Glial cell-derived neurotrophic factor (GDNF) promotes invasive bahavior in testicular seminoma cells

The neurotrophic factor GDNF has multiple functions that promote cell survival, proliferation and migration in different cell types. The experimental over-expression of GDNF in mouse testis leads to infertility and promotes seminomatous germ cell tumors in older animals, which suggests that although the underlying mechanisms are unknown, deregulation of the GDNF pathway may be implicated in germ cell carcinogenesis. To explore the involvement of the GDNF pathway in the onset and progression of testicular germ cell tumors, we firstly analyzed the expression pattern of GFRA1 and Ret, the two major co-receptors for GDNF, in seminoma samples. We report that GFRA1 was expressed more extensively in carcinoma in situ (CIS) cells and intratubular invasive seminoma compared with normal testis. Functional analysis of the GDNF biological activity was performed on TCam-2 human seminoma cell line. RT-PCR and immunohistochemical analyses demonstrate that TCam-2 cells express both GFRA1 and Ret mRNA, but only GFRA1 was detected at the protein level. It is well known that GDNF plays a central role in spermatogonial stem cell self-renewal and proliferation (Hoffman MC et al., 2008). Therefore, in order to evaluate if GDNF could act as a mitogenic factor in TCam-2 cells, we performed proliferation assays and cell cycle analyses. Interestingly we observed that GDNF doesn’t induce increase in total cell number or S-phase entry in TCam-2 cell line. Several evidences suggested that GDNF is able to induce cell migration and invasion in several normal and tumor cell types (Okada Y et al., 1999; Veit C et al., 2004; Su CM et al., 2009; Song H et al., 2006; Paratcha G et al., 2006; Tang MJ et al., 1998; Young HM et al., 2001). Therefore we have hypothesized that GDNF can act as a chemoattractant also in seminoma cells. On this purpose we stimulated directional TCam-2 cell migration and invasion in the presence of GDNF gradients and we investigated the downstream pathways responsible for the GDNF-induced invasive behaviour. We demonstrated that GDNF is able to induce migration, possibly through the Src and MEK pathways. It is documented that tumor cell invasion can occur through two modalities: a proteolytic mesenchymal-like or non-proteolytic amoeboid-like modality (Sahai E and Marshall CJ, 2003; Wolf K et al., 2003). GDNF is able to induce TCam-2 seminoma cell invasion in a mesenchymal-like metalloprotease-dependent manner. In conclusion, GFRA1 over-expression in CIS and seminoma cells, along with the functional analyses in TCam-2 cells, suggests an involvement of the GDNF pathway in the progression of testicular germ cell cancer

R-spondin 1/Dickkopf-1/beta-catenin machinery is involved in testicular embryonic angiogenesis

Testicular vasculogenesis is one of the key processes regulating male gonad morphogenesis. The knowledge of the molecular cues underlining this phenomenon is one of today's most challenging issues and could represent a major contribution toward a better understanding of the onset of testicular morphogenetic disorders. R-spondin 1 has been clearly established as a candidate for mammalian ovary determination. Conversely, very little information is available on the expression and role of R-spondin 1 during testicular morphogenesis. This study aims to clarify the distribution pattern of R-spondin 1 and other partners of its machinery during the entire period of testicular morphogenesis and to indicate the role of this system in testicular development. Our whole mount immunofluorescence results clearly demonstrate that R-spondin 1 is always detectable in the testicular coelomic partition, where testicular vasculature is organized, while Dickkopf-1 is never detectable in this area. Moreover, organ culture experiments of embryonic male UGRs demonstrated that Dickkopf-1 acted as an inhibitor of testis vasculature formation. Consistent with this observation, real-time PCR analyses demonstrated that DKK1 is able to slightly but significantly decrease the expression level of the endothelial marker Pecam1. The latter experiments allowed us to observe that DKK1 administration also perturbs the expression level of the Pdgf-b chain, which is consistent with some authors' observations relating this factor with prenatal testicular patterning and angiogenesis. Interestingly, the DKK1 induced inhibition of testicular angiogenesis was rescued by the co-administration of R-spondin 1. In addition, R-spondin 1 alone was sufficient to enhance, in culture, testicular angiogenesis

HIRO-NET.Heterogeneous intelligent robotic network for internet sharing in disaster scenarios

This article describes HIRO-NET, an Heterogeneous Intelligent Robotic Network. HIRO-NET is an emergency infrastructure-less network that aims to address the problem of providing connectivity in the immediate aftermath of a natural disaster, where no cellular or wide area network is operational and no Internet access is available. HIRO-NET establishes a two-tier wireless mesh network where the Lower Tier connects nearby survivors in a self-organized mesh via Bluetooth Low Energy (BLE) and the Upper Tier creates long-range VHF links between autonomous robots exploring the disaster stricken area. HIRO-NET’s main goal is to enable users in the disaster area to exchange text messages to share critical information and request help from first responders. The mesh network discovery problem is analyzed and a network protocol specifically designed to facilitate the exploration process is presented. We show how HIRO-NET robots successfully discover, bridge and interconnect local mesh networks. Results show that the Lower Tier always reaches network convergence and the Upper Tier can virtually extend HIRO-NET functionalities to the range of a small metropolitan area. In the event of an Internet connection still being available to some user, HIRO-NET is able to opportunistically share and provide access to low data-rate services (e.g., Twitter, Gmail) to the whole network. Results suggest that a temporary emergency network to cover a metropolitan area can be created in tens of minutes. Inde

Metatranscriptomics reveals temperature-driven functional changes in microbiome impacting cheese maturation rate

© The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Scientific Reports 6 (2016): 21871, doi:10.1038/srep21871.Traditional cheeses harbour complex microbial consortia that play an important role in shaping typical sensorial properties. However, the microbial metabolism is considered difficult to control. Microbial community succession and the related gene expression were analysed during ripening of a traditional Italian cheese, identifying parameters that could be modified to accelerate ripening. Afterwards, we modulated ripening conditions and observed consistent changes in microbial community structure and function. We provide concrete evidence of the essential contribution of non-starter lactic acid bacteria in ripening-related activities. An increase in the ripening temperature promoted the expression of genes related to proteolysis, lipolysis and amino acid/lipid catabolism and significantly increases the cheese maturation rate. Moreover, temperature-promoted microbial metabolisms were consistent with the metabolomic profiles of proteins and volatile organic compounds in the cheese. The results clearly indicate how processing-driven microbiome responses can be modulated in order to optimize production efficiency and product quality.F.D.F. was supported by a grant from Regione Campania within the program “POR CAMPANIA FSE 2007/2013” - project CARINA (Safety sustainability and competitiveness of the agro-food production in Campania) – CUP B25B09000080007

San Vincenzo, Isola di Stromboli (Lipari, Prov. Di Messina) - Campagna 2014

Presentazione sintetica dei risultati della campagne di scavo effettuate nel 2014 nel sito archeologico di San Vincenzo a Strombol

How Dual-Energy Contrast-Enhanced Spectral Mammography Can Provide Useful Clinical Information About Prognostic Factors in Breast Cancer Patients: A Systematic Review of Literature

Introduction: In the past decade, a new technique derived from full-field digital mammography has been developed, named contrast-enhanced spectral mammography (CESM). The aim of this study was to define the association between CESM findings and usual prognostic factors, such as estrogen receptors, progesterone receptors, HER2, and Ki67, in order to offer an updated overview of the state of the art for the early differential diagnosis of breast cancer and following personalized treatments. Materials and methods: According to the PRISMA guidelines, two electronic databases (PubMed and Scopus) were investigated, using the following keywords: breast cancer AND (CESM OR contrast enhanced spectral mammography OR contrast enhanced dual energy mammography) AND (receptors OR prognostic factors OR HER2 OR progesterone OR estrogen OR Ki67). The search was concluded in August 2021. No restriction was applied to publication dates. Results: We obtained 28 articles from the research in PubMed and 114 articles from Scopus. After the removal of six replicas that were counted only once, out of 136 articles, 37 articles were reviews. Eight articles alone have tackled the relation between CESM imaging and ER, PR, HER2, and Ki67. When comparing radiological characterization of the lesions obtained by either CESM or contrast-enhanced MRI, they have a similar association with the proliferation of tumoral cells, as expressed by Ki-67. In CESM-enhanced lesions, the expression was found to be 100% for ER and 77.4% for PR, while moderate or high HER2 positivity was found in lesions with non-mass enhancement and with mass closely associated with a non-mass enhancement component. Conversely, the non-enhancing breast cancer lesions were not associated with any prognostic factor, such as ER, PR, HER2, and Ki67, which may be associated with the probability of showing enhancement. Radiomics on CESM images has the potential for non-invasive characterization of potentially heterogeneous tumors with different hormone receptor status. Conclusions: CESM enhancement is associated with the proliferation of tumoral cells, as well as to the expression of estrogen and progesterone receptors. As CESM is a relatively young imaging technique, a few related works were found; this may be due to the "off-label" modality. In the next few years, the role of CESM in breast cancer diagnostics will be more thoroughly investigated

Microgravity Induces Transient EMT in Human Keratinocytes by Early Down-Regulation of E-Cadherin and Cell-Adhesion Remodeling

Abstract: Changes in cell–matrix and cell-to-cell adhesion patterns are dramatically fostered by the microgravity exposure of living cells. The modification of adhesion properties could promote the emergence of a migrating and invasive phenotype. We previously demonstrated that short exposure to the simulated microgravity of human keratinocytes (HaCaT) promotes an early epithelial– mesenchymal transition (EMT). Herein, we developed this investigation to verify if the cells maintain the acquired invasive phenotype after an extended period of weightlessness exposure. We also evaluated cells’ capability in recovering epithelial characteristics when seeded again into a normal gravitational field after short microgravity exposure. We evaluated the ultra-structural junctional features of HaCaT cells by Transmission Electron Microscopy and the distribution pattern of vinculin and E-cadherin by confocal microscopy, observing a rearrangement in cell–cell and cell–matrix interactions. These results are mirrored by data provided by migration and invasion biological assay. Overall, our studies demonstrate that after extended periods of microgravity, HaCaT cells recover an epithelial phenotype by re-establishing E-cadherin-based junctions and cytoskeleton remodeling, both being instrumental in promoting a mesenchymal–epithelial transition (MET). Those findings suggest that cytoskeletal changes noticed during the first weightlessness period have a transitory character, given that they are later reversed and followed by adaptive modifications through which cells miss the acquired mesenchymal phenotyp

Clinical and trichoscopic graded live visual scale for androgenetic alopecia

Introduction: Currently, the mostly used classifications of androgenetic alopecia (AGA), only provide a macroscopic and subjective description of this disorder, without evaluating trichoscopic features. Objective: The aim of this study is to elaborate a graded live visual AGA severity scale including macroscopic and microscopic (trichoscopic) pictures, and to determine the most frequent trichoscopic characteristics associated to each grade. Methods: A retrospective observational study was conducted on 122 patients (50 females and 72 males) affected by AGA. Macroscopic and trichoscopic photographs were taken at standardized scalp points. Results: Each picture was ranked from AGA stage I to VII, according to Hamilton scale for men and Savin scale for women, and the most representative images of each severity degree were collected to produce a graded live visual scale. In males, two live visual scales, one for the anterior and one for posterior region of the scalp were created. In females, only one scale of the anterior region was realized. For each stage of severity, the corresponding trichoscopic parameters were statistically analyzed. Conclusions: We realized new macroscopic and trichoscopic graded live visual scales for male and female patients affected by AGA, which could help physicians in giving an objective evaluation of the disease and in better managing it

Fasting glucose and body mass index as predictors of activity in breast cancer patients treated with everolimus-exemestane: the EverExt study

Evidence on everolimus in breast cancer has placed hyperglycemia among the most common high grade adverse events. Anthropometrics and biomarkers of glucose metabolism were investigated in a observational study of 102 postmenopausal, HR + HER2- metastatic breast cancer patients treated with everolimus-exemestane in first and subsequent lines. Best overall response (BR) and clinical benefit rate (CBR) were assessed across subgroups defined upon fasting glucose (FG) and body mass index (BMI). Survival was estimated by Kaplan-Meier method and log-rank test. Survival predictors were tested in Cox models. Median follow up was 12.4 months (1.0-41.0). The overall cohort showed increasing levels of FG and decreasing BMI (p < 0.001). Lower FG fasting glucose at BR was more commonly associated with C/PR or SD compared with PD (p < 0.001). We also observed a somewhat higher BMI associated with better response (p = 0.052). More patients in the lowest FG category achieved clinical benefit compared to the highest (p < 0.001), while no relevant differences emerged for BMI. Fasting glucose at re-assessment was also predictive of PFS (p = 0.037), as confirmed in models including BMI and line of therapy (p = 0.049). Treatment discontinuation was significantly associated with changes in FG (p = 0.014). Further research is warranted to corroborate these findings and clarify the underlying mechanisms