Bitter taste receptors expression in human granulosa and cumulus cells: New perspectives in female fertility

Bitter taste receptors (TAS2RS) expression is not restricted to the oral cavity and the presence of these receptors in the male reproductive system and sperm provides insights into their possible role in human reproduction. To elucidate the potential role of TAS2Rs in the female reproductive system, we investigated the expression and localization of bitter taste receptors and the components of signal transduction cascade involved in the pathway of taste receptors in somatic follicular cells obtained from women undergoing assisted reproductive techniques. We found that TAS2R genes are expressed in both cumulus (CCs) and granulosa (GCs) cells, with TAS2R14 being the most highly expressed bitter receptor subtype. Interestingly, a slight increase in the expression of TAS2R14 and TAS2R43 was shown in both GCs and CCs in young women (p < 0.05), while a negative correlation may be established between the number of oocytes collected at the pickup and the expression of TAS2R43. Regarding α-gustducin and α-transducin, two Gα subunits expressed in the taste buds on the tongue, we provide evidence for their expression in CCs and GCs, with α-gustducin showing two additional isoforms in GCs. Finally, we shed light on the possible downstream transduction pathway initiated by taste receptor activation in the female reproductive system. Our study, showing for the first time the expression of taste receptors in the somatic ovarian follicle cells, significantly extends the current knowledge of the biological role of TAS2Rs for human female fertility

Expression of taste receptor 2 subtypes in human testis and sperm

Taste receptors (TASRs) are expressed not only in the oral cavity but also throughout the body thus suggesting that they may play different roles in organ systems beyond the tongue. Recent studies showed the expression of several TASRs in mammalian testis and sperm indicating an involvement of these receptors in male gametogenesis and fertility. This notion is supported by an impaired reproductive phenotype of mouse carrying targeted deletion of taste receptor genes as well as by a significant correlation between human semen parameters and specific polymorphisms of taste receptor genes. To better understand the biological and thus clinical significance of these receptors for human reproduction we analyzed the expression of several members of the TAS2Rs family of bitter receptors in human testis and in ejaculated sperm before and after in vitro selection and capacitation. Our results provide evidence for the expression of TAS2R genes with TAS2R14 being the most expressed bitter receptor subtype in both testis tissue and sperm cells respectively. In addition it was observed that in vitro capacitation significantly affects both the expression and the subcellular localization of these receptors in isolated spermatozoa. Interestingly α-gustducin and α-transducin two Gα subunits expressed in taste buds on the tongue are also expressed in human spermatozoa; moreover a subcellular redistribution of both G protein α-subunits to different sub-compartments of sperm was registered upon in vitro capacitation. Finally we shed light on the possible downstream transduction pathway initiated upon taste receptor activation in the male reproductive system. Performing ultrasensitive droplets digital PCR assays to quantify RNA copy numbers of a distinct gene we found a significant correlation between the expression of TAS2Rs and TRPM5 (r = 0.87) the cation channel involved in bitter but also sweet and umami taste transduction in taste buds on the tongue. Even if further studies are needed to clarify the precise functional role of taste receptors for successful reproduction the presented findings significantly extend our knowledge of the biological role of TAS2Rs for human male fertility

'Once upon a time': A proof of concept augmented reality collaborative mobile application to discover city heritage

In this paper, we propose an innovative mobile application to discover and valorize city heritage and its identity considering the city as a outdoors virtual museum. The application, namely "Once upon a Time" Magic GLASS, aims to exploit digital images of city glimpse preserved at National Central Library of Florence to provide an immersive experience to the users, that through the smart devices can look at the city as it was. This project paves the way for a new class of applications able of exploiting long term preserved digital objects

Oxidative stress measurement in frozen/thawed human sperm: The protective role of an in vitro treatment with myo-inositol

Despite its widespread use, sperm cryopreservation induces serious detrimental alterations in sperm function; indeed, it is commonly associated with decreased sperm viability and motility, and DNA fragmentation. Mechanisms of human sperm cryodamage are thought to be multifactorial, but oxidative stress seems to have a prominent role. A huge amount of data supported the cryoprotective effect of different antioxidants able to minimize the detrimental effects of reactive oxygen species (ROS) and improve the quality of spermatozoa. Among others, myo-inositol is one of the most powerful and has been reported to be effective in improving sperm quality and motility when used both in vivo and in vitro. This study aimed to determine the in vitro impact of myo-inositol in ameliorating sperm oxidative status during sperm cryopreservation. In particular, we demonstrated a significant improvement of sperm parameters (vitality and motility) when myo-inositol was added after sperm thawing (p < 0.05). Moreover, we showed that myo-inositol induces a significant increase in oxygen consumption, the main index of oxidative phosphorylation efficiency and ATP production. Finally, by means of 2D-electrophoresis, we demonstrated a significant decrease in the level of carbonyl groups, the main structural changes occurring in conditions of oxidative stress (p < 0.05). In conclusion, the sperm cryopreservation procedure we developed, assuring the reduction of ROS-induced sperm modifications, may improve the in vitro procedure currently used in ART laboratory for sperm cryostorage

Soluble protein fraction of human seminal plasma

Human seminal plasma (SP) is a complex fluid where sperm cells are bathed. Until recently, SP was simply retained a spermatozoa transport medium with nourishing functions. Growing evidences are nowadays recognizing it as a main actor “on the stage” of reproduction: SP orchestrates the synchronized cascade of events that make spermatozoa able to fertilize and modulates male and female reproductive fitness in physiological and in pathological states. Despite all SP studies have focused on “whole” SP or on its extracellular vesicles, to obtain an in-depth functional comprehension of SP, also the merely vesicle-free SP (vf-SP) soluble fraction deserves consideration. Here we present the first vf-SP functional proteomic study applying 2-DE, MALDI-TOF MS, and cluster and pathway analyses. Our work evidenced the occurrence of few unique proteins in vf-SP and an unexpected high heterogeneity of their corresponding protein species, conceivably derived by massive co- and/or post-translational modification events. Generated nets and clusters revealed tight functional correlations among identified proteins as well as their involvement in key functions for spermatozoa support and fertilization. Moreover, since SP is released by urogenital tissues and glands, our work may pave the way to the evaluation of vf-SP expression-profiles in diagnosing pathological processes in the secreting tissues. Biological significance: Seminal plasma vesicles are emerging as rich reservoirs of biomarkers for male infertility and urogenital disorders. Our proteomic approach to human SP diverges from the general trend in SP characterization, investigating not sperm extracellular vesicles but rather the vesicle-depleted SP fraction. Here we discussed how also SP soluble fraction may be considered actively involved in spermatozoa maturation, and we suggested that vf-SP protein profile may offer a precious overview on spermatogenesis, epididymal maturation, ancillary gland functionality, and sperm quality, thus highlighting its potentiality as a biomarker source in man health and reproduction. The present work provided the first 2-DE reference map of the so far underestimated vf-SP and an innovative and comprehensive functional-overview of its proteins. Obtained data may concur to clarify biochemical and molecular processes that the in fieri and the ejaculated semen undergoes as well as how semen properties can affect fertility or may reflect genital disorders. This paper is dedicated to the memory of Dr Riccardo Focarelli, associated professor in Comparative Anatomy and Glycobiology at the Dpt. of Life Sciences, Siena University, to acknowledge his fundamental contribution to the design of this study and in recognition of his distinguished research legacy

Interoperability knowledge base for persistent identifiers interoperability framework

In this paper, we propose a semantic web based solution to implement the Interoperability Framework (IF) for Persistent Identifiers (PI) developed within the context of APARSEN EU project. The IF provides a comprehensive, semantics-aware solution for interoperability of heterogeneous Persistent Identifier systems. Such a solution aims to provide added-value services built on an Interoperability Knowledge Base. The IF ontology refinement and the related prototype specifics have been designed adopting a bottom-up approach that starts from a) the analysis of metadata provided by Content Providers and b) the collection of functional and semantic requirements of Persistent Identifier Domains (PID to fostering ontology-based metadata translation among different bodies. Conclusions and intended future work close the paper

Chitotriosidase and soluble IL-2 receptor: comparison of two markers of sarcoidosis severity

Abstract BACKGROUND: Sarcoidosis is a multisystemic granulomatous disease with an unpredictable clinical course characterized by accumulation of activated proliferating T lymphocytes and mononuclear phagocytes in affected organs. AIMS AND METHODS: The aims of this study were to describe the clinical, radiological and immunological features of a population of sarcoidosis patients followed at the Sarcoidosis Regional Centre in Siena and to analyse chitotriosidase and sIL-2R concentrations in serum of these patients in order to understand their potential as disease markers. RESULTS: Chitotriosidase and sIL-2R concentrations in serum of sarcoidosis patients were found to be significantly higher than in healthy controls (p<0.01) and a positive correlation between the two markers was documented for the first time. Moreover, chitotriosidase and sIL-2R were expressed differently in different radiographic stages of the disease. CONCLUSION: Chitotriosidase and sIL-2R are two markers of sarcoidosis of different origin, the values of which show a correlation in these patients; they are easily detectable in serum and could be useful clinical markers of progression. PMID:[PubMed 18609101 - indexed for MEDLINE