The chicken as a model organism to study heart development:Cold Spring Harbor Perspective in Biology

Heart development is a complex process and begins with the long-range migration of cardiac progenitor cells during gastrulation. This culminates in the formation of a simple contractile tube with multiple layers, which undergoes remodeling into a four-chambered heart. During this morphogenesis additional cell populations become incorporated. It is important to unravel the underlying genetic and cellular mechanisms to be able to identify the embryonic origin of diseases, including congenital malformations, which impair cardiac function and may affect life expectancy or quality. Owing to the evolutionary conservation of development, observations made in non-amniote and amniote vertebrate species allow us to extrapolate to human. This review will focus on the contributions made to a better understanding of heart development through studying avian embryos – mainly the chicken but also quail embryos. We will illustrate the classic and recent approaches used in the avian system, give an overview of the important discoveries made and summarize the early stages of cardiac development up to the establishment of the four chambered heart

Prospective Monitoring of Circulating Epithelial Tumor Cells (CETC) Reveals Changes in Gene Expression during Adjuvant Radiotherapy of Breast Cancer Patients

Circulating epithelial tumor cells (CETC) are considered to be responsible for the formation of metastases. Therefore, their importance as prognostic and/or predictive markers in breast cancer is being intensively investigated. Here, the reliability of single cell expression analyses in isolated and collected CETC from whole blood samples of patients with early-stage breast cancer before and after radiotherapy (RT) using the maintrac ® method was investigated. Single-cell expression analyses were performed with qRT-PCR on a panel of selected genes: GAPDH, EpCAM, NANOG, Bcl-2, TLR 4, COX-2, PIK3CA, Her-2/neu, Vimentin, c-Met, Ki-67. In all patients, viable CETC were detected prior to and at the end of radiotherapy. In 7 of the 9 (77.8%) subjects examined, the CETC number at the end of the radiotherapy series was higher than before. The majority of genes analyzed showed increased expression after completion of radiotherapy compared to baseline. Procedures and methods used in this pilot study proved to be feasible. The method is suitable for further investigation of the underlying molecular biological mechanisms occurring in cells surviving radiotherapy and possibly the development of radiation resistance

Einfluss der Bortezomib-induzierten p53-Reaktivierung auf die Strahlen- und Chemosensibilität HPV-assoziierter Plattenepithelkarzinome der Kopf-Hals-Region

Beim Plattenepithelkarzinom der Kopf-Hals-Region (HNSCC) hat sich die Einteilung in zwei ätiologisch verschiedene Subgruppen etabliert. Während Noxen-assoziierte HNSCC vorwiegend auf den Konsum von Tabak und Alkohol zurückzuführen sind, ist die Tumorprogression in mit Humanen Papillomaviren (HPV) assoziierten HNSCC durch das Wirken viraler Proteine vorangetrieben. Letztere zeigten in den vergangenen Jahren einen deutlichen Inzidenzanstieg, insbesondere im Bereich des Oropharynx. HPV-assoziierte HNSCC weisen eine deutlich bessere Prognose auf, was vor allem auf die höhere Sensibilität gegenüber Chemotherapeutika und Bestrahlung zurückgeführt wird. Die zugrundeliegenden Mechanismen hierfür sind bisher noch nicht genau be-kannt. Eine mögliche Ursache ist der in HPV-assoziierten HNSCC charakteristisch durch das Wirken viraler Proteine inhibierte Signalweg des Tumorsuppressorproteins p53, welches – im Gegensatz zu Noxen-assoziierten HNSCC – hier im Wildtyp vorliegt. Ziel war es, durch Behandlung mit dem Proteasom-Inhibitor Bortezomib in vitro eine Reaktivierung des p53-Signalwegs sowie dessen Auswirkungen auf die Therapiesensibilität von vier HPV-positiven und drei HPV-negativen HNSCC-Zelllinien zu untersuchen. Hierfür wurden die Zellviabilität, Proteinexpression, Zellzyklusprogression, Induktion von Apoptose sowie das klonogene Überleben nach Behandlung mit Bortezomib, Cisplatin und Bestrahlung analysiert. Bortezomib führte, entsprechend dem konstatierten Modell, nur in HPV-positiven HNSCC-Zelllinien zu einer funktionellen Reaktivierung von p53 mit Anstieg des Effektorproteins p21, was durch Kombination mit Bestrahlung verstärkt wurde. Konkordant zu diesen Ergebnissen zeigten sich zelluläre Veränderungen wie ein Zellzyklusarrest in der G1-Phase sowie die Induktion von Apoptose, die mit der Wirkung des reaktivierten p53 vereinbar sind, nur in HPV-positiven HNSCC-Zelllinien. Das klonogene Überleben wurde in allen Zelllinien durch Behandlung mit Bortezomib unabhängig vom HPV-Status reduziert. Eine Strahlensensibilisierung sowie Chemosensibilisierung ge-genüber Cisplatin war in beiden Gruppen nicht nachweisbar. Die Ergebnisse zeigen, dass der p53-Metabolismus in HPV-positiven HNSCC charakteristisch gestört ist, jedoch potentiell reaktiviert werden kann. Die divergenten Ergebnisse beim klonogenen Überleben und die ausbleibende Therapiesensibilisierung machen jedoch deutlich, dass die erhöhte Therapiesensibilität HPV-positiver HNSCC vermutlich durch andere Mechanismen determiniert ist, als nur durch das Wirken von intaktem p53

Unmasking a temperature-dependent effect of the P. anserina i-AAA protease on aging and development

Different molecular pathways involved in maintaining mitochondrial function are of fundamental importance to control cellular homeostasis. Mitochondrial i-AAA protease is part of such a surveillance system, and PaIAP is the putative ortholog in the fungal aging model Podospora anserina. Here, we investigate the role of PaIAP in aging and development. Deletion of the gene encoding PaIAP resulted in a specific phenotype. When incubated at 27°C, spore germination and fruiting body formation are not different from that of the corresponding wild-type strain. Unexpectedly, the lifespan of the deletion strain is strongly increased. In contrast, cultivation at an elevated temperature of 37°C leads to impairments in spore germination and fruiting body formation and to a reduced lifespan. The higher PaIAP abundance in wild-type strains of the fungus grown at elevated temperature and the phenotype of the deletion strain unmasks a temperature-related role of the protein. The protease appears to be part of a molecular system that has evolved to allow survival under changing temperatures, as they characteristically occur in nature

Cardiac injections of AntagomiRs as a novel tool for knockdown of miRNAs during heart development

Background: Studying microRNA networks during heart development is essential to obtain a better understanding of developmental defects and diseases associated with the heart and to identify novel opportunities for therapeutics. Here we highlight the advantages of chicken embryos as a vertebrate model for studying intermediate processes of heart development. Avians develop a four-chambered heart closely resembling human anatomy and they develop ex utero, which makes them easily accessible. Furthermore, embryos are available all year with a steady supply. Results: In this report we established a novel method for the knockdown of microRNA function by microinjecting AntagomiRs into the chicken heart in ovo. Our approach enables the targeted delivery of antagomirs into a locally restricted area and is not impacted by circulation. After further embryo development the successful miRNA knockdown was confirmed. Loss of function phenotypes can be evaluated rapidly, compared to more time-consuming genetic ablation experiments. The local application avoids potential systemic effects of microRNA knockdown, therefore allowing the assessment of impacts on heart development only. The method can be adjusted for different stages of chicken embryos (HH13-HH18) as well as for knockdown or targeted overexpression of coding genes. Conclusion: In conclusion our method allows targeted and locally restricted delivery of Antagomirs to the heart leading to successful knockdown of microRNA function. This method enables rapid phenotypic assessment, for example by gene expression analysis of multiple cardiac genes

Dual PI3K/mTOR inhibitor NVP-BEZ235 enhances radiosensitivity of head and neck squamous cell carcinoma (HNSCC) cell lines due to suppressed Double-Strand Break (DSB) repair by non-homologous end joining

The PI3K/Akt/mTOR pathway is frequently altered in human papillomavirus (HPV)-positive and negative squamous cell carcinoma of the head and neck (HNSCC) and overstimulation is associated with poor prognosis. PI3K drives Akt activation and constitutive signaling acts pro-proliferative, supports cell survival, DNA repair, and contributes to radioresistance. Since the small molecule NVP-BEZ235 (BEZ235) is a potent dual inhibitor of this pathway, we were interested whether BEZ235 could be an efficient radiosensitizer. The 50 nM BEZ235 was found to abrogate endogenous and irradiation-induced phosphorylation of Akt (Ser473). The anti-proliferative capacity of the drug resulted in an increase in G1-phase cells. Repair of radiation-induced DNA double-strand breaks (DSBs) was strongly suppressed. Reduction in DSB repair was only apparent in G1- but not in G2-phase cells, suggesting that BEZ235 primarily affects non-homologous end joining. This finding was confirmed using a DSB repair reporter gene assay and could be attributed to an impaired phosphorylation of DNA-PKcs (S2056). Cellular radiosensitivity increased strongly after BEZ235 addition in all HNSCC cell lines used, especially when irradiated in the G0 or G1 phase. Our data indicate that targeting the PI3K/Akt/mTOR pathway by BEZ235 with concurrent radiotherapy may be considered an effective strategy for the treatment of HNSCC, regardless of the HPV and Akt status

Early Mortality among Patients with Head and Neck Cancer Diagnosed in Thuringia, Germany, between 1996 and 2016—A Population-Based Study

Simple Summary When we consider the outcome of cancer treatment, we mostly focus on overall survival: studies on early mortality in head and neck cancer (HNC) are sparse. This retrospective population-based study investigated early mortality of HNC and the influence of patients’ tumor and treatment characteristics. All 8288 patients with primary HNC of the German federal state Thuringia from 1996 to 2016 were included. Statistics were performed to identify independent factors for 30-day, 90-day, and 180-day mortality. The 30-, 90-, and 180-day mortality risks were 1.8%, 5.1%, and 9.6%, respectively. Male sex, increasing age, larger tumor size, and distant metastasis, tumors of the cavity of mouth, oropharynx, and hypopharynx had a significantly greater 180-day mortality. Surgery, radiotherapy, and multimodal therapy were associated with decreased 180-day mortality. Typical factors associated with worse overall survival had the most important impact on early mortality in HNC patients in a population-based setting. Abstract Population-based studies on early mortality in head and neck cancer (HNC) are sparse. This retrospective population-based study investigated early mortality of HNC and the influence of patients’ tumor and treatment characteristics. All 8288 patients with primary HNC of the German federal state Thuringia from 1996 to 2016 were included. Univariate and multivariate analysis were performed to identify independent factors for 30-day, 90-day, and 180-day mortality. The 30-, 90-, and 180-day mortality risks were 1.8%, 5.1%, and 9.6%, respectively. In multivariable analysis, male sex (odds ratio (OR) 1.41; 95% confidence interval (CI) 1.08–1.84), increasing age (OR 1.81; CI 1.49–2.19), higher T (T4: OR 3.09; CI 1.96–4.88) and M1 classification (OR 1.97; CI 1.43–2.73), advanced stage (IV: OR 3.97; CI 1.97–8.00), tumors of the cavity of mouth (OR 3.47; CI 1.23–9.75), oropharynx (OR 3.01; CI 1.06–8.51), and hypopharynx (OR 3.27; CI 1.14–9.40) had a significantly greater 180-day mortality. Surgery (OR 0.51; CI 0.36–0.73), radiotherapy (OR 0.37; CI 0.25–0.53), and multimodal therapy (OR 0.10; CI 0.07–0.13) were associated with decreased 180-day mortality. Typical factors associated with worse overall survival had the most important impact on early mortality in a population-based setting

Role of Intraparotid and Neck Lymph Node Metastasis in Primary Parotid Cancer Surgery: A Population-Based Analysis

Simple Summary The prognostic role of intraparotid (PAR) and cervical lymph node (LN) metastasis on overall survival (OS) of primary parotid cancer is unclear. All 345 Thuringian patients with parotid cancer from 1996 to 2016 were included in a population-based study. OS was assessed in relation to the total number of removed PAR and cervical LN, number of positive intraparotid (PAR+), positive cervical LN, LN ratio, log odds of positive LN (LODDS), as well as including the PAR as LODDS-PAR. PAR was assessed in 42% of the patients (22% of these PAR+). T and N classification were not independent predictors of OS. When combining T with LODDS instead of N, higher T became a strong prognosticator, but not LODDS. When combining T classification with LODDS-PAR, both higher T classification and the classification with LODDS-PAR became independent predictors of worse OS. LODDS-PAR seems to be an optimal prognosticator for OS in primary parotid cancer. Abstract This population-based study investigated the prognostic role of intraparotid (PAR) and cervical lymph node (LN) metastasis on overall survival (OS) of primary parotid cancer. All 345 patients (median age: 66 years; 43% female, 49% N+, 31% stage IV) of the Thuringian cancer registries with parotid cancer from 1996 to 2016 were included. OS was assessed in relation to the total number of removed PAR and cervical LN, number of positive intraparotid (PAR+), positive cervical LN, LN ratio, log odds of positive LN (LODDS), as well as including the PAR as LODDS-PAR. PAR was assessed in 42% of the patients (22% of these PAR+). T and N classification were not independent predictors of OS. When combining T with LODDS instead of N, higher T (T3/T4) became a prognosticator (hazard ratio (HR) = 2.588; CI = 1.329–5.040; p = 0.005) but not LODDS ( p > 0.05). When combining T classification with LODDS-PAR, both higher T classification (HR = 2.256; CI = 1.288–3.950; p = 0.004) and the alternative classification with LODDS-PAR (≥median −1.11; HR 2.078; CI = 1.155–3.739; p = 0.015) became independent predictors of worse OS. LODDS-PAR was the only independent prognosticator out of the LN assessment for primary parotid cancer

Aporte del ejercicio físico y deportivo en la formación profesional y futuro laboral de los alumnos que cursan las carreras de pedagogía en la Universidad Andrés Bello sede Casona las Condes

Tesis (Profesor de Educación Física para la Enseñanza Básica, Licenciado en Educación)El presente estudio tiene por objetivo determinar si aporta el ejercicio físico y deportivo en la formación profesional y futuro laboral de los alumnos que cursan las carreras de Pedagogía en la Universidad Andrés Bello, sede Casona las Condes. La importancia del estudio radica en ser un aporte para futuras investigaciones en relación al ejercicio y la educación. Está dividido en cuatro capítulos, el primero aborda todo lo relacionado con la teoría que lo sustenta y se considerarán temas como el ejercicio físico y sus beneficios en el ámbito académico y laboral, motivaciones y razones por las que no se realiza, cualidades de un buen profesor, entre otros. El segundo versa sobre las características del estudio, la cual es una investigación cuantitativa-descriptiva, analizando una situación que ocurre en condiciones naturales. Además, es una investigación no experimental, por lo tanto, de campo, que consiste en una recolección de datos directamente de la muestra, sin manipulación. El tercero presenta los datos recopilados y los análisis que fueron realizados a través de una encuesta que fue aplicada a los alumnos que ejercen las carreras de Pedagogía en inglés, Educación Parvularia, Educación Física, Psicopedagogía, Educación Musical y Pedagogía General Básica de la Universidad Andrés Bello. Luego, con los datos se crearon gráficos para analizarlos y desarrollarlos en base al marco teórico. El cuarto y último corresponde a la conclusión, que se resume en que los estudiantes de pedagogía si creen que el ejercicio físico aportaría a la formación docente y al futuro laboral, sin embargo, esto no se ve reflejado en sus hábitos de ejercicio físico, ya que la mayoría de los estudiantes son sedentarios

Theranostics in Boron Neutron Capture Therapy

Boron neutron capture therapy (BNCT) has the potential to specifically destroy tumor cells without damaging the tissues infiltrated by the tumor. BNCT is a binary treatment method based on the combination of two agents that have no effect when applied individually: B-10 and thermal neutrons. Exclusively, the combination of both produces an effect, whose extent depends on the amount of B-10 in the tumor but also on the organs at risk. It is not yet possible to determine the B-10 concentration in a specific tissue using non-invasive methods. At present, it is only possible to measure the B-10 concentration in blood and to estimate the boron concentration in tissues based on the assumption that there is a fixed uptake of B-10 from the blood into tissues. On this imprecise assumption, BNCT can hardly be developed further. A therapeutic approach, combining the boron carrier for therapeutic purposes with an imaging tool, might allow us to determine the B-10 concentration in a specific tissue using a non-invasive method. This review provides an overview of the current clinical protocols and preclinical experiments and results on how innovative drug development for boron delivery systems can also incorporate concurrent imaging. The last section focuses on the importance of proteomics for further optimization of BNCT, a highly precise and personalized therapeutic approach