The Mechanism of Cleavage Under Basic Conditions of Succinyl-Anchored Oligonucleotides

Abstract

Studies with a model compound provide direct evidence that cleavage of succinyl-anchored oligonucleotides takes place by intramolecular nucleophilic attack of the conjugate base of the succinamide at the ester carbonyl group. An N-substituted succinimide is exclusively formed with piperidine, DBU and TBAF, but when ammonia is used-this general mechanism seems to coexist with ammonolysis of the succinate ester. Cleavage with TBAF is extremely fast.     

Concurrent Akt,ERK1/2 and AMPK Activation by Obestatin Inhibits Apoptotic Signaling Cascades on Nutrient-Deprived PC12 Cells

Targeting apoptosis in the ischemic penumbra is a rational therapeutic approach for restricting cerebral infarct volume after clinical stroke. The present work explored the capability of the obestatin peptide, as a novel approach to inhibit apoptotic signaling cascades on PC12 cells. According to the results, obestatin treatment significantly reduced nutrient deprivation-induced apoptotic cell death. The protective effects were related to the regulation of the anti-apoptotic protein, BCL-2, and the apoptotic protein caspase-3. This encompasses the control of apoptosis by the interplay between Akt, ERK1/2 and AMPK signaling pathways. The activation of Akt and AMPK was concomitant with the phosphorylation of their downstream targets, GSK3 and ACC, respectively. Besides, obestatin also causes FoxO1 nuclear export supporting the prevention of the apoptosome formation. The concurrent activation of Akt and AMPK by obestatin via the GPR39 receptor, supports a role for this system in the balance concerning the catabolic and the anabolic signaling to sustain cellular function and viability. Furthermore, these results provide both an insight into how the obestatin/GPR39 system regulates anti-apoptotic pathways, and a framework for ascertaining how this system can be optimally targeted in treatment of brain cell death after stroke.

     

Abundance,distribution, and expression of nematicidal crystal protein genes in Bacillus thuringiensis strains from diverse habitats

International Microbiology - Bacillus thuringiensis (Bt) is a Gram-positive bacterium that accumulates pesticidal proteins (Cry and Cyt) in parasporal crystals. Proteins from the Cry5, App6...     

Cytotoxicity studies of [PtCl2(H2bim)] (H2bim = 2,2′-biimidazole): Study of its interaction with a small protein PCI (potato carboxypeptidase inhibitor)

The effect of the platinum compound [PtCl₂(H₂bim)] (H₂bim = 2,2′-biimidazole) on the plasmid DNA conformation was previously studied by electrophoresis in agarose gel and on calf thymus DNA by circular dichroism spectroscopy. The effect of this compound on pBR322 plasmid DNA has now been visualized by atomic force microscopy, which shows that the complex modifies the DNA in the same way as cisplatin does. The cytotoxic activity of [PtCl₂(H₂bim)] in HeLa-229, HL-60, A2780 and A2780cisR cell lines has also been evaluated. Likewise, the interaction of [PtCl₂(H₂bim)] with the small protein potato carboxypeptidase inhibitor (PCI) and a PCI mutant in which glycine 39 was substituted by methionine has been followed by HPLC/mass spectrometry. The interaction with the mutant protein PCI showed the formation of monofunctional adducts that ultimately gave bifunctional adducts. PCI mutant protein could be a good carrier of this platinum compound to the tumour cells in which the antiproliferative behaviour was demonstrated.     

Survival and respiration of green abalone (Haliotis fulgens) facing very short-term marine environmental extremes

The frequency and strength of extreme events are increasing due to climate change. These events have the potential to cause mass mortalities and recruitment failure in very short time scales. Here, we explored three relevant basic questions using green abalone: how the lethal levels of environmental variables (i.e. temperature, oxygen, and pH) change trough time, what the short-term synergistic effects of stressors are, and what the metabolic responses and recovery capacity are at this timescale. We observed that very short-term events are less likely to cause mass mortalities than events lasting for several days, that the relative importance of hyperthermia, hypoxia, and combined effects change through time, and that the respiration rate increased under hyperthermia and decreased under hypoxia and the combined effects, while hemocyanin concentration increased under hypoxia and decreased under hyperthermia and combined effects. Independently of the stressor, green abalone re-established their respiratory rate after the stress.     

Responses of two Mediterranean seagrasses to experimental changes in salinity

The aim of this study is to examine the effects of variations in salinity levels on growth and survival of two fast-growing Mediterranean seagrasses, Cymodocea nodosa and Zostera noltii. We also tested the capacity of C. nodosa to acclimate to a gradual increase in salinity and to discover how it responds to a sharp rise in salinity in combination with other factors, such as increases in temperature, seasonality and different plant-population origins. Several short-term (10 days) experiments were conducted under controlled conditions. For each experiment, ten marked shoots were placed in 5-l aquaria, where they were exposed to different salinity treatments (ranging from 2 to 72 psu). Growth and survival of both species were significantly affected by salinity. A significant effect between salinity and temperature on the shoot growth rate of C. nodosa was also detected, but not on shoot mortality. When C. nodosa plants were acclimated by gradually increasing the salinity level, it was observed that acclimatisation improved tolerance to salinity changes. A different response to salinity variations, depending on the origin of the plants or the season of the year, was also detected. These results indicated that Z. noltii plants tolerate conditions of hyposalinity better than C. nodosa, and that the tolerance range of C. nodosa may change depending on the temperature, the season or the population.