than DNA sequence variations, which carry information content during cell division [1,2]. These heritable changes are ascribed to chromatin, which constitutes the ultrastructure of DNA and whose modifications affect the genetic material functionality. Differences in chromatin structure have been associated to transcription regulation [3-5] and chromosome stability [6,7], affecting both gene’s information, expression and heritability. Noteworthy, these epigenetic modifications are involved in both transcriptional activation and repression, indicating their widespread role as modulators of gene expression in numerous biological processes [8,9]. Chromatin is subjected to numerous modifications roughly classified in two groups: DNA and histone post-translational modifications (histone-PTMs). DNA methylation is the most studied epigenetic modification of DNA and corresponds to the covalent addition of a methyl (CH3) group to the nucleotide cytosine within CG dinucleotides or CNG trinucleotides where N can be C, A, G or T. Usually, DNA methylation induces decreased protein-DNA binding of transcription factors and leads to the repression of gene expression [10]. DNA “methylable” sequences are not uniform across the human genome but restricted in CpG rich DNA regions termed CpG islands (CGI). CGI are localized at repetitive sequences, heavy methylated, to prevent the reactivation of endoparasitic sequences such as transposons, and at gene promoter sequences, which are normally refractory to methylation in normal somatic cells [8,11].</br

The Role of the Customer in Building Design: A Literature Review

Abstract: Although nowadays building design as well as other design disciplines recognize the decisive role of the customer and the necessity of the early user needs identification to guarantee the effectiveness of solutions and design process, in building design this activity is not yet well-defined. In Italy the traditional approach based on dimensional standards and type solutions is widely applied even though a new approach based on customer requirements and corresponding performances is available. Besides, various experiences in the building sector show attempts to formalize the activity for the collection of the overall needs or requests for specific purposes. Building design may benefit from the introduction of well-established procedures for identifying customer needs. In order to evaluate contributions for the improvement of such activity, this study reviews the role of the customer in building design throughout the years and provides an overview on experiences and available design procedures developed in building and product design

Temperament and Depression After a First Acute Coronary Syndrome

Few studies assess the role of personality styles in predicting the onset of depression among cardiac patients. This study evaluates whether temperament and character can represent a risk factor for the development of incident first-ever depressive episodes in patients at their first acute coronary syndrome (ACS). Two hundred sixty-seven (72.1%) subjects (male) completed the Temperament and Character Inventory (TCI) a few days after the cardiac event. At baseline and after 1, 2, 4, 6, 9, 12, and 24 months of follow-up, the participants completed the Primary Care Evaluation of Mental Disorder (PRIME-MD) and the Hospital Anxiety and Depression Scale to establish the presence of a depressive episode and its severity. During the follow-up, 61 (22.8%) participants developed a depressive episode. Temperamental risk factors for incident depression were scored high on novelty seeking and harm avoidance at the TCI. Given the detrimental effect of depression on cardiac prognosis, clinicians should take temperament variables into account when determining the treatment plans of their patients with ACS

Hyaluronan Esters Drive Smad Gene Expression and Signaling Enhancing Cardiogenesis in Mouse Embryonic and Human Mesenchymal Stem Cells

BACKGROUND: Development of molecules chemically modifying the expression of crucial orchestrator(s) of stem cell commitment may have significant biomedical impact. We have recently developed hyaluronan mixed esters of butyric and retinoic acids (HBR), turning cardiovascular stem cell fate into a high-yield process. The HBR mechanism(s) remain still largely undefined. METHODOLOGY/PRINCIPAL FINDINGS: We show that in both mouse embryonic stem (ES) cells and human mesenchymal stem cells from fetal membranes of term placenta (FMhMSCs), HBR differentially affected the patterning of Smad proteins, one of the major conductors of stem cell cardiogenesis. Real-time RT-PCR and Western blot analyses revealed that in both cell types HBR enhanced gene and protein expression of Smad1,3, and 4, while down-regulating Smad7. HBR acted at the transcriptional level, as shown by nuclear run-off experiments in isolated nuclei. Immunofluorescence analysis indicated that HBR increased the fluorescent staining for Smad1,3, and 4, confirming that the transcriptional action of HBR encompassed the upregulation of the encoded Smad proteins. Chromatin immune precipitation and transcriptional analyses showed that HBR increased the transcription of the cardiogenic gene Nkx-2.5 through Smad4 binding to its own consensus Smad site. Treatment of mouse ES cells and FMhMSCs with HBR led to the concomitant overexpression of both Smad4 and α-sarcomeric actinin. Smad4 silencing by the aid of lentiviral-mediated Smad4 shRNA confirmed a dominant role of Smad4 in HBR-induced cardiogenesis. CONCLUSIONS/SIGNIFICANCE: The use of HBR may pave the way to novel combinatorial strategies of molecular and stem cell therapy based on fine tuning of targeted Smad transciption and signaling leading to a high-throughput of cardiogenesis without the needs of gene transfer technologies

OLENEKIAN TO EARLY LADINIAN STRATIGRAPHY OF THE WESTERN PART OF THE AGHDARBAND WINDOW (KOPEH-DAG, NE IRAN)

The structural setting and the stratigraphy of the Early to Middle Triassic sedimentary succession exposed in the western part of the Aghdarband window (Kopeh Dag, NE Iran) is described. Six stratigraphic sections in the Sefid-Kuh Limestone, Nazar-Kardeh Formation and Sina Formation have been studied in the tectonic units 1a and 2. The lithostratigraphy is revised, with bio-chronostratigraphic constrain provided by conodonts and ammonoids. The new Olenekian ammonoid genus Megatirolitesis erected. It is based on species thus far known only in Mangyshlak (West Kazakhstan) but it is occurs also in the Sefid-Kuh Limestone.The evolution of the Lower Triassic carbonate ramp of the Sefid-Kuh Limestone, persisted in the Middle Anisian, with a three-stage development (Sefid-Kuh 1, 2 and 3 members) separated by drowning and onset of siliciclastics. The last stage is in part coeval with the Middle Anisian basinal Nazarkardeh Formation.The unconformity-bounded, three-stage development of the carbonate ramp documents that in the Aghdarband Basin the tectonic control over sedimentation started already in the Olenekian, since the onset of the marine transgression. The transgression of the Ladinian Sina Formation sealed a complex morphology resulting from the uplift and erosion of the Middle Anisian units. A new paleogeographic position along the southern Laurasia margin is propsed for the Triassic Aghdarband Basin. Based on the paleobiogeographic affinity of the Olenekian ammonoid occurences, we suggest that the Aghdarband Basin was located in a back-arc position in close connection with Mangyshlak (West Kazakhstan) and Tuarkyr (Turkmenistan), passing northwestward to a large epicontinental basin extending to the Donbass area. At least during the Olenekian the Aghdarband Basin had no direct connection with the Nakhlak Basin, which was proably located in a different intra-arc or more probably fore-arc region with respect to the Palaeotethys subduction-related Triassic arc

Study protocol: The DOse REsponse Multicentre International collaborative initiative (DO-RE-MI)

INTRODUCTION: Current practices for renal replacement therapy in intensive care units (ICUs) remain poorly defined. The DOse REsponse Multicentre International collaborative initiative (DO-RE-MI) will address the issue of how the different modes of renal replacement therapy are currently chosen and performed. Here, we describe the study protocol, which was approved by the Scientific and Steering Committees. METHODS: DO-RE-MI is an observational, multicentre study conducted in ICUs. The primary end-point will be the delivered dose of dialysis, which will be compared with ICU mortality, 28-day mortality, hospital mortality, ICU length of stay and number of days of mechanical ventilation. The secondary end-point will be the haemodynamic response to renal replacement therapy, expressed as percentage reduction in noradrenaline (norepinephrine) requirement. Based on the the sample analysis calculation, at least 162 patients must be recruited. Anonymized patient data will be entered online in electronic case report forms and uploaded to an internet website. Each participating centre will have 2 months to become acquainted with the electronic case report forms. After this period official recruitment will begin. Patient data belong to the respective centre, which may use the database for its own needs. However, all centres have agreed to participate in a joint effort to achieve the sample size needed for statistical analysis. CONCLUSION: The study will hopefully help to collect useful information on the current practice of renal replacement therapy in ICUs. It will also provide a centre-based collection of data that will be useful for monitoring all aspects of extracorporeal support, such as incidence, frequency, and duration

The Interplay between Gut Microbiota and Parkinson's Disease: Implications on Diagnosis and Treatment

The bidirectional interaction between the gut microbiota (GM) and the Central Nervous System, the so-called gut microbiota brain axis (GMBA), deeply affects brain function and has an important impact on the development of neurodegenerative diseases. In Parkinson's disease (PD), gastrointestinal symptoms often precede the onset of motor and non-motor manifestations, and alterations in the GM composition accompany disease pathogenesis. Several studies have been conducted to unravel the role of dysbiosis and intestinal permeability in PD onset and progression, but the therapeutic and diagnostic applications of GM modifying approaches remain to be fully elucidated. After a brief introduction on the involvement of GMBA in the disease, we present evidence for GM alterations and leaky gut in PD patients. According to these data, we then review the potential of GM-based signatures to serve as disease biomarkers and we highlight the emerging role of probiotics, prebiotics, antibiotics, dietary interventions, and fecal microbiota transplantation as supportive therapeutic approaches in PD. Finally, we analyze the mutual influence between commonly prescribed PD medications and gut-microbiota, and we offer insights on the involvement also of nasal and oral microbiota in PD pathology, thus providing a comprehensive and up-to-date overview on the role of microbial features in disease diagnosis and treatment

Posaconazole and midostaurin in patients with FLT3-mutated acute myeloid leukemia: Pharmacokinetic interactions and clinical facts in a real life study

: Midostaurin is used in combination with chemotherapy to treat patients with newly diagnosed FLT3-mutated acute myeloid leukemia. Chemotherapy-induced neutropenia exposes these patients to a significant risk of invasive fungal infections (IFIs). International guidelines recommend primary antifungal prophylaxis with posaconazole (PCZ) but nested analysis of a phase III trial showed that strong PCZ inhibition of CYP3A4 diminished midostaurin metabolism and increased midostaurin plasma levels; however, midostaurin-related adverse events (AEs) were only moderately exacerbated. We conducted a prospective multicenter real-life study to evaluate (i) how often concerns around PCZ-midostaurin interactions made the hematologist prescribe antifungals other than PCZ, (ii) how remarkably PCZ increased midostaurin plasma levels, and (iii) how significantly PCZ-midostaurin interactions influenced hematologic and safety outcomes of induction therapy. Although the hematologists were blinded to pharmacokinetic findings, as many as 16 of 35 evaluable patients were prescribed antifungal prophylaxis with micafungin, weak CYP3A4 inhibitor, in place of PCZ (p &lt; 0.001 for deviation from guidelines). In the 19 patients managed as per guidelines, PCZ-midostaurin interactions were more remarkable than previously characterized, such that at the end of induction therapy midostaurin minimum plasma concentration (Cmin ) was greater than three times higher than reported; moreover, midostaurin Cmin , maximum plasma concentration, and area under the curve were more than or equal to four times higher with PCZ than micafungin. Hematologic outcomes (complete remission and duration of severe neutropenia) and safety outcomes (midostaurin-related any grade or grade ≥3 AEs) were nonetheless similar for patients exposed to PCZ or micafungin, as was the number of breakthrough IFIs. In waiting for randomized phase III trials of new prophylaxis regimens, these findings show that PCZ should remain the antifungal of choice for the midostaurin-treated patient

Antiproliferative and proapoptotic effects of Inula viscosa extract on Burkitt lymphoma cell line.

Burkitt lymphoma is a very aggressive B-cell non-Hodgkin lymphoma. Although remarkable progress has been made in the therapeutic scenario for patients with Burkitt lymphoma, search and development of new effective anticancer agents to improve patient outcome and minimize toxicity has become an urgent issue. In this study, the antitumoral activity of Inula viscosa, a traditional herb obtained from plants collected on the Asinara Island, Italy, was evaluated in order to explore potential antineoplastic effects of its metabolites on Burkitt lymphoma. Raji human cell line was treated with increasing Inula viscosa extract concentration for cytotoxicity screening and subsequent establishment of cell cycle arrest and apoptosis. Moreover, gene expression profiles were performed to identify molecular mechanisms involved in the anticancer activities of this medical plant. The Inula viscosa extract exhibited powerful antiproliferative and cytotoxic activities on Raji cell line, showing a dose- and time-dependent decrease in cell viability, obtained by cell cycle arrest in the G2/M phase and an increase in cell apoptosis. The treatment with Inula viscosa caused downregulation of genes involved in cell cycle and proliferation (c-MYC, CCND1) and inhibition of cell apoptosis (BCL2, BCL2L1, BCL11A). The Inula viscosa extract causes strong anticancer effects on Burkitt lymphoma cell line. The molecular mechanisms underlying such antineoplastic activity are based on targeting and downregulation of genes involved in cell cycle and apoptosis. Our data suggest that Inula viscosa natural metabolites should be further exploited as potential antineoplastic agents against Burkitt lymphoma