Profiling essential genes in human mammary cells by multiplex RNAi screening

By virtue of their accumulated genetic alterations, tumor cells may acquire vulnerabilities that create opportunities for therapeutic intervention. We have devised a massively parallel strategy for screening short hairpin RNA (shRNA) collections for stable loss-of-function phenotypes. We assayed from 6000 to 20,000 shRNAs simultaneously to identify genes important for the proliferation and survival of five cell lines derived from human mammary tissue. Lethal shRNAs common to these cell lines targeted many known cell-cycle regulatory networks. Cell line-specific sensitivities to suppression of protein complexes and biological pathways also emerged, and these could be validated by RNA interference (RNAi) and pharmacologically. These studies establish a practical platform for genome-scale screening of complex phenotypes in mammalian cells and demonstrate that RNAi can be used to expose genotype-specific sensitivities.     

The BRCT domain is a phospho-protein binding domain

The carboxyl-terminal domain (BRCT) of the Breast Cancer Gene 1 (BRCA1) protein is an evolutionarily conserved module that exists in a large number of proteins from prokaryotes to eukaryotes. Although most BRCT domain-containing proteins participate in DNA-damage checkpoint or DNA-repair pathways, or both, the function of the BRCT domain is not fully understood. We show that the BRCA1 BRCT domain directly interacts with phosphorylated BRCA1-Associated Carboxyl-terminal Helicase (BACH1). This specific interaction between BRCA1 and phosphorylated BACH1 is cell cycle regulated and is required for DNA damage-induced checkpoint control during the transition from G2 to M phase of the cell cycle. Further, we show that two other BRCT domains interact with their respective physiological partners in a phosphorylation-dependent manner. Thirteen additional BRCT domains also preferentially bind phospho-peptides rather than nonphosphorylated control peptides. These data imply that the BRCT domain is a phospho-protein binding domain involved in cell cycle control.     

The involvement of the anterior cingulate cortex in remote contextual fear memory

Although the molecular, cellular, and systems mechanisms required for initial memory processing have been intensively investigated, those underlying permanent memory storage remain elusive. We present neuroanatomical, pharmacological, and genetic results demonstrating that the anterior cingulate cortex plays a critical role in remote memory for contextual fear conditioning. Imaging of activity-dependent genes shows that the anterior cingulate is activated by remote memory and that this activation is impaired by a null alpha-CaMKII mutation that blocks remote memory. Accordingly, reversible inactivation of this structure in normal mice disrupts remote memory without affecting recent memory.     

Antarctic marine flora: uniquely devoid of kelps

The discovery of embryonic stages of the common large Antarctic brown seaweed Himantothallus has led to the conclusion that this plant, hitherto assigned equivocally to the Laminariales (kelps), is a member of the Desmarestiales. Moreover, field study of a large sample of Himantothallus and two other enigmatic brown algae, Phyllogigas and Phaeoglossum, has led to the merger of these three genera with the recognition of a single species, Himantothallus grandifolius. The correct placement of these kelp-like algae underscores the uniqueness of the Antarctic marine flora as the only cold-water flora without kelps.     

Monitoring of soil organic carbon over 10 years in a Mediterranean silvo-pastoral system: potential evaluation for differential management

Soil organic carbon (SOC) plays a vital role in determining the susceptibility to land degradation. The recommended procedure for the recovery of the characteristic poor soils of the Southern region of Portugal is the installation of grazed permanent pastures and increase of soil fertility. The objectives of this study were: (i) to identify the spatial and temporal patterns of soil nutrients at four points in time over a 10-year period in a perennial pastureland; (ii) to test new tools for survey of the spatial variability of soil nutrients; (iii) to evaluate the potential for differential organic management. A 6 ha permanent bio-diverse pasture field, grazed by sheep and improved by annual application of super phosphate fertilizer, was installed on a shallow soil in Mediterranean conditions. Spatial variability and temporal stability of topsoil macronutrients (phosphorus, nitrogen and potassium), SOC and pH were measured. The results indicate that SOC and pH have great potential for implementing differential management. In the case of SOC, the management classes map shows that over 80 % of the area has temporal stability, while more than 50 % of the area has low levels of SOC (<10 g kg^?1), justifying the potential for differential application of C-rich organic soil amendments. The geospatial measurements of apparent soil electrical conductivity (EC_a) and NDVI index showed significant correlation between these parameters and soil properties, revealing the potential of these tools for producing detailed soil maps, decisive for understanding the changes in soil properties under sustainable management systems.     

Effects of permanent magnetic fields on the proliferation of Phalaenopsis protocorm-like bodies using liquid medium

Magnetic fields (MFs) have been applied for the first time in orchid micropropagation. Protocorm-like bodies (PLBs) – approximately 3 mm in diameter – first derived from leaf segment culture of Phalaenopsis Gallant Beau ‘George Vazquez’, were subcultured every 2 months, and served as initial explants. The proliferation of Phalaenopsis PLBs in liquid medium in the Miracle Pack^® culture system was affected by the action of different intensities and polarities of MFs: 0.1, 0.15 and 0.2 Tesla (T) at North (N) and South (S) poles. The MF of 0.1 T – S resulted in the greatest fresh weight of regenerated PLBs. The average number of neo-PLBs formed per clusters in the PLB treated by MF: 0.1–0.2 T was decreased compared to the control exposed to natural MF (5 × 10⁻⁶ T). The proliferation of PLBs under 0.15 T – MF at both N and S poles for 2 and 7 weeks demonstrated that a longer duration of exposure to an MF of 0.15 T, regardless of the polarity, resulted in greater biomass of newly formed PLBs and smaller average number of newly formed PLBs. The S pole of MF had stronger effects on Phalaenopsis PLBs proliferation than the N pole did in all treatments.