A foraminiferal d18O record of sediment core GT90-3 covering the last 2,200 years

For many years the Torino Cosmogeophysics group has been studying sediment cores drilled from the Gallipoli Terrace in the Gulf of Taranto (Ionian Sea) and deposited in the last millennia. The gravity core GT90-3, in which the 18O series was measured, was drilled from the Gallipoli Terrace in the Gulf of Taranto (Ionian Sea) at 39°45'53''N, 17°53'33''E. It was extracted at a depth of 178 m and its length is 3.57 m. Thanks to its geographical location, the Gallipoli Terrace is a favourable site for climatic studies based on marine sediments, because of its closeness to the volcanically active Campanian area, a region that is unique in the world for its detailed historical documentation of volcanic eruptions. Tephra layers corresponding to historical eruptions were identified along the cores, thus allowing for accurate dating and determination of the sedimentation rate. The measurements performed in different cores from the same area showed that the sedimentation rate is uniform across the whole Gallipoli Terrace. We measured the oxygen isotope composition d18O of planktonic foraminifera. These measurements provided a high-resolution 2,200-year-long record. We sampled the core using a spacing of 2.5 mm corresponding to 3.87 years. Each sample of sediment (5 g) was soaked in 5% calgon solution overnight, then treated in 10% H2O2 to remove any residual organic material. Subsequently it was washed with a distilled-water jet through a sieve with a 150 µm mesh. The fraction > 150 µm was kept and oven-dried at 5°C. The planktonic foraminifera Globigerinoides ruber were picked out of the samples under a microscope. For each sample, 20-30 specimens were selected from the fraction comprised between 150 µm and 300 µm. The use of a relatively large number of specimens for each sample reduces the isotopic variability of individual organisms, giving a more representative d18O value. The stable isotope measurements were performed using a VG-PRISM mass spectrometer fitted with an automated ISO-CARB preparation device. Analytical precision based on internal standards was better than 0.1 per mil. Calibration of the mass spectrometer to VPDB scale was done using NBS19 and NBS18 carbonate standards. The strategic location of the drilling area makes this record a unique tool for climate and oceanographic studies of the Central Mediterranean.

Federalização dos crimes graves contra os direitos humanos : Estudo sobre a ponderação de princípios no controle abstrato de constitucionalidade / Ana Fabíola de Azevedo Ferreira. --

Inclui as Adins nºs 3.486 e 3.493.

Epistatic and independent functions of Caspase-3 and Bcl-XL in developmental programmed cell death

The number of neurons in the mammalian brain is determined by a balance between cell proliferation and programmed cell death. Recent studies indicated that Bcl-XL prevents, whereas Caspase-3 mediates, cell death in the developing nervous system, but whether Bcl-XL directly blocks the apoptotic function of Caspase-3 in vivo is not known. To examine this question, we generated bcl-x/caspase-3 double mutants and found that caspase-3 deficiency abrogated the increased apoptosis of postmitotic neurons but not the increased hematopoietic cell death and embryonic lethality caused by the bcl-x mutation. In contrast, caspase-3, but not bcl-x, deficiency changed the normal incidence of neuronal progenitor cell apoptosis, consistent with the lack of expression of Bcl-XL in the proliferative population of the embryonic cortex. Thus, although Caspase-3 is epistatically downstream to Bcl-XL in postmitotic neurons, it independently regulates apoptosis of neuronal founder cells. Taken together, these results establish a role of programmed cell death in regulating the size of progenitor population in the central nervous system, a function that is distinct from the classic role of cell death in matching postmitotic neuronal population with postsynaptic targets.

Stepwise in vitro affinity maturation of Vitaxin, an αvβ3-specific humanized mAb

A protein engineering strategy based on efficient and focused mutagenesis implemented by codon-based mutagenesis was developed. Vitaxin, a humanized version of the antiangiogenic antibody LM609 directed against a conformational epitope of the αvβ3 integrin complex, was used as a model system. Specifically, focused mutagenesis was used in a stepwise fashion to rapidly improve the affinity of the antigen binding fragment by greater than 90-fold. In the complete absence of structural information about the Vitaxin-αvβ3 interaction, phage-expressed antibody libraries for all six Ig heavy and light chain complementarity-determining regions were expressed and screened by a quantitative assay to identify variants with improved binding to αvβ3. The Vitaxin variants in these libraries each contained a single mutation, and all 20 amino acids were introduced at each complementarity-determining region residue, resulting in the expression of 2,336 unique clones. Multiple clones displaying 2- to 13-fold improved affinity were identified. Subsequent expression and screening of a library of 256 combinatorial variants of the optimal mutations identified from the primary libraries resulted in the identification of multiple clones displaying greater than 50-fold enhanced affinity. These variants inhibited ligand binding to receptor more potently as demonstrated by inhibition of cell adhesion and ligand competition assays. Because of the limited mutagenesis and combinatorial approach, Vitaxin variants with enhanced affinity were identified rapidly and required the synthesis of only 2,592 unique variants. The use of such small focused libraries obviates the need for phage affinity selection approaches typically used, permitting the use of functional assays and the engineering of proteins expressed in mammalian cell culture.

The 3′ untranslated region of a rice α-amylase gene functions as a sugar-dependent mRNA stability determinant

In plants, sugar feedback regulation provides a mechanism for control of carbohydrate allocation and utilization among tissues and organs. The sugar repression of α-amylase gene expression in rice provides an ideal model for studying the mechanism of sugar feedback regulation. We have shown previously that sugar repression of α-amylase gene expression in rice suspension cells involves control of both transcription rate and mRNA stability. The α-amylase mRNA is significantly more stable in sucrose-starved cells than in sucrose-provided cells. To elucidate the mechanism of sugar-dependent mRNA turnover, we have examined the effect of αAmy3 3′ untranslated region (UTR) on mRNA stability by functional analyses in transformed rice suspension cells. We found that the entire αAmy3 3′ UTR and two of its subdomains can independently mediate sugar-dependent repression of reporter mRNA accumulation. Analysis of reporter mRNA half-lives demonstrated that the entire αAmy3 3′ UTR and the two subdomains each functioned as a sugar-dependent destabilizing determinant in the turnover of mRNA. Nuclear run-on transcription analysis further confirmed that the αAmy3 3′ UTR and the two subdomains did not affect the transcription rate of promoter. The identification of sequence elements in the α-amylase mRNA that dictate the differential stability has very important implications for the study of sugar-dependent mRNA decay mechanisms.