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.     

Fullerene‐Promoted Singlet‐Oxygen Photochemical Oxygenations in Glass‐Polymer Microstructured Reactors

In this paper we report the fabrication of thiolene‐based microstructured reactors (MRs) that have been specifically designed to include solid‐supported reagents within the microchannels network. We propose a convenient solution to realize reversible press‐fit, leak‐proof interconnects that greatly simplify the MR coupling to the external environment such as capillary tubing, sample reservoirs and pumps. The MRs have been used to carry out the oxidation of α‐terpinene and methionine using [60]fullerene, covalently linked to Tentagel^® and silica gel matrices, as a singlet oxygen sensitizer. High conversions have been observed for both substrates although, in the case of α‐terpinene, a partial photodegradation of the endo‐peroxide product was detected. Interestingly, in the case of methionine, a quantitative conversion to the corresponding sulfoxides was achieved in about 40 seconds, using low‐power, white LED illumination. The reaction time is considerably shorter when compared to the batch procedure that requires, for the same process, about one hour illumination and the use of a 300‐W tungsten halogen lamp.     

Influence of microorganisms on retinol isomerization in milk

In order to have a deeper insight into the retinol isomerization phenomenon, in this work different milk samples have been analysed for their content of trans retinol and its cis-isomers, by means of reliable HPLC techniques. Levels of the different isomers and the degree of retinol isomerization (13-cis/all-trans ratio, %) have been monitored during milk storage at different temperatures and after addition of specific microorganisms. In raw milk stored at 4 degrees C for 96 h the degree of retinol isomerization shifted from 1.1 to 2.3%, while in raw milk stored at 22 degrees C for 24 h it increased from 1.1 to 12.7%. Among microorganisms tested in pasteurized milk, the most active in causing an increment in the 13-cis/all-trans ratio (%), from 3.4 to 33.4% in 8 h, was Streptococcus thermophilus. The results obtained demonstrated a relationship between microbial evolution and retinol isomerization. Therefore, the determination of retinol isomers is of importance not only for a more precise evaluation of vitamin A activity but also for the evaluation of the microbiological quality of milk.     

Alternative Methods to SO2 for Microbiological Stabilization of Wine

The use of sulfur dioxide (SO₂) as wine additive is able to ensure both antioxidant protection and microbiological stability. In spite of these undeniable advantages, in the last two decades the presence of SO₂ in wine has raised concerns about potential adverse clinical effects in sensitive individuals. The winemaking industry has followed the general trend towards the reduction of SO₂ concentrations in food, by expressing at the same time the need for alternative control methods allowing reduction or even elimination of SO_2. In the light of this, research has been strongly oriented toward the study of alternatives to the use of SO₂ in wine. Most of the studies have focused on methods able to replace the antimicrobial activity of SO₂. This review article gives a comprehensive overview of the current state‐of‐the‐art about the chemical additives and the innovative physical techniques that have been proposed for this purpose. After a focus on the chemistry and properties of SO₂ in wine_, as well as on wine spoilage and on the conventional methods used for the microbiological stabilization of wine, recent advances on alternative methods proposed to replace the antimicrobial activity of SO₂ in winemaking are presented and discussed. Even though many of the alternatives to SO₂ showed good efficacy, nowadays no other physical technique or additive can deliver the efficacy and broad spectrum of action as SO₂ (both antioxidant and antimicrobial), therefore the alternative methods should be considered a complement to SO₂ in low‐sulfite winemaking, rather than being seen as its substitutes.     

Nondegradative additive manufacturing of medical grade copolyesters of high molecular weight and with varied elastic response

Although additive manufacturing through melt extrusion has become increasingly popular as a route to design scaffolds with complex geometries the technique if often limited by the reduction in molecular weight and the viscoelastic response when degradable aliphatic polyesters of high molecular weight are used. Here we use a melt extruder and fused filament fabrication printer to produce a reliable nondegradative route for scaffold fabrication of medical grade copolymers of L-lactide, poly(ε-caprolactone-co-L-lactide), and poly(L-lactide-co-trimethylene carbonate). We show that degradation is avoided using filament extrusion and fused filament fabrication if the process parameters are deliberately chosen based upon the rheological behavior, mechanical properties, and polymer composition. Structural, mechanical, and thermal properties were assessed throughout the process to obtain comprehension of the relationship between the rheological properties and the behavior of the medical grade copolymers in the extruder and printer. Scaffolds with a controlled architecture were achieved using high-molecular-weight polyesters exhibiting a large range in the elastic response causing negligible degradation of the polymers. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48550.     

Inactivation of Staphylococcus aureus by the synergistic action of charged and reactive plasma particles

In this paper, a low-pressure capacitively coupled plasma discharge sustained in an argonoxygen mixture was studied in order to evaluate its properties in terms of inactivation of Staphylococcus aureus. The plasma parameters as electron temperature and plasma density were measured by the Langmuir probe (N_e ≈ 10¹⁵ m⁻³, T_e ≈ 1.5 eV), while the neutral atom density was in the range of 10²¹ m⁻³. In the plasma phase, oxygen radicals were taken as reference of the reactive species with antimicrobial activity, and oxygen spectral lines, over a range of plasma process parameters, were investigated by the optical emission spectroscopy. Optimal plasma conditions were found, and a count reduction of 4 log in a few minutes of the bacterium proves the potentiality of an industrial grade plasma reactor as a sterilization agent.     

Early CSF Biomarkers and Late Functional Outcomes in Spinal Cord Injury. A Pilot Study

Although, biomarkers are regarded as an important tool for monitoring injury severity and treatment efficacy, and for predicting clinical evolution in many neurological diseases and disorders including spinal cord injury, there is still a lack of reliable biomarkers for the assessment of clinical course and patient outcome. In this study, a biological dataset of 60 cytokines/chemokines, growth factorsm and intracellular and extracellular matrix proteins, analyzed in CSF within 24 h of injury, was used for correlation analysis with the clinical dataset of the same patients. A heat map was generated of positive and negative correlations between biomarkers and clinical rating scale scores at discharge, and between biomarkers and changes in clinical scores during the observation period. Using very stringent statistical criteria, we found 10 molecules which correlated with clinical scores at discharge, and five molecules, which correlated with changes in clinical scores. The proposed methodology may be useful for generating hypotheses regarding “predictive” and “treatment effectiveness” biomarkers, thereby suggesting potential candidates for disease-modifying therapies using a “bed-to-bench” approach.     

Conditioning of Rh/α-Al2O3 catalysts for H2 production via CH4 partial oxidation at high space velocity

Experimental evidence and literature indications suggest that the process of methane partial oxidation over Rh catalysts is structure sensitive. Crystal phases and Rh cluster size are thus expected to affect the final catalytic performance. In this work, it is observed that outstanding performances are obtained when the as-prepared catalysts are conditioned through repeated runs at increasing temperature and O₂/CH₄ = 0.56. Catalysts slowly activate, that is CH₄ conversion and synthesis gas selectivity progressively grow with time on stream. On the basis of TPO and CH₄ decomposition measurements, this phenomenon is herein explained as the result of a surface reconstruction driven by the repeated exposition to the reaction at high temperature; it is thought that such reconstruction tends to eliminate defect sites and disfavors C-deposition reactions (extremely fast over steps and kinks). Conditioning with O₂-enriched feed streams makes conditioning faster, since the accumulation of surface C-species is suppressed; however, the catalyst is eventually less active than a catalyst conditioned with standard feed mixtures. As an alternative, accumulation of carbon can be suppressed and surface reconstruction proceeds faster if the catalyst is directly exposed to the reaction at high temperature for several hours.