Along-Path Evolution of Biogeochemical and Carbonate System Properties in the Intermediate Water of the Western Mediterranean

A basin-scale oceanographic cruise (OCEANCERTAIN2015) was carried out in the Western Mediterranean (WMED) in summer 2015 to study the evolution of hydrological and biogeochemical properties of the most ubiquitous water mass of the Mediterranean Sea, the Intermediate Water (IW). IW is a relatively warm water mass, formed in the Eastern Mediterranean (EMED) and identified by a salinity maximum all over the basin. While it flows westward, toward and across the WMED, it gradually loses its characteristics. This study describes the along-path changes of thermohaline and biogeochemical properties of the IW in the WMED, trying to discriminate changes induced by mixing and changes induced by interior biogeochemical processes. In the first part of the path (from the Sicily Channel to the Tyrrhenian Sea), respiration in the IW interior was found to have a dominant role in determining its biogeochemical evolution. Afterward, when IW crosses regions of enhanced vertical dynamics (Ligurian Sea, Gulf of Lion and Catalan Sea), mixing with surrounding water masses becomes the primary process. In the final part of the investigated IW path (the Menorca-Mallorca region), the role of respiration is further masked by the effects of a complex circulation of IW, indicating that short-term sub-regional hydrological processes are important to define IW characteristics in the westernmost part of the investigated area. A pronounced along-path acidification was detected in IW, mainly due to remineralization of organic matter. This induced a shift of the carbonate equilibrium toward more acidic species and makes this water mass increasingly less adequate for an optimal growth of calcifying organisms. The carbonate buffering capacity also decreases as IW flows through the WMED, making it more exposed to the adverse effects of a decreasing pH. The present analysis indicates that IW evolution in the sub-basins of the WMED is currently driven by complex hydrological and biogeochemical processes, which could be differently impacted by coming climate changes, in particular considering expected increases of extreme meteorological events, mainly due to the warming of the Mediterranean basin

Local injection of bone marrow progenitor cells for the treatment of anal sphincter injury: in-vitro expanded versus minimally-manipulated cells

Background: Anal incontinence is a disabling condition that adversely affects the quality of life of a large number of patients, mainly with anal sphincter lesions. In a previous experimental work, in-vitro expanded bone marrow (BM)-derived mesenchymal stem cells (MSC) were demonstrated to enhance sphincter healing after injury and primary repair in a rat preclinical model. In the present article we investigated whether unexpanded BM mononuclear cells (MNC) may also be effective. Methods: Thirty-two rats, divided into groups, underwent sphincterotomy and repair (SR) with primary suture of anal sphincters plus intrasphincteric injection of saline (CTR), or of in-vitro expanded MSC, or of minimally manipulated MNC; moreover, the fourth group underwent sham operation. At day 30, histologic, morphometric, in-vitro contractility, and functional analysis were performed. Results: Treatment with both MSC and MNC improved muscle regeneration and increased contractile function of anal sphincters after SR compared with CTR (p < 0.05). No significant difference was observed between the two BM stem cell types used. GFP-positive cells (MSC and MNC) remained in the proximity of the lesion site up to 30 days post injection. Conclusions: In the present study we demonstrated in a preclinical model that minimally manipulated BM-MNC were as effective as in-vitro expanded MSC for the recovery of anal sphincter injury followed by primary sphincter repair. These results may serve as a basis for improving clinical applications of stem cell therapy in human anal incontinence treatment

Systemic Sclerosis Sera Impair Angiogenic Performance of Dermal Microvascular Endothelial Cells: Therapeutic Implications of Cyclophosphamide

In systemic sclerosis (SSc), dermal capillaries are progressively lost with consequent chronic tissue hypoxia insufficiently compensated by angiogenesis. Clinical studies reported that intravenous cyclophosphamide (CYC) may improve SSc-related peripheral microvascular damage. Recently, we showed that CYC treatment may normalize SSc sera-induced abnormalities in endothelial cell-matrix interactions. Our objective was to evaluate in vitro the effects of sera from treatment-naïve or CYC-treated SSc patients on dermal blood microvascular endothelial cell (dMVEC) angiogenesis, migration, proliferation and apoptosis. dMVECs were challenged with sera from 21 SSc patients, treatment-naïve (n = 8) or under CYC treatment (n = 13), and 8 healthy controls. Capillary morphogenesis on Geltrex matrix was significantly reduced upon challenge with sera from naïve SSc patients compared with healthy controls. When dMVECs were challenged with sera from CYC-treated SSc patients, their angiogenic capacity was comparable to that of cells treated with healthy sera. Wound healing capacity and chemotaxis in Boyden chamber were both significantly decreased in the presence either of naïve or CYC-treated SSc sera compared with healthy sera. WST-1 assay revealed that cell proliferation was significantly decreased in dMVECs challenged with sera from naïve SSc patients compared with healthy sera. Conversely, dMVEC proliferation was not impaired in the presence of sera from CYC-treated SSc patients. Accordingly, the percentage of TUNEL-positive apoptotic dMVECs was significantly higher in the presence of sera from naïve SSc patients than healthy controls, while CYC-treated SSc sera did not induce dMVEC apoptosis. Levels of the angiostatic mediators endostatin, pentraxin 3, angiostatin and matrix metalloproteinase-12 were all significantly elevated in sera from naïve SSc patients compared with sera from both healthy controls and CYC-treated SSc patients. In SSc, CYC treatment might boost angiogenesis and consequently improve peripheral microangiopathy through the normalization of the endothelial cell-matrix interactions, reduction of endothelial cell apoptosis and rebalance of dysregulated angiostatic factors

Morphometric analysis of lymphatics vessels in fibrotic human lung

In pulmonary fibrosis, the usual interstitial pneumonia (UIP) pattern is characterised by heterogeneous, patchy fibrosis, with areas of normal lung adjacent to areas of complete destruction (honeycombing) and by fibroblastic foci (FF). The NSIP pattern which is characteristic of systemic sclerosis, is characterised by a more homogeneous involvement of the lung without honeycombing and FF. Little is known on lymphatic vessels in lung fibrosis. Defective lymphatic clearance could lead to prolonged exposure to pathogenic antigens and/or pro-inflammatory/pro-fibrotic mediators. We evaluated the distribution and morphology of lymphatic vessels in lung biopsies of 6 patients with UIP, 6 NSIP and 5 controls. Consecutive sections were stained with Movat’s pentachrome and with double immunostaining for von Willebrand factor and podoplanin (D2-40). Area, perimeter and position were recorded for vessels with a diameter &gt; 5µm. We investigated separately in lintralobular, sub-pleural, and interlobular spaces. Lymphatics were consistently larger in subpleural spaces and in interlobular septa than in intralobular tissue. In the latter, the density of lymphatic vessels was significantly reduced in NSIP and in UIP (both 21±1 mm-2) compared to controls (35±4 mm-2) . In controls, 85±6% of the intralobular lymphatics were close (&lt; 100 µm) to a blood vessel, and only 5±4% were in the proximity of bronchoalveolar spaces, while in the disease groups they were less frequently perivascular (NSIP 55 ±3%, UIP 56 ±2%) and more frequently associated with the bronchoalveolar lumen (NSIP 85 ±3%, UIP 69 ±2%). By contrast, in interlobular septa, lymphatic density was significantly increased in NSIP (303±28 mm-2) and in UIP (286±124 mm-2) compared to controls (96±69 mm-2). No differences in lymphatic density was seen in subpleural spaces. Thus, our data show a marked redistribution of lymphatic vessels within the lung in pulmonary fibrosis, without noticeable differences between the NSIP and UIP patterns

New Insights into the Pathophysiology of Primary and Secondary Lymphedema: Histopathological Studies on Human Lymphatic Collecting Vessels

Background:Lymphedema is characterized by an accumulation of interstitial fluids due to inefficient lymphatic drainage. Primary lymphedema is a rare condition, including congenital and idiopathic forms. Secondary lymphedema is a common complication of lymph node ablation in cancer treatment. Previous studies on secondary lymphedema lymphatic vessels have shown that after an initial phase of ectasia, worsening of the disease is associated with wall thickening accompanied by a progressive loss of the endothelial marker podoplanin. Methods and Results:We enrolled 17 patients with primary and 29 patients with secondary lymphedema who underwent lymphaticovenous anastomoses surgery. Histological sections were stained with Masson's trichrome, and immunohistochemistry was performed with antibodies to podoplanin, smooth muscle alpha-actin (alpha-SMA), and myosin heavy chain 11 (MyH11). In secondary lymphedema, we found ectasis, contraction, and sclerosis vessel types. In primary lymphedema, the majority of vessels were of the sclerosis type, with no contraction vessels. In both primary and secondary lymphedema, not all alpha-SMA-positive cells were also positive for MyH11, suggesting transformation into myofibroblasts. The endothelial marker podoplanin had a variable expression unrelatedly with the morphological vessel type. Conclusions:Secondary lymphedema collecting vessels included all the three types described in literature, that is, ectasis, contraction, and sclerosis, whereas in primary lymphedema, we found the ectasis and the sclerosis but not the contraction type. Some cells in the media stained positively for alpha-SMA but not for MyH11. These cells, possibly myofibroblasts, may contribute to collagen deposition

Antimicrobial activity against Helicobacter pylori strains and antioxidant properties of blackberry leaves (Rubus ulmifolius) and isolated compounds

8noRubus spp. (Rosaceae) provide extracts used in traditional medicine as antimicrobial, anticonvulsant, muscle relaxant and radical scavenging agents. Resistance to antibiotics used to treat Helicobacter pylori infection as well as their poor availability in developing countries prompted us to test the antimicrobial activity of Rubus ulmifolius leaves and isolated polyphenols against two H. pylori strains with different virulence (CagA+ strain 10K and CagA(-) strain G21). The antioxidant activity (TEAC values) of the tested compounds ranged from 4.88 (gallic acid) to 1.60 (kaempferol), whilst the leaf extract gave a value of 0.12. All the isolated polyphenols as well as the leaf extract showed antibacterial activity against both of the H. pylori strains. The minimum bactericidal concentrations (MBCs) of the extract for H. pylori strains G21 and 10K, respectively, were 1200 microg/mL and 1500 microg/mL after 24h of exposure and 134 microg/mL and 270 microg/mL after 48 h exposure. Ellagic acid showed very low MBC values towards both of the H. pylori strains after 48 h (2 microg/mL and 10 microg/mL for strains G21 and 10K, respectively) and kaempferol toward G21 strain (MBC=6 microg/mL). A relationship between antimicrobial activity and antioxidant capacity was found only for H. pylori strain G21 CagA(-) strain.reservedmixedMartini, Silvia; D'Addario, C.; Colacevich, Andrea; Focardi, Silvia; Borghini, Francesca; Santucci, Annalisa; Figura, Natale; Rossi, ClaudioMartini, Silvia; D'Addario, C.; Colacevich, Andrea; Focardi, Silvia; Borghini, Francesca; Santucci, Annalisa; Figura, Natale; Rossi, Claudi

The carbonate chemistry of the Western Mediterranean during the OCEAN CERTAIN 2015 cruise

In Summer 2015 (4-31 August), CNR-ISMAR carried out an oceanographic field-study in the Western Mediterranean Sea, on board of R/V MINERVA UNO. Sampling stations consisted in 7 transects, that spanned from Sicily Channel to Ligurian sea, Catalan sea, and Balearic Basin, dividing the area in sub-basins. 92 stations were visited totally. The dataset includes 550 discrete data of carbonate chemistry (pH-total scale and Total Alkalinity), concentrations of dissolved oxygen, and basic hydrological data (temperature, salinity and density). Methods: At each station, pressure (dbar), temperature (°C), and conductivity(mS/cm) were measured with a CTD SBE 911 plus General Oceanics Rosette System, equipped with 24 12-litres Niskin Bottles. Salinity (S, psu) and depth (m) were calculated by Sea-Bird Scientific routines. Seawater samples (n = 550) for the determination of biogeochemical parameters were collected from the Niskin bottles. Samples for dissolved oxygen (DO) were drawn in 60-mL BOD bottles and treated with Winkler reagents immediately after collection. For the determination of pH on the total hydrogen ion scale at 25 °C (pHT25), the samples were drawn after DO samples into 10 cm long cylindrical glass cells and analyzed spectrophotometrically. For the determination of total alkalinity (TA; μmol kg-1), the samples were collected in 300 ml borosilicate bottles, poisoned with mercuric chloride, tightly closed and stored in the dark at a temperature similar to the in situ one (4-25 °C). DO samples were analyzed by the Winkler method (Grasshoff et al., 1999) using an automated Metrohm 798 MPT Titrino potentiometric titration system (CV = 0.17 % at 210 µmol L-1). pHT25 was measured on board, within 24 h after the sampling, using the spectrophotometric method with m-cresol purple as indicator (Clayton and Byrne, 1993). The precision was ±0.002 units (n = 3), accuracy and stability of the method were checked daily with reference seawater certified for TA and TCO2 (n = 34, CRM batch 146 provided by Prof A. G. Dickson, Scripps, 210 California). TA was determined by potentiometric titration in an open cell with a difference derivative readout (Hernandez-Ayon et al., 1999). The average precision was ±2.0 μmol kg-1 (n = 86 duplicate samples) and the accuracy was checked daily by the titration of certified reference seawater (n = 59, CRM batch 146

Estimating Lagrangian transport blending drifters with HF radar data and models: Results from the TOSCA experiment in the Ligurian Current (North Western Mediterranean Sea)

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Systemic sclerosis sera affect fibrillin-1 deposition and focal adhesion molecule expression by dermal blood microvascular endothelial cells: Therapeutic implications of cyclophosphamide

Systemic sclerosis (SSc, scleroderma) is characterized by fibrosis of the skin and internal organs, autoantibodies and diffuse microangiopathy. In SSc, fibrillin-1 microfibrils are disorganized and unstable. Mutations in FBN1 gene and anti-fibrillin- 1 autoantibodies have also been reported in SSc patients. Fibrillin-1 microfibrils sequester in the extracellular matrix the latent form of the potent pro-fibrotic cytokine transforming growth factor-β (TGF-β). We herein report the effects of SSc sera on the deposition of fibrillin-1 and microfibril-associated glycoprotein-1 (MAGP-1) and the expression of focal adhesion molecules by dermal blood (B-MVECs) and lymphatic (Ly-MVECs) microvascular endothelial cells. Cells were challenged with sera from SSc patients, naïve or under cyclophosphamide (CYC) treatment, and healthy controls. Fibrillin-1/MAGP-1 synthesis and deposition and the expression of αvβ3 integrin/ phosphorylated-FAK and vinculin/actin were evaluated by immunofluorescence and quantified by morphometric analysis. Fibrillin-1 and MAGP-1 co-localized in all experimental conditions, forming a honeycomb pattern in B-MVECs and a dense mesh of short segments in Ly-MVECs. In B-MVECs, fibrillin-1/MAGP-1 production and αvβ3 integrin expression significantly decreased upon challenge with sera from naïve SSc patients compared with healthy controls. Vinculin was overexpressed in the cells exposed to the serum of naïve patients with the diffuse form of SSc. B-MVECs challenged with sera from CYC-treated SSc patients showed fibrillin-1, αvβ3 integrin and vinculin levels comparable to those of controls. Ly-MVECs challenged with SSc sera did not differ from controls in the expression of any of the molecules assayed. Due to the critical role of fibrillin-1 in sequestering TGF-β in the extracellular matrix, its decreased deposition by B-MVECs challenged with SSc sera might contribute to dermal fibrosis. CYC treatment might limit fibrosis through the maintenance of physiologic fibrillin-1 synthesis and deposition by B-MVECs. CYC-induced normalization of αvβ3 integrin expression may restore effective interaction of endothelial cells with fibrillin. Since vinculin is necessary for angiogenesis, its normalization upon CYC treatment may contribute to restore the impaired angiogenesis found in SSc