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The αs2-casein is a phosphoprotein secreted in the milk of most mammals, and it is the most hydrophilic of all caseins. Contrary to genes found in ruminants, in donkeys two different encoding genes for donkey αs2-casein (CSN1S2 I and CSN1S2 II) have been identified. However, unlike in ruminants, the variability at these loci has not been characterized in detail in donkeys until now. In this study, we analyze the transcript profile of the donkey CSN1S2 I and CSN1S2 II genes, and we identify and describe the variability of these loci in the Ragusana and Amiatina breeds reared in Italy. The analysis of the CSN1S2 I Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) products and subsequent sequencing showed, in addition to correctly spliced mRNA, seven other minor mRNAs resulting from differential splicing events involving, in various combinations, entire exons (4, 5, 6, and 11), parts of exons (5′ or 3′ end of exon 17), or the recognition of intronic sequences as an exon (exon 12′). Similarly, the transcription analysis of the CSN1S2 II gene revealed a remarkable variability in splicing events, mainly concerning the alternative insertion of an extra exon 7 (named 7′); the first 33 bp of exon 13; or the alternative skipping of exons 9, 10, 11, 12, and 15, and their combinations. At the mRNA level for CSN1S2 I, seven SNPs were observed, five of which led to amino acid changes: p.T73>A, p.I109>V, p.I130>V, p.I146>T, and p.D217>Y. Similarly, nine SNPs were observed at the CSN1S2 II locus, seven of which are non-synonymous: p.L63>F, p.H70>Q, p.D90>N, p.129A>T, p.H131>Y, p.E144>G, and p.F157>S. In addition, the DNA sequencing of exon 17 and flanking introns of the CSN1S2 I gene revealed a G>A transition at the splice acceptor site of CSN1S2 I exon 17 (FM946022.1:c.375-1G>A), resulting in an allele-specific skipping of the first 15 nucleotides of this exon, which encode the peptide 176NKINQ180, and the recognition of an in-frame cryptic splicing acceptor site: arAACAAAATCAACCAG. A genotyping method based on restriction fragment length polymorphism (XbaI PCR-RFLP) was set up for this SNP. In the total population studied (105 Ragusana and 14 Amiatina donkeys), the A allele had a frequency of 0.2437 with no evidence of deviation from the Hardy–Weinberg equilibrium. This study adds new knowledge regarding the genetic variability of αs2-caseins in donkeys and may contribute significantly to the genetic improvement of milk production for this species.

The sea bottom acts as a key natural archive where the memory of long-term timescale environmental changes is recorded. This study discusses some ecological and chemical features of fjord sediments that were explored during the AREX cruise carried out in the Svalbard archipelago in the summer of 2021. The activity rates of the enzymes leucine aminopeptidase (LAP), beta-glucosidase (GLU), and alkaline phosphatase (AP) and community-level physiological profiles (CLPPs) were studied with the aim of determining the functional diversity of the benthic microbial community, while bacterial isolates were screened for their susceptibility to antibiotics in order to explore the role of these extreme environments as potential reservoirs of antibiotic resistance. Enzyme activity rates were obtained using fluorogenic substrates, and CLPPs were obtained using Biolog Ecoplates; antibiotic susceptibility assays were performed through the standard disk diffusion method. Spatial trends observed in the functional profiles of the microbial community suggested variability in the microbial community’s composition, presumably related to the patchy distribution of organic substrates. Complex carbon sources, carbohydrates, and amino acids were the organic polymers preferentially metabolized by the microbial community. Multi-resistance to enrofloxacin and tetracycline was detected in all of the examined samples, stressing the role of sediments as a potential reservoir of chemical wastes ascribable to antibiotic residuals. This study provides new insights on the health status of fjord sediments of West Spitsbergen, applying a dual ecological and biochemical approach. Microbial communities in the fjord sediments showed globally a good functional diversity, suggesting their versatility to rapidly react to changing conditions. The lack of significant diversification among the three studied areas suggests that microbial variables alone cannot be suitable descriptors of sediment health, and that additional measures (i.e., physical–chemical characteristics) should be taken to better define environmental status.

The β-thalassemias are a group of monogenic hereditary hematological disorders caused by deletions and/or mutations of the β-globin gene, leading to low or absent production of adult hemoglobin (HbA). For β-thalassemia, sirolimus has been under clinical consideration in two trials (NCT03877809 and NCT04247750). A reduced immune response to anti-SARS-CoV-2 vaccination has been reported in organ recipient patients treated with the immunosuppressant sirolimus. Therefore, there was some concern regarding the fact that monotherapy with sirolimus would reduce the antibody response after SARS-CoV-2 vaccination. In the representative clinical case reported in this study, sirolimus treatment induced the expected increase of fetal hemoglobin (HbF) but did not prevent the production of anti-SARS-CoV-2 IgG after vaccination with mRNA-1273 (Moderna). In our opinion, this case report should stimulate further studies on β-thalassemia patients under sirolimus monotherapy in order to confirm the safety (or even the positive effects) of sirolimus with respect to the humoral response to anti-SARS-CoV-2 vaccination. In addition, considering the extensive use of sirolimus for the treatment of other human pathologies (for instance, in organ transplantation, systemic lupus erythematosus, autoimmune cytopenia, and lymphangioleiomyomatosis), this case report study might be of general interest, as large numbers of patients are currently under sirolimus treatment.

The search for DNA polymorphisms useful for the genetic improvement of dairy farm animals has spanned more than 40 years, yielding relevant findings in cattle for milk traits, where the best combination of alleles for dairy processing has been found in casein genes and in DGAT1. Nowadays, similar results have not yet been reached in river buffaloes, despite the availability of advanced genomic technologies and accurate phenotype records. The aim of the present study was to investigate and validate the effect of four single nucleotide polymorphisms (SNP) in the CSN1S1, CSN3, SCD and LPL genes on seven milk traits in a larger buffalo population. These SNPs have previously been reported to be associated with, or affect, dairy traits in smaller populations often belonging to one farm. A total of 800 buffaloes were genotyped. The following traits were individually recorded, monthly, throughout each whole lactation period from 2010 to 2021: daily milk yield (dMY, kg), protein yield (dPY, kg) and fat yield (dFY, kg), fat and protein contents (dFP, % and dPP, %), somatic cell count (SCC, 103 cell/mL) and urea (mg/dL). A total of 15,742 individual milk test day records (2496 lactations) were available for 680 buffalo cows, with 3.6 ± 1.7 parities (from 1 to 13) and an average of 6.1 ± 1.2 test day records per lactation. Three out four SNPs in the CSN1S1, CSN3 and LPL genes were associated with at least one of analyzed traits. In particular, the CSN1S1 (AJ005430:c.578C>T) gave favorable associations with all yield traits (dMY, p = 0.022; dPY, p = 0.014; dFY, p = 0.029) and somatic cell score (SCS, p = 0.032). The CSN3 (HQ677596: c.536C>T) was positively associated with SCS (p = 0.005) and milk urea (p = 0.04). Favorable effects on daily milk yield (dMY, p = 0.028), fat (dFP, p = 0.027) and protein (dPP, p = 0.050) percentages were observed for the LPL. Conversely, the SCD did not show any association with milk traits. This is the first example of a confirmation study carried out in the Mediterranean river buffalo for genes of economic interest in the dairy field, and it represents a very important indication for the preselection of young bulls destined for breeding programs aimed at more sustainable dairy production.

Background: In atherothrombotic acute ischemic stroke (AIS), when the atheroma breaks down, the clot can incorporate foam cells (FCs). The identification of the correct etiology is paramount for secondary stroke prevention. This study aims to evaluate the presence of the FC in the arterial clot, and to determine whether patients with FCs and patients without FCs (NFCs) had different cerebrovascular risk factors, haemato-chemical parameters, and atherosclerotic disease incidence, in order to predict the etiological diagnosis. Methods: We collected 100 clots retrieved by mechanical thrombectomy from 495 consecutive AIS patients with large vessel occlusion. An expert pathologist evaluated the FC presence by histological examination stained with hematoxylin and eosin. Results: We observed FCs in 29/100 (29%) of retrieved clots and divided the patients into two groups, with/without FCs. The two groups had similar clinical and laboratory features, with a discrepancy between the FC presence in the clot and the clinical etiological diagnosis, even if not statistically significant. Conclusions: Our study showed the presence of FCs in approximately one-third of the retrieved clots, but the identification of the clot that presumably comes from the atheromatous plaque rupture tended to disagree with the clinical diagnosis. Future studies may reveal their potential to disclose clot origin or specific patient characteristics, guiding treatment options.

Induction of fetal hemoglobin (HbF) is highly beneficial for patients carrying β-thalassemia, and novel HbF inducers are highly needed. Here, we describe a new class of promising HbF inducers characterized by an isoxazole chemical skeleton and obtained through modification of two natural molecules, geldanamycin and radicicol. After preliminary biological assays based on benzidine staining and RT-qPCR conducted on human erythroleukemic K562 cells, we employed erythroid precursors cells (ErPCs) isolated from β-thalassemic patients. ErPCs weretreated with appropriate concentrations of isoxazole derivatives. The accumulation of globin mRNAs was studied by RT-qPCR, and hemoglobin production by HPLC. We demonstrated the high efficacy of isozaxoles in inducing HbF. Most of these derivatives displayed an activity similar to that observed using known HbF inducers, such as hydroxyurea (HU) or rapamycin; some of the analyzed compounds were able to induce HbF with more efficiency than HU. All the compounds were active in reducing the excess of free α-globin in treated ErPCs. All the compounds displayed a lack of genotoxicity. These novel isoxazoles deserve further pre-clinical study aimed at verifying whether they are suitable for the development of therapeutic protocols for β-thalassemia.

Botulinum toxin is a protein deriving from the bacteria Clostridium botulinum and it is widely used for the treatment of a variety of muscle hyperactivity syndromes and for cosmetic indications. Having a long-lasting effect, Botulinum toxin type A (BTA) is one of the most botulin toxin products used. Even if BTA has shown benefits in reducing the vertical lines between the eyebrows, Adverse Drug Reactions (ADRs) have been experienced as well, of which the most common ones are headache and drooping eyelids. In addition, since other local and systemic risks have been identified, a non-interventional post-authorization safety study (PASS) has been started. The aim of the present study was to report cases of skin toxicity associated with this drug, considering Individual Case Safety Reports (ICSRs) existing on the Eudravigilance website. Among 1464 ICSRs sent to the EV database, 718 ICSRs, including 5154 PTs, reported BTA as a suspected drug associated with cutaneous toxicity. The majority of patients experiencing BTA-induced skin toxicity were female (92.1%) belonging mostly to the age group of 18–64 years. The most serious criteria, when reported, were “Other Medically Important Condition” and “Caused/prolonged hospitalization”, although the outcome was mainly reported as “Unknown”. The most reported PTs, related to skin disorders, were: “Erythema”, “Rash”, “Pruritus”, “Urticaria”, “Swelling face”, “Brow ptosis”, “Eyelid ptosis”, “Injection site pain”, and “Angioedema”. Considering that in most ICSRs, ADRs related to skin disorders were symptoms of hypersensitivity reactions which in some conditions could be life-threatening, further studies are required to better define the safety profile of BTA used for aesthetic procedures.

The β-thalassemias are hereditary monogenic diseases characterized by a low or absent production of adult hemoglobin and excess in the content of α-globin. This excess is cytotoxic for the erythroid cells and responsible for the β-thalassemia-associated ineffective erythropoiesis. Therefore, the decrease in excess α-globin is a relevant clinical effect for these patients and can be realized through the induction of fetal hemoglobin, autophagy, or both. The in vivo effects of sirolimus (rapamycin) and analogs on the induction of fetal hemoglobin (HbF) are of key importance for therapeutic protocols in a variety of hemoglobinopathies, including β-thalassemias. In this research communication, we report data showing that a decrease in autophagy-associated p62 protein, increased expression of ULK-1, and reduction in excess α-globin are occurring in erythroid precursors (ErPCs) stimulated in vitro with low dosages of sirolimus. In addition, increased ULK-1 mRNA content and a decrease in α-globin content were found in ErPCs isolated from β-thalassemia patients recruited for the NCT03877809 clinical trial and treated with 0.5–2 mg/day sirolimus. Our data support the concept that autophagy, ULK1 expression, and α-globin chain reduction should be considered important endpoints in sirolimus-based clinical trials for β-thalassemias.

In this review article, we present the fascinating story of rapamycin (sirolimus), a drug able to induce γ-globin gene expression and increased production of fetal hemoglobin (HbF) in erythroid cells, including primary erythroid precursor cells (ErPCs) isolated from β-thalassemia patients. For this reason, rapamycin is considered of great interest for the treatment of β-thalassemia. In fact, high levels of HbF are known to be highly beneficial for β-thalassemia patients. The story of rapamycin discovery began in 1964, with METEI, the Medical Expedition to Easter Island (Rapa Nui). During this expedition, samples of the soil from different parts of the island were collected and, from this material, an antibiotic-producing microorganism (Streptomyces hygroscopicus) was identified. Rapamycin was extracted from the mycelium with organic solvents, isolated, and demonstrated to be very active as an anti-bacterial and anti-fungal agent. Later, rapamycin was demonstrated to inhibit the in vitro cell growth of tumor cell lines. More importantly, rapamycin was found to be an immunosuppressive agent applicable to prevent kidney rejection after transplantation. More recently, rapamycin was found to be a potent inducer of HbF both in vitro using ErPCs isolated from β-thalassemia patients, in vivo using experimental mice, and in patients treated with this compound. These studies were the basis for proposing clinical trials on β-thalassemia patients.

The CSN1S2 gene encodes αs2-casein, the third most abundant protein in camel milk. Despite its importance in foals, human nutrition, and dairy processing, the CSN1S2 gene in camels has received little attention. This study presents the first complete characterization of the CSN1S2 gene sequence in Old-World camels (Camelus bactrianus and Camelus dromedarius). Additionally, the gene promoter, consisting of 752 bp upstream of exon 1, was analyzed. The entire gene comprises 17 exons, ranging in length from 24 bp (exons 4, 8, 11, and 13) to 280 bp (exon 17). Interesting was the identification of the exon 12 in both species. The promoter analysis revealed 24 putative binding sites in the Bactrian camel and 22 in dromedary camel. Most of these sites were typical elements associated with milk protein, such as C/EBP-α, C/EBP-β, Oct-1, and AP1. The SNP discovery showed relatively high genetic diversity compared to other camel casein genes (CSN1S1, CSN2, and CSN3), with a total of 34 polymorphic sites across the two species. Particularly noteworthy is the transition g.311G>A in the CSN1S2 promoter, creating a new putative consensus binding site for a C/EBP-β in the Bactrian camel. At the exon level, two novel variants were found. One was detected in exon 6 of the Bactrian camel (g.3639C>G), resulting in an amino acid replacement, p.36Ile>Met. The second variant was found in noncoding exon 17 of dromedary CSN1S2 (g.1511G>T). Although this mutation occurs in the 3′-UnTranslated Region, it represents the first example of exonic polymorphism in the CSN1S2 for this species. This SNP also affects the binding sites of different microRNAs, including the seed sequence of the miRNA 4662a-3p, highlighting its role as a regulatory factor for CSN1S2 gene. A PCR-RFLP was set up for genotyping a dromedary Tunisian population (n = 157), and the minor allele frequency was found to be 0.27 for the G allele, indicating a potential yield improvement margin. The interspersed elements (INEs) analysis revealed 10 INEs covering 7.34% and 8.14% of the CSN1S2 sequence in the Bactrian and dromedary camels, respectively. Furthermore, six elements (A, B, F, H, I, and L) are shared among cattle and camels and are partially found in other ruminants, suggesting a common ancestral origin of these retrotransposons. Conversely, elements C, D, E, and G are specific to camels.

Over the past decade, the fields of civil engineering, i.e., structural engineering, have increasingly used the building information modelling (BIM) approach in both professional practice and as the focus of research. However, the field of structural engineering, which can be seen as a sub-discipline of civil engineering, misses, as far as the authors are aware, a real state-of-the-art on the use of BIM in this regard. The aim of this paper, therefore, is to start bridging that gap. In particular, the authors have conducted a traditional literature review on the utilisation of BIM in structural engineering, enabling them to perform a detailed content analysis of publications. The qualitative investigation of the literature that the authors have conducted has highlighted six main BIM uses in structural engineering: (1) structural analyses; (2) production of shop drawings; (3) optimized structural design, early identification of constructability issues, and a comparison of different structural solutions; (4) seismic risk assessments; (5) existing-condition modelling and retrofitting of structures; and (6) structural health monitoring. Each of these is discussed in relation to their reference workflows; use of information models; information exchanges; and main limitations. In the conclusions, the authors identify current gaps in knowledge, as well as likely developments and improvements in the utilization of BIM in structural engineering. The authors also outline the possible significance of this work more broadly.

A new understanding of plankton ecology has been obtained by studying the phenotypic traits of free-living prokaryotes in the Sicily Channel (Central Mediterranean Sea), an area characterised by oligotrophic conditions. During three cruises carried out in July 2012, January 2013 and July 2013, the volume and morphology of prokaryotic cells were assessed microscopically using image analysis in relation to environmental conditions. The study found significant differences in cell morphologies among cruises. The largest cell volumes were observed in the July 2012 cruise (0.170 ± 0.156 µm3), and the smallest in the January 2013 cruise (0.060 ± 0.052 µm3). Cell volume was negatively limited by nutrients and positively by salinity. Seven cellular morphotypes were observed among which cocci, rods and coccobacilli were the most abundant. Cocci, although they prevailed numerically, always showed the smallest volumes. Elongated shapes were positively related to temperature. Relationships between cell morphologies and environmental drivers indicated a bottom-up control of the prokaryotic community. The morphology/morphometry-based approach is a useful tool for studying the prokaryotic community in microbial ecology and should be widely applied to marine microbial populations in nature.

(1) Background: MicroRNAs are involved in the expression of the gene encoding the chloride channel CFTR (Cystic Fibrosis Transmembrane Conductance Regulator); the objective of this short report is to study the effects of the treatment of bronchial epithelial Calu-3 cells with molecules mimicking the activity of pre-miR-145-5p, pre-miR-335-5p, and pre-miR-101-3p, and to discuss possible translational applications of these molecules in pre-clinical studies focusing on the development of protocols of possible interest in therapy; (2) Methods: CFTR mRNA was quantified by Reverse Transcription quantitative Polymerase Chain Reaction (RT-qPCR). The production of the CFTR protein was assessed by Western blotting; (3) Results: The treatment of Calu-3 cells with agomiR-145-5p caused the highest inhibition of CFTR mRNA accumulation and CFTR production; (4) Conclusions: The treatment of target cells with the agomiR pre-miR-145-5p should be considered when CFTR gene expression should be inhibited in pathological conditions, such as polycystic kidney disease (PKD), some types of cancer, cholera, and SARS-CoV-2 infection.

Pectus excavatum is a deformity of the thorax characterized by ventrodorsal narrowing of the sternum bone and costal cartilages, which can lead to compression and cardiopulmonary alterations in dogs, presenting a high prevalence in brachycephalic breeds. The aim of this report was to describe two types of management for the noninvasive treatment of pectus excavatum in newborn puppies of the breeds French Bulldog and American Bully. The puppies presented dyspnea, cyanosis and substernal retraction during inspiration. The diagnosis was performed by physical examination and confirmed by chest X-ray. Two types of splints were performed (a circular splint with plastic pipe and a paper box splint on the chest), aiming at thoracic lateral compression and frontal chest remodeling. The management was effective for the conservative treatment of mild-grade pectus excavatum, resulting in the repositioning of the thorax and improvement of the respiratory pattern.

In this work, cyclic-load tests on reduced-scale corroded reinforced-concrete hollow cross-section bridge piers have been experimentally performed and compared to the results of similar non-corroded piers. Piers were aged by using an imposed electric current and sodium chloride water solution before performing a mechanical cyclic-load test. The corrosion process has been detected with Non-Destructive Evaluation techniques by means of SonReb method (to check concrete degradation) and by measuring corrosion potential (to check steel degradation). The crack pattern was recorded by dedicated cameras, and an LVDT system was set up to monitor the cyclic-load test. Experimental results focused on degradation monitoring and mechanical performance under cyclic loads. During the cyclic-load mechanical test, the first cracks on the piers surface occurred diagonally, inclined at about 45°. This is the consequence of the failure mode change from ductile failure, as expected for slender designed piers, to brittle shear failure. The flexural failure occurred in the case of non-corroded piers. Presented tests can provide a useful contribution of experimental data to analyse the behaviour of corroded reinforced concrete hollow bridge piers, scarcely tested. In particular, the cyclic response can be a useful reference for the proposition/validation of nonlinear capacity models for the evaluation of the seismic capacity of corroded bridge piers.

Airborne Laser Scanners (ALS) and Terrestrial Laser Scanners (TLS) are two lidar systems frequently used for remote sensing forested ecosystems. The aim of this study was to compare crown metrics derived from TLS, ALS, and a combination of both for describing the crown structure and fuel attributes of longleaf pine (Pinus palustris Mill.) dominated forest located at Eglin Air Force Base (AFB), Florida, USA. The study landscape was characterized by an ALS and TLS data collection along with field measurements within three large (1963 m2 each) plots in total, each one representing a distinct stand condition at Eglin AFB. Tree-level measurements included bole diameter at breast height (DBH), total height (HT), crown base height (CBH), and crown width (CW). In addition, the crown structure and fuel metrics foliage biomass (FB), stem branches biomass (SB), crown biomass (CB), and crown bulk density (CBD) were calculated using allometric equations. Canopy Height Models (CHM) were created from ALS and TLS point clouds separately and by combining them (ALS + TLS). Individual trees were extracted, and crown-level metrics were computed from the three lidar-derived datasets and used to train random forest (RF) models. The results of the individual tree detection showed successful estimation of tree count from all lidar-derived datasets, with marginal errors ranging from −4 to 3%. For all three lidar-derived datasets, the RF models accurately predicted all tree-level attributes. Overall, we found strong positive correlations between model predictions and observed values (R2 between 0.80 and 0.98), low to moderate errors (RMSE% between 4.56 and 50.99%), and low biases (between 0.03% and −2.86%). The highest R2 using ALS data was achieved predicting CBH (R2 = 0.98), while for TLS and ALS + TLS, the highest R2 was observed predicting HT, CW, and CBD (R2 = 0.94) and HT (R2 = 0.98), respectively. Relative RMSE was lowest for HT using three lidar datasets (ALS = 4.83%, TLS = 7.22%, and ALS + TLS = 4.56%). All models and datasets had similar accuracies in terms of bias (

Polar marine environments host a complex assemblage of cold-adapted auto- and heterotrophic microorganisms that affect water biogeochemistry and ecosystem functions. However, due to logistical difficulties, remote regions like those in close proximity to glaciers have received little attention, resulting in a paucity of microbiological data. To fill these gaps and obtain novel insights into microbial structure and function in Arctic regions, a survey of microbial communities in an area close to the Blomstrandbreen glacier in Kongsfjorden (Svalbard Archipelago; Arctic Ocean) was carried out during an early summer period. An Unmanned Autonomous Vehicle designed to safely obtain seawater samples from offshore-glacier transects (PROTEUS, Portable RObotic Technology for Unmanned Surveys) was equipped with an automatic remotely-controlled water multi-sampler so that it could sample just beneath the glacier, where access from the sea is difficult and dangerous. The samples were analysed by image analysis for the abundance of total prokaryotes, viable and respiring cells, their morphological traits and biomass; by flow cytometry for autotrophic and prokaryotic cells (with high and low nucleic acid contents) as well as virus-like particle counts; by BIOLOG ECOPLATES for potential community metabolism; and by fluorimetry for potential enzymatic activity rates on organic polymers. Contextually, the main physical and chemical (temperature, salinity, pH, dissolved oxygen and nutrients) parameters were detected. Altogether, besides the PROTEUS vehicle’s suitability for collecting samples from otherwise inaccessible sites, the multivariate analysis of the overall dataset allowed the identification of three main sub-regions differently affected by the haline gradient (close to the glacier) or terrigenous inputs coming from the coast. A complex microbiological scenario was depicted by different patterns of microbial abundance and metabolism among the transects, suggesting that ice melting and Atlantic water inflow differently supported microbial growth.

Calcium-sensitive caseins are the main protein component of milk. In the goat, they are encoded by three genes (CSN1S1, CSN2, and CSN1S2) located on chromosome 6. A high number of alleles has been discovered for these genes in the goat species, responsible for changes in the milk’s qualitative and quantitative characteristics. This study aimed to develop an Allele-Specific PCR (AS-PCR), which allowed us to unequivocally detect goat carriers of the CSN201 allele. Subsequently, the calcium-sensitive casein loci genotype was investigated in three native goat breeds of the Lazio Region (Bianca Monticellana, Capestrina, and Ciociara Grigia). No individuals were carriers of the CSN1S101, CSN1S1E, CSN201, CSN1S2D, and CSN1S20 alleles, while a high frequency of the alleles CSN1S1F and CSN1S1A*,B* was observed. Association analyses between the different genotypes at the CSN1S1 locus and some milk traits, namely the fat and protein yielded and the fat, protein, solids-not-fat, and casein percentages without an effect on the milk yield, were observed.

Background: Balance impairment is a common disability in post-stroke survivors, leading to reduced mobility and increased fall risk. Robotic gait training (RAGT) is largely used, along with traditional training. There is, however, no strong evidence about RAGT superiority, especially on balance. This study aims to determine RAGT efficacy on balance of post-stroke survivors. Methods: PubMed, Cochrane Library, and PeDRO databases were investigated. Randomized clinical trials evaluating RAGT efficacy on post-stroke survivor balance with Berg Balance Scale (BBS) or Timed Up and Go test (TUG) were searched. Meta-regression analyses were performed, considering weekly sessions, single-session duration, and robotic device used. Results: A total of 18 trials have been included. BBS pre-post treatment mean difference is higher in RAGT-treated patients, with a pMD of 2.17 (95% CI 0.79; 3.55). TUG pre-post mean difference is in favor of RAGT, but not statistically, with a pMD of −0.62 (95%CI − 3.66; 2.43). Meta-regression analyses showed no relevant association, except for TUG and treatment duration (β = −1.019, 95% CI − 1.827; −0.210, p-value = 0.0135). Conclusions: RAGT efficacy is equal to traditional therapy, while the combination of the two seems to lead to better outcomes than each individually performed. Robot-assisted balance training should be the focus of experimentation in the following years, given the great results in the first available trials. Given the massive heterogeneity of included patients, trials with more strict inclusion criteria (especially time from stroke) must be performed to finally define if and when RAGT is superior to traditional therapy.

The human homologue of mouse Ly-1 antibody reactive clone protein (LYAR) is a putative novel regulator of γ-globin gene transcription. The LYAR DNA-binding motif (5′-GGTTAT-3′) is located within the 5′-UTR of the Aγ-globin gene. The LYAR rs368698783 (G>A) polymorphism is present in β-thalassemia patients and decreases the LYAR binding efficiency to the Aγ-globin gene. The objective of this study was to stratify β-thalassemia patients with respect to the rs368698783 (G>A) polymorphism and to verify whether their erythroid precursor cells (ErPCs) differentially respond in vitro to selected fetal hemoglobin (HbF) inducers. The rs368698783 (G>A) polymorphism was detected by DNA sequencing, hemoglobin production by HPLC, and accumulation of globin mRNAs by RT-qPCR. We found that the LYAR rs368698783 (G>A) polymorphism is associated with high basal and induced production of fetal hemoglobin in β-thalassemia patients. The most striking association was found using rapamycin as an HbF inducer. The results presented here could be considered important not only for basic biomedicine but also in applied translational research for precision medicine in personalized therapy of β-thalassemia. Accordingly, our data suggest that the rs368698783 polymorphism might be considered among the parameters useful to recruit patients with the highest probability of responding to in vivo hydroxyurea (HU) treatment.

(1) Background: Treated water distributed in a Latin American megapolis has been the target of criticism in recent years. During the summers of 2020 and 2021, treated water had a taste and smell of mud in the metropolitan area of Rio de Janeiro city, affecting ~10 million people. (2) Methods: To evaluate the SRJ water quality, a comprehensive dataset was analyzed. In total, 194,821 samples were analyzed between 2018 and 2021, corresponding to three parameters (chlorine concentration, n = 67,723; turbidity, n = 55,186; and coliform abundance, n = 71,912). The 92 SRJ cities were analyzed using two approaches according to the relevant law: a quantitative and qualitative approach. (3) Results: In the qualitative analyses, four groups of cities were found (Group A: three parameters analyzed; 49 to 51 cities; Group B, two parameters analyzed, 9 to 14 cities; Group C, one parameter analyzed, 17 cities; Group D, no parameters analyzed, 12 to 16 cities). (4) Conclusions: In 2021, the top ten cities in Group A showed 100% performance in both the qualitative and quantitative rankings. However, several large cities, such as Duque de Caxias and São Gonçalo, did not have the minimum required number of samples or analyses, which poses serious risks for water quality control and public health.

A regression-based approach was used to test the suitability of a range of parameters, including total prokaryotic cell abundance and biomass, as lipopolysaccharide (LPS) content, and exoenzymatic activities (leucine aminopeptidase, LAP, beta-glucosidase, ß-G, and alkaline phosphatase, AP) as predictors of heterotrophic prokaryotic production (HPP) in the Ross Sea epipelagic waters. A close association between HPP and protein hydrolysis mediated by enzymatic activity (LAP), and to a lower significance level with the other variables, was recorded. Three multiple regression equations were developed from two microbial datasets collected during middle austral summer periods. All showed a good predictive ability for HPP, and this was further validated through a comparison between the predicted and the observed HPP values. The obtained regression equations proved to represent a promising example of empirical models for further predictive studies in the Ross Sea where—through the incorporation of additional microbiological and environmental parameters—the developed models could find a practical application to cover the entire austral summer period.

A series of new-generation TMA (4,6,4′-trimethyl angelicin) analogues was projected and synthetized in order to ameliorate anti-inflammatory activity, with reduced or absent toxicity. Since the NF-κB transcription factor (TF) plays a critical role in the expression of IL-8 (Interluekin 8), a typical marker of lung inflammation in Cystic Fibrosis (CF), the use of agents able to interfere with the NF-κB pathway represents an interesting therapeutic strategy. Through preliminary EMSA experiments, we identified several new TMA derivatives able to inhibit the NF-κB/DNA complex. The selected active molecules were then analyzed to evaluate the anti-inflammatory effect using both Pseudomonas aeruginosa (PAO1) infection and TNF-alpha stimulus on the CF IB3-1 cell line. It was demonstrated that mainly two TMA analogues, GY971a mesylate salt (6-p-minophenyl-4,4′-dimethyl-angelicin) and GY964 (4-phenyl-6,4′-dimethyl-angelicin), were able to decrease the IL-8 gene expression. At the same time, these molecules were found to have no pro-apoptotic, mutagenic and phototoxic effects, facilitating our decision to test the efficacy in vivo by using a mouse model of acute P. aeruginosa lung infection. The anti-inflammatory effect of GY971a was confirmed in vivo; this derivative was able to deeply decrease the total number of inflammatory cells, the neutrophil count and the cytokine/chemokine profile in the P. aeruginosa acute infection model, without evident toxicity. Considering all the obtained and reported in vitro and in vivo pre-clinical results, GY971a seems to have interesting anti-inflammatory effects, modulating the NF-κB pathway, as well as the starting lead compound TMA, but without side effects.

In this study, the effects of different operating conditions on excess sludge minimization in an oxic-settling-anaerobic (OSA) process were evaluated. The experiment involved two systems operating in parallel, one implementing the OSA process and a conventional activated sludge (CAS) system as control, both configured according to a pre-denitrification scheme. Five periods (P1–P5) were studied, during which the OSA was operated under different layouts, which differed from the returned sludge to the anoxic (A) or aerobic (B) mainstream reactors and the hydraulic retention time in the anaerobic reactor of the OSA system (8–12 h). The excess sludge production in the OSA plant was lower in all the investigated configurations, indicating that successful sludge minimization was achieved. Specifically, the sludge production was lowered by approximately 12% (P1), 29% (P2), 40% (P3), 26% (P4) and 41% (P5). Scheme A enabled the establishment of the uncoupling metabolism and the extracellular polymeric substance (EPS) destructuration. In contrast, scheme B enabled the establishment of the maintenance metabolism in addition to the uncoupling metabolisms, whereas cell lysis and EPS destruction were minimized. This allowed for obtaining higher sludge reduction yield (26–40%) without compromising the effluent quality.

Several studies were conducted previously on fuel and biofuel performance of micro-turbines. The present paper combines experimental and statistical approaches to study the vibrational behavior of a gas micro-turbine supplied with different pure fuels and admixed with rapeseed oils. Experimental tests carried out at different operating conditions have allowed us to build a classification model through using discriminant analysis. The classification model can distinguish the vibrational behavior occurring when the turbine is fueled with kerosene, or pure and admixed diesel with rapeseed oil. Moreover, the methodology has even allowed us to highlight differences in vibrational behavior caused by small amounts of rapeseed oil admixed in the fuel. The model reliability, in terms of Cohen’s kappa, results in optimal data classification.

Conversion of wastewater treatment plants into biorefineries is a sustainable alternative for obtaining valuable compounds, thus reducing pollutants and costs and protecting the environment and human health. Under specific operating conditions, microbial fermentative products of sewage sludge are volatile fatty acids (VFA) that can be precursors of polyhydroxyalkanoate thermoplastic polyesters. The role of various operating parameters in VFA production has yet to be elucidated. This study aimed to correlate the levels of VFA yields with prokaryotic microbiota structures of sewage sludge in two sets of batch fermentations with an initial pH of 8 and 10. The sewage sludge used to inoculate the batch fermentations was collected from a Sicilian WWTP located in Marineo (Italy) as a case study. Gas chromatography analysis revealed that initial pH 10 stimulated chemical oxygen demands (sCOD) and VFA yields (2020 mg COD/L) in comparison with initial pH 8. Characterization of the sewage sludge prokaryotic community structures—analyzed by next-generation sequencing of 16S rRNA gene amplicons—demonstrated that the improved yield of VFA paralleled the increased abundance of fermenting bacteria belonging to Proteobacteria, Bacteroidetes, Chloroflexi, and Firmicutes phyla and, conversely, the reduced abundance of VFA-degrading strains, such as archaeal methanogens.

We investigate the effects of external and autonomous global interaction fields on an adaptive network of social agents with an opinion formation dynamics based on a simple imitation rule. We study the competition between global fields and adaptive rewiring on the space of parameters of the system. The model represents an adaptive society subject to global mass media such as a directed opinion influence or feedback of endogenous cultural trends. We show that, in both situations, global mass media contribute to consensus and to prevent the fragmentation of the social network induced by the coevolutionary dynamics. We present a discussion of these results in the context of dynamical systems and opinion formation dynamics.

Botulinum Neurotoxin type-A (BoNT-A) is the treatment of choice for focal post-stroke spasticity (PSS). Due to its mechanism of action and the administration method, some authors raised concern about its possible systemic diffusion leading to contralateral muscle weakness and autonomic nervous system (ANS) alterations. Stroke itself is a cause of motor disability and ANS impairment; therefore, it is mandatory to prevent any source of additional loss of strength and adjunctive ANS disturbance. We enrolled 15 hemiparetic stroke survivors affected by PSS already addressed to BoNT-A treatment. Contralateral handgrip strength and ANS parameters, such as heart rate variability, impedance cardiography values, and respiratory sinus arrythmia, were measured 24 h before (T0) and 10 days after (T1) the ultrasound (US)-guided BoNT-A injection. At T1, neither strength loss nor modification of the basal ANS patterns were found. These findings support recent literature about the safety profile of BoNT-A, endorsing the importance of the US guide for a precise targeting and the sparing of “critical” structures as vessels and nerves.

Wastewaters generated by crude oil extraction processes, called “produced waters” (PWs), are complex solutions that contain organic compounds, mainly hydrocarbons, and often exhibit high salinity. The large amounts of PWs represent a global issue because of their environmental impact. An approach widely used in the oil industry is the reinjection of this wastewater into the extraction wells after a suitable treatment. The high salt concentration of such solutions may be used in salinity gradient technologies to produce green electricity. Among these technologies, reverse electrodialysis (RED) is one of the most promising. In this work, the application of RED for energy generation from two different real oil industry brines was investigated. An experimental campaign was performed by testing 10 × 10 cm2 units in long-run continuous operations, monitoring the performance for more than 25 days. Fouling phenomena, occurring during the continuous operation, decrease the unit performance and several anti-fouling strategies were adopted to tackle this issue. As a result, a positive net power density for up to 18 days of continuous operation was obtained. A maximum power density of about 2.5 W/m2 was observed, demonstrating how the RED technology could be an important strategy to harvest energy from an industrial waste.

The employment of vehicles such as rovers equipped with automictic and robotic systems in agriculture is an emerging field. The development of suitable simulation models can aid in the design and testing of agricultural rovers before prototyping. Here, we propose a simulation test rig based on a multibody model to investigate the main issues connected with agricultural rover designs. The results of the simulations show significant differences between the two structures, especially regarding the energy savings, which is a key aspect for the applicability of a rover in field operations. The modular structure of the proposed simulation model can be easily adapted to other vehicle structures.

The current exploitation of freshwater, as well as the significant increase in sewage sludge production from wastewater treatment plants (WWTPs), represent nowadays a critical issue for the implementation of sustainable development consistent with the circular economy concept. There is an urgent need to rethink the concept of WWTPs from the conventional approach consisting in pollutant removal plants to water resource recovery facilities (WRRFs). The aim of this paper is to provide an overview of the demonstration case studies at the Marineo and Corleone WRRFs in Sicily (IT), with the final aim showing the effectiveness of the resources recovery systems, as well as the importance of plant optimization to reduce greenhouse gas (GHG) emissions from WRRFs. This study is part of the H2020 European Project “Achieving wider uptake of water-smart solutions—Wider-Uptake”, which final aim is to demonstrate the water-smart solution feasibility in the wastewater sector. The main project goal is to overcome the existing barriers that hamper the transition to circularity through the implementation of a governance analysis tool. The preliminary actions in the two demonstration cases are first presented, while, subsequently, the water-smart solutions to be implemented are thoroughly described, highlighting their roles in the transition process. The achieved preliminary results underlined the significant potential of WRRF application, a great chance to demonstrate the feasibility of innovative solutions in the wastewater sector to overcome the existing social, administrative and technical barriers.

The wastewater sector paradigm is shifting from wastewater treatment to resource recovery. In addition, concerns regarding sustainability during the operation have increased. In this sense, there is a need to break barriers (i.e., social, economic, technological, legal, etc.) for moving forward towards water resource recovery facilities and demonstration case studies can be very effective and insightful. This paper presents a new water resource recovery case study which is part of the Horizon 2020 EU Project “Achieving wider uptake of water-smart solutions—Wider Uptake”. The final aim is to demonstrate the importance of a resource recovery system based on the circular economy concept. The recovery facilities at Palermo University (Italy) are first presented. Afterwards, the resource recovery pilot plants are described. Preliminary results have underlined the great potential of the wastewater treatment plant in terms of resources recovery and the central role of the University in fostering the transition towards circular economy. The fermentation batch test highlighted a volatile fatty acids (VFAs) accumulation suitable for polyhydroxyalkanoates (PHAs) production. The results of static adsorption and desorption tests showed that the highest amount of adsorbed NH4+ was recorded for untreated and HCl-Na treated clinoptilolite.

β-thalassemias are among the most common inherited hemoglobinopathies worldwide and are the result of autosomal mutations in the gene encoding β-globin, causing an absence or low-level production of adult hemoglobin (HbA). Induction of fetal hemoglobin (HbF) is considered to be of key importance for the development of therapeutic protocols for β-thalassemia and novel HbF inducers need to be proposed for pre-clinical development. The main purpose on this study was to analyze Cinchona alkaloids (cinchonidine, quinidine and cinchonine) as natural HbF-inducing agents in human erythroid cells. The analytical methods employed were Reverse Transcription quantitative real-time PCR (RT-qPCR) (for quantification of γ-globin mRNA) and High Performance Liquid Chromatography (HPLC) (for analysis of the hemoglobin pattern). After an initial analysis using the K562 cell line as an experimental model system, showing induction of hemoglobin and γ-globin mRNA, we verified whether the two more active compounds, cinchonidine and quinidine, were able to induce HbF in erythroid progenitor cells isolated from β-thalassemia patients. The data obtained demonstrate that cinchonidine and quinidine are potent inducers of γ-globin mRNA and HbF in erythroid progenitor cells isolated from nine β-thalassemia patients. In addition, both compounds were found to synergize with the HbF inducer sirolimus for maximal production of HbF. The data obtained strongly indicate that these compounds deserve consideration in the development of pre-clinical approaches for therapeutic protocols of β-thalassemia.

Equinovarus foot is one of the most commonly spasticity related conditions in stroke survivors, leading to an impaired gait and poor functional performances. Notably, spastic muscles undergo a dynamic evolution following typical pathophysiological patterns. Botulinum Neurotoxin Type A (BoNT-A) is the gold standard for focal spasticity treatment, and ultrasound (US) imaging is widely recommended to guide injections and monitor muscle evolution. The role of BoNT-A in influencing muscle fibroadipose degeneration is still unclear. In this study, we analyzed medial gastrocnemius (MG) and soleus (SOL) US characteristics (cross-sectional area, muscle thickness, pennation angle, and mean echo intensity) in 53 patients. MG and SOL alterations, compared to the unaffected side, depend on the spasticity only and not on the BoNT-A treatment. In functionally preserved patients (functional ambulation classification, FAC > 3; modified Ashworth scale, MAS ≤ 2), the ultrasonographic changes of MG compared to ipsilateral SOL observed in the paretic limb alone seems to be due to histological, anatomical, pathophysiological, and biomechanical differences between the two muscles. In subjects with poor walking capability and more severe spasticity, such ipsilateral difference was found in both calves. In conclusion, BoNT-A does not seem to influence muscle degeneration. Similar muscles undergo different evolution depending on the grade of walking deficit and spasticity.

Hand prostheses partially restore hand appearance and functionalities. In particular, 3D printers have provided great opportunities by simplifying the manufacturing process and reducing costs. The “Federica” hand is 3D-printed and equipped with a single servomotor, which synergically actuates its five fingers by inextensible tendons; no springs are used for hand opening. A differential mechanical system simultaneously distributes the motor force on each finger in predefined portions. The proportional control of hand closure/opening is achieved by monitoring muscle contraction by means of a thin force sensor, as an alternative to EMG. The electrical current of the servomotor is monitored to provide sensory feedback of the grip force, through a small vibration motor. A simple Arduino board was adopted as the processing unit. A closed-chain, differential mechanism guarantees efficient transfer of mechanical energy and a secure grasp of any object, regardless of its shape and deformability. The force sensor offers some advantages over the EMG: it does not require any electrical contact or signal processing to monitor muscle contraction intensity. The activation speed (about half a second) is high enough to allow the user to grab objects on the fly. The cost of the device is less then 100 USD. The “Federica” hand has proved to be a lightweight, low-cost and extremely efficient prosthesis. It is now available as an open-source project (CAD files and software can be downloaded from a public repository), thus allowing everyone to use the “Federica” hand and customize or improve it.

Background: Autism spectrum disorders (ASD) are a heterogeneous group of neurodevelopmental conditions whose etiopathogenesis derives from a complex interaction between genetic liability and environmental factors. In this framework, mounting evidence suggests that immune system dysfunction could be a risk factor contributing to the development of ASD in at least a subpopulation of individuals. In particular, some studies suggest an association between celiac disease (CD)—a long-term autoimmune disorder that primarily affects the small intestine triggered by the ingestion of gluten—and ASD, while others hypothesized a random link. This investigation aimed to evaluate the prevalence of CD in a large sample of school-aged children with ASD and to characterize their clinical profile. Methods: Medical records of 405 children with ASD aged 5–11 years (mean age: 7.2 years; SD: 1.8 years) consecutively referred to a tertiary-care university hospital between January 2014 and December 2018 were reviewed; among them, 362 had carried out serological testing for CD. Results: Nine patients with positive CD serology were identified, eight of which satisfied the criteria for CD diagnosis. The estimated CD prevalence in ASD children was 2.18% (95% CI, 0.8–3.7), which was not statistically different (1.58%; p = 0.36) from that of an Italian population, matched for age range, considered as a control group (95% CI, 1.26–1.90). Three out of the eight ASD patients with CD did not have any symptoms suggestive of CD. Conclusions: Our findings did not show a higher prevalence of CD in ASD children than in the control population, but could suggest the utility of routine CD screening, given its frequent atypical clinical presentation in this population.

Masseter Muscle Hypertrophy (MMH) is a well-known clinical benign condition that is not gender-specific and it can be monolateral or bilateral. Botulinum Toxin type A (BoNTA) injection has been widely described for MMH treatment and non-surgical facial slimming. BoNTA masseter injections have high efficacy and safety profile, but the risks of side effects remain. Muscular bulging during mastication is a complication due to the superficial overcompensation of masseteric fibers in response to neurotoxic weakening of the deep masseter. We present a biphasic-injection technique for BoNTA administration, based following anatomical concept and developed in order to prevent paradoxical bulging. A total of 98 treatments from 2015 to 2020 were performed with this technique. No remarkable complications occurred in our study. No cases of loss of full smile, difficulty in mouth opening, dizziness, headache, neurapraxia, and xerostomia were reported. A case of asymmetric smiling was self-resolved within a week. No patient claimed transient muscle weakness as distressing. No cases of paradoxical bulging were observed. Extensive knowledge of muscular anatomy and appropriate injection technique are key factors in achieving the desired result and avoiding complications. We feel that sharing this tip could be helpful for all the physicians involved in MMH treatment with BoNTA.

Cytokines are a broad group of small regulatory proteins with many biological functions involved in regulating the hematopoietic and immune systems. However, in pathological conditions, hyperactivation of the cytokine network constitutes the fundamental event in cytokine release syndrome (CRS). During the last few decades, the development of therapeutic monoclonal antibodies and T-cell therapies has rapidly evolved, and CRS can be a serious adverse event related to these treatments. CRS is a set of toxic adverse events that can be observed during infection or following the administration of antibodies for therapeutic purposes and, more recently, during T-cell-engaging therapies. CRS is triggered by on-target effects induced by binding of chimeric antigen receptor (CAR) T cells or bispecific antibody to its antigen and by subsequent activation of bystander immune and non-immune cells. CRS is associated with high circulating concentrations of several pro-inflammatory cytokines, including interleukins, interferons, tumor necrosis factors, colony-stimulating factors, and transforming growth factors. Recently, considerable developments have been achieved with regard to preventing and controlling CRS, but it remains an unmet clinical need. This review comprehensively summarizes the pathophysiology, clinical presentation, and treatment of CRS caused by T-cell-engaging therapies utilized in the treatment of hematological malignancies.