Valorization of Side Stream Products from Sea Cage Fattened Bluefin Tuna (Thunnus thynnus): Production and In Vitro Bioactivity Evaluation of Enriched ω-3 Polyunsaturated Fatty Acids.

The valorization of side streams from fishery and aquaculture value-chains is a valuable solution to address one of the challenges of the circular economy: turning wastes into profit. Side streams produced after filleting of sea cage fattened bluefin tuna (Thunnus thynnus) were analyzed for proximate composition and fatty acid profile to evaluate the possibility of producing tuna oil (TO) as a valuable source of ω-3 polyunsaturated fatty acids (PUFA) and testing its bioactivity in vitro. Ethyl esters of total fatty acids (TFA), obtained from TO, were pre-enriched by urea complexation (PUFA-Ue) and then enriched by short path distillation (SPD) up to almost 85% of the PUFA fraction (PUFA-SPe). The bioactivity of TFA, PUFA-SPe, and ethyl esters of depleted PUFA (PUFA-SPd) were tested in vitro, through analysis of lipid metabolism genes, in gilthead sea bream (Sparus aurata) fibroblast cell line (SAF-1) exposed to oils. TFA and PUFA-SPd upregulated transcription factors (pparß and pparγ) and lipid metabolism-related genes (D6D, fas, fabp, fatp1, and cd36), indicating the promotion of adipogenesis. PUFA-SPe treated cells were similar to control. PUFA-SPe extracted from farmed bluefin tuna side streams could be utilized in fish feed formulations to prevent excessive fat deposition, contributing to improving both the sustainability of aquaculture and the quality of its products.

Vitamin D receptor, STAT3, and TET2 cooperate to establish tolerogenesis.

The active form of vitamin D, 1,25-dihydroxyvitamin D3, induces a stable tolerogenic phenotype in dendritic cells (DCs). This process involves the vitamin D receptor (VDR), which translocates to the nucleus, binds its cognate genomic sites, and promotes epigenetic and transcriptional remodeling. In this study, we report the occurrence of vitamin D-specific DNA demethylation and transcriptional activation at VDR binding sites associated with the acquisition of tolerogenesis in vitro. Differentiation to tolerogenic DCs associates with activation of the IL-6-JAK-STAT3 pathway. We show that JAK2-mediated STAT3 phosphorylation is specific to vitamin D stimulation. VDR and the phosphorylated form of STAT3 interact with each other to form a complex with methylcytosine dioxygenase TET2. Most importantly, pharmacological inhibition of JAK2 reverts vitamin D-induced tolerogenic properties of DCs. This interplay among VDR, STAT3, and TET2 opens up possibilities for modulating DC immunogenic properties in clinics.

Effect of Cold Smoking and Natural Antioxidants on Quality Traits, Safety and Shelf Life of Farmed Meagre (Argyrosomus regius) Fillets, as a Strategy to Diversify Aquaculture Products.

Aquaculture has been playing a leading role over the years to satisfy the global growing demand for seafood. Moreover, innovative techniques are necessary to increase the competitiveness, sustainability and profitability of the seafood production chain, exploiting new species from the aquaculture, such as meagre (Argyrosomus regius), to develop value-added products and diversify their production. In the present work, the effectiveness of cold smoking combined with antioxidants (SA) compared to cold smoking alone (S) on meagre fillets, the quality and shelf life were investigated. Sensory, biochemical, physical-chemical and microbiological analyses were performed on the smoked fillets during vacuum-packaged storage for 35 days at 4 ± 0.5 °C. The results showed positive effects of the SA treatment on the biochemical parameters of meagre fillets. The total volatile basic nitrogen (TVB-N) in smoked meagre fillets was significantly lower in the SA treatment at the end of storage compared to the S treatment. Moreover, SA had a positive effect on lipid peroxidation. Lower values of malondialdehyde (mg MDA/kg) were observed in the SA treatment during preservation compared to the S treatment. This work will contribute to the growth of the fish production chain, producing a value-added fish product by exploiting meagre, whose production has been increasing over decades.

Farmed Gilthead Sea Bream (Sparus aurata) by-Products Valorization: Viscera Oil ω-3 Enrichment by Short-Path Distillation and In Vitro Bioactivity Evaluation.

This study shows a pilot scale protocol aimed to obtain an omega 3-enriched oil after the processing of farmed gilthead sea bream viscera (SBV); this was oil was tested in vitro for bioactivity, attesting to the possibility to turn waste into profit The quality of the oil, in terms of requirements for animal and human consumption, was assessed by determining some chemical parameters, such as peroxide value (PV), thiobarbituric acid reactive substances (TBARS), ρ-anisidine (ρ-AV) content, total oxidation value (TOTOX), and phospholipids and free fatty acid (%), both in crude viscera oil (CVO) and refined viscera oil (RVO). Among the extraction conditions, the higher CVO yields were obtained at 60 °C for 10 min (57.89%) and at 80 °C for 10 min (67.5%), and the resulting oxidation levels were low when utilizing both extraction conditions. RVO, obtained from CVO extracted at 60 °C, showed the highest quality on the basis of the assessed parameters. The ethyl esters of the total fatty acid (TFA) contents extracted from RVO were enriched in the ω-3 polyunsaturated fatty acid fraction (PUFAE) up to almost 56% via short path distillation (SPD). Antioxidant activities and adipogenic properties were tested in vitro. PUFAE protected 3T3 L1 cells from oxidative stress and exerted an anti-adipogenic effect in Dicentrarchus labrax pre-adipocytes, attesting to the beneficial properties for both farmed fish and human health. These results could stimulate the adoption of solutions aimed to recover and utilize aquaculture by-products at a higher scale, turning "waste into profit" and indicating a strategy to reach more sustainable business models in aquaculture resource utilization according to the principles of the circular economy.

Combination of Freezing, Low Sodium Brine, and Cold Smoking on the Quality and Shelf-Life of Sea Bass (Dicentrarchus labrax L.) Fillets as a Strategy to Innovate the Market of Aquaculture Products.

Aquaculture is playing a leading role in both meeting the growing demand for seafood and increasing the sustainability of the fish production sector. Thus, innovative technologies that improve its sustainability, competitiveness, and safety are necessary for growth in the sector. This study aimed to develop cold smoked sea bass fillets from aquaculture. The aptitude of frozen and fresh fillets for cold smoking was investigated by processing both fresh and thawed fillets kept previously at -20 °C for 15, 30, 60, and 90 days. Moreover, to develop a low-salt product, fillets were immersed in low-sodium or standard brine. Sensory, biochemical, and physical-chemical analyses were performed on both the raw fillets and the smoked fillets during vacuum packaged storage for 35 days at 1 ± 0.5 °C. Young modulus values, representative of texture and sensory evaluation, showed that the quality of fresh fillets was better compared to the thawed ones, thus affecting the quality of the final product as the correlation between parameters showed (principal component analysis). Cold smoking was effective in both maintaining the total volatile basic nitrogen (TVB-N) below the threshold for spoilage and preventing lipid peroxidation. Moreover, partial sodium replacement by potassium did not alter the sensory attributes of smoked fillets, which maintained high scores up to 21 days.

Robot-assisted re-do sacrohysteropexy after anterior abdominal wall hysteropexy.

INTRODUCTION AND HYPOTHESIS: We report the case of a 49-year-old woman who came to our institution complaining of bowel constipation and bladder outlet obstruction due to the recurrence of a pelvic prolapse after anterior abdominal wall hysteropexy performed 1 year before. METHODS: The patient was counseled to undergo robot-assisted re-do sacrohysteropexy performed using the da Vinci Xi surgical system. The step-by-step technique used for the herein reported clinical case of robot-assisted re-do sacrohysteropexy is shown in the accompanying video. RESULTS: Operative time was 120 min. There was virtually no blood loss. No intraoperative complications occurred. Use of analgesic drugs was stopped on the 1st postoperative day. The patient was mobilized on postoperative day 2. The catheter was removed on the 3rd postoperative day, and the patient got back to spontaneous micturition with no post-voiding residual. On the same day, the patient was discharged. At 3-month follow-up, the patient was well with neither bowel nor bladder symptoms. Uroflowmetry was normal. CONCLUSION: We demonstrated the feasibility of robot-assisted re-do sacrohysteropexy. In the surgical technique herein reported, we showed how we were able to partially re-use a previously implanted mesh together with a new one to ensure fixation of a multi-compartment pelvic organ prolapse.

ERBB2 mRNA Expression and Response to Ado-Trastuzumab Emtansine (T-DM1) in HER2-Positive Breast Cancer.

Trastuzumab emtansine (T-DM1) is approved for the treatment of human epidermal growth factor receptor 2 (HER2)-positive (HER2+) metastatic breast cancer (BC) and for residual disease after neoadjuvant therapy; however, not all patients benefit. Here, we hypothesized that the heterogeneity in the response seen in patients is partly explained by the levels of human epidermal growth factor receptor 2 gene (ERBB2) mRNA. We analyzed ERBB2 expression using a clinically applicable assay in formalin-fixed paraffin-embedded (FFPE) tumors (primary or metastatic) from a retrospective series of 77 patients with advanced HER2+ BC treated with T-DM1. The association of ERBB2 levels and response was further validated in 161 baseline tumors from the West German Study (WGS) Group ADAPT phase II trial exploring neoadjuvant T-DM1 and 9 in vitro BC cell lines. Finally, ERBB2 expression was explored in 392 BCs from an in-house dataset, 368 primary BCs from The Cancer Genome Atlas (TCGA) dataset and 10,071 tumors representing 33 cancer types from the PanCancer TCGA dataset. High ERBB2 mRNA was found associated with better response and progression-free survival in the metastatic setting and higher rates of pathological complete response in the neoadjuvant setting. ERBB2 expression also correlated with in vitro response to T-DM1. Finally, our assay identified 0.20-8.41% of tumors across 15 cancer types as ERBB2-high, including gastric and esophagus adenocarcinomas, urothelial carcinoma, cervical squamous carcinoma and pancreatic cancer. In particular, we identified high ERBB2 mRNA in a patient with HER2+ advanced gastric cancer who achieved a long-lasting partial response to T-DM1. Our study demonstrates that the heterogeneity in response to T-DM1 is partly explained by ERBB2 levels and provides a clinically applicable assay to be tested in future clinical trials of breast cancer and other cancer types.

The combined impact of cold smoking and natural antioxidants on quality and shelf life of dolphinfish (Coryphaena hippurus) fillets.

This study aimed to produce new value-added products from dolphinfish (Coryphaena hippurus) as fillets when using cold smoking combined with natural antioxidants, obtained from Halocnemum strobilaceum a plant belonging to halophytes. The fillet treatments were controlled (untreated), immersed in standard brine (STD), treated with NaCI and antioxidant before freezing (Aox-pre) and treated with NaCl and antioxidant after thawing (Aox-post). The quality of dolphinfish fillets was assessed using sensory, biochemical, microbiological, and technological parameters. Treating fillets with antioxidants before freezing and cold smoking (Aox-pre) enhanced significantly the shelf life, as well as improved the biochemical, microbiological, and sensory aspects of the product. Aox-pre fillets had the lowest peroxide values (PV) and malondialdehyde (MDA) content, demonstrating that the immediate treatment of fillets with the polyphenols protected lipids from peroxidation. The smoking process, together with antioxidant treatment, significantly reduced microbial counts after 15 days of storage, compared to the control. Overall, combining antioxidant treatments with cold smoking has a positive effect on the quality of Aox-pre fillets with respect to both sensory components and biochemical parameters associated with shelf life. Consequently, Aox-pre treatment enhances the marketability of this species, promoting local and seasonal products, contributing to costal fisheries sustainability.

Redox stress in Marfan syndrome: Dissecting the role of the NADPH oxidase NOX4 in aortic aneurysm.

Marfan syndrome (MFS) is characterized by the formation of ascending aortic aneurysms resulting from altered assembly of extracellular matrix fibrillin-containing microfibrils and dysfunction of TGF-ß signaling. Here we identify the molecular targets of redox stress in aortic aneurysms from MFS patients, and investigate the role of NOX4, whose expression is strongly induced by TGF-ß, in aneurysm formation and progression in a murine model of MFS. Working models included aortae and cultured vascular smooth muscle cells (VSMC) from MFS patients, and a NOX4-deficient Marfan mouse model (Fbn1C1039G/+-Nox4-/-). Increased tyrosine nitration and reactive oxygen species levels were found in the tunica media of human aortic aneurysms and in cultured VSMC. Proteomic analysis identified nitrated and carbonylated proteins, which included smooth muscle α-actin (αSMA) and annexin A2. NOX4 immunostaining increased in the tunica media of human Marfan aorta and was transcriptionally overexpressed in VSMC. Fbn1C1039G/+-Nox4-/- mice aortas showed a reduction of fragmented elastic fibers, which was accompanied by an amelioration in the Marfan-associated enlargement of the aortic root. Increase in the contractile phenotype marker calponin in the tunica media of MFS mice aortas was abrogated in Fbn1C1039G/+-Nox4-/- mice. Endothelial dysfunction evaluated by myography in the Marfan ascending aorta was prevented by the absence of Nox4 or catalase-induced H2O2 decomposition. We conclude that redox stress occurs in MFS, whose targets are actin-based cytoskeleton members and regulators of extracellular matrix homeostasis. Likewise, NOX4 have an impact in the progression of the aortic dilation in MFS and in the structural organization of the aortic tunica media, the VSMC phenotypic modulation, and endothelial function.

Differences in the Thoracic Aorta by Region and Sex in a Murine Model of Marfan Syndrome.

Marfan syndrome (MFS) is a hereditary disorder of the connective tissue that causes life-threatening aortic aneurysm, which initiates at the aortic root and can progress into the ascending portion. However, analysis of ascending aorta reactivity in animal models of MFS has remained elusive. Epidemiologic evidence suggests that although MFS is equally prevalent in men and women, men are at a higher risk of aortic complications than non-pregnant women. Nevertheless, there is no experimental evidence to support this hypothesis. The aim of this study was to explore whether there are regional and sex differences in the thoracic aorta function of mice heterozygous for the fibrillin 1 (Fbn1) allele encoding a missense mutation (Fbn1C1039G/+), the most common class of mutation in MFS. Ascending and descending thoracic aorta reactivity was evaluated by wire myography. Ascending aorta mRNA and protein levels, and elastic fiber integrity were assessed by qRT-PCR, Western blotting, and Verhoeff-Van Gieson histological staining, respectively. MFS differently altered reactivity in the ascending and descending thoracic aorta by either increasing or decreasing phenylephrine contractions, respectively. When mice were separated by sex, contractions to phenylephrine increased progressively from 3 to 6 months of age in MFS ascending aortas of males, whereas contractions in females were unchanged. Endothelium-dependent relaxation was unaltered in the MFS ascending aorta of either sex; an effect related to augmented endothelium-dependent hyperpolarization-type dilations. In MFS males, the non-selective cyclooxygenase (COX) inhibitor indomethacin prevented the MFS-induced enhancement of phenylephrine contractions linked to increased COX-2 expression. In MFS mice of both sexes, the non-selective nitric oxide synthase inhibitor L-NAME revealed negative feedback of nitric oxide on phenylephrine contractions, which was associated with upregulation of eNOS in females. Finally, MFS ascending aortas showed a greater number of elastic fiber breaks than the wild-types, and males exhibited more breaks than females. These results show regional and sex differences in Fbn1C1039G/+ mice thoracic aorta contractility and aortic media injuries. The presence of more pronounced aortic alterations in male mice provides experimental evidence to support that male MFS patients are at increased risk of suffering aortic complications.

Role of foetal MRI in the evaluation of ischaemic-haemorrhagic lesions of the foetal brain.

OBJECTIVE: The purpose of this study is to define the role of foetal magnetic resonance imaging (MRI) in evaluating cerebral ischaemic-haemorrhagic lesions and the extension of parenchymal injuries. STUDY DESIGN: From September 2006 to September 2010, 271 foetal MRI have been performed on cases referred to us for ultrasound suspect of brain abnormalities or cytomegalovirus infection and Toxoplasma serum conversion. Foetal MRI was performed with a 1.5-T magnet system without mother sedation. RESULTS: Foetal MRI detected ischaemic-haemorrhagic lesions in 14 of 271 foetuses, consisting of 5% incidence. MRI confirmed the diagnosis in three of 14 cases with ultrasonography (US) suspect of ischaemic-haemorrhagic lesions associated with ventriculomegaly. In one of 14 cases with US findings of cerebellar haemorrhage, MRI confirmed the diagnosis and provided additional information regarding the parenchymal ischaemic injury. In eight of 14 cases with US suspect of ventriculomegaly (3), corpus callosum agenesis (2), hypoplasia of cerebellar vermis (1), holoprosencephaly (1) and spina bifida (1), MRI detected ischaemic and haemorrhagic lesions unidentified at US examination. In two of 14 foetuses with US suspect of intracerebral space-occupying lesion, MRI modified the diagnosis to extra-axial haematoma associated with dural sinus malformation. Results were compared with post-mortem findings or afterbirth imaging follow-up. CONCLUSIONS: Foetal MRI is an additional imaging modality in the diagnosis of cerebral ischemic-haemorrhagic lesions, and it is useful in providing further information on the extension of the parenchymal injury and associated abnormalities, thus improving delivery management.

MRI and DWI: feasibility of DWI and ADC maps in the evaluation of placental changes during gestation.

OBJECTIVE: To establish if a correlation exists between apparent diffusion coefficient (ADC) values, obtained by diffusion-weighted imaging (DWI), and placental aging. METHOD: The study is divided into a retrospective phase and a prospective one.In the first phase, 145 pregnant women underwent fetal magnetic resonance imaging (MRI) for suspected disorders in several organs. We performed DWI (b value 0, 200 and 700 s/mm(2)) in all the fetuses, evaluating the patients in whom the whole placenta was visible.In the prospective phase, 50 women (52 fetuses) underwent MRI. We performed, in the same patient, two echo-planar sequences with b values of 0, 200 and 700, and 50, 200 and 700 s/mm(2), including the whole placenta.The ADC maps were calculated for all fetuses, divided into three groups based on gestational age (GA): group I: 20-26 weeks' gestation, II: 27-33, III: 34-40. RESULTS: In the retrospective phase, ADC values had a range from 1 to 2.4 mm(2)/s, showing a significant correlation between ADC values and GA.ADC values obtained by DWI with b value 0, 200 and 700 s/mm(2) had a range from 0.8 to 2.5 mm(2)/s, with an inverse correlation between ADC values and GA, whereas the ADC values with b value 50, 200 and 700 s/mm(2) did not show any statistical correlation (range: 1.5-1.7 mm(2)/s). CONCLUSION: DWI with ADC maps can not be considered markers for placental aging because they are affected by perfusional and circulatory motion.

Diffusion-weighted MR imaging and apparent diffusion coefficient of the normal fetal lung: preliminary experience.

OBJECTIVE: To assess if a correlation is present between apparent diffusion coefficient (ADC) values and normal lung maturation during gestation to define potential reference values as indicators of the lung development. METHODS: Our study included 50 pregnant women (gestational age, GA: 18-36 weeks), with normal fetal development of lungs assessed by a previous obstetric ultrasound (US), and then confirmed by our magnetic resonance (MR) examination. We used T2-weighted sequences, diffusion-weighted imaging sequences (DWI) and ADC maps for studying pulmonary tissue. In all cases the resulting ADC values were related to GA using Pearson correlation. RESULTS: ADC values ranged from 1,2 microm(2)/ms at 18 weeks' gestation to 3,9 microm(2)/ms at 36 weeks' gestation with a mean value, regardless for the gestational age, of 2, 352 +/- 0,623106 microm(2)/ms. We found a significant correlation between ADC and gestational age (Pearson correlation = 0,816). CONCLUSION: The ADC values correlate with gestational age since alveolar fluid secretion and angiogenesis increase gradually. Therefore, ADC can be considered as a new parameter for studying lung maturity.

MRI with diffusion-weighted imaging (DWI) and apparent diffusion coefficient (ADC) assessment in the evaluation of normal and abnormal fetal kidneys: preliminary experience.

OBJECTIVE: To investigate apparent diffusion coefficient (ADC) mapping with the measurement of renal tissue ADC value of normal and pathological fetal kidneys at various gestational ages (GAs). METHODS: Fifty pregnant women underwent magnetic resonance images (MRI) after ultrasound (US) for suspected fetal genitourinary disorders (16) or for suspected disorders in other organs (34). A multiplanar study of urinary system was obtained by using conventional T2-weighted sequences and echo planar imaging (EPI); Diffusion-weighted images and ADC maps were evaluated. The renal tissue ADC value was measured for all normal and abnormal fetuses and related to GA. RESULTS: MRI confirmed urinary anomalies in 15 fetuses [2 renal developmental variants, 2 nephropathies, 4 multicystic dysplastic kidneys (MCDK), 7 renal tract dilatations] and detected normal kidneys in the remaining 35 fetuses. Normal renal parenchyma showed bright signal on diffusion-weighted images with ADC values ranging from 1,065 to 1,327 microm(2)/s with a tendency to decrease over GA. A pathological ADC was detected in cases of bilateral MCDK, huge dilatations and in cases of nephropathies. CONCLUSION: Diffusion-weighted imaging (DWI) with ADC mapping can be used in the evaluation of fetal renal parenchyma and may become a tool of assessing function of the fetal kidney by means of measurement of renal tissue ADC values.

Use of systemic cyclosporin A in a case of severe Toxocara uveitis.

The report describes the case of an adult patient presenting a severe, serologically proven, Toxocara endophthalmitis, unresponsive to the common surgical and medical approach (vitrectomy, anti-parasitic treatment followed by high-dose corticosteroids). The association of oral cyclosporin A to a lower dose of prednisone was successful in achieving a long-term control of the ocular inflammation without systemic side effects.