SILICON AND PLATELET CONCENTRATES IN TISSUE REGENERATION:AN ¿IN VITRO¿ STUDY.

Tissue regeneration is a complex process of healing and tissue growth, which involves different biological elements and strategies, including the use of autologous cells, growth factors and scaffolds. All of these elements are present in platelet concentrates, such as Concentrated Growth Factors (CGF). Moreover, some trace elements play an important role in enhancing cell growth and proliferation and so tissue development and regeneration. Among these, Silicon seems to be beneficial for different organs and tissues, such as: bone and cartilage, brain, skin, nails and hair, cardiovascular system and immune system. For this reason, in the present study we evaluated the in vitro effect of Silicon (in the soluble form of Sodium Orthosilicate) and CGF on three different human cell lines of fibroblasts (NHDF), endothelial cells (HUVEC) and osteoblasts (HOBs). Each cell type, was treated with Sodium Orthosilicate at the final concentration of 0,5 mM and 1 mM, CGF and CGF supplemented with Sodium Orthosilicate, for 72 hours. At the end of the experimental period, the effect of the different treatments, on cell growth, proliferation and metabolic activity was evaluated by performing a simple cell count, using an automated cell counter and by evaluating the expression of the intracellular proliferation marker Ki-67, using FACS. Moreover, the expression of other cell markers and active molecules such as Collagen type I (Col I), Osteopontin (OPN), Vascular Endothelial Growth Factor (VEGF) and endothelial Nitric Oxide Synthase (eNOS), was evaluated, through immunohistochemical analyses on fixed cells. Results obtained showed that the use of CGF in combination with Sodium Orthosilicate stimulates cell growth, proliferation and metabolic activity. Overall, these findings suggest that in vitro treatment with CGF and Sodium Orthosilicate seems to be promised in promoting cell growth and proliferation and so in tissue regeneration

Optimization of pixel size and propagation distance in X-ray phase-contrast virtual histology

X-ray phase-contrast coupled to high-spatial resolution imaging systems provides a high sensitivity for distinguishing soft tissue structures in small samples, thus being suited for X-ray virtual histology. Propagation-based phase-contrast tomography can deliver a considerable gain in signal-to-noise ratio (SNR) at small pixel sizes when it is combined to a suitable phase retrieval filter. We optimized acquisition parameters, namely the propagation distance and the pixel size, with the aim of providing adequate spatial resolution and sensitivity for virtual histology of breast surgery specimens, scanned with a phase-contrast microtomography (mu CT) system employing a commercial sCMOS detector at the SYRMEP beamline of the Italian synchrotron facility Elettra (Trieste, Italy). A pathological breast tissue sample was embedded in paraffin and imaged using a polychromatic synchrotron beam at an average energy of 24 keV. The high numerical optical aperture of the imaging system enabled to adjust the pixel size to 1, 2.5 and 4 mu m. The scans were acquired at five sample-to-detector distances: 4.5, 150, 250, 500 and 1000 mm. SNR was measured in an homogeneous region portion of the mu CT image for each combination of pixel size and propagation distance. Experimental results were compared to a theoretical model taking into account the actual point spread function of the employed imaging system. The measured gain of SNR associated with the application of the phase-retrieval matched the predictions for large Fresnel numbers (N-F > 2). For each pixel size, an optimal range of propagation distances was found. Optimal mu CT reconstructions were then compared with their respective histopatological images, showing an excellent visibility of relevant structures. The optimization performed in this study will allow to select the most appropriate geometrical configurations for future acquisitions of virtual histology images of different specimens via phase-contrast microtomography

Atmospheric synoptic conditions of snow precipitation in East Antarctica using ice core and reanalysis data

In the framework of the International Partnerships in Ice Core Sciences (IPCS) initiatives the GV7 site (70°41’ S – 158°51’ E) in East Antarctica was chosen as the new drilling site for the Italian contribution to the understanding of the climatic variability in the last 2000 years (IPICS 2k Array). Water stable isotopes and snow accumulation (SMB) values from a shallow firn core, obtained at GV7 during the 2001-2002 International Trans-Antarctic Scientific Expedition (ITASE) traverse, are analyzed and compared with different meteorological model output in order to characterize the atmospheric synoptic conditions driving precipitation events at the site. On annual basis, ECMWF +24h forecasted snowfalls (SF) seem to well reproduce GV7 SMB values trend for the period from 1980 to 2005. Calculated air mass back-trajectories show that Eastern Indian - Western Pacific oceans represent the main moisture path toward the site during autumn - winter season. Analysis of the ECMWF 500 hPa Geopotential height field (GP500) anomalies shows that atmospheric blocking events developing between 130°E and 150°W at high latitudes drive the GV7 SMB by blocking zonal flow and conveying warm and moist deep air masses from ocean into the continental interior. On inter-annual basis, The SF variability over GV7 region follows the temporal oscillation of the third CEOF mode (CEOF3 10% of the total explained variance) of a combined complex empirical orthogonal function (CEOF) performed over GP500 and SF field. The CEOF3 highlights an oscillating feature, with wavenumber 2, in GP500 field over the Western Pacific-Eastern Indian Oceans and propagating westward. The pattern is deeply correlated with the Indian Dipole Oscillation and ENSO and their associated quasi-stationary Rossby waves propagating from the lower toward the higher latitudes

Acute Esophageal Necrosis as a Rare Complication of Metabolic Acidosis in a Diabetic Patient: A Case Report

Objective: Challenging differential diagnosisBackground: Acute esophageal necrosis, or Gurvits syndrome, is a rare clinical process often secondary to a systemic low -flow state. It can be caused by several medical conditions, and it is thought to arise from a combination of impaired mucosal barrier and chemical and ischemic insults to the esophagus. Acute esophageal necrosis usually presents with severe complications due to delayed diagnosis and only rarely has surgical indications. We present a case of Gurvits syndrome, presumably triggered by metabolic acidosis in a diabetic patient.Case Report: A 61-year-old man with history of hypertension and type 2 diabetes mellitus treated with metformin, canagliflozin, glimepiride, and pioglitazone came to our attention with persistent vomiting, odynophagia, chest pain after each meal, and progressive weight loss. Arterial blood analysis showed mild metabolic acidosis, while the first esophagogastroduodenoscopy performed revealed a circumferential black appearance of the esophageal mucosa, as in concentric necrosis of the distal esophagus with possible fungal superinfection. Brushing cytology confirmed the infection by Candida spp. and the patient was treated with intravenous fluconazole. The second esophagogastroduodenoscopy, performed after 2 weeks, showed almost complete healing of the esophageal mucosa; in this case, biopsy confirmed mucosal ischemia and necrosis, without showing deep impairment of the mucosa by fungal agents.Conclusions: Due to its high lethality, often caused by the underlying medical diseases, acute esophageal disease should be considered in the differential diagnosis of digestive symptoms, even without upper gastrointestinal bleeding. Prompt diagnosis and treatment of contextual collateral conditions can help clinicians to avoid the worst out-comes of the disease. Among the causative factors of metabolic acidosis leading to esophageal necrosis we recognized metformin and dapagliflozin

Prognostic Implications of the Complement Protein C1q in Gliomas

The contribution of the complement system in the pathophysiology of brain cancers has been recently considered in light of its well-known involvement in carcinogenesis. Complement system represents an important component of the inflammatory response, which acts as a functional bridge between the innate and adaptive immune response. C1q, the first recognition subcomponent of the complement classical pathway, has recently been shown to be involved in a range of pathophysiological functions that are not dependent on complement activation. C1q is expressed in the microenvironment of various types of human tumors, including melanoma, prostate, mesothelioma, and ovarian cancers, where it can exert a protective or a harmful effect on cancer progression. Despite local synthesis of C1q in the central nervous system, the involvement of C1q in glioma pathogenesis has been poorly investigated. We, therefore, performed a bioinformatics analysis, using Oncomine dataset and UALCAN database in order to assess whether the expression of the genes encoding for the three chains of C1q (C1qA, C1qB, and C1qC) could serve as a potential prognostic marker for gliomas. The obtained results were then validated using an independent glioma cohort from the Chinese Glioma Genome Atlas datasets. Our bioinformatics analysis, coupled with immunohistochemistry and fluorescence microscopy, appears to suggest a positive correlation between higher levels of C1q expression and unfavorable prognosis in a diverse grade of gliomas

THE CLINICOPATHOLOGICAL AND PROGNOSTIC SIGNIFICANCES OF C1Q EXPRESSION IN GLIOMAS: A BIOINFORMATICS ANALYSIS

Introduction. The complement system represents an important component of the inflammatory response and acts as a functional bridge between the innate and adaptive immune response. The contribution of the complement component C1q in the pathophysiology of brain cancers has been recently considered in light of its well-known involvement in carcinogenesis. Brain malignancies arise from cells of the CNS and are classified according to the tissue of phylogenetic origin. Gliomas represent the most common and aggressive form of brain tumours in adults. They derive from glial cells that help to support the functions of the other main brain cells type, the neurons (1). These are a heterogeneous group of diseases with multiple subtypes (1, 2). Glioblastoma multiforme (GBM) is the most common and fatal form of a primary brain tumour, accounting for approximately 60% of all glioma cases (3), whereas grade-II and -III gliomas are the second most common type of glioma in adults (~30%) (3). C1q molecule, together with other complement components, can be locally produced within the CNS by microglia and astrocytes, rendering it an attractive player in primary brain tumour development (4). The role of C1q in gliomas microenvironment is still poorly characterized and it is still quite puzzling whether it exerts a beneficial or a harmful activity for cancer progression. In the present study we performed a bioinformatics analysis aimed at investigating if C1q can serve as a potential prognostic marker for gliomas. Methods. The expression levels of C1qA, C1qB and C1qC genes in gliomas were analysed using Oncomine analysis. Available genomics data from The Cancer Genome Atlas project was used for Kaplan–Meier survival analysis to generate survival probability plots, using UALCAN analysis. Results. From the analysis performed on several data- sets using Oncomine, we showed a significantly higher mRNA expression levels for C1qA, C1qB and C1qC chains were detected in gliomas (different histotypes and grades) as compared to normal brain tissue (Fig. 1). We observed a positive correlation between the mRNA expression of C1qA, C1qB and C1qC mRNA poly- peptide chains and the unfavorable prognosis only in gliomas grade-II and -III, where the survival probability is indeed reduced (P <0.05) (Fig. 2). No correlation was observed in glioblastoma multiforme (Fig. 2). By immu- nohistochemical approaches we detected a high depo- sition of C1q in the tumor microenvironment of both in grade-II and -III gliomas and in GBMs examined (Fig. 3a glioma, 3b glioblastoma multiforme; 20x Magnification). Moreover, in double immunocytochemical experiments we demonstrated that CD68 positive infiltrating cells are actively synthesizing C1q in the tumor micro-envi- ronment. CD68 expression is characteristic of tumor- associated macrophages, whose enrichment in glioma has been associated with poor prognosis (5). Conclusion. In our study C1q expression was significantly correlated with poor survival probability in gliomas grade-II and -III while this is not the case for GBM. These data altogether underline how complex, multifaceted and still poorly understood is the role C1q can exert on tumor progression, and how the very same molecule can differentially affect the outcome depending on the biological context it comes to act

Effects of different sources of air pollution on the carbonate stone surface of relevant European monuments

This contribution focuses on spectrometric analyses carried out on black crust samples, collected from buildings and churches belonging to the European built Heritage, i.e., the Corner Palace in Venice (Italy), the Cathedral of St. Rombouts in Mechelen (Belgium), the Church of St. Eustache in Paris (France) and the Tower of London (United Kingdom). Such monuments, all built in carbonate stones, were selected for their historic and artistic relevance, as well as for their location in different urban contexts (exposed to intense vehicular traffic or pedestrian areas). For a complete characterization of the black crusts, an approach integrating complementary techniques was used, including optical (OM) and scanning electron microscopy coupled with energy-dispersive X-ray spectrometry (SEM-EDS), Fourier transform infrared spectroscopy (FT-IR) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). The complete characterization of the damage layers provided information on their chemical composition, the state of conservation of the underlying substrates and the interactions between crusts and stones. In particular, the geochemical study in terms of trace elements revealed that all crusts are enriched in heavy metals (As, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb, Sb, Sn, Ti, V, and Zn) compared to substrates. The different concentrations of such elements in all analyzed crust samples can be ascribed to several factors, such as: height of sampling, morphology of the sampled surfaces (vertical or horizontal), exposure to atmospheric agents, exposure to direct (road or boat traffic) or indirect (industries) sources of pollution, accumulation time of pollutants on the surfaces, wash out and particulate air pollution. Specifically, the crusts collected at lower heights (some samples of the Corner Palace, Cathedral of St. Rombouts and Tower of London) resulted to be mainly influenced by mobile sources of pollution (vehicular or boat traffic), while samples taken at higher heights (Church of St. Eustache and some samples of the Corner Palace) are generally mostly affected by stationary combustion sources. In some cases, the detailed analysis of multilayered crusts (Palazzo Corner) contributed to recognize the variation of combustion sources responsible for the deterioration of surfaces over time. In addition, the possibility of analyzing altered portions of the substrate (Tower of London) permitted to observe that some elements (Zn, Cu and Ni) show concentrations similar and, sometimes, higher than the overlying crusts. This result can be explained by the geochemical mobility of such elements (at specific environmental conditions), which accelerate the process of sulfating, rapidly creating new layers of crust In conclusion, the study of black crusts and altered substrates in terms of trace elements may provide information useful to understand the influence of the pollutants in the genesis of such degradation forms

TGS1 mediates 2,2,7-trimethyl guanosine capping of the human telomerase RNA to direct telomerase dependent telomere maintenance

Pathways that direct the selection of the telomerase-dependent or recombination-based, alternative lengthening of telomere (ALT) maintenance pathway in cancer cells are poorly understood. Using human lung cancer cells and tumor organoids we show that formation of the 2,2,7-trimethylguanosine (TMG) cap structure at the human telomerase RNA 5′ end by the Trimethylguanosine Synthase 1 (TGS1) is central for recruiting telomerase to telomeres and engaging Cajal bodies in telomere maintenance. TGS1 depletion or inhibition by the natural nucleoside sinefungin impairs telomerase recruitment to telomeres leading to Exonuclease 1 mediated generation of telomere 3′ end protrusions that engage in RAD51-dependent, homology directed recombination and the activation of key features of the ALT pathway. This indicates a critical role for 2,2,7-TMG capping of the RNA component of human telomerase (hTR) in enforcing telomerase-dependent telomere maintenance to restrict the formation of telomeric substrates conductive to ALT. Our work introduces a targetable pathway of telomere maintenance that holds relevance for telomere-related diseases such as cancer and aging

Prevalence of echinococcosis in humans, livestock and dogs in northern Italy

AbstractThe presence of Echinococcus sp. cysts was investigated in 822 sheep, 123 goats and 112,521 cattle from Lombardy region, North Italy. Faecal samples from 40 sheepdogs were also analyzed, with 9 samples containing taeniid eggs (22.5 %), 8 samples being coproantigen-positive (20 %), and one dog from a northern province (Lecco) positively confirmed by PCR. Cystic Echinococcosis (CE) was detected in 0.36 % of sheep and in 0.29 % of cattle in 2004. No goat resulted to be infected. Data from CE patients treated in Lombardy were collected by inspecting hospital discharge records. In 2004, 156 CE-related admissions (62 % male and 38 % female) were reported in Lombardy. Total hospital stay was 1,372 days (1,286 for inpatients, 86 for outpatients). Most patients (72.4 %) were residents in Lombardy and 1.9 % were from Piedmont; the remaining patients were from central and southern Italy. According to acquired data CE resulted hypoendemic in animals in Lombardy. Prevalence rates in humans were higher than expected in this region, usually considered as non-endemic. Assessment of the prevalence of CE in humans remains a difficult, costly, time-consuming and labourintensive task. The present study suggests establishing a National Registry of Cystic Echinococcosis with the aim to highlight regional risk factors and to benefit from its matching both clinical and epidemiological data

Prognostic implications of the complement protein C1q in gliomas

Associazione Italiana per la Ricerca sul Cancro (AIRC); Program Innovative Tools for Cancer Risk Assessment and Diagnosis