Different domains cooperate to target the human ribosomal L7a protein to the nucleus and to the nucleoli.

The human ribosomal protein L7a is a component of the major ribosomal subunit. We transiently expressed in HeLa cells L7a-β-galactosidase fusion proteins and studied their subcellular localization by indirect immunofluorescence staining with anti-β-galactosidase antibodies. We have identified three distinct domains responsible for the nuclear targeting of the protein: domain I, amino acids 23-51; domain II, amino acids 52-100; domain III, amino acids 101-220, each of which contains at least one nuclear localization signal (NLS). Through subcellular localization analysis of deletion mutants of L7a-β-galactosidase chimeras, we demonstrate that domain II plays a special role because it is necessary, although not sufficient, to target the chimeric β-galactosidase to the nucleoli. In fact, we demonstrate that the nucleolar targeting process requires the presence of domain II plus an additional basic domain that can be represented by an NLS or a basic stretch of amino acids without NLS activity. Thus, when multiple NLS are present, each NLS exerts distinct functions. Domain II drives nucleolar accumulation of a reporter protein with the cooperative action of a short basic amino acid sequence, suggesting a mechanism requiring protein-protein or protein-nucleic acid interactions

Feasibility analysis for floating offshore wind energy

Purpose The assessment of the economic feasibility of foating ofshore wind farms (FOWFs) plays an important role in the future possible spreading of this challenging technology in the wind power industry. The use of specifc economic analyses is fundamental to point out the potential of FOWFs and to sustain their technical value. Within this topic, the implementation of the FOWF life cycle cost model and producibility analysis in a geographic information system is developed, with the aim of carrying out a feasibility analysis at the territorial scale, for diferent types of foater. Moreover, a simplifed model for a quick life cycle cost assessment is proposed and calibrated. Methods The available cost model is frst validated comparing the costs of FOWFs based on diferent foaters (Semi-Submersible Platform—SSP, Spar Buoy—SB and Tension Leg Platform—TLP) with corresponding results available in the literature. Then, it is implemented in QGIS to be used for territorial-scale analyses and sensitivity analyses of the cost parameters. A feasibility analysis is developed through the main fnancial parameters. Finally, the results are then used to calibrate a simplifed version of the cost model that depends on three main parameters, namely distance to shore, distance from the port of operation and bathymetry. Results and discussion The FOWF cost values are found to be in good agreement with those coming from analytical methods similar to the one from the authors. However, some discrepancies with those based on average costs are observed. Then, the results of the sensitivity analysis are presented as life cycle cost maps, giving an overall picture of the variation of the total cost of FOWF installations on a reference domain. The results show that among the three types of foaters considered here, the SSP proved to be the most promising one, giving lower costs than the SB and the TLP. Moreover, a good agreement between the results in terms of total cost of FOWFs calculated with the analytical and simplifed models for SSPs, SBs and TLPs is observed. Finally, the feasibility analysis showed that the fnancial parameters are more infuenced by the wind speed than by the cost of the farm. Conclusions The paper aims to provide guidance on how to carry out feasibility analyses of a specifc site for FOWF installation, thus supporting decision-making procedures. The approach and the results presented here are meant for use in the early stage of the decision-making process, as a tool for the assessment of the economic feasibility of FOWFs installation

Sensitivity analysis of cost parameters for floating offshore wind farms: An application to Italian waters

Floating offshore wind farms represent the next frontier in wind power industry. However, the development of this technology is strongly dependent on its economic feasibility. There follows that the development of economic analyses is crucial to highlight the possible greater potential of floating offshore wind farms and to support their sustainability and technical value. In this context, the purpose of this paper is to present a sensitivity analysis of the main cost parameters for floating offshore wind farms, namely the distance from the coast, the distance from the closest port and the sea depth. It can give specific information on which parameters are more important, and how much they affect the total cost. To this aim, a comprehensive life cycle cost assessment of floating offshore wind farms has been developed. In this study the cost model has been applied to the Italian waters. The results shown should provide guidance on how to preliminary assess the quality of a given site for floating offshore wind farm installation, and should be helpful for future development of decision-making procedures in the offshore wind sector

Annexin A2 is regulated by ovarian cancer-peritoneal cell interactions and promotes metastasis

Our recent research identified the protein annexin A2 to be regulated by ovarian cancer-peritoneal cell interactions. This study investigated the role of annexin A2 in ovarian cancer metastasis and its potential utility as a novel therapeutic target, using in vitro and in vivo ovarian cancer models. Annexin A2 expression was examined by qRT-PCR and western blotting in ovarian cancer cell lines and immunohistochemistry in serous ovarian carcinoma tissues. Annexin A2 siRNAs were used to evaluate the effects of annexin A2 suppression on ovarian cancer cell adhesion, motility, and invasion. Furthermore, annexin A2 neutralizing antibodies were used to examine the role of annexin A2 in tumor invasion and metastasis in vivo using a chick chorioallantoic membrane assay and an intraperitoneal xenograft mouse model. Strong annexin A2 immunostaining was observed in 90% (38/42) of the serous ovarian cancer cells and was significantly increased in the cancer-associated stroma compared to non-malignant ovarian tissues. Annexin A2 siRNA significantly inhibited the motility and invasion of serous ovarian cancer cells and adhesion to the peritoneal cells. Annexin A2 neutralizing antibodies significantly inhibited OV-90 cell motility and invasion in vitro and in vivo using the chick chorioallantoic membrane assay. The growth of SKOV-3 cells and their peritoneal dissemination in nude mice was significantly inhibited by annexin A2 neutralizing antibodies. Annexin A2 plays a critical role in ovarian cancer metastasis and is therefore a potential novel therapeutic target against ovarian cancer.Noor A. Lokman, Alison SF. Elder, Miranda P Ween, Carmen E. Pyragius, Peter Hoffmann, Martin K. Oehler, and Carmela Ricciardell

Keratin 5 overexpression is associated with serous ovarian cancer recurrence and chemotherapy resistance

This study investigated the clinical significance of keratin 5 and 6 expression in serous ovarian cancer progression and chemotherapy resistance. KRT5 and KRT6 (KRT6A, KRT6B & KRT6C) gene expression was assessed in publically available serous ovarian cancer data sets, ovarian cancer cell lines and primary serous ovarian cancer cells. Monoclonal antibodies which detect both K5/6 or only K5 were used to assess protein expression in ovarian cancer cell lines and a cohort of high grade serous ovarian carcinomas at surgery (n = 117) and after neoadjuvant chemotherapy (n = 21). Survival analyses showed that high KRT5 mRNA in stage III/IV serous ovarian cancers was significantly associated with reduced progression-free (HR 1.38, P < 0.0001) and overall survival (HR 1.28, P = 0.013) whilst high KRT6 mRNA was only associated with reduced progression-free survival (HR 1.2, P = 0.031). Both high K5/6 (≥ 10%, HR 1.78 95% CI; 1.03−2.65, P = 0.017) and high K5 (≥ 10%, HR 1.90, 95% CI; 1.12−3.19, P = 0.017) were associated with an increased risk of disease recurrence. KRT5 but not KRT6C mRNA expression was increased in chemotherapy resistant primary serous ovarian cancer cells compared to chemotherapy sensitive cells. The proportion of serous ovarian carcinomas with high K5/6 or high K5 immunostaining was significantly increased following neoadjuvant chemotherapy. K5 can be used to predict serous ovarian cancer prognosis and identify cancer cells that are resistant to chemotherapy. Developing strategies to target K5 may therefore improve serous ovarian cancer survival.Carmela Ricciardelli, Noor A Lokman, Carmen E Pyragius, Miranda P Ween, Anne M Macpherson, Andrew Ruszkiewicz, Peter Hoffmann, Martin K Oehle

Transfer of gene corrected T cells corrects humoral and cytotoxic defects in X-linked lymphoproliferative disease (XLP1)

BACKGROUND: XLP1 arises from mutations in the SH2D1A gene encoding SAP, an adaptor protein expressed in T, NK and NKT cells. Defects lead to abnormalities of T and NK cell cytotoxicity and T cell dependent humoral function. Clinical manifestations include haemophagocytic lymphohistiocytosis (HLH), lymphoma and dysgammaglobulinaemia. Curative treatment is limited to haematopoietic stem cell transplant with outcome reliant on a good donor match. OBJECTIVES: As most symptoms arise from defective T cell function, we investigated whether transfer of SAP gene corrected T cells could reconstitute known effector cell defects. METHODS: CD3+ lymphocytes from sap-deficient mice were transduced with a gammaretroviral vector encoding human SAP cDNA before transfer into sub-lethally irradiated sap-deficient recipients. Following immunisation with the T-dependent antigen NP-CGG, recovery of humoral function was evaluated through germinal centre formation and antigen specific responses. To efficiently transduce patient CD3+ cells, we generated an equivalent lentiviral SAP vector. Functional recovery was demonstrated using in vitro cytotoxicity and TFH cell function assays, alongside tumour clearance in an in vivo LCL lymphoma xenograft model. RESULTS: In sap-deficient mice, 20-40% engraftment of gene modified T cells led to significant recovery of germinal centre formation and NP-specific antibody responses. Gene corrected patient T cells demonstrated improved cytotoxicity and TFH cell function in vitro. Adoptive transfer of gene corrected patient CTLs reduced tumour burden to a level comparable with healthy donor CTLs in an in vivo lymphoma model. CONCLUSIONS: These data demonstrate that autologous T cell gene therapy corrects SAP dependent defects and may offer an alternative therapeutic option for XLP1 patients

Pentadecanal and pentadecanoic acid coatings reduce biofilm formation of Staphylococcus epidermidis on PDMS

Staphylococcus epidermidis is well known to be one of the major causes of infections related to medical devices, mostly due to its strong capacity to form device-associated biofilms. Nowadays, these infections represent a severe burden to the public health system and the necessity of novel antibacterial strategies for the treatment of these difficult-to-eradicate infections is urgent. The Antarctic marine bacterium Pseudoalteromonas haloplanktis TAC125 was found to be able to produce an anti-biofilm molecule, the pentadecanal, active against S. epidermidis. In this work, we modified one of the most widely used silicone-based polymers, polydimethylsiloxane (PDMS), by adsorption of pentadecanal and its most promising derivative, pentadecanoic acid, on the PDMS surface. The biofilm formation of S. epidermidis RP62A on both untreated and modified PDMS was performed in a parallel plate flow chamber system, demonstrating the capability of the proposed anti-biofilm coatings to strongly reduce the biofilm formation. Furthermore, drug-release capacity and long-term efficacy (21 days) were also proven for the pentadecanoic acid coating

Sustainable water use for rice agro-ecosystems in northern Italy

I n the Mediterranean basin, rice is cultivated over an area of 1,300,000 hectares. The most important rice-producing countries are Italy and Spain in Europe (72% of the EU production; 345,000 ha), and Egypt and Turkey among the extra-EU countries (almost totality of the production; 789,000 ha). Traditionally, rice is grown under continuous flooding; thus, it requires much more irrigation than non-ponded crops. The MEDWATERICE project (PRIMA-Section 2-2018; https://www.medwaterice.org/) aims at exploring sustainability of innovative rice irrigation management solutions, in order to reduce rice water consumption and environmental impacts, and to extend rice cultivation outside of traditional paddy areas to meet the escalating demand. Within the MEDWATERICE project, irrigation management options to address the main site-specific problems are being tested for each rice areas involved in the project (IT, ES, PT, EG, TR). Case studies are being conducted in pilot farms, with the involvement of Stake-Holder Panels (SHPs) in each country. Data collected at the farm level will be extrapolated to the irrigation district level, to support water management decisions and policies. Moreover, indicators for quantitative assessment of environmental, economic and social sustainability of the irrigation options will be defined. This work illustrates the first year of results for the Italian Case Study (Lomellina area, Pavia) at the pilot farm scale. This area is characterized by a growing water scarcity in drought years in many districts. Within the farm managed by the National Rice Research Center (CRR), in the agricultural season 2019 the experimentation was conducted in six plots of about 20 m x 80 m each, with two replicates for each of the following water regimes: i) water-seeded rice with continuous flooding (WFL), ii) dry-seeded rice with continuous flooding from the 3-4 leaf stage (DFL), and iii) water seeded-rice with alternate wetting and drying from fertilization at the tillering stage (AWD). One out of the two replicates of each treatment was instrumented with: water inflow and outflow meters, set of piezometers, set of tensiometers and water tubes for the irrigation management in the AWD plots. A soil survey was conducted before the agricultural season (EMI sensor and physico-chemical analysis of soil samples). Periodic measurements of crop biometric parameters (LAI, crop height, crop rooting depth) were performed. Moreover, nutrients (TN, NO3, PO4, K) and two widely used pesticides (Sirtaki \u2013 a.i. Clomazone; Tripion E \u2013 a.i. MCPA) were measured in irrigation water (inflow and outflow), groundwater, and porous cups installed at two soil depths (20 and 70 cm, above and below the plough pan). Finally, rice grain yields and quality (As and Cd in the grain) were determined. First results in terms of cumulative water balance components (rainfall, irrigation inflow and outflow, difference in soil and ponding water storage, evapotranspiration, net percolation), water application efficiency (evapotranspiration over net water input), and water productivity (grain production over net water input), will be presented and discussed. Results of a 1D Richard-equation-based numerical simulation model applied to generalize results obtained under the different irrigation regimes will be moreover illustrated

A Feedforward Neural Network Approach for the Detection of Optically Thin Cirrus From IASI-NG

The identification of optically thin cirrus is crucial for their accurate parameterization in climate and Earth's system models. This study exploits the characteristics of the infrared atmospheric sounding interferometer-new generation (IASI-NG) to develop an algorithm for the detection of optically thin cirrus. IASI-NG has been designed for the European Organization for the Exploitation of Meteorological Satellites (EUMETSAT) polar system second-generation program to continue the service of its predecessor IASI from 2024 onward. A thin-cirrus detection algorithm (TCDA) is presented here, as developed for IASI-NG, but also in parallel for IASI to evaluate its performance on currently available real observations. TCDA uses a feedforward neural network (NN) approach to detect thin cirrus eventually misidentified as clear sky by a previously applied cloud detection algorithm. TCDA also estimates the uncertainty of "clear-sky" or "thin-cirrus" detection. NN is trained and tested on a dataset of IASI-NG (or IASI) simulations obtained by processing ECMWF 5-generation reanalysis (ERA5) data with the s-IASI radiative transfer model. TCDA validation against an independent simulated dataset provides a quantitative statistical assessment of the improvements brought by IASI-NG with respect to IASI. In fact, IASI-NG TCDA outperforms IASI TCDA by 3% in probability of detection (POD), 1% in bias, and 2% in accuracy, and the false alarm ratio (FAR) passes from 0.02 to 0.01. Moreover, IASI TCDA validation against state-of-the-art cloud products from Cloudsat/CPR and CALIPSO/Cloud-Aerosol Lidar with Orthogonal Polarization (CALIOP) real observations reveals a tendency for IASI TCDA to underestimate the presence of thin cirrus (POD = 0.47) but with a low FAR (0.07), which drops to 0.0 for very thin cirrus

A comprehensive molecular and clinical analysis of the piRNA pathway genes in ovarian cancer

Ovarian cancer (OC) is one of the most lethal gynecological malignancies, yet molecular mechanisms underlying its origin and progression remain poorly understood. With increasing reports of piRNA pathway deregulation in various cancers, we aimed to better understand its role in OC through a comprehensive analysis of key genes: PIWIL1-4, DDX4, HENMT1, MAEL, PLD6, TDRD1,9 and mutants of PIWIL1 (P1∆17) and PIWIL2 (PL2L60). High-throughput qRT-PCR (n = 45) and CSIOVDB (n = 3431) showed differential gene expression when comparing benign ovarian tumors, low grade OC and high grade serous OC (HGSOC). Significant correlation of disparate piRNA pathway gene expression levels with better progression free, post-progression free and overall survival suggests a complex role of this pathway in OC. We discovered PIWIL3 expression in chemosensitive but not chemoresistant primary HGSOC cells, providing a potential target against chemoresistant disease. As a first, we revealed that follicle stimulating hormone increased PIWIL2 expression in OV-90 cells. PIWIL1, P1∆17, PIWIL2, PL2L60 and MAEL overexpression in vitro and in vivo decreased motility and invasion of OVCAR-3 and OV-90 cells. Interestingly, P1∆17 and PL2L60, induced increased motility and invasion compared to PIWIL1 and PIWIL2. Our results in HGSOC highlight the intricate role piRNA pathway genes play in the development of malignant neoplasms.Eunice Lee, Noor A. Lokman, Martin K. Oehler, Carmela Ricciardelli and Frank Grutzne