Radiation-mediated induction of apoptotic cell death in rat hippocampus

Ionizing radiation is commonly used in the treatment of brain tumors but it can impair cognitive functions, such as learning and memory. Since cognitive dysfunctions are predominantly result of cell death by apoptosis in hippocampal cells, in this study we analyzed acute effects of cranial gamma-irradiation (10 Gy) on expression of proapoptotic molecules (p53, Bax) and antiapoptotic molecule Bcl-2, as well as caspase-3 activation and cytochrome c redistribution in the hippocampus of young rats. The selected regimen of irradiation resembles the established animal model for childhood prophylactic cranial radiotherapy. Our results demonstrated that p53 mRNA expression was unchanged after irradiation, while induction of p53 protein was rapid. In parallel, Bax mRNA and protein levels were also increased following irradiation, whereas Bcl-2 expression was not changed during the examined post-irradiation period. These changes were accompanied with early hallmarks of apoptosis, such as increased cytochrome c release and stimulated activation of caspase-3. Overall, this study demonstrates that cranial irradiation is associated with the augmented apoptotic pathway in the rat hippocampus, which could be related to the cognitive decline observed in patients after prophylactic cranial radiotherapy, but also opens perspective in finding radioprotectors that can mitigate radiation injury of normal brain tissue.Jonizujuće zračenje se često koristi u tretmanu tumora mozga ali ono može da ošteti kognitivne funkcije, kao što su učenje i pamćenje. Pošto je smanjenje kognitivnih funkcija većinom rezultat ćelijske smrti putem apoptoze u hipokampalnim ćelijama u ovoj studiji su analizirani efekti kranijalnog gama-zračenja (10 Gy) na ekspresiju proapoptotskih (p53, Bax) i antiapoptotskih molekula (Bcl-2), kao i na aktivaciju kaspaze-3 i redistribuciju citohroma c u hipokampusu mladih pacova. Odabrani režim zračenja odgovara uspostavljenom životinjskom modelu za dečiju profilaktičku kranijalnu radioterapiju. Naši rezultati pokazuju da je ekspresija p53 i RNK nepromenjena nakon zračenja, dok je indukcija p53 proteina veoma brza. Istovremeno Bax iRNK i protein su povećani nakon zračenja, dok je ekspresija Bcl-2 nepromenjena tokom ispitivanog perioda. Ove promene su praćene ranim znacima apoptoze, kao što su povećano oslobađanje citohroma c i aktivacija kaspaze-3. Generalno, ova studija pokazuje da je kranijalno zračenje povezano sa stimulisanim apoptotskim putem u hipokampusu pacova, što se može povezati sa kognitivnim oštećenjima uočenih kod pacijenata nakon profilaktičke kranijalne radioterapije, ali takođe otvara perspektive u pronalaženju radioprotektora koji smanjuju oštećenje normalnog nervnog tkiva nakon zračenja.nul

Radiation-mediated induction of apoptotic cell death in rat hippocampus

Ionizing radiation is commonly used in the treatment of brain tumors but it can impair cognitive functions, such as learning and memory. Since cognitive dysfunctions are predominantly result of cell death by apoptosis in hippocampal cells, in this study we analyzed acute effects of cranial gamma-irradiation (10 Gy) on expression of proapoptotic molecules (p53, Bax) and antiapoptotic molecule Bcl-2, as well as caspase-3 activation and cytochrome c redistribution in the hippocampus of young rats. The selected regimen of irradiation resembles the established animal model for childhood prophylactic cranial radiotherapy. Our results demonstrated that p53 mRNA expression was unchanged after irradiation, while induction of p53 protein was rapid. In parallel, Bax mRNA and protein levels were also increased following irradiation, whereas Bcl-2 expression was not changed during the examined post-irradiation period. These changes were accompanied with early hallmarks of apoptosis, such as increased cytochrome c release and stimulated activation of caspase-3. Overall, this study demonstrates that cranial irradiation is associated with the augmented apoptotic pathway in the rat hippocampus, which could be related to the cognitive decline observed in patients after prophylactic cranial radiotherapy, but also opens perspective in finding radioprotectors that can mitigate radiation injury of normal brain tissue.Jonizujuće zračenje se često koristi u tretmanu tumora mozga ali ono može da ošteti kognitivne funkcije, kao što su učenje i pamćenje. Pošto je smanjenje kognitivnih funkcija većinom rezultat ćelijske smrti putem apoptoze u hipokampalnim ćelijama u ovoj studiji su analizirani efekti kranijalnog gama-zračenja (10 Gy) na ekspresiju proapoptotskih (p53, Bax) i antiapoptotskih molekula (Bcl-2), kao i na aktivaciju kaspaze-3 i redistribuciju citohroma c u hipokampusu mladih pacova. Odabrani režim zračenja odgovara uspostavljenom životinjskom modelu za dečiju profilaktičku kranijalnu radioterapiju. Naši rezultati pokazuju da je ekspresija p53 i RNK nepromenjena nakon zračenja, dok je indukcija p53 proteina veoma brza. Istovremeno Bax iRNK i protein su povećani nakon zračenja, dok je ekspresija Bcl-2 nepromenjena tokom ispitivanog perioda. Ove promene su praćene ranim znacima apoptoze, kao što su povećano oslobađanje citohroma c i aktivacija kaspaze-3. Generalno, ova studija pokazuje da je kranijalno zračenje povezano sa stimulisanim apoptotskim putem u hipokampusu pacova, što se može povezati sa kognitivnim oštećenjima uočenih kod pacijenata nakon profilaktičke kranijalne radioterapije, ali takođe otvara perspektive u pronalaženju radioprotektora koji smanjuju oštećenje normalnog nervnog tkiva nakon zračenja.nul

Host Genetic Background Strongly Influences the Response to Influenza A Virus Infections

The genetic make-up of the host has a major influence on its response to combat pathogens. For influenza A virus, several single gene mutations have been described which contribute to survival, the immune response and clearance of the pathogen by the host organism. Here, we have studied the influence of the genetic background to influenza A H1N1 (PR8) and H7N7 (SC35M) viruses. The seven inbred laboratory strains of mice analyzed exhibited different weight loss kinetics and survival rates after infection with PR8. Two strains in particular, DBA/2J and A/J, showed very high susceptibility to viral infections compared to all other strains. The LD50 to the influenza virus PR8 in DBA/2J mice was more than 1000-fold lower than in C57BL/6J mice. High susceptibility in DBA/2J mice was also observed after infection with influenza strain SC35M. In addition, infected DBA/2J mice showed a higher viral load in their lungs, elevated expression of cytokines and chemokines, and a more severe and extended lung pathology compared to infected C57BL/6J mice. These findings indicate a major contribution of the genetic background of the host to influenza A virus infections. The overall response in highly susceptible DBA/2J mice resembled the pathology described for infections with the highly virulent influenza H1N1-1918 and newly emerged H5N1 viruses

Fumarate is cardioprotective via activation of the Nrf2 antioxidant pathway

The citric acid cycle (CAC) metabolite fumarate has been proposed to be cardioprotective; however, its mechanisms of action remain to be determined. To augment cardiac fumarate levels and to assess fumarate's cardioprotective properties, we generated fumarate hydratase (Fh1) cardiac knockout (KO) mice. These fumarate-replete hearts were robustly protected from ischemia-reperfusion injury (I/R). To compensate for the loss of Fh1 activity, KO hearts maintain ATP levels in part by channeling amino acids into the CAC. In addition, by stabilizing the transcriptional regulator Nrf2, Fh1 KO hearts upregulate protective antioxidant response element genes. Supporting the importance of the latter mechanism, clinically relevant doses of dimethylfumarate upregulated Nrf2 and its target genes, hence protecting control hearts, but failed to similarly protect Nrf2-KO hearts in an in vivo model of myocardial infarction. We propose that clinically established fumarate derivatives activate the Nrf2 pathway and are readily testable cytoprotective agents. © 2012 Elsevier Inc

Identification and validation of novel EST-SSR markers in olives

ABSTRACT: The olive (Olea europaea L.) is a leading oil crop in the Mediterranean area. Limited information on the inheritance of agronomic significant traits hinders progress in olive breeding programs, which encourages the development of markers linked to the traits. In this study, we report on the development of 46 olive simple sequence repeat (SSR) markers, obtained from 577,025 expressed sequence tags (ESTs) in developing olive fruits generated in the framework of the Slovenian national olive transcriptome project. Sequences were de novo assembled into 98,924 unigenes, which were then used as a source for microsatellites searching. We identified 923 unigenes that contained 984 SSRs among which dinucleotide SSRs (36 %) were the most abundant, followed by tri- (33 %) and hexa- (21 %) nucleotides. Microsatellite repeat motif GA (37 %) was the most common among dinucleotides, while microsatellite repeat motif GAA was the most abundant trinucleotide SSR motif (16 %). Gene ontology annotations could be assigned to 27 % of the unigenes. A hundred and ten expressed sequence tag-derived-simple sequence repeats (EST-SSRs) with annotated genes were selected for primer designing and finally, 46 (42 %) polymorphic EST-SSRs were successfully amplified and used to validate genetic diversity among 24 olive varieties. The average number of alleles per locus, observed heterozygosity, expected heterozygosity, and polymorphic information content were 4.5, 0.649, 0.604 and 0.539, respectively. Twenty-seven EST-SSRs showed good diversity properties and were recommended for further olive genome investigation

Separation of mineral oil droplets using polypropylene fibre bed coalescence

This paper investigates the separation possibilities of model emulsion oil-in-water using polypropylene fibre bed coalescence. Experiments were carried out over a wide range of physico-chemical characteristics of mineral oils, bed permeability and operating fluid velocities. The aim of this study was to analyse the influence of the dispersed oil phase nature and of the bed geometry on the separation efficiency. From the obtained results, it can be concluded that polypropylene fibres in the broadest studied range of bed permeabilities and fluid velocities, effectively separate oil that is highly polar. On the contrary, for the other two investigated oils at low values of bed permeability a region was detected in which the coalescer is incapable to operate. It has to be emphasized that the polypropylene fibres efficiently separate all three investigated oils at the highest studied bed permeability. [Projekat Ministarstva nauke Republike Srbije, br. 172022