Effects of different sevoflurane concentrations on Akt isoforms in normal and cancer breast cells. An experimental model

Grigore T. Popa University of Medicine and Pharmacy, Iași, România, Regional Institute of Oncology, Anaesthesia and Intensive Care Department, Iași, România, Regional Institute of Oncology, Department of Molecular Biology, TRANSCEND, Iași, România,The 5th International Congress of the Society of Anesthesiology and Reanimatology of the Republic of Moldova, 16th Edition of the International Course of Guidelines and Protocols in Anesthesia, Intensive Care and Emergency Medicine, 28th Meeting of the European Society for Computing and Technology in Anesthesia and Intensive Care September 27-29, 2018, Chisinau, the Republic of MoldovaIntroduction: Multiple perioperative factors influence cancer patient evolution and outcome. Microenvironmental factors activate different gene programs that enable tumor cell to invade, survive and promote drug resistance and metastasis. The effects of anesthetic drugs on cancer progression is under scrutiny, but published data are controversial and the involved mechanisms unclear. Tumor development implies PI3K/AKT pathway activation. Akt isoforms (1,2,3) are frequently amplified in various malignant tumors and associated with malignant cell survival, proliferation and invasion. Their activation is often observed in human cancers and is associated with decreased survival rate. Objective: Identification of Akt isoforms activated in sevoflurane exposed breast tumor cells. Material and methods: Normal breast cells MCF10A (ATCC®) and breast cancer cells MDA-MB-231 (ATCC®) were cultured 2D (standard adhesive culture plastic plates) and 3D (matrigel). Study groups were exposed to different sevoflurane concentrations (0.5, 2, 3, 4 mM) compared to control unexposed groups. Unexposed and sevoflurane exposed cells (2D and 3D) were evaluated by optic microscopy and viability tests. Akt isoforms were assessed by immunofluorescence. Results: Sevoflurane alters tumor cell proliferation and Akt isoforms expression in a dose-dependent manner. The phenotype of 3D 2mM sevoflurane exposed cells show an increased migration capacity which indicates increased aggressivity. Conclusions: Sevoflurane exposure of breast cancer cells influences cell proliferation, phenotype and Akt isoform expression. Increased sevoflurane concentrations activate different Akt isoforms, putatively related to epithelial-mesenchimal transition and promote cancer cell invasion, migration and metastasis

Hyperoxia influences cancer growth and metastasis. A pilot experimental model

Grigore T. Popa University of Medicine and Pharmacy, Iași, România, Regional Institute of Oncology, Anaesthesia and Intensive Care Department, Iași, România, Regional Institute of Oncology, Department of Molecular Biology, TRANSCEND, Iași, România, CEMEX Research Center, Grigore T. Popa University of Medicine and Pharmacy, Iași, România, The 5th International Congress of the Society of Anesthesiology and Reanimatology of the Republic of Moldova, 16th Edition of the International Course of Guidelines and Protocols in Anesthesia, Intensive Care and Emergency Medicine, 28th Meeting of the European Society for Computing and Technology in Anesthesia and Intensive Care September 27-29, 2018, Chisinau, the Republic of MoldovaIntroduction: Perioperative care of cancer patients is under scrutiny. Among many factors promoting cancer recurrence and metastasis, high oxygen concentration exposure is underevaluated. While oxygen toxicity is documented in several circumstances, its implication in tumor cell growth and progression is poorly understood. Objective: To characterize high oxygen concentration exposure effects on tumor progression using a breast cancer murine model. Material and methods: A highly aggressive breast tumor cell line 4T1 (ATCC®) was injected in mammary gland in 8 week old females BALB/c mice. We divided the animals into 3 groups, each including 6 individuals: G1 – tumor bearing mice with no intervention post inoculation; G2 – primary tumor removal at 2 weeks post inoculation; G3 - primary tumor removal at 2 weeks post inoculation followed by 6 hours of 75% oxygen exposure. In all groups cancer evolution was assessed at 6 weeks by standard pathomorphological evaluation: specimens from the primary tumor, locally recurrent tumor and target organ metastasis were assessed by hematoxylin-eosin staining, and digital microscopy. Results: Surgically removed primary tumors in G3 group had similar characteristics with those in G2 group and previously described models. At study endpoint, compared with both G2 and G1 groups, G3 animals showed significantly higher tumor burden: larger local recurrence and more metastasis (larger number and dimensions) in liver and lungs, associated with significantly enlarged spleen. Conclusions: Short term (6 hours) high oxygen (75%) concentration exposure results in significantly more aggressive progression of a 4T1-BALB/c murine breast cancer model

Selective Survival and Maturation of Adult-Born Dentate Granule Cells Expressing the Immediate Early Gene Arc/Arg3.1

Progenitor cells in the adult dentate gyrus provide a constant supply of neuronal precursors, yet only a small fraction of these cells survive and develop into mature dentate granule cells (DGCs). A major challenge of current research is thus to understand the stringent selection process that governs the maturation and functional integration of adult-born DGCs. In mature DGCs, high-frequency stimulation (HFS) of the perforant path input elicits robust expression of the immediate early gene Arc/Arg3.1, trafficking of its mRNA to dendrites, and local synthesis of the protein necessary for consolidation of long-term potentiation (LTP). Given the synaptic commitment inherent in LTP consolidation, we considered that HFS-evoked expression of Arc could be used to timemap the functional integration of newborn DGCs. Dividing cells were birthmarked by BrdU-labeling at 1, 7, 14, 21, or 28 days prior to induction of LTP and expression of Arc was examined by confocal microscopy. Contrary to expectation, LTP did not induce Arc expression in newborn cells at any age, suggesting they might be refractory to synaptically-evoked Arc expression for at least one month. Importantly, however, spontaneous expression of Arc was detected in BrdU-labeled cells and strongly associated with the survival and maturation of NeuN-positive DGCs. Moreover, Arc expression at the earliest ages (1 and 7 days), clearly precedes the formation of glutamatergic synapses on new neurons. These results suggest an unexpected early role for Arc in adult-born DGCs, distinct from its functions in LTP, LTD, and homeostatic synaptic plasticity

La question de la vérité chez Hans Urs von Balthasar : la vérité du monde

Aujourd’hui, réfléchir sur la vérité n’est pas un acte intellectuel simple ; plusieurs registres de vérité peuvent être réfléchis dans notre univers intellectuel. Avec Hans Urs von Balthasar, (l’auteur que nous étudions) nous approchons les vérités du monde (les réalités sensibles ainsi que l’être en tant qu’étant fini). Notre auteur construit sa réflexion à partir d’une ontologie transcendantale ; son interlocuteur principal est M. Heidegger. Néanmoins, Balthasar ne reste pas dans la ligne de réflexion phénoménologique de Heidegger, car il réalise un tournant de la méthode phénoménologique vers des réflexions théologiques : Balthasar comprend Dieu comme source de toute vérité. Notre auteur souhaite surprendre la vérité, il pense découvrir son mystère en faisant une lecture de phénomène et en le questionnant. Son but est de faire passer le phénomène d’un côté caché à un côté plus perceptible. Le phénomène de la vérité se dévoile. Balthasar est à la recherche d’un processus de la connaissance. Ce processus est construit autour de l’être qui se dévoile et autour du sujet qui accueille l’être et son essence (le fond). Autrement dit l’apparition (l’image) de l’être exerce deux mouvements : 1° l’apparition éclaire l’essence de l’être qui se manifeste ; 2° l’apparition se retire et se rend superflue en dirigeant l’attention sur l’essence de l’être (le fond). Du côté du sujet, celui-ci construit une signification de l’être en tant qu’être. L’être impose un double mouvement au sujet conscient : 1° renoncer à la variété du sensible pour construire un concept général ; 2° ne jamais utiliser ce concept sans le monde des images, car le concept pur n’a pas de vie ni de vérité s’il reste un concept abstrait. Ce qui signifie que le sujet connait un double mouvement : l’intuition va au fond dans le cœur du concept et le concept revient, il sort de toute abstraction pour rejoindre l’intuition. H.U. von Balthasar a pensé la vérité une et plurielle à partir de la philosophie (phénoménologie transcendantale) et de la théologie chrétienne. L’apport théologique à la réflexion de notre auteur souhaite donner des ailes et des perspectives supplémentaires à cette existence et lui donner un sens qui ne s’épuisera pas.Master [120] en philosophie, Université catholique de Louvain, 201

Exploring Hyperoxia Effects in Cancer—From Perioperative Clinical Data to Potential Molecular Mechanisms

Increased inspiratory oxygen concentration is constantly used during the perioperative period of cancer patients to prevent the potential development of hypoxemia and to provide an adequate oxygen transport to the organs, tissues and cells. Although the primary tumours are surgically removed, the effects of perioperative hyperoxia exposure on distal micro-metastases and on circulating cancer cells can potentially play a role in cancer progression or recurrence. In clinical trials, hyperoxia seems to increase the rate of postoperative complications and, by delaying postoperative recovery, it can alter the return to intended oncological treatment. The effects of supplemental oxygen on the long-term mortality of surgical cancer patients offer, at this point, conflicting results. In experimental studies, hyperoxia effects on cancer biology were explored following multiple pathways. In cancer cell cultures and animal models, hyperoxia increases the production of reactive oxygen species (ROS) and increases the oxidative stress. These can be followed by the induction of the expression of Brain-derived neurotrophic factor (BDNF) and other molecules involved in angiogenesis and by the promotion of various degrees of epithelial mesenchymal transition (EMT)

Enhancing Anti-Tumoral Potential of CD-NHF by Modulating PI3K/Akt Axis in U87 Ex Vivo Glioma Model

Background: In the latest years, there has been an increased interest in nanomaterials that may provide promising novel approaches to disease diagnostics and therapeutics. Our previous results demonstrated that Carbon-dots prepared from N-hydroxyphthalimide (CD-NHF) exhibited anti-tumoral activity on several cancer cell lines such as MDA-MB-231, A375, A549, and RPMI8226, while U87 glioma tumor cells were unaffected. Gliomas represent one of the most common types of human primary brain tumors and are responsible for the majority of deaths. In the present in vitro study, we expand our previous investigation on CD-NHF in the U87 cell line by adding different drug combinations. Methods: Cell viability, migration, invasion, and immunofluorescent staining of key molecular pathways have been assessed after various treatments with CD-NHF and/or K252A and AKTVIII inhibitors in the U87 cell line. Results: Association of an inhibitor strongly potentiates the anti-tumoral properties of CD-NHF identified by significant impairment of migration, invasion, and expression levels of phosphorylated Akt, p70S6Kinase, or by decreasing expression levels of Bcl-2, IL-6, STAT3, and Slug. Conclusions: Using simultaneously reduced doses of both CD-NHF and an inhibitor in order to reduce side effects, the viability and invasiveness of U87 glioma cells were significantly impaired

ROMANIAN DOCTORAL RESEARCH – PERSPECTIVES ON INTERNATIONAL ACCOUNTING HARMONIZATION

The purpose of this study is to analyze the interest of Romanian accounting doctoral researchers regarding the process of international accounting harmonization. We analyzed the research outcomes, doctoral dissertations, with the subtopic on international harmonization for the period 2000-2010, from the most important Romanians research and doctoral schools of accounting. The results of our quantitative study confirm our hypothesis, showing that there is an increasing trend concerning the interest in accounting harmonization, and the results of our qualitative study confirm our second hypothesis showing that the research topics represent sensitive issues of the accounting harmonization process. Our findings conclude that the Romanian accounting research environment is in line with the international accounting research environment on the topic of accounting harmonization and its subtopics

Long-Term Deleterious Effects of Short-term Hyperoxia on Cancer Progression—Is Brain-Derived Neurotrophic Factor an Important Mediator? An Experimental Study

Perioperative factors promoting cancer recurrence and metastasis are under scrutiny. While oxygen toxicity is documented in several acute circumstances, its implication in tumor evolution is poorly understood. We investigated hyperoxia long-term effects on cancer progression and some underlying mechanisms using both in vitro and in vivo models of triple negative breast cancer (TNBC). We hypothesized that high oxygen exposure, even of short duration, may have long-term effects on cancer growth. Considering that hyperoxic exposure results in reactive oxygen species (ROS) formation, increased oxidative stress and increased Brain-Derived Neurotrophic Factor (BDNF) expression, BDNF may mediate hyperoxia effects offering cancer cells a survival advantage by increased angiogenesis and epithelial mesenchymal transition (EMT). Human breast epithelial MCF10A, human MDA-MB-231 and murine 4T1 TNBC were investigated in 2D in vitro system. Cells were exposed to normoxia or hyperoxia (40%, 60%, 80% O2) for 6 h. We evaluated ROS levels, cell viability and the expression of BDNF, HIF-1α, VEGF-R2, Vimentin and E-Cadherin by immunofluorescence. The in vivo model consisted of 4T1 inoculation in Balb/c mice and tumor resection 2 weeks after and 6 h exposure to normoxia or hyperoxia (40%, 80% O2). We measured lung metastases and the same molecular markers, immediately and 4 weeks after surgery. The in vitro study showed that short-term hyperoxia exposure (80% O2) of TNBC cells increases ROS, increases BDNF expression and that promotes EMT and angiogenesis. The in vivo data indicates that perioperative hyperoxia enhances metastatic disease and this effect could be BDNF mediated