Status of the ACCULINNA-2 project at FLNR

The project of a new and more powerful in-flight fragment separator ACCULINNA-2 at U-400M cyclotron in FLNR, JINR planned to build in addition to the existing separator ACCULINNA is presented. The new separator will provide high intensity RIBs in the lowest energy range (5÷50 MeV/nucleon) which is attainable for in-flight separators. The possibilities for the astrophysics studies at the proposed device are presented. ACCULINNA-2 separator is planned to be constructed in the years 2010-2015. The current status of the project is reported

Sec12 Binds to Sec16 at Transitional ER Sites

COPII vesicles bud from an ER domain known as the transitional ER (tER). Assembly of the COPII coat is initiated by the transmembrane guanine nucleotide exchange factor Sec12. In the budding yeast Pichia pastoris, Sec12 is concentrated at tER sites. Previously, we found that the tER localization of P. pastoris Sec12 requires a saturable binding partner. We now show that this binding partner is Sec16, a peripheral membrane protein that functions in ER export and tER organization. One line of evidence is that overexpression of Sec12 delocalizes Sec12 to the general ER, but simultaneous overexpression of Sec16 retains overexpressed Sec12 at tER sites. Additionally, when P. pastoris Sec12 is expressed in S. cerevisiae, the exogenous Sec12 localizes to the general ER, but when P. pastoris Sec16 is expressed in the same cells, the exogenous Sec12 is recruited to tER sites. In both of these experimental systems, the ability of Sec16 to recruit Sec12 to tER sites is abolished by deleting a C-terminal fragment of Sec16. Biochemical experiments confirm that this C-terminal fragment of Sec16 binds to the cytosolic domain of Sec12. Similarly, we demonstrate that human Sec12 is concentrated at tER sites, likely due to association with a C-terminal fragment of Sec16A. These results suggest that a Sec12–Sec16 interaction has a conserved role in ER export

Integrated high-content quantification of intracellular ROS levels and mitochondrial morphofunction

Oxidative stress arises from an imbalance between the production of reactive oxygen species (ROS) and their removal by cellular antioxidant systems. Especially under pathological conditions, mitochondria constitute a relevant source of cellular ROS. These organelles harbor the electron transport chain, bringing electrons in close vicinity to molecular oxygen. Although a full understanding is still lacking, intracellular ROS generation and mitochondrial function are also linked to changes in mitochondrial morphology. To study the intricate relationships between the different factors that govern cellular redox balance in living cells, we have developed a high-contentmicroscopy-based strategy for simultaneous quantification of intracellular ROS levels and mitochondrial morphofunction. Here, we summarize the principles of intracellular ROS generation and removal, and we explain the major considerations for performing quantitative microscopy analyses of ROS and mitochondrial morphofunction in living cells. Next, we describe our workflow, and finally, we illustrate that a multiparametric readout enables the unambiguous classification of chemically perturbed cells as well as laminopathy patient cells

Practical guidelines for rigor and reproducibility in preclinical and clinical studies on cardioprotection

The potential for ischemic preconditioning to reduce infarct size was first recognized more than 30 years ago. Despite extension of the concept to ischemic postconditioning and remote ischemic conditioning and literally thousands of experimental studies in various species and models which identified a multitude of signaling steps, so far there is only a single and very recent study, which has unequivocally translated cardioprotection to improved clinical outcome as the primary endpoint in patients. Many potential reasons for this disappointing lack of clinical translation of cardioprotection have been proposed, including lack of rigor and reproducibility in preclinical studies, and poor design and conduct of clinical trials. There is, however, universal agreement that robust preclinical data are a mandatory prerequisite to initiate a meaningful clinical trial. In this context, it is disconcerting that the CAESAR consortium (Consortium for preclinicAl assESsment of cARdioprotective therapies) in a highly standardized multi-center approach of preclinical studies identified only ischemic preconditioning, but not nitrite or sildenafil, when given as adjunct to reperfusion, to reduce infarct size. However, ischemic preconditioning—due to its very nature—can only be used in elective interventions, and not in acute myocardial infarction. Therefore, better strategies to identify robust and reproducible strategies of cardioprotection, which can subsequently be tested in clinical trials must be developed. We refer to the recent guidelines for experimental models of myocardial ischemia and infarction, and aim to provide now practical guidelines to ensure rigor and reproducibility in preclinical and clinical studies on cardioprotection. In line with the above guideline, we define rigor as standardized state-of-the-art design, conduct and reporting of a study, which is then a prerequisite for reproducibility, i.e. replication of results by another laboratory when performing exactly the same experiment

The suggested new fragment separator acculinna-2

We present new project of fragment separator ACCULINNA-2 that is being planned to be constructed in Flerov Laboratory of Nuclear Reaction, JINR. The ACCULINNA-2 facility is not intended to compete with the new large in-flight RIB facilities. It should complement the existing/constructed facilities in certain fields. Namely, ACCULINNA-2 should provide high intensity RIBs in the lowest energy range attainable for in-flight separators

Long range plan with radioactive beams at Dubna

A program for upgrade of existing radioactive ion beams facilities at Flerov Laboratory of Nuclear Reactions, JINR Dubna is presented. A project of a new in-flight fragment separator ACCULINNA-2 is proposed. It is expected the new instrument will be more universal and powerful than the existing nowadays. The beam intensity should be increased by factor 10-15, its optical quality greatly improved and the range of the accessible secondary radioactive beams broadened up to Z∼20. Main ion-optical characteristics, operating principles and a tentative plan for the project realization are included. An extensive research program based on local experiments made so far and international proposals for these equipments is discussed

Long range plan with radioactive beams at Dubna

A program for upgrade of existing radioactive ion beams facilities at Flerov Laboratory of Nuclear Reactions, JINR Dubna is presented. A project of a new in-flight fragment separator ACCULINNA-2 is proposed. It is expected the new instrument will be more universal and powerful than the existing nowadays. The beam intensity should be increased by factor 10-15, its optical quality greatly improved and the range of the accessible secondary radioactive beams broadened up to Z∼20. Main ion-optical characteristics, operating principles and a tentative plan for the project realization are included. An extensive research program based on local experiments made so far and international proposals for these equipments is discussed