First insights into the microbiology of three antarctic briny systems of the northern Victoria land

Different polar environments (lakes and glaciers), also in Antarctica, encapsulate brine pools characterized by a unique combination of extreme conditions, mainly in terms of high salinity and low temperature. Since 2014, we have been focusing our attention on the microbiology of brine pockets from three lakes in the Northern Victoria Land (NVL), lying in the Tarn Flat (TF) and Boulder Clay (BC) areas. The microbial communities have been analyzed for community structure by next generation sequencing, extracellular enzyme activities, metabolic potentials, and microbial abundances. In this study, we aim at reconsidering all available data to analyze the influence exerted by environmental parameters on the community composition and activities. Additionally, the prediction of metabolic functions was attempted by the phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt2) tool, highlighting that prokaryotic communities were presumably involved in methane metabolism, aromatic compound biodegradation, and organic compound (proteins, polysaccharides, and phosphates) decomposition. The analyzed cryoenvironments were different in terms of prokaryotic diversity, abundance, and retrieved metabolic pathways. By the analysis of DNA sequences, common operational taxonomic units ranged from 2.2% to 22.0%. The bacterial community was dominated by Bacteroidetes. In both BC and TF brines, sequences of the most thermally tolerant and methanogenic Archaea were detected, some of them related to hyperthermophiles

La psichiatria di consultazione e collegamento nell’ospedale generale: l’esperienza perugina

Objective - This study describes the Consultation-Liaison Service of the Perugia University and investigates the significant associations between a many variables of the assessed population. Results - During the time from July 2008 to June 2009, 722 consultations were performed at the general hospital in Perugia. First examinations were 605. Most consultations involved European patients (95,2%) of female gender (56.3%); mean age was 55.77 (SD ± 21.27). Emergencies were 22.5%; one fifth of patients were not informed of having been referred to our service and half of interventions were requested by departments of internal medicine. The primary reasons for the referral were depression (18.6%), unexplained physical symptoms (12.3%) and anxiety (10.4%); most patients were already taking psychotropic medication before our intervention (58.8%).The significant associations are the following: associations between gender and social status (p < 0.01), social condition (p < 0.01), work (p < 0.01) and advice about the need of the consultation (p < 0.05). The area (medical, surgical and specialized area) are related with the advice (p < 0.05), the reason (p < 0.01) and the type of the consultation (p < 0.01), the diagnostic explanations (p < 0.01), the liaison investigations (p < 0.01) and, at last, with the longrange plan after discharge (p < 0.01)

Modelling approach to the assessment of biogenic fluxes at a selected Ross Sea site, Antarctica

Several biogeochemical data have been collected in the last 10 years of Italian activity in Antarctica (ABIOCLEAR, ROSSMIZE, BIOSESO-I/II). A comprehensive 1-D biogeochemical model was implemented as a tool to link observations with processes and to investigate the mechanisms that regulate the flux of biogenic material through the water column. The model is ideally located at station B (175° E–74° S) and was set up to reproduce the seasonal cycle of phytoplankton and organic matter fluxes as forced by the dominant water column physics over the period 1990–2001. Austral spring-summer bloom conditions are assessed by comparing simulated nutrient drawdown, primary production rates, bacterial respiration and biomass with the available observations. The simulated biogenic fluxes of carbon, nitrogen and silica have been compared with the fluxes derived from sediment traps data. The model reproduces the observed magnitude of the biogenic fluxes, especially those found in the bottom sediment trap, but the peaks are markedly delayed in time. Sensitivity experiments have shown that the characterization of detritus, the choice of the sinking velocity and the degradation rates are crucial for the timing and magnitude of the vertical fluxes. An increase of velocity leads to a shift towards observation but also to an overestimation of the deposition flux which can be counteracted by higher bacterial remineralization rates. Model results suggest that the timing of the observed fluxes depends first and foremost on the timing of surface production and on a combination of size-distribution and quality of the autochtonous biogenic material. It is hypothesized that the bottom sediment trap collects material originated from the rapid sinking of freshly-produced particles and also from the previous year's production period

Modelling approach to the assessment of biogenic fluxes at a selected Ross Sea site, Antarctica

Abstract Several biogeochemical data have been collected in the last 10 years of Italian activity in Antarctica (ABIOCLEAR, ROSSMIZE, BIOSESO-I/II). A comprehensive 1-D biogeochemical model was implemented as a tool to link observations with processes and to investigate the mechanisms that regulate the flux of biogenic material through the water column. The model is ideally located at station B (175^{o}E - 74^{o}S) and was set up to reproduce the seasonal cycle of phytoplankton and organic matter fluxes as forced by the dominant water column physics over the period 1990-2001. Austral spring-summer bloom conditions are assessed by comparing simulated nutrient drawdown, primary production rates, bacterial respiration and biomass with the available observations. The simulated biogenic fluxes of carbon, nitrogen and silica have been compared with the fluxes derived from sediment traps data. The model reproduces quite well the magnitude of the biogenic fluxes, expecially those observed in the bottom sediment trap, but the peaks are delayed in time. Sensitivity experiments have shown that the characterization of detritus, the choice of the sinking velocity and the degradation rates are crucial for the timing and magnitude of the vertical fluxes. An increase of velocity leads to a shift towards observation but also to an overestimation of the deposition flux which can be counteracted by higher bacterial remineralization rates. Model results suggest that observed fluxes could be explained by the size-distribution and quality of the locally-produced biogenic material. It is hypothesized that the bottom sediment trap collects material originated from rapid sinking of particles and also from previous years production periods, likely modulated by advective and aggregation mechanisms which are still not resolved by the model

An Efficient Molecular Dynamics Scheme for the Calculation of Dopant Profiles due to Ion Implantation

We present a highly efficient molecular dynamics scheme for calculating the concentration depth profile of dopants in ion irradiated materials. The scheme incorporates several methods for reducing the computational overhead, plus a rare event algorithm that allows statistically reliable results to be obtained over a range of several orders of magnitude in the dopant concentration. We give examples of using this scheme for calculating concentration profiles of dopants in crystalline silicon. Here we can predict the experimental profile over five orders of magnitude for both channeling and non-channeling implants at energies up to 100s of keV. The scheme has advantages over binary collision approximation (BCA) simulations, in that it does not rely on a large set of empirically fitted parameters. Although our scheme has a greater computational overhead than the BCA, it is far superior in the low ion energy regime, where the BCA scheme becomes invalid.Comment: 17 pages, 21 figures, 2 tables. See: http://bifrost.lanl.gov/~reed

Microbial assemblages in pressurized antarctic brine pockets (Tarn flat, northern Victoria land): A hotspot of biodiversity and activity

Two distinct pressurized hypersaline brine pockets (named TF4 and TF5), separated by a thin ice layer, were detected below an ice-sealed Antarctic lake. Prokaryotic (bacterial and archaeal) diversity, abundances (including virus-like particles) and metabolic profiles were investigated by an integrated approach, including traditional and new-generation methods. Although similar diversity indices were computed for both Bacteria and Archaea, distinct bacterial and archaeal assemblages were observed. Bacteroidetes and Gammaproteobacteria were more abundant in the shallowest brine pocket, TF4, and Deltaproteobacteria, mainly represented by versatile sulphate-reducing bacteria, dominated in the deepest, TF5. The detection of sulphate-reducing bacteria and methanogenic Archaea likely reflects the presence of a distinct synthrophic consortium in TF5. Surprisingly, members assigned to hyperthermophilic Crenarchaeota and Euryarchaeota were common to both brines, indicating that these cold habitats host the most thermally tolerant Archaea. The patterns of microbial communities were different, coherently with the observed microbiological diversity between TF4 and TF5 brines. Both the influence exerted by upward movement of saline brines from a sub-surface anoxic system and the possible occurrence of an ancient ice remnant from the Ross Ice Shelf were the likely main factors shaping the microbial communities

po 259 inhibition of the hexosamine biosynthetic pathway by targeting pgm3 causes breast cancer growth arrest and apoptosis

Introduction Cancer aberrant N - and O -linked protein glycosylation, frequently resulting from an augmented flux through the Hexosamine Biosynthetic Pathway (HBP), play different roles in tumour progression. Recent studies reported an association between the tumorigenic potential, metastasis and chemoresistance of several type of breast cancer cells and tumours, among which the Triple Negative Breast Cancer (TNBC), and the alteration of their membrane glycans composition and ramification as well as of their level of protein O -Glc N Ac. However, the low specificity and toxicity of the existing HBP inhibitors prevented their use for cancer treatment. Material and methods In order to identify a novel inhibitor of HBP pathway and in particular of the PGM3 enzyme, we performed a virtual screening by using computational approaches. These approaches lead us to the identification of a lead compound. This compound, named FR054, has been synthetized and in vitro and in vivo tested by using several biophysical methods (NMR, LC/MS, HPLC) and biochemical assay (CETSA, ITDRF, FACS analysis) as well as tested in TNBC xenograft mice model. Results and discussions Here we report the preclinical evaluation of FR054, a novel inhibitor of the HBP enzyme PGM3, with a remarkable anti-breast cancer effect. In fact, FR054 induces in different breast cancer cells a dramatic decrease in cell proliferation and survival. In particular, in a model of Triple Negative Breast Cancer (TNBC) cells, MDA-MB-231, we show that these effects are correlated to FR054-dependent reduction of both N - and O -glycosylation level that cause also to a strong reduction of cancer cell adhesion and migration. Moreover we show that impaired survival of cancer cells upon FR054 treatment is associated with activation of the Unfolded Protein Response (UPR) and accumulation of intracellular ROS. Finally, we show that FR054 suppresses cancer growth in MDA-MB-231 xenograft mice. Conclusion Our data support the advantage of targeting HBP for therapeutic purpose and encourage further investigation about the use of this small-molecule as promising compound for breast cancer therapy

Molecular analysis of TP53, Ki-Ras and P16 methylation status in tissue and plasma of subjects affected by gastrointestinal cancer (GIC)

BACKGROUND: Despite the improvement in detection and surgical therapy in the last years, the outcome of patients affected by colorectal carcinoma (CRC) remains limited by metastatic relapse. The aim of this study was to investigate the presence of free tumor DNA in the plasma of CRC patients in order to understand its possible prognostic role. PATIENTS AND METHODS: Ki-Ras, TP53 mutations and p16(INK4A) methylation status were prospectively evaluated in tumor tissues and plasma of 66 CRC patients. RESULTS: In 50 of the 66 primitive tumor cases (76%) at least one significant alteration was identified in Ki-Ras and/or TP53 and/or p16(INK4A) genes. Eighteen of the 50 patients presented the same alteration both in the plasma and in the tumor tissue. At univariate analysis, Ki-Ras mutations proved to be significantly related to quicker relapse (P <0.01), whereas only a trend towards statistical significance (P = 0.083) was observed for the TP53 mutations CONCLUSIONS: Detection of Ki-Ras and TP53 mutation in plasma should be significantly related to disease recurrence. These data suggest that patients with a high risk of recurrence can be identified by means of the analysis of tumor-derived plasma DNA with the use of fairly non-invasive techniques

Interaction of Chandipura Virus N and P Proteins: Identification of Two Mutually Exclusive Domains of N Involved in Interaction with P

The nucleocapsid protein (N) and the phosphoprotein (P) of nonsegmented negative-strand (NNS) RNA viruses interact with each other to accomplish two crucial events necessary for the viral replication cycle. First, the P protein binds to the aggregation prone nascent N molecules maintaining them in a soluble monomeric (N0) form (N0-P complex). It is this form that is competent for specific encapsidation of the viral genome. Second, the P protein binds to oligomeric N in the nucleoprotein complex (N-RNA-P complex), and thereby facilitates the recruitment of the viral polymerase (L) onto its template. All previous attempts to study these complexes relied on co-expression of the two proteins in diverse systems. In this study, we have characterised these different modes of N-P interaction in detail and for the first time have been able to reconstitute these complexes individually in vitro in the chandipura virus (CHPV), a human pathogenic NNS RNA virus. Using a battery of truncated mutants of the N protein, we have been able to identify two mutually exclusive domains of N involved in differential interaction with the P protein. An unique N-terminal binding site, comprising of amino acids (aa) 1–180 form the N0-P interacting region, whereas, C-terminal residues spanning aa 320–390 is instrumental in N-RNA-P interactions. Significantly, the ex-vivo data also supports these observations. Based on these results, we suggest that the P protein acts as N-specific chaperone and thereby partially masking the N-N self-association region, which leads to the specific recognition of viral genome RNA by N0

Inhibition of nuclear factor kappa-B signaling reduces growth in medulloblastoma in vivo

Abstract Background Medulloblastoma is a highly malignant pediatric brain tumor that requires surgery, whole brain and spine irradiation, and intense chemotherapy for treatment. A more sophisticated understanding of the pathophysiology of medulloblastoma is needed to successfully reduce the intensity of treatment and improve outcomes. Nuclear factor kappa-B (NFκB) is a signaling pathway that controls transcriptional activation of genes important for tight regulation of many cellular processes and is aberrantly expressed in many types of cancer. Methods To test the importance of NFκB to medulloblastoma cell growth, the effects of multiple drugs that inhibit NFκB, pyrrolidine dithiocarbamate, diethyldithiocarbamate, sulfasalazine, curcumin and bortezomib, were studied in medulloblastoma cell lines compared to a malignant glioma cell line and normal neurons. Expression of endogenous NFκB was investigated in cultured cells, xenograft flank tumors, and primary human tumor samples. A dominant negative construct for the endogenous inhibitor of NFκB, IκB, was prepared from medulloblastoma cell lines and flank tumors were established to allow specific pathway inhibition. Results We report high constitutive activity of the canonical NFκB pathway, as seen by Western analysis of the NFκB subunit p65, in medulloblastoma tumors compared to normal brain. The p65 subunit of NFκB is extremely highly expressed in xenograft tumors from human medulloblastoma cell lines; though, conversely, the same cells in culture have minimal expression without specific stimulation. We demonstrate that pharmacological inhibition of NFκB in cell lines halts proliferation and leads to apoptosis. We show by immunohistochemical stain that phosphorylated p65 is found in the majority of primary tumor cells examined. Finally, expression of a dominant negative form of the endogenous inhibitor of NFκB, dnIκB, resulted in poor xenograft tumor growth, with average tumor volumes 40% smaller than controls. Conclusions These data collectively demonstrate that NFκB signaling is important for medulloblastoma tumor growth, and that inhibition can reduce tumor size and viability in vivo. We discuss the implications of NFκB signaling on the approach to managing patients with medulloblastoma in order to improve clinical outcomes.</p