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

Prokaryotic abundance and heterotrophic metabolism in the deep Mediterranean Sea

A synthesis of field data carried out in the Mediterranean Sea are presented, aimed at contributing to the knowledge of three prokaryotic-mediated processes and their implications on the Carbon cycle. The distribution of exoenzymatic activities, secondary production and respiration rates was studied together with the prokaryotic abundances. Particular attention was paid to the meso- and bathypelagic layers which play an important role in the Mediterranean carbon cycle. The study is noteworthy because of its large spatial scale spanning the entire Mediterranean Sea over 4 years. In addition, two Atlantic stations in front of the Gibraltar Strait were investigated. The longitudinal distribution of prokaryotic activities and abundance along the MED showed different trends along the depthlayers. In particular, higher exoenzymatic rates were detected in the Eastern basin compared to the Western one; carbon respiration rate showed patterns variable with the sampling periods in the epipelagic and bathypelagic layers, while a consistent Westwards decreasing trend at the mesopelagic layers occurred. Specific enzyme activities per cell showed high values in the deepest layers for leucine aminopeptidase. Comparison with Carbon respiration rate data collected before the 2000s showed changing patterns of microbial heterotrophic processes in the Mediterranean Sea

Complete Acid Ceramidase ablation prevents cancer-initiating cell formation in melanoma cells

Acid ceramidase (AC) is a lysosomal cysteine hydrolase that catalyzes the conversion of ceramide into fatty acid and sphingosine. This reaction lowers intracellular ceramide levels and concomitantly generates sphingosine used for sphingosine-1-phosphate (S1P) production. Since increases in ceramide and consequent decreases of S1P reduce proliferation of various cancers, AC might offer a new target for anti-tumor therapy. Here we used CrispR-Cas9-mediated gene editing to delete the gene encoding for AC, ASAH1, in human A375 melanoma cells. ASAH1-null clones show significantly greater accumulation of long-chain saturated ceramides that are substrate for AC. As seen with administration of exogenous ceramide, AC ablation blocks cell cycle progression and accelerates senescence. Importantly, ASAH1-null cells also lose the ability to form cancer-initiating cells and to undergo self-renewal, which is suggestive of a key role for AC in maintaining malignancy and self-renewal of invasive melanoma cells. The results suggest that AC inhibitors might find therapeutic use as adjuvant therapy for advanced melanoma

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

Investigating molecular alterations to profile short- and long-term recurrence-free survival in patients with primary glioblastoma

Glioblastoma (GB) is the most aggressive type of primary brain tumor. Despite the progress in recent years regarding the diagnosis and treatment of GB, the recurrence rate remains high, due to the infiltrative and dispersive nature of the tumor, which typically results in poor patient prognosis. In the present study, 19 formalin-fixed, paraffin-embedded GB samples were selected from patients with GB tumors. The samples were classified into a short or long recurrence-free survival (RFS) group, based on the time of first recurrence of the disease in the patients. The 19 samples were molecularly characterized for mutations in the isocitrate dehydrogenase 1 (IDH1) gene, amplification of the epidermal growth factor receptor (EGFR) gene, presence of the EGFR variant III, and methylation of the promoter region of the O6-methylguanine-DNA methyltransferase (MGMT) gene. Then, the expression of 84 genes involved in cell-cell and cell-matrix interactions, and that of 84 microRNAs (miRNAs) associated with brain cancer, was profiled. In addition, a copy number variation analysis of 23 genes reported to undergo frequent genomic alterations in human glioma was also performed. Differences in the expression levels of a number of genes were detected across the short and long RFS groups. Among these genes, 5 in particular were selected, and a 5-genes combination approach was developed, which was able to differentiate between patients with short and long RFS outcome. The high levels of sensitivity and precision displayed by this 5-genes combination approach, which were confirmed with a cross-validation method, provide a strong foundation for further validation of the involvement of the aforementioned genes in GB in a larger patient population. In conclusion, the present study has demonstrated how the expression pattern of miRNAs and mRNAs in patients with GB defines a particular molecular hallmark that may increase or reduce the aggressive behavior of GB tumors, thus influencing the survival rates of patients with GB, their response to therapy and their tendency to suffer a relapse

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