Alessandra Ordinelli

Bio: Alessandra Ordinelli is an academic researcher from University of Teramo. The author has contributed to research in topics: Capacitation & Citation analysis. The author has an hindex of 6, co-authored 12 publications receiving 81 citations.

Clinically relevant radioresistant rhabdomyosarcoma cell lines: functional, molecular and immune-related characterization.

Abstract: The probability of local tumor control after radiotherapy (RT) remains still miserably poor in pediatric rhabdomyosarcoma (RMS). Thus, understanding the molecular mechanisms responsible of tumor relapse is essential to identify personalized RT-based strategies. Contrary to what has been done so far, a correct characterization of cellular radioresistance should be performed comparing radioresistant and radiosensitive cells with the same isogenic background. Clinically relevant radioresistant (RR) embryonal (RD) and alveolar (RH30) RMS cell lines have been developed by irradiating them with clinical-like hypo-fractionated schedule. RMS-RR cells were compared to parental isogenic counterpart (RMS-PR) and studied following the radiobiological concept of the “6Rs”, which stand for repair, redistribution, repopulation, reoxygenation, intrinsic radioresistance and radio-immuno-biology. RMS-RR cell lines, characterized by a more aggressive and in vitro pro-metastatic phenotype, showed a higher ability to i) detoxify from reactive oxygen species; ii) repair DNA damage by differently activating non-homologous end joining and homologous recombination pathways; iii) counteract RT-induced G2/M cell cycle arrest by re-starting growth and repopulating after irradiation; iv) express cancer stem-like profile. Bioinformatic analyses, performed to assess the role of 41 cytokines after RT exposure and their network interactions, suggested TGF-β, MIF, CCL2, CXCL5, CXCL8 and CXCL12 as master regulators of cancer immune escape in RMS tumors. These results suggest that RMS could sustain intrinsic and acquire radioresistance by different mechanisms and indicate potential targets for future combined radiosensitizing strategies.

Capacitation-Related Lipid Remodeling of Mammalian Spermatozoa Membrane Determines the Final Fate of Male Gametes: A Computational Biology Study.

Abstract: To become fully fertile, mammalian spermatozoa must undergo a complex process of biochemical maturation within the female genital tract, which determines a marked lipid remodeling (LR) of membranes. Here, we represent this process as a biological network, which is a graph constituted by nodes (the molecules involved in LR) and by edges (their interactions). As a result, we found that LR network has a scale-free and small world topology. This implies that it is robust against random damage and that it allows a fast and specific transmission of information. In addition, the hubs in the network allow identification of the control mechanisms involved in membrane-related signaling, which could concur in determining the fate of ejaculated spermatozoa. Interestingly, different pathways involved in LR (maintenance of functional incompetence, reaching of fertilizing ability, apoptosis) are overlapped and some molecules take part in different signalling cascades; thus their role in sperm biology needs to be interpreted in a more large context. In addition, it was possible to differentiate, either based on their topological and biological characteristics, the molecules acting as global or local controller in LR. These findings may contribute to the understanding of capacitation-related signaling and of sperm physiopathology.

Network Analyses of Sperm–Egg Recognition and Binding: Ready to Rethink Fertility Mechanisms?

Abstract: The rapid growth of published literature makes biomedical text mining increasingly invaluable for unpacking implicit knowledge hidden in unstructured text. We employed biomedical text mining and biological networks analyses to research the process of sperm egg recognition and binding (SERB). We selected from the literature the molecules expressed either on spermatozoa or on oocytes thought to be involved in SERB and, using an automated literature search software (Agilent Literature Search), we realized a network, SERBN, characterized by a hierarchical scale free and a small world topology. We used an integrated approach, either based on selection of hubs or by a cluster analysis, to discern the key molecules of SERB. We found that in most cases some of them are not directly situated on spermatozoa and oocyte, but are dispersed in oviductal fluid or embedded in exosomes present in the perivitelline space. To confirm and validate our results, we performed further analyses using STRING and Reactome FI software. Our findings underscore that the fertility is not a property of gametes in isolation, but rather depends on the functional integrity of the entire reproductive system. These observations collectively underscore the importance of integrative biology in exploring biological systems and in rethinking of fertility mechanisms in the light of this innovative approach.

Nuclear survivin expression as a potentially useful tool for the diagnosis of canine cutaneous sebaceous lesions

Abstract: Background – Sebaceous glands are specialized cutaneous adnexal glands, which work under constant hormonal control to produce sebum. They can give rise to several proliferative lesions, such as hamartoma, hyperplasia and neoplasms (adenoma, epithelioma and carcinoma). Their nomenclature is currently confusing, both in veterinary and in human medicine, owing to the difficulty of differentiating between some of these lesions. Methods – The present study used immunohistochemistry to determine the expression levels and patterns of survivin and Ki67 in five samples of normal canine skin and 44 cases of canine cutaneous lesions with sebaceous differentiation (10 hamartomas, nine hyperplasia, eight adenomas, eight epitheliomas and nine carcinomas). Results – In normal glands, survivin, as well as Ki67, was expressed in scattered reserve cells. In hamartomas, survivin was more highly expressed than in normal skin, indicating a possible role of this molecule in the pathogenesis of these congenital lesions. In tumours, a moderate or high level of survivin and Ki67 expression (more than two and four and more than two positive cells, respectively) were significantly correlated with a malignant histotype, infiltrative growth and a moderate or high number of mitoses (more than two). Conclusions and clinical importance – The level of survivin expression increased with increasing malignancy, designating survivin as a new diagnostic marker in the assessment of malignancy of sebaceous tumours. Resume Contexte – Les glandes sebacees sont des glandes cutanees specialisees dont la production de sebum est sous controle hormonal constant. Elles peuvent etre a l’origine de plusieurs lesions proliferatives telles que hamartomes, hyperplasies et neoplasmes (adenomes, epitheliomes et carcinomes). Leur actuelle nomenclature est confuse, en medecine humaine comme en medecine veterinaire, ce qui est liea la difficulte de differencier certaines de ces lesions entre elles. Methodes – Cette etude utilise l’immunohistochimie pour determiner les niveaux et les patrons d’expression de la survivine et du Ki67 dans cinq echantillons de peau normale de chien et 44 cas de lesions cutanees canines avec differentiation cutanee (10 hamartomes, neuf hyperplasies, huit adenomes, huit epitheliomes et neuf carcinomes). Resultats – Dans les glandes normales, la survivine ainsi que le Ki67, etaient exprimes uniquement au sein des cellules de reserve. Dans les hamartomes, la survivine etait exprimee plus intensement que dans la peau normale, indiquant un role possible de cette molecule dans la pathogenie de ces lesions congenitales. Dans les tumeurs, un niveau d’expression modere ou eleve de survivine et de Ki67 (respectivement plus de deux et quatre et plus que deux cellules positives) etait correle avec un histotype tumoral, une infiltration et un nombre modere ou eleve de mitoses (plus que deux). Conclusions et importance clinique – Le niveau d’expression de survivine augmentant avec la malignite, permet de designer la survivine comme un nouveau marqueur diagnostic dans l evaluation de malignite des tumeurs sebacees. Resumen Introduccion – las glandulas sebaceas son glandulas anejas cutaneas especializadas, que funcionan bajo control hormonal constante para producir el sebo. Pueden dar lugar a varias lesiones proliferativas, tales como hamartoma, hiperplasia y neoplasias (adenoma, epitelioma y carcinoma). Su nomenclatura es actualmente confusa, tanto en medicina veterinaria como en humana, debido a la dificultad de distinguir entre algunas de estas lesiones. Metodos – el presente estudio utiliza la inmunohistoquimica para determinar los niveles de expresion y la distribucion de survivina y de Ki67 en cinco muestras de piel canina normal y 44 casos de lesiones cutaneas caninas con diferenciacion sebacea (10 hamartomas, nueve hiperplasias, ocho adenomas, ocho epiteliomas y nueve carcinomas). Resultados – en glandulas normales, la survivina, asi como el Ki67, se expresaron en celulas dispersas de reserva. En hamartomas, la survivina tenia una mayor expresion comparada con piel normal, indicando un posible papel de esta molecula en la patogenia de estas lesiones congenitas. En tumores, la presencia de un nivel moderado a alto nivel de survivina y de Ki67 (mas de dos y cuatro y mas de dos celulas positivas, respectivamente) fue correlacionado de forma significativa con un histotipo maligno, un crecimiento infiltrativo y un numero moderado a alto de mitosis (mas de dos). Conclusiones e importancia clinica – el nivel de expresion de survivina aumento con el aumento de la malignidad, lo cual indica que la survivina puede ser un marcador de importancia diagnostica en la valoracion de la malignidad de los tumores sebaceos. Zusammenfassung Hintergrund – Die Talgdrusen sind spezialisierte Hautanhangsdrusen, die unter standiger hormoneller Kontrolle stehen, um Talg zu produzieren. Sie konnen die Ursache fur verschiedene proliferative Veranderungen, wie Hamartom, Hyperplasien und Neoplasien (Adenome, Epitheliome und Karzinome) sein. Die Nomenklatur ist momentan sowohl in der Veterinar- als auch in der Humanmedizin verwirrend, was auf die Schwierigkeit einige dieser Veranderungen zu differenzieren, zuruckzufuhren ist. Methoden – In der gegenwartigen Studie wurde die Immunhistochemie angewandt, um die Exprimierungslevels und Muster von Survivan und Ki67 in funf Proben von normaler Hundehaut und in 44 Fallen von Hautveranderungen von Hunden, bei denen als Ursprung die Talgdrusen differenziert werden konnten (10 Hamartome, neun Hyperplasien, acht Adenome, acht Epitheliome und neun Karzinome). Ergebnisse – In normalen Drusen wurde sowohl Survivin als auch Ki67 in vereinzelten Reservezellen exprimiert. In Hamartomen wurde Survivin haufiger exprimiert als in normaler Haut, was auf eine mogliche Rolle dieser Molekule in der Pathogenese dieser angeborenen Zellen hinweist. In Tumoren wurde ein moderates bis hohes Level an Survivin und Ki67 exprimiert (an mehr als zwei und vier bzw an mehr als zwei positiven Zellen), was signifikant mit einem malignen Histiotyp, infiltrativem Wachstum und einer moderaten bis hohen Anzahl an Mitosen (mehr als zwei) korrelierte. Schlussfolgerung und klinische Bedeutung – Das Ausmas der Exprimierung von Survivin nahm mit zunehmender Malignitat zu, was bedeutet, dass Survivin als neuer diagnostischer Marker bei der Feststellung der Malignitat von Tumoren der Talgdrusen designiert werden kann.

Scientometric study of the effects of exposure to non-ionizing electromagnetic fields on fertility: A contribution to understanding the reasons of partial failure.

Abstract: The exposure to Non-Ionizing-Electromagnetic Fields (NI-EMFs) is often indicated as a cofactor responsible for the fertility reduction, which has been described in recent years. Despite the great interest in this topic and the research effort in exploring it, to date, there are no reliable data. Therefore, we carried out a scientometric analysis of the scientific literature published in peer reviewed Journals concerning this topic to better understand the reasons of this partial failure. To this aim, we identified and analysed 104 papers, published in last 26 years in peer-reviewed Journals, present in ISI Web of Knowledge Core Collection. Then, we analysed the impact of the Journals in which the papers were published as well as that of the single papers, the paper citation dynamics, the keywords citation busts, the geographical localization of citations and the co-authorship dynamics of the Authors. As a result, we found that different animal models (rodent, rabbit, guinea pig, and swine) and different experimental approaches (epidemiological vs. experimental studies) have the same impact, highlighting the lack of universally adopted standard in research activity. The analysis of the temporal trend in keywords and the high differences in citations between the different countries (also in those belonging to the same geographical and socio-economical area) pointed out the difficulties in approaching this branch of study. Lastly, it was evident that the Authors did not behave as a connected community, but as unconnected clusters of very small size. In conclusion, based on the results of our analysis, we think that important efforts must be undertaken to adopt more standardized models and to improve the research quality and the information exchange within the scientific community, with the aim of improving the reliability and usefulness of the results of research regarding the effect of NI-EMFs on fertility.

SPAdes, a new genome assembly algorithm and its applications to single-cell sequencing ( 7th Annual SFAF Meeting, 2012)

Abstract: The lion's share of bacteria in various environments cannot be cloned in the laboratory and thus cannot be sequenced using existing technologies. A major goal of single-cell genomics is to complement gene-centric metagenomic data with whole-genome assemblies of uncultivated organisms. Assembly of single-cell data is challenging because of highly non-uniform read coverage as well as elevated levels of sequencing errors and chimeric reads. We describe SPAdes, a new assembler for both single-cell and standard (multicell) assembly, and demonstrate that it improves on the recently released E+V-SC assembler (specialized for single-cell data) and on popular assemblers Velvet and SoapDeNovo (for multicell data). SPAdes generates single-cell assemblies, providing information about genomes of uncultivatable bacteria that vastly exceeds what may be obtained via traditional metagenomics studies. SPAdes is available online ( http://bioinf.spbau.ru/spades ). It is distributed as open source software.

Role of Mitochondria in the Oxidative Stress Induced by Electromagnetic Fields: Focus on Reproductive Systems.

Abstract: Modern technologies relying on wireless communication systems have brought increasing levels of electromagnetic field (EMF) exposure. This increased research interest in the effects of these radiations on human health. There is compelling evidence that EMFs affect cell physiology by altering redox-related processes. Considering the importance of redox milieu in the biological competence of oocyte and sperm, we reviewed the existing literature regarding the effects of EMFs on reproductive systems. Given the role of mitochondria as the main source of reactive oxygen species (ROS), we focused on the hypothesis of a mitochondrial basis of EMF-induced reproductive toxicity. MEDLINE, Web of Science, and Scopus database were examined for peer-reviewed original articles by searching for the following keywords: “extremely low frequency electromagnetic fields (ELF-EMFs),” “radiofrequency (RF),” “microwaves,” “Wi-Fi,” “mobile phone,” “oxidative stress,” “mitochondria,” “fertility,” “sperm,” “testis,” “oocyte,” “ovarian follicle,” and “embryo.” These keywords were combined with other search phrases relevant to the topic. Although we reported contradictory data due to lack of uniformity in the experimental designs, a growing body of evidence suggests that EMF exposure during spermatogenesis induces increased ROS production associated with decreased ROS scavenging activity. Numerous studies revealed the detrimental effects of EMFs from mobile phones, laptops, and other electric devices on sperm quality and provide evidence for extensive electron leakage from the mitochondrial electron transport chain as the main cause of EMF damage. In female reproductive systems, the contribution of oxidative stress to EMF-induced damages and the evidence of mitochondrial origin of ROS overproduction are reported, as well. In conclusion, mitochondria seem to play an important role as source of ROS in both male and female reproductive systems under EMF exposure. Future and more standardized studies are required for a better understanding of molecular mechanisms underlying EMF potential challenge to our reproductive system in order to improve preventive strategies.

Impact of male fertility status on the transcriptome of the bovine epididymis.

Abstract: Study question Can region-specific transcriptional profiling of the epididymis from fertile and sub-fertile bulls predict the etiology of fertility/sub-fertility in males? Summary answer The highly regulated gene expression along the bovine epididymis is affected by the fertility status of bulls used for artificial insemination. What is known already In mammals, sperm maturation and storage occur in the epididymis. Each epididymal segment has his own transcriptomic signature that modulates the intraluminal composition and consequently governs sequential modifications of the maturing male gamete. Study design, size, duration Epididymides from six Holstein bulls with documented fertility were used. These bulls were divided into two groups: high fertility (n = 3), and medium-low fertility (n = 3) and their epididymal transcriptomic profiles were analyzed. Participants/materials, setting, methods Bovine cDNA microarray probing and bioinformatic tools were used to identify genes that are differentially expressed in caput, corpus and cauda epididymidal tissues of bulls with the documented fertility index. Main results and the role of chance Hierarchical clustering and principal component analysis revealed a clear separation between caput, corpus and cauda epididymides. Some transcripts characterize a particular anatomical segment, whereas others are expressed in two out of three epididymal segments. Gene ontology analysis allowed deduction of specific functions played by each epididymal segment. The transcriptional profiles between fertile versus sub-fertile conditions clustered most closely in the corpus and cauda segments, whereas the profiles in the caput segment were distinct between fertile and sub-fertile bulls. Of the differently expressed genes, 10 (AKAP4, SMCP, SPATA3, TCP11, ODF1, CTCFL, SPATA18, ADAM28, SORD and FAM161A) were found to exert functions related to reproductive systems and 5 genes (DEAD, CYST11, DEFB119, DEFB124 and MX1) were found to be associated with the defense response. Large scale data The GEO number for public access of bovine epididymis microarray data is GSE96602. Limitations, reasons for caution Further work is required to link these modulations of epididymal functions with sperm fertilizing ability in order to understand the etiology of certain cases of idiopathic infertility in livestock and men. Wider implications of the findings As fertility can be quantified in bulls used for artificial insemination, this species is a unique model to aid in the understanding of male fertility/sub-fertility in man. Our data provide a molecular characterization that will facilitate advances in understanding the involvement of epididymal physiology in sub/infertility etiology. Study funding/competing interest(s) This work was supported by a grant to R.S. from the Natural Sciences and Engineering Research Council (NSERC) of Canada. C.L., A.A., E.C. and R.S. have no conflict of interest to declare. P.B. is R&D director at Alliance Boviteq Inc., a bovine artificial insemination company.