Detection of Germline Variants in 450 Breast/Ovarian Cancer Families with a Multi-Gene Panel Including Coding and Regulatory Regions

With the progress of sequencing technologies, an ever-increasing number of variants of unknown functional and clinical significance (VUS) have been identified in both coding and non-coding regions of the main Breast Cancer (BC) predisposition genes. The aim of this study is to identify a mutational profile of coding and intron-exon junction regions of 12 moderate penetrance genes (ATM, BRIP1, CDH1, CHEK2, NBN, PALB2, PTEN, RAD50, RAD51C, RAD51D, STK11, TP53) in a cohort of 450 Italian patients with Hereditary Breast/Ovarian Cancer Syndrome, wild type for germline mutation in BRCA1/2 genes. The analysis was extended to 5′UTR and 3′UTR of all the genes listed above and to the BRCA1 and BRCA2 known regulatory regions in a subset of 120 patients. The screening was performed through NGS target resequencing on the Illumina platform MiSeq. 8.7% of the patients analyzed is carriers of class 5/4 coding variants in the ATM (3.6%), BRIP1 (1.6%), CHEK2 (1.8%), PALB2 (0.7%), RAD51C (0.4%), RAD51D (0.4%), and TP53 (0.2%) genes, while variants of uncertain pathological significance (VUSs)/class 3 were identified in 9.1% of the samples. In intron-exon junctions and in regulatory regions, variants were detected respectively in 5.1% and in 32.5% of the cases analyzed. The average age of disease onset of 44.4 in non-coding variant carriers is absolutely similar to the average age of disease onset in coding variant carriers for each proband’s group with the same cancer type. Furthermore, there is not a statistically significant difference in the proportion of cases with a tumor onset under age of 40 between the two groups, but the presence of multiple non-coding variants in the same patient may affect the aggressiveness of the tumor and it is worth underlining that 25% of patients with an aggressive tumor are carriers of a PTEN 3′UTR-variant. This data provides initial information on how important it might be to extend mutational screening to the regulatory regions in clinical practice.     

Placental Complement Activation in Fetal and Neonatal Alloimmune Thrombocytopenia: An Observational Study

Fetal and neonatal alloimmune thrombocytopenia (FNAIT) is a disease that causes thrombocytopenia and a risk of bleeding in the (unborn) child that result from maternal alloantibodies directed against fetal, paternally inherited, human platelet antigens (HPA). It is hypothesized that these alloantibodies can also bind to the placenta, causing placental damage. This study aims to explore signs of antibody-mediated placental damage in FNAIT. We performed a retrospective study that included pregnant women, their newborns, and placentas. It comprised 23 FNAIT cases, of which nine were newly diagnosed (14 samples) and 14 were antenatally treated with intravenous immune globulins (IVIg) (21 samples), and 20 controls, of which 10 had anti-HLA-class I antibodies. Clinical information was collected from medical records. Placental samples were stained for complement activation markers (C1q, C4d, SC5b-9, and mannose-binding lectin) using immunohistochemistry. Histopathology was examined according to the Amsterdam criteria. A higher degree of C4d deposition was present in the newly diagnosed FNAIT cases (10/14 samples), as compared to the IVIg-treated FNAIT cases (2/21 samples, p = 0.002) and anti-HLA-negative controls (3/20 samples, p = 0.006). A histopathological examination showed delayed maturation in four (44%) placentas in the newly diagnosed FNAIT cases, five (36%) in the IVIg-treated FNAIT cases, and one in the controls (NS). C4d deposition at the syncytiotrophoblast was present in combination with low-grade villitis of unknown etiology in three newly diagnosed FNAIT cases that were born SGA. We conclude that a higher degree of classical pathway-induced complement activation is present in placentas from pregnancies with untreated FNAIT. This may affect placental function and fetal growth.     

Long-Term Efficacy of Intranasal Esketamine in Treatment-Resistant Major Depression: A Systematic Review

Esketamine (ESK) has been approved as a rapid-acting intranasal treatment for treatment-resistant depression (TRD). Although existing studies have investigated the efficacy of ESK in the 4-week induction phase, our knowledge about long-term ESK efficacy remains poor. The aim of this systematic review was to summarize the available data on long-term ESK efficacy for TRD. A systematic search was performed including articles in English, up to 31 March 2021. The search found 7 relevant studies, involving 1024 adult TRD patients. Continuing treatment with ESK after the 4-week induction phase may be associated with stable efficacy in relapse prevention among TRD patients. Conversely, the long-term antidepressant effectiveness upon discontinuation of ESK might be limited, although data from three studies had a moderate to high risk of bias. Overall, the results on the effectiveness of this compound in the long term are mixed. According to our findings, ESK treatment should be continued following the induction phase to reach a stable efficacy in relapse prevention, while the long-term antidepressant and anti-suicidal effects of ESK after discontinuation are inconsistent. Currently, the level of proof of ESK efficacy in long-term TRD treatment remains low and more RCTs with larger sample sizes and active comparators are needed.     

Neoantigen-Specific T-Cell Immune Responses: The Paradigm of NPM1-Mutated Acute Myeloid Leukemia

The C-terminal aminoacidic sequence from NPM1-mutated protein, absent in normal human tissues, may serve as a leukemia-specific antigen and can be considered an ideal target for NPM1-mutated acute myeloid leukemia (AML) immunotherapy. Different in silico instruments and in vitro/ex vivo immunological platforms have identified the most immunogenic epitopes from NPM1-mutated protein. Spontaneous development of endogenous NPM1-mutated-specific cytotoxic T cells has been observed in patients, potentially contributing to remission maintenance and prolonged survival. Genetically engineered T cells, namely CAR-T or TCR-transduced T cells, directed against NPM1-mutated peptides bound to HLA could prospectively represent a promising therapeutic approach. Although either adoptive or vaccine-based immunotherapies are unlikely to be highly effective in patients with full-blown leukemia, these strategies, potentially in combination with immune-checkpoint inhibitors, could be promising in maintaining remission or preemptively eradicating persistent measurable residual disease, mainly in patients ineligible for allogeneic hematopoietic stem cell transplant (HSCT). Alternatively, neoantigen-specific donor lymphocyte infusion derived from healthy donors and targeting NPM1-mutated protein to selectively elicit graft-versus-leukemia effect may represent an attractive option in subjects experiencing post-HSCT relapse. Future studies are warranted to further investigate dynamics of NPM1-mutated-specific immunity and explore whether novel individualized immunotherapies may have potential clinical utility in NPM1-mutated AML patients.     

High-performance liquid chromatography evaluation of phenols in olive fruit, virgin olive oil, vegetation waters, and pomace and 1D- and 2D-nuclear magnetic resonance characterization

Phenolic compounds are the most important antioxidants of virgin olive oil. This paper reports on the application of solid phase extraction (SPE) in the separation of phenolic compounds from olive fruit, olive oil, and by-products of the mechanical extraction of the oil and the complete spectroscopic characterization by nuclear magnetic resonance of demethyloleuropein and verbascoside extracted from olive fruit. SPE led to a higher recovery of phenolic compounds from olives than did liquid/liquid extraction. SPE also was used to separate phenolic compounds from pomaces and vegetation waters. Phenylacid and phenyl-alcohol concentrations in extracts obtained from SPE and liquid/liquid extraction were not significantly different (P<0.05). The recovery of the dialdehydic form of elenolic acid linked to 3,4-(dihydroxyphenyl)ethanol and an isomer of oleuropein aglycon, however, was low.     

Energy saving in existing buildings by an intelligent use of interoperable ICTs

In this paper, we report a methodology, developed in the context of Smart Energy Efficient Middleware for Public Spaces European Project, aimed at exploiting ICT monitoring and control services to reduce energy usage and CO₂ footprint in existing buildings. The approach does not require significant construction work as it is based on commercial-off-the-shelf devices and, where present, it exploits and integrates existing building management systems with new sensors and actuator networks. To make this possible, the proposed approach leverages upon the following main contributions: (a) to develop an integrated building automation and control system, (b) to implement a middleware for the energy-efficient buildings domain, (c) to provide a multi-dimensional building information modelling-based visualisation, and (d) to raise people’s awareness about energy efficiency. The research approach adopted in the project started with the selection, as case studies, of representative test and reference rooms in modern and historical buildings chosen for having different requirements and constraints in term of sensing and control technologies. Then, according to the features of the selected rooms, the strategies to reduce the energy consumptions were defined, taking into account the potential savings related to lighting, heating, ventilation, and air conditioning (HVAC) systems and other device loads (PC, printers, etc.). The strategies include both the control of building services and devices and the monitoring of environmental conditions and energy consumption. In the paper, the energy savings estimated through simulation, for both HVAC and lighting, are presented to highlight the potential of the designed system. After the implementation of the system in the demonstrator, results will be compared with the monitored data.     

Process simulation of a hybrid SOFC/mGT and enriched air/steam fluidized bed gasifier power plant

The aim of this work is to experimentally and numerically analyze the performance of a integrated power plant composed by a steam oxygen fluidized bed biomass gasifier fed by woods, a Solid Oxide Fuel Cell (SOFC) and a micro Gas Turbine (mGT). The numerical analysis is carried out by using ChemCAD software. In particular, SOFC and gasifier were modeled using proper developed Fortran subroutines interfaced to the basic software. The adopted SOFC model was already validated by the authors in previous works, while the gasifier model was here developed and validated by means of experimental activities carried out by using a bench scale gasifier. Different compounds (Benzene, Toluene, Naphthalene, Phenols) were chosen to analyze the tar evolution in the gaseous stream during the gasification process. Hot gas cleaning (based on catalytic ceramic filter candles inserted in the freeboard of the gasifier – UNIQUE concept) was adopted to remove tar and particulates from the fuel hot gas stream. Different moisture contents in the range between 10 and 30% (i.e. in a deviation of 10% around the usual wood moisture content of 20%) were numerically simulated as well as the degree of purity of the oxygen utilized in the power plant (between 25% and 95%, the rest being N₂). The power requirement for pure oxygen production leads to a reduction of the electrical efficiency of the whole power plant. For this reason, a sensitivity analysis was conducted to find the optimal operation conditions in order to maximise the syngas (H₂, CO) content in the produced gas, while maintaining a high overall electrical efficiency.     

Shell-and-Tube Minichannel Condenser for Low Refrigerant Charge

The design of an innovative shell-and-tube heat pump condenser using 2 mm ID minichannels is presented. This condenser has been designed aiming at the minimization of the charge, which can be required by safety or environmental restrictions. Nevertheless, minichannels represent also a solution to the high-pressure challenge when using carbon dioxide as a refrigerant. The present prototype was realized for the use with propane in the framework of the European project SHERHPA, concerning the development of heat pumps working with natural fluids. Experimental data for heat transfer and pressure drop are reported in the present paper. The measurements have been obtained using refrigerant R22, which displays a temperature versus pressure saturation curve pretty close to the one of propane. The data have also been compared against a computational procedure for shell-and-tube heat exchangers design. The refrigerant charge has been computed by means of different void fraction correlations, showing that the expected charge is less than half the quantity required by a brazed plate condenser giving the same capacity.