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Abstract Background Homozygous familial hypercholesterolemia (HoFH) is a rare life-threatening condition that represents a therapeutic challenge. The vast majority of HoFH patients fail to achieve LDL-C targets when treated with the standard protocol, which associates maximally tolerated dose of lipid-lowering medications with lipoprotein apheresis (LA). Lomitapide is an emerging therapy in HoFH, but its place in the treatment algorithm is disputed because a comparison of its long-term efficacy versus LA in reducing LDL-C burden is not available. We assessed changes in long-term LDL-C burden and goals achievement in two independent HoFH patients’ cohorts, one treated with lomitapide in Italy (n = 30) and the other with LA in France (n = 29). Results The two cohorts differed significantly for genotype (p = 0.004), baseline lipid profile (p padj = 0.004). Yearly on-treatment LDL-C padj = 0.002). A trend towards fewer total cardiovascular events was observed in the Lomitapide than in the LA cohort. Conclusions In comparison with LA, lomitapide appears to provide a better control of LDL-C in HoFH. Further studies are needed to confirm this data and establish whether this translates into a reduction of cardiovascular risk.

The expansion of the electric vehicle market is driving the request for efficient and reliable power electronic systems for electric energy conversion and processing. The efficiency, size, and cost of a power system is strongly related to the performance of power semiconductor devices, where massive industrial investments and intense research efforts are being devoted to new wide bandgap (WBG) semiconductors, such as silicon carbide (SiC) and gallium nitride (GaN). The electrical and thermal properties of SiC and GaN enable the fabrication of semiconductor power devices with performance well beyond the limits of silicon. However, a massive migration of the power electronics industry towards WBG materials can be obtained only once the corresponding fabrication technology reaches a sufficient maturity and a competitive cost. In this paper, we present a perspective of power electronics based on WBG semiconductors, from fundamental material characteristics of SiC and GaN to their potential impacts on the power semiconductor device market. Some application cases are also presented, with specific benchmarks against a corresponding implementation realized with silicon devices, focusing on both achievable performance and system cost.

Background NEPA is an oral fixed-dose combination of netupitant, a new highly selective neurokinin-1 receptor antagonist, and palonosetron. This study was conducted to evaluate whether the efficacy of NEPA against chemotherapy-induced nausea and vomiting (CINV) in cycle 1 would be maintained over subsequent chemotherapy cycles in breast cancer patients receiving adjuvant anthracycline plus cyclophosphamide (AC). The study also describes the relationship between efficacy on day 1 through 5 (overall period) and control of CINV on day 6 through 21 (very late period) in each cycle. Methods In this multicentre, phase II study, patients received both NEPA and dexamethasone (12 mg intravenously) just before chemotherapy. The primary efficacy endpoint was overall complete response (CR; no emesis and no rescue medication use) in cycle 1. Sustained efficacy was evaluated during the subsequent cycles by calculating the rate of CR in cycles 2–4 and by assessing the probability of sustained CR over multiple cycles. The impact of both overall CR and risk factors for CINV on the control of very late events (vomiting and moderate-to-severe nausea) were also examined. Results Of the 149 patients enrolled in the study, 139 were evaluable for a total of 552 cycles; 97.8% completed all 4 cycles. The proportion of patients with an overall CR was 70.5% (90% CI, 64.1 to 76.9) in cycle 1, and this was maintained in subsequent cycles. The cumulative percentage of patients with a sustained CR over 4 cycles was 53%. NEPA was well tolerated across cycles. In each cycle, patients with CR experienced a significantly better control of very late CINV events than those who experienced no CR. Among the patients with CR, the only predictor for increased likelihood of developing very late CINV was pre-chemotherapy (anticipatory) nausea (adjusted odds ratio = 0.65–0.50 for no CINV events on cycles 3 and 4). Conclusion The high anti-emetic efficacy seen with the NEPA regimen in the first cycle was maintained over multiple cycles of adjuvant AC for breast cancer. Preliminary evidence also suggests that patients achieving a CR during the overall period gain high protection even against very late CINV events in each chemotherapy cycle. Trial registration This trial was retrospectively registered at Clinicaltrials.gov identifier (NCT03862144) on 05/Mar/2019.

Background In the last years, functional imaging has given a significant contribution to the clinical decision-making of biochemically relapsed prostate cancer (PCa). Hereby, we present a prospective study aiming to validate the role of [18F]Fluoro-Methyl Choline ([18F]FMCH) PET/CT in the selection of PCa patients suitable for stereotactic body radiotherapy (SBRT). Methods Patients with biochemical recurrence limited up to three lesions revealed by [18F]FMCH PET/CT were enrolled in the present study and treated with SBRT on all active lesions. Systemic therapy-free survival since the [18F]FMCH PET/CT was considered as the primary endpoint. Results Forty-six patients were evaluated, and a total of 67 lesions were treated. After a median follow-up of 28.9 months, systemic therapy was started in 30 patients (65.2%) and median systemic therapy-free survival was 39.1 months (95% CI 6.5–68.6); 6, 12, and 24-month ratios were 93.5%, 73.9%, and 63.1%, respectively. At univariate Cox regression analysis, Delta PSA demonstrated an impact on systemic therapy-free survival (p Conclusions Based on our findings, [18F]FMCH PET/CT can identify oligometastatic prostate cancer patients suitable for SBRT, resulting in a systemic therapy-free survival of 39.1 months.

In vitro cell cultures are often proposed as “Alternatives” to animal models, but they are still inadequate to reproduce the human pathophysiology. This is mainly due to the technological limitations of the standard equipment used in cell culture laboratories, such as the lack of a 3D micro-architecture, the static environment and the absence of cross talk between different tissues. These limitations cause the poor predictivity of a standard in-vitro model, if compared with the human reality.

The development of cannabinoid receptor type-1 (CB1R) modulators has been implicated in multiple pathophysiological events ranging from memory deficits to neurodegenerative disorders among others, even if their central psychiatric side effects such as depression, anxiety, and suicidal tendencies, have limited their clinical use. Thus, the identification of ligands which selectively act on peripheral CB1Rs, is becoming more interesting. A recent study reported a class of peripheral CB1R selective antagonists, characterized by a 5-aryl substituted nicotinamide core. These derivatives have structural similarities with the biphenyl compounds, endowed with CB2R antagonist activity, previously synthesized by our research group. In this work we combined the pharmacophoric portion of both classes, in order to obtain novel CBR antagonists. Among the synthesized compounds rather unexpectedly two compounds of this series, C7 and C10, did not show the radioligand ([

Background: Real-world data on chemotherapy-naïve patients with metastatic castration-resistant prostate cancer (mCRPC) treated with abiraterone plus prednisone are limited, largely deriving from small retrospective studies. Methods: ABitude is an Italian, observational, prospective, multicenter study of mCRPC patients receiving abiraterone plus prednisone in clinical practice. Chemotherapy-naïve mCRPC patients were consecutively enrolled at abiraterone start (February 2016 to June 2017) and are being followed for 3 years, with evaluation approximately every 6 months. Several clinical and patients reported outcomes were examined. Results: In this second interim analysis, among 481 enrolled patients, 453 were evaluable for analyses. At baseline, the median age was 77 years and ~69% of patients had comorbidities (mainly cardiovascular diseases). Metastases were located mainly at bones and lymph nodes; 8.4% of patients had visceral metastases. During a median follow-up of 18 months, 1- and 2-year probability of radiographic progression-free survival were 73.9% and 56.2%, respectively; the corresponding rates for overall survival were 87.3% and 70.4%. In multivariable analyses, the number of bone metastases significantly affected radiographic progression-free survival and overall survival. During abiraterone plus prednisone treatment, 65% of patients had a ⩾50% prostate-specific antigen decline, and quality of life remained appreciably high. Among symptomatic patients according to the Brief Pain Inventory) (32%), scores significantly declined after 6 months of treatment. Overall, eight patients (1.7%) had serious adverse reactions to abiraterone. Conclusions: Abiraterone plus prednisone is effective and safe for chemotherapy-naïve mCRPC patients in clinical practice.

Androgen deprivation therapy (ADT) is a cornerstone of treatment for prostate cancer and, in recent years, androgen receptor (AR)-targeted therapies (abiraterone and enzalutamide) have both been used for the treatment of castration-resistant prostate cancer (CRPC). In our study, we sought to investigate the association between ADT and immune disorders, considering a potential role of androgens in the immune modulation. We retrospectively evaluated CRPC patients treated with abiraterone/enzalutamide between July 2011 and December 2018. We assessed the risk of developing immune alterations and their impact on outcome. We included 844 CRPC patients receiving AR-directed therapies, of whom 36 (4.3%) had autoimmune diseases and 47 (5.6%) second tumors as comorbidities. Median age was 70 years [interquartile range (IQR) = 63&ndash, 75)]. We showed higher significant incidence of autoimmune diseases during their hormone sensitive status (p = 0.021) and the presence of autoimmune comorbidities before starting treatment with abiraterone/enzalutamide was significantly associated with worse overall survival (OS) (10.1 vs. 13.7 months, HR = 1.59, 95% CI 1.03&ndash, 2.27, p = 0.038). In a multivariate analysis, the presence of autoimmune disorders was an independent predictor of OS (HR = 1.65, 95% CI 1.05&ndash, 2.60, p = 0.031). In conclusion, CRPC patients with autoimmune alterations before starting AR-directed therapies may have worse prognosis. Further prospective studies are warranted to assess the role of immune modulation in the management of prostate cancer patients.

Holotomographic (HT) microscopy, combines two techniques, holography and tomography, and, in this way, it allows to quantitatively and noninvasively investigate cells and thin tissue slices, by obtaining three-dimensional (3D) images and by monitoring inner morphological changes. HT has indeed two significant advantages: it is label-free and low-energy light passes through the specimen with minimal perturbation. Using quantitative phase imaging with optical diffraction tomography, it can produce 3D images by measuring the refraction index (RI). Therefore, based on RI values, HT can provide structural and chemical cell information, such as dry mass values, morphological changes, or cellular membrane dynamics. In this study, suspended and adherent culture cells have been processed for HT analyses. Some of them have been treated with known apoptotic drugs or pro-oxidant agents and cell response has been investigated both by conventional microscopic approaches and by HT. The ultrastructural and fluorescence images have been compared to those obtained by HT and their congruence has been discussed, with particular attention to apoptotic cell death and on correlated plasma membrane changes. HT appears a valid approach to further characterize well-known apoptotic features such as cell blebbing, chromatin condensation, micronuclei, and apoptotic bodies. Taken together, our data demonstrate that HT appears suitable to highlight suspended or adherent cell behavior under different conditions. In particular, this technique appears an important new tool to distinguish healthy cells from the apoptotic ones, as well as to monitor outer and inner cell changes in a rapid way and with a noninvasive, label-free, approach.

Background: Exercise may affect lipid profile which in turn is related to inflammation, although changes of ceramides, diacylglycerols-DAG and sphingomyelin-SM and their relationship with inflammatory parameters following a half-marathon have never been examined. Methods: Ceramides, DAG and SM, and markers of inflammation (soluble fractalkine-CX3CL1, vascular endothelial growth factor-VEGF, interleukin6-IL-6 and tumor necrosis factorα-TNFα) were evaluated in trained half-marathoners before, post-race (withdrawal within 20 min after the race end) and 24 h after. Results: IL-6 and CX3CL1 increased immediately after the race, returning to baseline after 24 h. Total ceramides and total DAG significantly decreased post-race. Several ceramide classes decreased after exercise, while only one of the DAG (36:3) changed significantly. Total SM and specific species did not significantly change. Conclusion: Some inflammatory parameters (IL-6 and CX3CL1) transiently increased after the race, and, being reversible, these changes might represent a physiological response to acute exercise rather than a damage-related response. The decrease of specific lipid classes, i.e., DAGs and ceramides, and the lack of their relationship with inflammatory parameters, suggest their involvement in beneficial training effects, opening promising research perspectives to identify additional mechanisms of aerobic exercise adaptation.

A novel green method for graphene oxide (GO) reduction via ascorbic acid has been adopted to realize bio-friendly reduced graphene oxide (RGO)/polycaprolactone (PCL) nanofibrous meshes, as substrates for bone tissue engineering applications. PCL fibrous mats enriched with either RGO or GO (0.25 wt%) were fabricated to recapitulate the fibrillar structure of the bone extracellular matrix (ECM) and the effects of RGO incorporation on the structural proprieties, biomechanics and bioactivity of the nano-composites meshes were evaluated. RGO/PCL fibrous meshes displayed superior mechanical properties (i.e. Young's Modulus and ultimate tensile strength) besides supporting noticeably improved cell adhesion, spreading and proliferation of fibroblasts and osteoblast-like cell lines. Furthermore, RGO-based electrospun substrates enhanced in vitro calcium deposition in the ECM produced by osteoblast-like cells, which was paralleled, in human mesenchymal stem cells grown onto the same substrates, by an increased expression of the osteogenic markers mandatory for mineralization. In this respect, the capability of graphene-based materials to adsorb osteogenic factors cooperates synergically with the rougher surface of RGO/PCL-based materials, evidenced by AFM analysis, to ignite mineralization of the neodeposited matrix and to promote the osteogenic commitment of the cultured cell in the surrounding microenvironment.

Innate immune cells such as monocytes and macrophages are activated in response to microbial and other challenges and mount an inflammatory defensive response. Exposed cells develop the so-called innate memory, which allows them to react differently to a subsequent challenge, aiming at better protection. In this study, using human primary monocytes in vitro, we have assessed the memory-inducing capacity of two antigenic molecules of Schistosoma mansoni in soluble form compared to the same molecules coupled to outer membrane vesicles of Neisseria lactamica. The results show that particulate challenges are much more efficient than soluble molecules in inducing innate memory, which is measured as the production of inflammatory and anti-inflammatory cytokines (TNFα, IL-6, IL-10). Controls run with LPS from Klebsiella pneumoniae compared to the whole bacteria show that while LPS alone has strong memory-inducing capacity, the entire bacteria are more efficient. These data suggest that microbial antigens that are unable to induce innate immune activation can nevertheless participate in innate activation and memory when in a particulate form, which is a notion that supports the use of nanoparticulate antigens in vaccination strategies for achieving adjuvant-like effects of innate activation as well as priming for improved reactivity to future challenges.

Objectives The aim of our study was to collect data about of the outcome of metastatic renal cell carcinoma patients who progressed after immune checkpoint inhibitors in order to enhance data about efficacy and safety of treatment beyond immune-oncology (IO). Materials and methods A total of 162 eligible patients, progressing to IO, were enrolled from 16 Italian referral centers adhering to the Meet-Uro association. Baseline characteristics, outcome data and toxicities were retrospectively collected. Descriptive analysis was made using median values and ranges. Kaplan-Meier method and Mantel-Haenszel log-rank test were performed to compare differences between groups. Results A total of 111 patients (68.5%) were treated after IO progression. In all, 51 patients (31.5%) did not receive further treatment for clinical deterioration. Median IO progression free survival (PFS) was 4 months (95% confidence interval [CI]: 3.1-4.8). IO-PFS tends to be longer in patients reporting adverse events (AE) of any grade (5.03 [95% CI: 3.8-6.1] vs. 2.99 [95% CI: 2.4-3.5] months P=0.004). Subsequent therapies included cabozantinib (n=79, 48%), everolimus (n=11, 6.7%), and others (n=21, 12.9%).Median PFS post-IO was 6.5 months (95% CI: 5.1-7.8). Cabozantinib showed longer PFS compared with everolimus (7.6 mo [95% CI: 5.2-10.1] vs. 3.2 mo [95% CI: 1.8-4.5]) (hazard ratio: 0.2; 95% CI: 0.1026-0.7968) and other drugs (4.3 mo [95% CI: 1.3-7.4]) (hazard ratio: 0.6; 95% CI: 0.35-1.23). All grade AE were reported in 83 patients (74%) and G3 to G4 AE in 39 patients (35%). Target therapies post-IO showed median overall survival of 14.7 months (95% CI: 0.3-21.4). Conclusions In our real world experience after progression to IO, vascular endotelial groth factor-tyrosine kinase inhibitors, given to patients, proved to be active and safe choices. Cabozantinib was associated with a better outcome in terms of median PFS.

BACKGROUND Modulation of oxidative stress/inflammation during exercise may have both positive and negative health effects, depending by a number of factors (e.g. training status, and exercise type, intensity and duration) and the oxidative stress or inflammation-related biomarkers considered, which may reflect different levels of the oxidative stress/inflammatory multi entities. The aim of this study was to evaluate oxidative stress and inflammatory multi-biomarker panel in response to a half-marathon during early and delayed recovery. METHODS Blood samples (baseline, postrace within 20 min after the race end, and 24 h and 48 h after the run) from runners (N.=31, 20 males, mean age 47±6 years) were assessed for reactive oxygen species (ROM assay) and total antioxidant capacity (OXY test), leukocyte telomere length (LTL), procoagulant activity of circulating microparticles (MP-PCA), inflammatory parameters obtained by hemocrome, and irisin. RESULTS A significant decrease for OXY (from 375±71 to 280±66, 239±54, 239±45 µmolHClO/mL) after the half-marathon and during recovery was observed. A reduction for ROMs was also evidenced respect to baseline (from 328±46 to 301±39, 290±56, 320±55 AU). Instead, MP-PCA increased after the race (from 6.2±6 to 10.5±6, 7±4.3 and 5.8±2.1 nmol/L), whereas the other biomarkers did not significantly change. CONCLUSIONS The oxidant counterpart did not increase in response to the half-marathon, likely counteracted by antioxidants, which appeared greatly worn out. MP-PCA and WBC increase, always within the normality range, may represent an adaptation to regular chronic endurance training. In any case, antioxidant supply could be considered and tailored for each athlete in this exercise setting.

The spatial and temporal changes of morphological and mechanical properties of living cells reflect complex functionally-associated processes. Monitoring these modifications could provide a direct information on the cellular functional state. Here we present an integrated biophysical approach to the quantification of the morphological and mechanical phenotype of single cells along a maturation pathway. Specifically, quantitative phase microscopy and single cell biomechanical testing were applied to the characterization of the maturation of human foetal osteoblasts, demonstrating the ability to identify effective label-free biomarkers along this fundamental biological process.

Chronic lymphocytic leukemia (CLL) cells disseminate into supportive tissue microenvironments. To investigate the mechanisms involved in leukemic cell tissue retention we developed a threedimensional bone marrow (BM) microenvironment that recreates the interactions between CLL and BM stromal cells inside a scaffold within a bioreactor. Our system allows the parallel analysis of CLL cells retained inside the scaffold and those released in the presence/absence of pharmacological agents, mimicking tissue and circulating cell compartments, respectively. CLL cells can be retained within the scaffold only in the presence of microenvironmental elements, which through direct contact downregulate the expression of HS1 cytoskeletal protein in CLL cells. Consistent with this, the expression of HS1 was lower in CLL cells obtained from patients’ BM than in CLL cells circulating in the peripheral blood. Moreover, we demonstrate that CLL cells with inactive HS1, impaired cytoskeletal activity and a more aggressive phenotype are more likely to be retained within the scaffold despite the presence of ibrutinib, whose mobilizing effect is mainly exerted on those with active HS1, ensuing dynamic cytoskeletal activity. This differential effect would not otherwise be assessable in a traditional two-dimensional system and may underlie a distinctive resistance of single CLL clones. Notably, CLL cells mobilized in the peripheral blood of patients during ibrutinib therapy exhibited activated HS1, underscoring that our model reliably mirrors the in vivo situation. The three-dimensional model described herein is suitable for reproducing and identifying critical CLL-BM interactions, opening the way to pathophysiological studies and the evaluation of novel targeted therapies in an individualized manner.

Microplastic presence in the marine environment has generated considerable concern. Many procedures for microplastics detection in fish gastrointestinal tract have been recently developed. In this study, we compared efficiencies of two common procedures applied for the digestion of organic matter (10% KOH; 15% H2O2) with a new proposal (mixture of 5% HNO3 and 15% H2O2). We considered ecological diversity among species and differences in their diet compositions as factors that could affect the efficiency and feasibility of analytical approaches. Our aim was to understand whether either one of the three protocols might be suitable for all species or it might be more advisable to select a method according to the gut content determined by different food preferences. The results showed that the trophic level and feeding habits should be considered for protocol selection. Finally, we applied the best protocols on samples from the Tyrrhenian sea.

For the first time, we report a correspondence between microplastics (MP) ingestion and ecotoxicological effects in gelatinous zooplankton (Cnidarian jellyfish). The ephyra stage of the jellyfish Aurelia sp. was exposed to both environmental and high concentrations of fluorescent 1–4 μm polyethylene MP (0.01–10 mg/L). After 24 and 48 h, MP accumulation, acute (Immobility) and behavioral (Frequency pulsation) endpoints were investigated. MP were detected by confocal and tomographic investigations on gelatinous body and mouth, either attached on the surface or ingested. This interaction was responsible for impairing ephyrae survival and behavior at all tested concentrations after 24 h. Acute and behavioral effects were also related to mechanical disturbance, caused by MP, triggering a loss of radial symmetry. Contaminated ephyrae exposed to clean seawater showed full recovery after 72 h highlighting the organisms without the microspheres, attached on body jellyfish surface around the mouth and lappets. In conclusion, short-term exposure to MP affects ephyrae jellyfish health, impairing both their survival and behavior. Polyethylene MP temporarily affect both Immobility and Frequency of pulsation of Aurelia sp. jellyfish. This study provides a first step towards understanding and clarifying the potential impacts of MP contamination in gelatinous zooplankton.

The emerging outbreak of the coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread worldwide. We prescribed some promising medication to our patients with mild to moderate pneumonia due to SARS-CoV-2, however such drugs as chloroquine, hydrossichloroquine, azithromycin, antivirals (lopinavir/ritonavir, darunavir/cobicistat) and immunomodulating agents (steroids, tocilizumab) were not confirmed as effective against SARS-CoV2. We, therefore, started to use auto-hemotherapy treated with an oxygen/ozone (O2/O3) gaseous mixture as adjuvant therapy. In Udine University Hospital (Italy) we performed a case–control study involving hospitalized adult patients with confirmed COVID-19 with mild to moderate pneumonia. Clinical presentations are based upon clinical phenotypes identified by the Italian Society of Emergency and Urgency Medicine (SIMEU—Società Italiana di Medicina di Emergenza-Urgenza) and patients that met criteria of phenotypes 2 to 4 were treated with best available therapy (BAT), with or without O3-autohemotherapy. 60 patients were enrolled in the study: 30 patients treated with BAT and O2/O3 mixture, as adjuvant therapy and 30 controls treated with BAT only. In the group treated with O3-autohemotherapy plus BAT, patients were younger but with more severe clinical phenotypes. A decrease of SIMEU clinical phenotypes was observed (2.70 ± 0.67 vs. 2.35 ± 0.88, p = 0.002) in all patients during hospitalization but this clinical improvement was statistically significant only in O3-treated patients (2.87 ± 0.78 vs. 2.27 ± 0.83, p

The concept of food quality, traditionally based on nutritional and sensory properties, has recently acquired an additional meaning, referring to the health-promoting properties of plant products, that are ascribed to plant secondary metabolites called phytochemicals, primarily represented by polyphenolic compounds and glucosinolates. The diversity and content of phytochemicals in plant products are affected by different variables, such as plant genotype, agronomic factors, and arbuscular mycorrhizal fungi (AMF), which establish mycorrhizal symbioses with most crops, including cereals, legumes, vegetables, fruit trees, sunflower, cotton, and sugarcane. AMF and associated bacteria enhance plant growth and health, and affect the production of polyphenols and carotenoids, and the activity of antioxidant enzymes. The production of health-promoting phytochemicals was shown to be differentially modulated by different AMF isolates and bacterial strains, in several food plants, i.e., tomato, lettuce, strawberry, artichoke, maize, grapevine, sunflower. Here, we provide an overview of recent studies concerning the multiple roles played by AMF and associated bacteria in the modulation of the biosynthesis of plant secondary metabolites with health-promoting activity, and discuss the development of designed multifunctional consortia to be used in sustainable agriculture.

We describe the case of a male subject affected by retroperitoneal advanced, anthracycline-pretreated liposarcoma, who experienced a long, beneficial clinical effect from eribulin treatment. In March 2013, a left, paraortic, retroperitoneal mass was surgically removed and diagnosed as Mdm2-positive dedifferentiated liposarcoma. In June 2015, a CT scan revealed disease progression and first-line epirubicin/ifosfamide treatment was started, followed by epirubicin in monotherapy. In January 2017, following a new disease progression, the patient started a second-line eribulin treatment that went on for about 1 year with no major adverse events. The CT scans performed every 3–4 months showed stable disease. After 13 months of treatment, a CT scan revealed disease progression and 10 days later, the patient died of bowel perforation and peritonitis.

Chondrosarcoma (CS) is a cartilage malignancy of adulthood that is treated by surgery alone, since chemotherapy is considered ineffective. Unfortunately, a large proportion of patients with CS develop lung metastases, and several die of the disease. In this study, we compared 3D-spheroid cultures and conventional cell monolayer models in order to identify the best way to select anticancer agents that could be effective for the systemic control of CS. Using SW1353 cells, we developed a three-dimensional (3D) in vitro culture model to mimic in vivo features of CS microenvironment and evaluated the efficacy of different drugs to modulate CS cell proliferation and survival in 2D versus 3D-cultures. Doxorubicin (DXR) and cisplatin, that are widely employed in sarcomas, were less effective on 3D-CS spheroids when compared to standard monolayer models, whereas treatment with the ionophore salinomycin (SAL) had a strong cytotoxic effect both on 2D and 3D-cultures. Furthermore, as demonstrated by the reduced viability and the enhanced DXR nuclear localization, SAL enhanced DXR cytotoxicity in 3D-CS spheroids also at sub-lethal doses. SAL activity on 3D-CS spheroids was mediated by a significant induction of apoptosis via caspase activation. This study demonstrates that preclinical tests significantly differ in monolayer and 3D cultures of CS cells. Using this approach, SAL, alone or, at sub-lethal concentrations, in combination with DXR, represents a promising agent for the systemic treatment of CS. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.

Purpose This study evaluated the effect of low-level laser therapy (LLLT) on the healing of bone defects filled with autogenous bone or bioactive glass. Materials and methods A critical size defect with 5-mm diameter was created on the calvaria of 60 adult male rats divided into 6 groups (n = 10): group C (control), group LLLT (LLLT - GaAlAs, wavelength of 780 nm, power of 100 mW, energy density of 210 J/cm2 per point during 60 seconds/point, in five points, only once, after creation of the surgical defect), group AB (autogenous bone), group AB+LLLT (autogenous bone + LLLT), group BG (bioactive glass), group BG+LLLT (bioactive glass + LLLT). All animals were sacrificed at 30 days after surgery. The areas of newly formed bone (ANFB) and areas of remaining particles (ARP) were calculated in relation to the total area (TA). Results The highest mean ± SD ANFB was observed for group LLLT (47.67% ± 8.66%), followed by groups AB+LLLT (30.98% ± 16.59%) and BG+LLLT (31.13% ± 16.98%). There was a statistically significant difference in relation to ANFB between group C and the other groups, except for comparison with group BG (Tukey test, P > .05). There was no statistically significant difference in ANFB values between group AB and the other study groups (Tukey test, P > .05), group AB+LLLT and groups BG and BG+LLLT (Tukey test, P > .05), and between groups BG and BG+LLLT (Tukey test, P > .05). The highest mean ± SD ARP was found for group BG (25.15% ± 4.82%), followed by group BG+LLLT (17.06% ± 9.01%), and there was no significant difference between groups (t test, P > .05). Conclusion The LLLT, in the present application protocol, did not increase the area of new bone formation when associated with autogenous bone or bioactive glass.

Sustained autophagy contributes to the metabolic adaptation of cancer cells to hypoxic and acidic microenvironments. Since cells in such environments are resistant to conventional cytotoxic drugs, inhibition of autophagy represents a promising therapeutic strategy in clinical oncology. We previously reported that the efficacy of hydroxychloroquine (HCQ), an autophagy inhibitor under clinical investigation is strongly impaired in acidic tumor environments, due to poor uptake of the drug, a phenomenon widely associated with drug resistance towards many weak bases. In this study we identified salinomycin (SAL) as a potent inhibitor of autophagy and cytotoxic agent effective on several cancer cell lines under conditions of transient and chronic acidosis. Since SAL has been reported to specifically target cancer-stem cells (CSC), we used an established model of breast CSC and CSC derived from breast cancer patients to examine whether this specificity may be associated with autophagy inhibition. We indeed found that CSC-like cells are more sensitive to autophagy inhibition compared to cells not expressing CSC markers. We also report that the ability of SAL to inhibit mammosphere formation from CSC-like cells was dramatically enhanced in acidic conditions. We propose that the development and use of clinically suitable SAL derivatives may result in improved autophagy inhibition in cancer cells and CSC in the acidic tumor microenvironment and lead to clinical benefits.

Homozygous familial hypercholesterolaemia (HoFH) is a rare form of inherited dyslipidemia resistant to conventional cholesterol-lowering medications so that lipoprotein apheresis (LA) is usually required. Lomitapide has been approved for the treatment of HoFH. The aim of this study was to evaluate the benefits of lomitapide in HoFH patients followed with the usual clinical care. Homozygous familial hypercholesterolaemia (HoFH) is a rare form of inherited dyslipidemia resistant to conventional cholesterol-lowering medications so that lipoprotein apheresis (LA) is usually required. Lomitapide has been approved for the treatment of HoFH. The aim of this study was to evaluate the benefits of lomitapide in HoFH patients followed with the usual clinical care.Clinical and biochemical data were retrospectively collected in 15 HoFH patients (10 with mutations in the LDLR gene and 5 in the LDLRAP1 gene) treated for at least 6 months with lomitapide in addition to lipid-lowering therapies (LLT) in different Lipid Clinics across Italy.The mean follow-up period was 32.3 +/- 29.7 months. During background therapies, HoFH patients showed a mean LDL-C level of 426.0 +/- 204.0 mg/dl. The addition of lomitapide at the average dosage of 19 mg/day lowered LDL-C levels by 68.2 +/- 24.8%. At their last visit, 60% of patients showed LDL-C < 100 mg/dl and 46.6% < 70 mg/dl. During follow-up, 8 of 10 patients receiving LA (80%) stopped this treatment due to marked LDL-C reduction. A wide range (13-95%) of individual LDL-C reduction was observed, but this was not related to genotype. During follow-up, 53.3% of patients reported at least one episode of diarrhea, but none was referred as severe; none had liver transaminase > 5x ULN or had to stop treatment due to side effects. A subset of patients was evaluated by liver ultrasound and fibroscan (n = 5) or nuclear magnetic resonance with spectroscopy (MRS) (n = 1) not showing clinical evidence of liver damage.In this real-world experience, lomitapide was confirmed to be a very powerful cholesterol-lowering agent in HoFH showing a good safety profile.

Trawling pressure and environmental changes may affect the composition of fish assemblages. Our knowledge on large spatio-temporal patterns of demersal fish composition remains incomplete for the Mediterranean Sea. We investigated (1) the spatio-temporal stability of demersal assemblages, (2) the relationships between these assemblages and potential structuring factors (trawling pressure and environmental conditions) in order to assess the dynamic of the assemblage structure at the scale of the northern Mediterranean Sea. We analysed a dataset of 18062 hauls from 10 to 800 m depth performed annually during the last two decades across 17 Geographical Sub-Areas (GSAs) (MEDITS program). A multi-table analysis (STATICO-CoA) evidenced a strong inter-GSAs stability in the organization of assemblages, with specificities for some GSAs. The most stable structuring factors were linked to combined gradients of chlorophyll a, phytoplancton carbon biomass and temperature, inversely correlated with depth, salinity and nutrient gradients (axis 1 of the STATICO-CoA compromise, 93.74% of the total variability). A common pattern linking the distribution of species to these environmental gradients was evidenced for most of the 17 GSAs. Estimate of trawling pressure showed a minor role in the organization of the assemblages for the spatial scale and years investigated (axis 2, 4.67%). La presión pesquera y los cambios ambientales pueden afectar a la composición de las agrupaciones de peces. En el Mediterráneo, nuestro conocimiento a gran escala sobre los patrones espacio-temporales de la composición de especies de peces demersales sigue siendo incompleto. Investigamos (1) la estabilidad espacio-temporal de las agrupaciones demersales (2) las relaciones entre dichas agrupaciones y los posibles factores estructurantes (presión pesquera de arrastre y condiciones ambientales) para evaluar la dinámica de la estructura de las agrupaciones en el norte del mar Mediterráneo. Analizamos un conjunto de 18062 lances entre 10 y 800 m. de profundidad realizados anualmente, durante las últimas dos décadas, en 17 sub-áreas geográficas (GSAs) (Proyecto MEDITS). El análisis multi-tablas (STATICO-CoA) evidenció una fuerte estabilidad inter-GSAs en la organización de agrupaciones, con especificidades para algunas GSAs. Los factores estructurantes más estables se relacionaron con los gradientes combinados de clorofila a, fitoplancton, carbono y temperatura, correlacionados inversamente con los gradientes de profundidad, salinidad y nutrientes (eje 1 del STATICO-CoA, 93.74% de la variabilidad total). En la mayoría de las GSAs encontramos un patrón común que vincula la distribución de las especies a estos gradientes ambientales. La estimación de la presión pesquera de arrastre mostró un papel menor en la organización de las agrupaciones para la escala espacial y los años investigados (eje 2, 4.67%).

Arbuscular mycorrhizal fungi (AMF) play a key role in plant nutrition and in the maintenance of soil fertility in agroecosystems, but their survival can be affected by different agricultural practices, including the use of pesticides. In this work, we assessed the impact of chemicals with herbicidal and fungicidal activity on mycelial growth and structure of the worldwide distributed AMF Funneliformis mosseae. Results showed that mycelial growth and interconnectedness of three different F. mosseae lineages were affected by the chemicals tested at concentrations lower than those indicated for agricultural use. Indeed, benomyl reduced mycelial growth in one lineage and fenhexamid negatively affected germlings growth in the three lineages, while both fungicides significantly affected mycelial viability and induced abnormal hyphal branching. The three F. mosseae lineages showed sensitivity to herbicides, with significant growth and viability decreases in the presence of the active ingredient glufosinate ammonium and a low viability at increasing concentrations of dicamba. Anastomosis rates were reduced by the two fungicides and by very low levels of the herbicide glufosinate, whose impact on AMF networks should be further investigated, given its wide use in agriculture. Our results suggest that the use of some pesticides in agroecosystems may lead to growth reduction and disruption of AMF mycelial structural traits, even in AMF species able to tolerate soil disturbance, posing serious threats to AMF symbiotic performance and to the maintenance of soil mycorrhizal potential.

The extraradical mycelium (ERM) produced by arbuscular mycorrhizal fungi is fundamental for the maintenance of biological fertility in agricultural soils, representing an important inoculum source, together with spores and mycorrhizal root fragments. Its viability and structural traits, such as density, extent and interconnectedness, which are positively correlated with the growth and nutrition of host plants, may be affected by different agronomic practices, including the use of pesticides and by different mycorrhizospheric communities. This work, carried out using a whole-plant experimental model system, showed that structural traits of ERM, such as length and density, were strongly decreased by the herbicides dicamba and glufosinolate and the fungicides benomyl and fenhexamid, while anastomosis frequency and hyphal branching were differentially modulated by singly inoculated mycorrhizospheric bacteria, depending on their identity.

319 Background: Nivolumab is approved in the second or further line of treatment for patients with metastatic renal cell carcinoma (mRCC); cabozantinib is approved in a similar setting of patients. Unfortunately, no evidence is currently available regarding the best treatment option after disease progression to both nivolumab and cabozantinib. The aim of this study is to compare the treatment choices after progression to nivolumab and cabozantinib including patients followed in best supportive care (BSC) or active therapy. Methods: In this retrospective observational study, we selected 42 patients from 8 Italian cancer centers. Enrolled patients had progressed to both nivolumab and cabozantinib and subsequently referred to active treatment or BSC. Primary endpoint of the study was the OS of patients on active treatment versus BSC. Secondary endpoints were ORR, PFS and OS of patients on active treatment who received sorafenib versus everolimus. Results: The median age was 65 years, 76.2% were male. The majority of patients had undergone nephrectomy (78.6%), had clear cell histology (83%) and were at intermediate-poor risk at the diagnosis (85.7%). The most frequent site of metastatic disease in the general population and in patients referred to BSC was the lung (73.8% and 88.9%, respectively). For patients referred to active treatment, the most frequent site of metastasis was bone (70.8%). Sunitinib (71.4%), nivolumab (64.3%), and cabozantinib (54.7%) were the most commonly used drugs in the I, II and III lines of treatment, respectively. After progression to both nivolumab and cabozantinib 42.9% of patients were referred to BSC, while 57.1% received active treatment (28.6% everolimus, 16.7% sorafenib, 4.8% sunitinib, 4.8% IL2-HD, 2.4% lenvatinib + everolimus). Median OS was 13 (95% CI: 4-NR) and 3 months (95% CI: 2-4) in patients on active treatment versus BSC ( p=0.001). Patients treated with sorafenib had better disease control when compared with those treated with everolimus (SD 71.4% versus 16.7%, PD 14.3% versus 58.3%; p=0.03), but no significant advantage in terms of PFS (5 versus 3 months, 95% CI: 2-6 versus 2-5; p= 0.5) and OS (NR versus 13 months, 95% CI: 3-NR versus 2-NR; p=0.2) was observed. Conclusions: After treatment with both nivolumab and cabozantinib, when possible, the choice of an active treatment seems to produce an OS advantage when compared with BSC. However, although sorafenib seems to demonstrate better results, we cannot indicate which is the drug of choice, as no significant advantage was shown in terms of OS or PFS from the comparison between sorafenib and everolimus. The limitations of this study are given by the size of the sample examined and its retrospective nature. Further studies are needed to confirm whether active treatment choice is associated with improved OS.

We read with great interest the article by Ma et al.1 on the relationship between reduced low-density lipoprotein cholesterol (LDL-C) levels and intracerebral hemorrhage (ICH). In our opinion, this possibly causal association needs to be better investigated to avoid ICH secondary to therapeutic LDL-C lowering. Indeed, international guidelines on acute coronary syndromes recommend early administration of high-intensity statins to satisfy “the lower, the faster, the better” concept.2 Nowadays, new lipid-lowering therapies—such as proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors—allow patients to reach very low LDL-C levels. Unfortunately, long-term follow-up for these drugs is not available, but it has been suggested that in patients with preexisting ischemic heart disease already on statin therapy, LDL-C levels do not necessarily need to be lowered below 70 mg/dL.3 For these reasons, we cautiously suggest that in patients with chronic ischemic heart disease, PCSK9 inhibitor therapy may be administered at greater intervals4 so as to obtain an optimal balance between cardiovascular risk reduction, hemorrhagic risk avoidance, and mitigation of side effects related to lipid-lowering therapy.

The increasing demand for products based from Stevia rebaudiana Bertoni (both leaves and purified steviol glycosides) makes of interest the research on sustainable production systems, in order to guarantee secure availability and high quality of agricultural raw materials. Arbuscular mycorrhizal fungi (AMF) symbiosis represents an interesting tool for increasing crop production and quality, especially thanks to improved nutrient absorption, particularly phosphorus (P). In the present study, Stevia rebaudiana Bert. plants were exposed to different levels of P fertilization (0, 25 and 50 mg P2O5 kg−1 soil) with or without Rhizoglomus irregulare inoculation, in order to evaluate root colonization, plant growth and productive parameters, steviol glycosides (SVglys) yield, as well as nitrogen (N) and P concentrations and uptake. A nutrient balance was also carried out and the nutrient use efficiency was evaluated. Stevia roots were highly colonized by Rhizoglomus irregulare, especially in the absence of P fertilization. During the whole vegetative growth, the AMF symbiosis, in association with the P supply, benefitted stevia growth, especially with regard to leaf dry biomass production and SVglys yield. Arbuscular mycorrhizal fungi symbiosis was able to modify the growth habit of stevia plants, with increased branching and a reduced plant height. At the end of the vegetative growth, mycorrhizal plants reached the highest leaf dry yield, together with the highest SVglys production. The application of 25 mg P2O5 kg−1 soil in association with AMF symbiosis seemed to be the most effective treatment in improving stevia SVglys yield and P uptake together with P nutrient use efficiency.

Background Androgen deprivation therapy (ADT) is a cornerstone of treatment for prostate cancer, but limited studies have been performed to investigate the association between ADT and immune alterations, such as autoimmune diseases and risk of second tumours, considering that androgens may also play a role in the immune modulation. Methods We retrospectively evaluated patients (pts) treated with abiraterone (abi) and enzalutamide (enza) in 12 Italian Institutes between July 2011 and December 2018. In particular, we assessed the risk of systemic or single-organ autoimmune diseases [according to the International Classification of Diseases 10th Revision (ICD-10)] and second tumors, by performing a logistic regression analysis. Results We included 844 pts receiving abi/enza, whose 36 (4.3%) had diagnosis of autoimmune diseases at baseline [13 (1.5%) arthritis (rheumatoid or psoriatic), 12 (1.4%) autoimmune thyroiditis, 4 (0.5%) gastrointestinal autoimmune disease, 4 (0.5%) psoriasis, and 3 (0.4%) vasculitis] and 58 (6.9%) had second tumors (45 solid and 13 hematological). Median age was 70 years [interquartile range (IQR) 63-75]. Median duration of hormone sensitive of prostate cancer was 29 months (IQR 14-59), while median CRPC duration was 22 months (IQR 13-39). Most pts (N = 764, 90.5%) did ≤ 2 therapeutic lines, whose 477 (56.5%) and 367 (43.5%) were treated with abi and enza, respectively, and 359 (42.5%) were chemotherapy naive. We observed a significant increase in the risk of autoimmune diseases and a trend for higher incidence of second tumors in pts showing a shorter time to CRPC [odd ratio (OR) 0.98 (95% CI 0.96-0.99) p = 0.01, and OR 0.99 (95% CI 0.98-1.01) p = 0.09, respectively], whereas no association was reported between CRPC duration and the incidence of immune disorders. Clinical outcome associated with abi/enza in CRPC was independent from pre-treatment presence of immune diseases, but a worse overall survival was observed from diagnosis of prostate cancer in pts developing autoimmune diseases (HR 1.69, 95% CI 1.10-2.61, p = 0.016) . Conclusions Time to CRPC may represent a risk factor of developing immune alterations with a negative prognostic role in the overall survival of prostate cancer pts. Further larger prospective studies are warranted. Legal entity responsible for the study The authors. Funding Has not received any funding. Disclosure V. Conteduca: Advisory / Consultancy, Travel / Accommodation / Expenses: Bayer; Travel / Accommodation / Expenses: Sanofi; Travel / Accommodation / Expenses: Janssen-Cilag; Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Astellas. O. Caffo: Advisory / Consultancy, Speaker Bureau / Expert testimony: Janssen-Cilag; Advisory / Consultancy, Speaker Bureau / Expert testimony: Astellas; Advisory / Consultancy, Speaker Bureau / Expert testimony: Bayer; Advisory / Consultancy, Speaker Bureau / Expert testimony: Pfizer; Advisory / Consultancy, Speaker Bureau / Expert testimony: Sanofi; Advisory / Consultancy, Speaker Bureau / Expert testimony: AstraZeneca. E. Zanardi: Advisory / Consultancy: Janssen. G. Procopio: Travel / Accommodation / Expenses: Bayer; Travel / Accommodation / Expenses: Ipsen; Travel / Accommodation / Expenses: Novartis; Travel / Accommodation / Expenses: MSD; Travel / Accommodation / Expenses: Pfizer; Travel / Accommodation / Expenses: BMS. U.F.F. De Giorgi: Advisory / Consultancy, Speaker Bureau / Expert testimony: Janssen; Advisory / Consultancy, Speaker Bureau / Expert testimony: Sanofi; Advisory / Consultancy, Speaker Bureau / Expert testimony: Bayer; Advisory / Consultancy, Speaker Bureau / Expert testimony: Astellas; Advisory / Consultancy, Speaker Bureau / Expert testimony: Janssen; Advisory / Consultancy, Speaker Bureau / Expert testimony: BMS. All other authors have declared no conflicts of interest.

We investigated the ability of quercetin and apigenin to modulate platelet activation and aggregation, and compared the observed efficacy with that displayed by their synthetic analogues 2-phenyl-4H-pyrido[1,2-a]pyrimidin-4-ones, 1-4, and 2,3-diphenyl-4H-pyrido[1,2-a]pyrimidin-4-ones, 5-7. Platelet aggregation was explored through a spectrophotometric assay on platelet-rich plasma (PRP) treated with the thromboxane A2 mimetic U46619, collagen and thrombin in presence/absence of various bioisosteres of flavonoids (12.5-25-50-100 μM). The platelet density, (mean platelet component, MPC), was measured by the Advia 120 Hematology System as a marker surrogate of platelet activation. The induced P-selectin expression, which reflects platelet degranulation/activation, was quantified by flow cytometry on PRP. Our synthetic compounds modulated significantly both platelet activation and aggregation, thus turning out to be more effective than the analogues quercetin and apigenin when tested at a concentration fully consistent with their use in vivo. Accordingly, they might be used as food supplements to increase the efficacy of natural flavonoids.

Arbuscular mycorrhizal fungi (AMF) establish beneficial mutualistic symbioses with land plants, receiving carbon in exchange for mineral nutrients absorbed by the extraradical mycelium (ERM). With the aim of obtaining in vivo produced ERM for gene expression analyses, a whole-plant bi-dimensional experimental system was devised and tested with three host plants and three fungal symbionts. In such a system, Funneliformis mosseae in symbiosis with Cichorium intybus var. foliosum, Lactuca sativa, and Medicago sativa produced ERM whose lengths ranged from 9.8 ± 0.8 to 20.8 ± 1.2 m per plant. Since ERM produced in symbiosis with C. intybus showed the highest values for the different structural parameters assessed, this host was used to test the whole-plant system with F. mosseae, Rhizoglomus irregulare, and Funneliformis coronatus. The whole-plant system yielded 1–7 mg of ERM fresh biomass per plant per harvest, and continued producing new ERM for 6 months. Variable amounts of high-quality and intact total RNA, ranging from 15 to 65 μg RNA/mg ERM fresh weight, were extracted from the ERM of the three AMF isolates. Ammonium transporter gene expression was successfully determined in the cDNAs obtained from ERM of the three fungal symbionts by RT-qPCR using gene-specific primers designed on available (R. irregulare) and new (F. mosseae and F. coronatus) ammonium transporter gene sequences. The whole-plant experimental system represents a useful research tool for large production and easy collection of ERM for morphological, physiological, and biochemical analyses, suitable for a wide variety of AMF species, for a virtually limitless range of host plants and for studies involving diverse symbiotic interactions., Financial support of University of Pisa and CNR is gratefully acknowledged. This paper was part of A. Pepe doctoral thesis work at the University of Pisa.

Nuclear aggregates of polyamines (NAPs) are supramolecular compounds generated by the self-assembly of protonated nuclear polyamines (spermine, spermidine and putrescine) and phosphate ions. In the presence of genomic DNA, the hierarchical process of self-structuring ultimately produces nanotube-like polymers that envelop the double helix. Because of their modular nature and their aggregation-disaggregation dynamics, NAPs confer plasticity and flexibility to DNA. Through the disposition of charges, NAPs also enable a bidirectional stream of information between the genome and interacting moieties. High mobility group (HMG) B1 is a non-histone chromosomal protein that binds to DNA and that influences multiple nuclear processes. Because genomic DNA binds to either NAPs or HMGB1 protein, we explored the ability of in vitro self-assembled NAPs ( iv NAPs) to mediate the DNA-HMGB1 interaction. To this end, we structured DNA-NAPs-HMGB1 and DNA-HMGB1-NAPs ternary complexes in vitro through opportune sequential incubations. Mobility shift electrophoresis and atomic force microscopy showed that the DNA- iv NAPs-HGMB1 complex had conformational assets supposedly more suitable those of the DNA-HGMB1- iv NAPs to comply with the physiological and functional requirements of DNA. Our findings indicated that iv NAPs act as mediators of the DNA-HMGB1 interaction.

Permeability studies across biological barriers are of primary importance in drug delivery as well as in toxicology when investigating the absorption and translocation of a substance. The study of nanomaterial interaction with epithelial barriers is of particular interest given their growing use in nanomedicine as well as concerns about their potential hazard. Here we describe the design and fabrication of a new bioreactor with an ultrathin microporous sensing support for the study of nanoparticle toxicity in intestinal epithelial cells in conditions which better recapitulate the physiological environment. Thanks to the integration of 4 electrodes in the microporous membrane, the system allows real-time and continuous sensing of TEER (trans epithelial electrical resistance) during flow without interruption or perturbation of experiments. The TEER bioreactor was tested using Caco-2 cells as an in vitro model of intestinal epithelia. When exposed to silver nanoparticles, which are known to be toxic, the embedded electrodes enabled non-invasive evaluation of barrier impairment over time. This device can be used to study barrier integrity and the kinetics of nanomaterial induced damage to epithelial barriers in physiologically relevant conditions.

We investigated the biophysical properties of the transport mediated by ion channels in hemocytes from the hemolymph of the bivalve Mytilus galloprovincialis. Besides other transporters, mytilus hemocytes possess a specialized channel sensitive to the osmotic pressure with functional properties similar to those of other transport proteins present in vertebrates. As chloride fluxes may play an important role in the regulation of cell volume in case of modifications of the ionic composition of the external medium, we focused our attention on an inwardly-rectifying voltage-dependent, chloride-selective channel activated by negative membrane potentials and potentiated by the low osmolality of the external solution. The chloride channel was slightly inhibited by micromolar concentrations of zinc chloride in the bath solution, while the antifouling agent zinc pyrithione did not affect the channel conductance at all. This is the first direct electrophysiological characterization of a functional ion channel in ancestral immunocytes of mytilus, which may bring a contribution to the understanding of the response of bivalves to salt and contaminant stresses.

The energy from sugarcane is one of the most important in Brazil’s energy matrix and the efficiency of extraction and processing is fully dependent on the quality of the raw material. The soil present in sugarcane was investigate here as a factor that can affect the production of energy. Chemical elements (Fe, Hf, Sc and Th) determined by instrumental neutron activation analysis were used for tracing soil in sugarcane and its derived bagasse. The lower calorific value (LCV) of bagasse demonstrated a good negative correlation (r = −0.9727) with the ash content of the bagasse, which in turn was positively correlated to the amount of soil in the sugarcane. Therefore, the presence of soil reduces the production of energy from burning bagasse. The proportion of loss in the LCV was just slightly higher than the soil content, i.e. for an soil content of 10 % a reduction of 12.7 % was observed in the LCV.

Objective To estimate the effects of prostaglandin E1 (PGE1) and E2 (PGE2) on myometrial contractility and structure in vitro.

High LDL-cholesterol (LDL-C) characterizes familial hypercholesterolemia (FH) and familial combined hyperlipidemia (FCH). LDL-apheresis, used in these patients to reduce LDL-C levels, has been shown to also affect HDL levels and composition. We studied LDL-apheresis effects on six FH and nine FCH subjects’ serum capacity to modulate cellular cholesterol efflux, an index of HDL functionality, and to load macrophages with cholesterol. Serum cholesterol efflux capacity (CEC) and macrophage cholesterol loading capacity (CLC) were measured before, immediately after, and two days after LDL-apheresis. The procedure reduced total cholesterol (TC), LDL-C, and apoB plasma levels (−69%, −80% and −74%, respectively), parameters only partially restored two days later. HDL-C and apoA-I plasma levels, reduced after LDL-apheresis (−27% and −16%, respectively), were restored to almost normal levels two days later. LDL-apheresis reduced serum aqueous diffusion (AD) CEC, SR-BI-CEC, and ABCA1-CEC. AD and SR-BI were fully restored whereas ABCA1-CEC remained low two days later. Sera immediately and two days after LDL-apheresis had a lower CLC than pre-LDL-apheresis sera. In conclusion, LDL-apheresis transiently reduces HDL-C levels and serum CEC, but it also reduces also serum capacity to deliver cholesterol to macrophages. Despite a potentially negative effect on HDL levels and composition, LDL-apheresis may counteract foam cells formation.

Nanostructured polymeric capsules are regarded as highly promising systems with different potential applications ranging from drug delivery, biosensing and artificial cells. To fully exploit this potential, it is required to produce bio-activated stable and biocompatible capsules. To this purpose, in present work we proposed the combination of the layer-by-layer self assembly method with bacterial S-layer technology to fabricate stable and biocompatible polymeric capsules having a well defined arrangement of functional groups allowing the covalent attachment of antibody molecules. Hollow microcapsules were obtained by the layer-by-layer self assembly of oppositely charged polyelectrolytes onto colloidal particles, followed by removal of the cores at acidic pH. S-layers were crystallized onto the shell of the obtained capsules. Quartz crystal microbalance was used to characterize the crystallization process onto planar surfaces. S-layer containing capsules were investigated by atomic force microscopy. Immunoenzymatic tests were performed to assess the effective modification of the S-layer with antibody molecules both on planar surfaces and on hollow capsules. Fluorescent microscopy was employed to visualize the presence of the antibody molecules onto the capsule shell and immunological tests used to assess the bioactivity of the immobilized antibodies. Finally, the in vitro cytotoxicity of fabricated S-layer containing capsules was studied. The obtained results demonstrated the possibility to fabricate bio-activated S-layer containing capsules with improved features in terms of biocompatibility.