Osteoprotegerin: a pancreatic islets dysfunction and vascular injury modulator

Background. Osteoprotegerin (OPG) is a soluble glycoprotein of the tumor necrosis factor (TNF) receptor superfamily, which was initially identified as a key regulator in bone turnover. It acts as a decoy receptor for the receptor activator of nuclear factor kB ligand (RANKL) and for the TNF-related apoptosis-inducing ligand (TRAIL), counterbalancing their biological effects. OPG is produced by a wide range of tissues, including the cardiovascular system, and its levels are particularly high in aortic and renal arteries. Several studies have clearly demonstrated that the serum levels of OPG are elevated in diabetic and nondiabetic patients affected by cardiovascular diseases, and increased levels of OPG represent a risk factor for cardiovascular mortality, especially in diabetic patients. However, in spite of the reported findings, the physiopathological role of elevated serum levels of OPG in vascular biology and in pancreatic islet function are not well understood. Aim of the study. The aims of our studies were: Study 1. Evaluate the potential role of OPG in the pathogenesis of diabetes associated atherosclerosis. Study 2. Investigate OPG effects on pancreatic islet function and its interaction with local pancreatic renin-angiotensin system (RAS). Materials and Methods. Study 1.A. In vivo study: 80 apoE knockout male mice were further randomized into 4 groups (n=20) and followed for 3 months. One group of non diabetic animals received an intraperitoneal (i.p.) injection of vehicle and served as a control; another group of non-diabetic animals received every 3 weeks an i.p. injection of human recombinant OPG (OPG). The other two groups, rendered diabetic by 5 daily i.p. injections of streptozotocin (55mg/Kg/die), received injections of OPG or an equivalent volume of vehicle. At the end of the study, animals were culled, the blood was collected for biochemical analysis, and the entire aorta was excised out to study the total plaques extent and to evaluate the lesion composition and complexity of the aortic plaques. B. In vitro study: Murine vascular smooth muscle cells (VMSC) were treated with different concentrations of OPG, TGFβ and SB431542 (TGFβ- type 1 receptor inhibitor). Subsequently, cellular proliferation and pro-fibrotic markers gene expression were evaluated at different time points. OPG protein release was measured in growth media (ELISA technique). Study 2. 40 male mice C57Bl/6J, aged 10 weeks, were randomized into 4 groups (n=10) and studied for 3 months. Group 1 received every 3 weeks an i.p. injection of vehicle and served as a control. Group 2 received every 3 weeks an i.p. injection of OPG. Group 3 received the ACE inhibitor ramipril at the dose of 10mg/Kg/die in drinking water in co-treatment with i.p. injections of vehicle. Group 4 received ramipril in co-treatment with i.p. injections of OPG. At the end of the study, animals were culled, the blood was collected for biochemical analysis, and the pancreas was dissected out for subsequent quantitative RT-PCR measurements and immunohistochemical analysis. Results. Study 1.A. At the end of the study, diabetic animals injected with OPG presented a significant increase in total plaques extent, with an increase of smooth muscle cells content in aortic plaques. Moreover OPG treated animals showed an increase in the collagen content in aortic media in respect to control mice. B. OPG promoted VSMC proliferation and pro-fibrotic markers gene expression. TGFβ treatment of VSMC induced a dose-dependent increase of OPG gene and protein expression, that was completed prevented by pre-treatment with the SB431542 inhibitor. Study 2. OPG-treated animals showed increased islet monocyte-macrophage infiltration, fibrosis and apoptosis with reduction of islet function. The remodeling of islet architecture was associated with increased pancreatic expression of components of the RAS, growth factor genes (TGFβ and CTGF) and inflammatory molecules (MCP-1 and VCAM-1). Prevention of these changes with improvement of insulin secretion was observed in ramipril treated animals. Conclusion. Study 1.A-B OPG seems to play an important pathogenetic role in the development and progression of diabetic atherosclerosis. Study 2. Our data suggest that OPG might play an important role in promoting beta cell dysfunction and the upregulation of the local RAS represents one possible mechanism responsible for the OPG-induced beta cell dysfunction

Roles and Clinical Applications of OPG and TRAIL as Biomarkers in Cardiovascular Disease

Cardiovascular diseases (CVD) remain the major cause of death and premature disability in Western societies. Assessing the risk of CVD is an important aspect in clinical decision-making. Among the growing number of molecules that are studied for their potential utility as CVD biomarkers, a lot of attention has been focused on osteoprotegerin (OPG) and its ligands, which are receptor activator of nuclear factor \u3baB ligand (RANKL) and TNF-related apoptosis-inducing ligand. Based on the existing literature and on our experience in this field, here we review what the possible roles of OPG and TRAIL in CVD are and their potential utility as CVD biomarkers

The risk of cancer progression in women with gynecological malignancies and thrombophilic polymorphisms: a pilot case-control study.

Cancer produces a hypercoagulable state, which might lead to thrombosis, and on contrary, unprovoked venous thromboembolism might be the manifestation of an occult cancer. In this pilot case-control study, we assessed the risk of gynecological malignant diseases related to the presence of the factor V Leiden and prothrombin G20210A polymorphisms. Fifty-two women underwent an operation for gynecological malignancy and were enrolled in the study. Women who underwent an operation for gynecological nonmalignant disease in the same days of cases were considered as controls. The presence of factor V Leiden and prothrombin G20210A was assessed in case and control groups. In all, 7 out of 52 cases were carriers of the 2 polymorphisms compared with 20 out of 198 controls (odds ratio = 1.3; 95% confidence interval, 0.6-3.0). The results were also similar when the risk was considered separately for the site of cancer. As for advanced and metastatic malignancies, the odds ratios were 2.3 (95% confidence interval, 0.9-6.0) and 3.3 (95% confidence interval, 1.0-11), respectively, compared to noncancer patients. When these 2 groups were compared to nonadvanced cancer group, the odds ratios for carriers of polymorphisms were 2.7 (95%confidence interval, 0.7-11.0) and 3.9 (95%confidence interval, 0.8-18.6) for advanced cancer and metastatic malignancies, respectively. Women with factor V Leiden or prothrombin G20210A polymorphisms who developed gynecological malignancy might present with a higher stage of cancer at the time of surgery. Larger case-control studies in similar cohort of patients are needed to confirm these findings

Children With Short Stature Display Reduced ACE2 Expression in Peripheral Blood Mononuclear Cells

Background: The cause of short stature remains often unknown. The renin-angiotensin system contributes to growth regulation. Several groups reported that angiotensin-converting enzyme 2 (ACE2)-knockout mice weighed less than controls. Our case-control study aimed to investigate if children with short stature had reduced ACE2 expression as compared to controls, and its significance. Materials and Methods: children aged between 2 and 14 years were consecutively recruited in a University Hospital pediatric tertiary care center. Cases were children with short stature defined as height SD ≤ −2 diagnosed with growth hormone deficiency (GHD) or idiopathic short stature (ISS), before any treatment. Exclusion criteria were: acute diseases, kidney disease, endocrine or autoimmune disorders, precocious puberty, genetic syndromes, SGA history. ACE and ACE2 expression were measured in peripheral blood mononuclear cells, angiotensins were measured by ELISA. Results: Children with short stature displayed significantly lower ACE2 expression, being 0.40 fold induction (0.01-2.27) as compared to controls, and higher ACE/ACE2, with no differences between GHD and ISS. ACE2 expression was significantly and inversely associated with the risk of short stature, OR 0.26 (0.07-0.82), and it had a moderate accuracy to predict it, with an AUC of 0.73 (0.61-0.84). The cutoff of 0.45 fold induction of ACE2 expression was the value best predicting short stature, identifying correctly 70% of the children. Conclusions: Our study confirms the association between the reduction of ACE2 expression and growth retardation. Further studies are needed to determine its diagnostic implications

Clinical characterization of neonatal and pediatric enteroviral infections: an Italian single center study

Enteroviruses (EVs) are an important cause of illness, especially in neonates and young infants. Clinical and laboratory findings at different ages, brain imaging, and outcomes have been inadequately investigated.Background Enteroviruses (EVs) are an important cause of illness, especially in neonates and young infants. Clinical and laboratory findings at different ages, brain imaging, and outcomes have been inadequately investigated. Methods We retrospectively investigated EV infections occurring at an Italian tertiary care center during 2006-2017. Cases were confirmed with a positive polymerase chain reaction on blood or cerebrospinal fluid. Clinical and laboratory findings according to age at presentation were analyzed. Results Among 61 cases of EV infection, 56 had meningitis, 4 had encephalitis, and 1 had unspecific febrile illness. Forty-seven cases (77.0%) presented at less than 1 year of age, and most were less than 90 days of age (n = 44). Presentation with fever (p < 0.01), higher median temperature (p < 0.01), and irritability (p < 0.01) were significantly more common among infants aged less than 90 days, who also had significantly higher peak temperatures during the course of the disease (p < 0.01). In contrast, gastrointestinal symptoms were more common in infants and children aged over 90 days (p = 0.02). Only 4 of 61 infections (6.5%) were severe and all affected younger infants (p < 0.01). Conclusions We detail epidemiological, clinical, and laboratory findings in a cohort of 61 children. Infants aged less than 90 days have more severe disease; they are more likely to present with fever, higher median temperature, and irritability and less likely to develop gastrointestinal symptoms

Structural analysis of sulfate vein networks in Gale crater (Mars)

The Curiosity rover's campaign in the Gale crater on Mars provides a large set of close-up images of sedimentary formations outcrops displaying a variety of diagenetic features such as light-toned veins, nodules and raised ridges. Through 2D and 3D analyses of Mastcam images we herein reconstruct the vein network of a sample area and estimated the stress field. Assessment of the spatial distribution of light-toned veins shows that the basin infillings, after burial and consolidation, experienced a sub-vertical compression and lateral extension coupled with fluid overpressure and cracking. Overall, rock failure and light-toned veins formations could have been generated by an overload produced by infilling material within the basin

Characterization and significance of ACE2 and Mas receptor in human colon adenocarcinoma

Introduction: A new arm of the renin–angiotensin system (RAS) has been recently characterized; this includes angiotensin converting enzyme (ACE)2 and angiotensin (Ang)1-7, a heptapeptide acting through the Mas receptor (MasR). Recent studies show that Ang1-7 has an antiproliferative action on lung adenocarcinoma cells. The aim of this study was to characterize RAS expression in human colon adenocarcinoma and to investigate whether Ang1-7 exerts an antiproliferative effect on human colon adenocarcinoma cells. Materials and methods: Gene, protein expression and enzymatic activity of the main components of the RAS were determined on non-neoplastic colon mucosa as well as on the tumor mass and the mucosa taken 5 cm distant from it, both collected from patients with colon adenocarcinoma. Two different human colon cancer cell lines were treated with AngII and Ang1-7. Results: The novel finding of this study was that MasR was significantly upregulated in colon adenocarcinoma compared with non-neoplastic colon mucosa, which showed little or no expression of it. ACE gene expression and enzymatic activity were also increased in the tumors. However, AngII and Ang1-7 did not have any pro-/antiproliferative effects in the cell lines studied. Conclusions: The data suggest that upregulation of the MasR could be used as a diagnostic marker of colon adenocarcinoma

TRAIL reduces impaired glucose tolerance and NAFLD in the high-fat diet-fed mouse

Recent studies suggest that a circulating protein called TRAIL (TNF-related apoptosis inducing ligand) may have an important role in the treatment of type 2 diabetes. It has been shown that TRAIL deficiency worsens diabetes and that TRAIL delivery, when it is given before disease onset, slows down its development. The present study aimed at evaluating whether TRAIL had the potential not only to prevent, but also to treat type 2 diabetes. Thirty male C57BL/6J mice were randomized to a standard or a high-fat diet (HFD). After 4 weeks of HFD, mice were further randomized to receive either placebo or TRAIL, which was delivered weekly for 8 weeks. Body weight, food intake, fasting glucose, and insulin were measured at baseline and every 4 weeks. Tolerance tests were performed before drug randomization and at the end of the study. Tissues were collected for further analyses. Parallel in vitro studies were conducted on HepG2 cells and mouse primary hepatocytes. TRAIL significantly reduced body weight, adipocyte hypertrophy, free fatty acid levels, and inflammation. Moreover, it significantly improved impaired glucose tolerance, and ameliorated non-alcoholic fatty liver disease (NAFLD). TRAIL treatment reduced liver fat content by 47% in vivo as well as by 45% in HepG2 cells and by 39% in primary hepatocytes. This was associated with a significant increase in liver peroxisome proliferator-activated receptor (PPAR) \u3b3 (PPAR\u3b3) co-activator-1 \u3b1 (PGC-1\u3b1) expression both in vivo and in vitro, pointing to a direct protective effect of TRAIL on the liver. The present study confirms the ability of TRAIL to significantly attenuate diet-induced metabolic abnormalities, and it shows for the first time that TRAIL is effective also when administered after disease onset. In addition, our data shed light on TRAIL therapeutic potential not only against impaired glucose tolerance, but also against NAFLD

Hydrothermal Alteration of Ultramafic Rocks in Ladon Basin, Mars—Insights From CaSSIS, HiRISE, CRISM, and CTX

The evolution of the Ladon basin has been marked by intense geological activity and the discharge of huge volumes of water from the Martian highlands to the lowlands in the late Noachian and Hesperian. We explore the potential of the ExoMars Trace Gas Orbiter/Color and Stereo Surface Imaging System color image data set for geological interpretation and show that it is particularly effective for geologic mapping in combination with other data sets such as HiRISE, Context, and Compact Reconnaissance Imaging Spectrometer for Mars. The study area displays dark lobate flows of upper Hesperian to early Amazonian age, which were likely extruded from a regional extensional fault network. Spectral analysis suggests that these flows and the underlying rocks are ultramafic. Two distinct altered levels are observed below the lobate flows. The upper, yellow-orange level shows hundreds of structurally controlled narrow ridges reminiscent of ridges of listwanite, a suite of silicified, fracture-controlled silica-carbonate rocks derived from an ultramafic source and from serpentine. In addition to serpentinite, the detected mineral assemblages may include chlorite, carbonates, and talc. Kaolin minerals are detected in the lower, white level, which could have formed by groundwater alteration of plagioclase in the volcanic pile. Volcanism, tectonics, hydrothermal activity, and kaolinization are interpreted to be coeval, with hydrothermal activity and kaolinization controlled by the interactions between the aquifer and the hot, ultramafic lobate flows. Following our interpretations, East Ladon may host the first listwanite ridges described on Mars, involving a hydrothermal system rooted in a Hesperian aquifer and affecting ultramafic rocks from a magmatic source yet to be identified

QUALIDADE PÓS COLHEITA DE GOIABAS ‘PALUMA’ DE CULTIVO ORGÂNICO ARMAZENADAS EM FRIO NA SAFRA 2018

A goiaba é uma fruta que apresenta um período de vida muito curto pós-colheita, e poucas informações existem sobre a armazenagem de goiabas de cultivo orgânico. O trabalho avaliou a qualidade de goiabas ‘Paluma’ produzidas em cultivo orgânico e armazenadas em frio sob atmosfera normal (AN) e modificada (AM). Em AN os frutos foram armazenados em bandejas alveoladas, dentro de caixas de papelão e em câmara fria a 7ºC. Em AM os frutos foram armazenados em bandejas alveoladas envoltas por bolsas plásticas fechadas, dentro de caixas de papelão em câmara fria a 7ºC. Avaliações ocorreram aos 0, 10, 20, 30 e 40 dias de armazenamento refrigerado para perda de massa fresca, firmeza de polpa externa e interna, conteúdo de suco, diâmetro e sólidos solúveis. A perda de massa e a firmeza interna e externa não apresentaram diferenças entre os tratamentos. O conteúdo de suco foi superior nos frutos em AM após 20 e 40 dias em frio. Os sólidos solúveis foram maiores aos 30 dias em frio nos frutos em AM