AUTOPHAGY AND APOPTOSIS MODULATION BY AQUEOUS EXTRACTS FROM LEAVES AND RHIZOMES OF Posidonia oceanica ON HEPG2 HEPATOCARCINOMA CELLS

2023

Toxicity of vanadium during development of sea urchin embryos: bioaccumulation, calcium depletion, ERK modulation and cell-selective apoptosis

2022

Vanadium toxicology is a topic of considerable importance as this metal is widely used in industrial and biomedical fields. However, it represents a potential emerging environmental pollutant because wastewater treatment plants do not adequately remove metal compounds that are subsequently released into the environment. Vanadium applications are limited due to its toxicity, so it is urgent to define this aspect. This metal is associated with sea urchin embryo toxicity as it perturbs embryogenesis and skeletogenesis, triggering several stress responses. Here we investigated its bioaccumulation and the correlation with cellular and molecular developmental pathways. We used cytotoxic concentra…

Methylation of cytokines gene promoters in IL-1β-treated human intestinal epithelial cells

2017

Objective and design: Epigenetic regulation is important in the activation of inflammatory cells. In the present study, we evaluated if DNA-methylation variations are involved in Interleukin-1β (IL-1β)-induced intestinal epithelial cells activation. Materials and methods: Differentiated Caco-2 cells were exposed to IL-1β or to 5-azadeoxycytidine (5-azadC) for 24 or 48 h. Genome-wide methylation status was evaluated, while DNA methylation status at the promoter region of the gene encoding interleukin-6, 8 and 10 (IL-6, 8 and 10) was estimated. The levels of the corresponding gene products as well as DNA methyltransferases (DNMTs) quantity were assessed. Results: IL-1β decreased genomic m…

Vanadium Toxicity Monitored by Fertilization Outcomes and Metal Related Proteolytic Activities in Paracentrotus lividus Embryos

2022

Metal pharmaceutical residues often represent emerging toxic pollutants of the aquatic environment, as wastewater treatment plants do not sufficiently remove these compounds. Recently, vanadium (V) derivatives have been considered as potential therapeutic factors in several diseases, however, only limited information is available about their impact on aquatic environments. This study used sea urchin embryos (Paracentrotus lividus) to test V toxicity, as it is known they are sensitive to V doses from environmentally relevant to very cytotoxic levels (50 nM; 100 nM; 500 nM; 1 µM; 50 µM; 100 µM; 500 µM; and 1 mM). We used two approaches: The fertilization test (FT) and …

Heavy Metals and Metalloids as Autophagy Inducing Agents: Focus on Cadmium and Arsenic

2012

In recent years, research on the autophagic process has greatly increased, invading the fields of biology and medicine. Several markers of the autophagic process have been discovered and various strategies have been reported studying this molecular process in different biological systems in both physiological and stress conditions. Furthermore, mechanisms of metalloid- or heavy metal-induced toxicity continue to be of interest given the ubiquitous nature and distribution of these contaminants in the environment where they often play the role of pollutants of numerous organisms. The aim of this review is a critical analysis and correlation of knowledge of autophagic mechanisms studied under …

Relationship between apoptosis and survival molecules in human cumulus cells as markers of oocyte competence

2017

SummaryTo select from a single patient the best oocytes able to reach the blastocyst stage, we searched for valuable markers for oocytes competence. We evaluated the DNA fragmentation index (DFI) and the level of some survival molecules, such as AKT, pAKT and pERK1/2, in individual cumulus cell–oocyte complexes (COC). The study included normo-responder women. The average age of the patients was 34.3. DFI in cumulus cells was evaluated using the terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labelling (TUNEL) assayin situ. AKT, pAKT and pERK1/2 were measured by immunological assay and densitometric analysis of fluorescent signals using NIS-Elements BR 3.10 image software. Statisti…

Toxic effects induced by vanadium on sea urchin embryos

2020

Vanadium, a naturally occurring element widely distributed in soil, water and air, has received considerable interest because its compounds are often used in different applications, from industry to medicine. While the possible medical use of vanadium compounds is promising, its potential harmful effects on living organisms are still unclear. Here, for the first time, we provide a toxicological profile induced by vanadium on Paracentrotus lividus sea urchin embryos, reporting an integrated and comparative analysis of the detected effects reflecting vanadium-toxicity. At the morphological level we found a dose-dependent induction of altered phenotypes and of skeletal malformations. At the mo…

Autophagy is required for sea urchin oogenesis and early development.

2016

SummaryAutophagy is a major intracellular pathway for the degradation and recycling of cytosolic components. Emerging evidence has demonstrated its crucial role during the embryo development of invertebrates and vertebrates. We recently demonstrated a massive activation of autophagy in Paracentrotus lividus embryos under cadmium stress conditions, and the existence of a temporal relationship between induced autophagy and apoptosis. Although there have been numerous studies on the role of autophagy in the development of different organisms, information on the autophagic process during oogenesis or at the start of development in marine invertebrates is very limited. Here we report our recent …

Sea urchin embryos as a model system for studying autophagy induced by cadmium stress

2011

It is well known that sea urchin embryos are able to activate different defense strategies against stress. We previously demonstrated that cadmium treatment triggers the accumulation of metal in embryonic cells and the activation of defense systems depending on concentration and exposure time, through the synthesis of heat shock proteins and/or the initiation of apoptosis. Here we show that Paracentrotus lividus embryos exposed to Cd adopt autophagy as an additional stratagem to safeguard the developmental program. At present, there are no data focusing on the role of this process in embryo development of marine organisms. In this paper we utilized different techniques to detect autophagy i…

Cytotoxic effects of Jay Amin hydroxamic acid (JAHA), a ferrocene-based class I histone deacetylase inhibitor, on triple-negative MDA-MB231 breast ca…

2012

The histone deacetylase inhibitors (HDACis) are a class of chemically heterogeneous anticancer agents of which suberoylanilide hydroxamic acid (SAHA) is a prototypical member. SAHA derivatives may be obtained by three-dimensional manipulation of SAHA aryl cap, such as the incorporation of a ferrocene unit like that present in Jay Amin hydroxamic acid (JAHA) and homo-JAHA [ Spencer , et al. ( 2011 ) ACS Med. Chem. Lett. 2 , 358 - 362 ]. These metal-based SAHA analogues have been tested for their cytotoxic activity toward triple-negative MDA-MB231 breast cancer cells. The results obtained indicate that of the two compounds tested, only JAHA was prominently active on breast cancer cells with a…

MITOCHONDRIAL MASS, DISTRIBUTION AND ACTIVITY DURING SEA URCHIN OOGENESIS

2015

The sea urchin egg is a favourite model for studies of the molecular biology and physiology of fertilization and early development, yet we know sparingly little of its oocytes and of mitochondria behaviour during oogenesis. The process of oogenesis in most echinoderms is asynchronous so each ovary lobe has hundreds of oocytes at all stages of development. At the beginning of oogenesis, the oocyte is about 10 µm in diameter. During the vitellogenic phase of oogenesis, the oocyte accumulate yolk proteins and grow to ten times their original size to 80 to 100 µm in sea urchins. The oocyte, arrested at the prophase of the first meiotic division, is apparent with its large nucleus, the germinal …

Interactive effects of increased temperature and gadolinium pollution in Paracentrotus lividus sea urchin embryos: a climate change perspective

2021

Gradual ocean warming and marine heatwaves represent major threats for marine organisms already facing other anthropogenic-derived hazards, such as chemical contamination in coastal areas. In this study, the combined effects of thermal stress and exposure to gadolinium (Gd), a metal used as a contrasting agent in medical imaging which enters the aquatic environment, were investigated in the embryos and larvae of the sea urchin Paracentrotus lividus. Embryos were exposed to six treatments of three temperatures (18 °C, 21 °C, 24 °C) and two Gd concentrations (control: 0 μM; treated: 20 μM). With respect to developmental progression, increased temperature accelerated development and achievemen…

APOPTOTIC ANALYSIS OF CUMULUS CELLS FOR THE SELECTION OF COMPETENT OOCYTES TO BE FERTILIZED BY INTRACYTOPLASMIC SPERM INJECTION (ICSI)

2015

Oocyte quality is one of the main factors for the success of in vitro fertilization protocols. Apoptosis is known to affect oocyte quality and may impair subsequent embryonic development and implantation. The aim of this study was to investigate the apoptosis rate of single and pooled cumulus cells of cumulus cell–oocyte complexes (COCs), as markers of oocyte quality, prior to intracytoplasmatic sperm injection (ICSI).We investigated the apoptosis rate by TUNEL assay (DNA fragmentation) and caspase-3 immunoassay of single and pooled cumulus cells of COCs. The results showed that DNA fragmentation in cumulus cells was remarkably lower in patients who achieved a pregnancy than in those who di…

The Histone Deacetylase Inhibitor JAHA Down-Regulates pERK and Global DNA Methylation in MDA-MB231 Breast Cancer Cells

2015

The histone deacetylase inhibitor N-1-(ferrocenyl)-N-8-hydroxyoctanediamide (JAHA) down-regulates extracellular-signal-regulated kinase (ERK) and its activated form in triple-negative MDA-MB231 breast cancer cells after 18 h and up to 30 h of treatment, and to a lesser extent AKT and phospho-AKT after 30 h and up to 48 h of treatment. Also, DNA methyltransferase 1 (DNMT1), 3b and, to a lesser extent, 3a, downstream ERK targets, were down-regulated already at 18 h with an increase up to 48 h of exposure. Methylation-sensitive restriction arbitrarily-primed (MeSAP) polymerase chain reaction (PCR) analysis confirmed the ability of JAHA to induce genome-wide DNA hypomethylation at 48 h of expos…

Vanadium Modulates Proteolytic Activities and MMP-14-Like Levels during Paracentrotus lividus Embryogenesis

2022

The increasing industrial use of vanadium (V), as well as its recent medical use in various pathologies has intensified its environmental release, making it an emerging pollutant. The sea urchin embryo has long been used to study the effects induced by metals, including V. In this study we used an integrated approach that correlates the biological effects on embryo development with proteolytic activities of gelatinases that could better reflect any metal-induced imbalances. V-exposure caused morphological/morphometric aberrations, mainly concerning the correct distribution of embryonic cells, the development of the skeleton, and the embryo volume. Moreover, V induced a concentration change …

The Role of Autophagy and Apoptosis During Embryo Development

2015

Programmed cell death (PCD) and cell survival are two sides of the same coin. Autopha‐ gy and apoptosis are crucial processes during embryo development of Invertebrates and Vertebrates organisms, as they are necessary for the formation of a new organism, start‐ ing from a fertilized egg. Fertilization triggers cell remodeling from each gamete to a toti‐ potent zygote. During embryogenesis, the cells undergo various processes, thus allowing the transformation of the embryo into an adult organism. In particular, cells require the appropriate tools to suddenly modify their morphology and protein content in order to respond to intrinsic and external stimuli. Autophagy and apoptosis are involved…

Autophagy

2012

Klionsky, Daniel J. et al.

An In Vitro Model of Glioma Development

2023

Gliomas are the prevalent forms of brain cancer and derive from glial cells. Among them, astrocytomas are the most frequent. Astrocytes are fundamental for most brain functions, as they contribute to neuronal metabolism and neurotransmission. When they acquire cancer properties, their functions are altered, and, in addition, they start invading the brain parenchyma. Thus, a better knowledge of transformed astrocyte molecular properties is essential. With this aim, we previously developed rat astrocyte clones with increasing cancer properties. In this study, we used proteomic analysis to compare the most transformed clone (A-FC6) with normal primary astrocytes. We found that 154 proteins are…

Cadmium stress effects indicating marine pollution in different species of sea urchin employed as environmental bioindicators

2019

In recent years, researches about the defense strategies induced by cadmium stress have greatly increased, invading several fields of scientific research. Mechanisms of cadmium-induced toxicity continue to be of interest for researchers given its ubiquitous nature and environmental distribution, where it often plays the role of pollutant for numerous organisms. The presence in the environment of this heavy metal has been constantly increasing because of its large employment in several industrial and agricultural activities. Cadmium does not have any biological role and, since it cannot be degraded by living organisms, it is irreversibly accumulated into cells, interacting with cellular comp…

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

2016

Seuls les 100 premiers auteurs dont les auteurs INRA ont été entrés dans la notice. La liste complète des auteurs et de leurs affiliations est accessible sur la publication.; International audience; In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues…

Phytochemical profile and antioxidant properties of the edible and non-edible portions of black sapote (Diospyros digyna Jacq.)

2022

This study evaluated the phytochemical profile and antioxidative properties of the edible and non-edible portions of black sapote. The phytochemical analysis highlighted the presence of several bioactive compounds, differently distributed among peel, pulp and seeds. In particular, the peel resulted rich of flavan-3-ols and proanthocyanidins, whereas seeds contained high amount of organic acids, including ferulic, citric and sinapic acids. Concerning functional properties, both edible and non-edible portions showed a significant prevention of lipid peroxidation in a cell-based model. Moreover, the results suggested that the antioxidant protection involved both redox active properties and gen…

In Vitro Cytotoxic Effect of Aqueous Extracts from Leaves and Rhizomes of the Seagrass Posidonia oceanica (L.) Delile on HepG2 Liver Cancer Cells: Fo…

2023

Aqueous extracts from Posidonia oceanica’s green and brown (beached) leaves and rhizomes were prepared, submitted to phenolic compound and proteomic analysis, and examined for their potential cytotoxic effect on HepG2 liver cancer cells in culture. The chosen endpoints related to survival and death were cell viability and locomotory behavior, cell-cycle analysis, apoptosis and autophagy, mitochondrial membrane polarization, and cell redox state. Here, we show that 24 h exposure to both green-leaf- and rhizome-derived extracts decreased tumor cell number in a dose–response manner, with a mean half maximal inhibitory concentration (IC50) estimated at 83 and 11.5 μg of dry extract/mL, respecti…

Effects of magnesium deprivation on development and biomineralization in the sea urchin Arbacia lixula

2019

Echinoderms have an extensive endoskeleton composed of magnesian calcite and occluded matrix proteins. As biomineralization in sea urchin larvae is sensitive to the Magnesium:Calcium ratio of seawater, we investigated the effects of magnesium deprivation on development and skeletogenesis in the sea urchin Arbacia lixula. We focused on the localization of the skeletogenic cells (primary mesenchyme cells) and the spatial expression of associated genes. Embryos reared in Mg-free seawater exhibited developmental delay from 6-h post-fertilization and at 24 h embryos showed complete lack of biomineral formation. Larvae (48–72 h) exhibited severe skeleton malformations. Fluorescent labelling revea…

Vanadium induces calcium depletion and cell selective apoptosis during development of sea urchin embryos

2021

Vanadium (V) is a metal widely distributed in soil, water and air. It has recently received growing interest because its compounds are often used in different applications, from industry to medicine.1 Here, using atomic absorption spectrometry, we demonstrate the predisposition of V to accumulate directly into embryonic cells, interfering with Ca uptake. At the morphological level, we observed dose- and time-dependent effects on phenotypes and on skeletal malformations. At the molecular level, V-exposed embryos showed the activation of the cellular stress response, inducing Hsp 60 and Hsp 70 synthesis and the activation of autophagy and apoptosis. The Hsps-mediated stress response to V appe…

Toxicological Impact of Rare Earth Elements (REEs) on the Reproduction and Development of Aquatic Organisms Using Sea Urchins as Biological Models

2022

The growing presence of lanthanides in the environment has drawn the attention of the scientific community on their safety and toxicity. The sources of lanthanides in the environment include diagnostic medicine, electronic devices, permanent magnets, etc. Their exponential use and the poor management of waste disposal raise serious concerns about the quality and safety of the ecosystems at a global level. This review focused on the impact of lanthanides in marine organisms on reproductive fitness, fertilization and embryonic development, using the sea urchin as a biological model system. Scientific evidence shows that exposure to lanthanides triggers a wide variety of toxic insults, includi…

Autophagy as a defense strategy against stress: focus on Paracentrotus lividus sea urchin embryos exposed to cadmium

2015

Autophagy is used by organisms as a defense strategy to face environmental stress. This mechanism has been described as one of the most important intracellular pathways responsible for the degradation and recycling of proteins and organelles. It can act as a cell survival mechanism if the cellular damage is not too extensive or as a cell death mechanism if the damage/stress is irreversible; in the latter case, it can operate as an independent pathway or together with the apoptotic one. In this review, we discuss the autophagic process activated in several aquatic organisms exposed to different types of environmental stressors, focusing on the sea urchin embryo, a suitable system recently in…

Induction of skeletal abnormalities and autophagy in Paracentrotus lividus sea urchin embryos exposed to gadolinium.

2017

Abstract Gadolinium (Gd) concentration is constantly increasing in the aquatic environment, becoming an emergent environmental pollutant. We investigated the effects of Gd on Paracentrotus lividus sea urchin embryos, focusing on skeletogenesis and autophagy. We observed a delay of biomineral deposition at 24 hours post fertilization (hpf), and a strong impairment of skeleton growth at 48 hpf, frequently displayed by an asymmetrical pattern. Skeleton growth was found partially resumed in recovery experiments. The mesodermal cells designated to biomineralization were found correctly migrated at 24 hpf, but not at 48 hpf. Western blot analysis showed an increase of the LC3-II autophagic marker…

Erratum

2016

Author(s): Klionsky, DJ; Abdelmohsen, K; Abe, A; Abedin, MJ; Abeliovich, H; Arozena, AA; Adachi, H; Adams, CM; Adams, PD; Adeli, K; Adhihetty, PJ; Adler, SG; Agam, G; Agarwal, R; Aghi, MK; Agnello, M; Agostinis, P; Aguilar, PV; Aguirre-Ghiso, J; Airoldi, EM; Ait-Si-Ali, S; Akematsu, T; Akporiaye, ET; Al-Rubeai, M; Albaiceta, GM; Albanese, C; Albani, D; Albert, ML; Aldudo, J; Algul, H; Alirezaei, M; Alloza, I; Almasan, A; Almonte-Beceril, M; Alnemri, ES; Alonso, C; Altan-Bonnet, N; Altieri, DC; Alvarez, S; Alvarez-Erviti, L; Alves, S; Amadoro, G; Amano, A; Amantini, C; Ambrosio, S; Amelio, I; Amer, AO; Amessou, M; Amon, A; An, Z; Anania, FA; Andersen, SU; Andley, UP; Andreadi, CK; Andrieu-Ab…

Sea urchin embryos exposed to cadmium as an experimental model for studying the relationship between autophagy and apoptosis

2014

The sea urchin embryo is a suitable model that offers an excellent opportunity to investigate different defence strategies activated in stress conditions. We previously showed that cadmium accumulates in a dose- and time-dependent manner into embryonic cells, activating different stress and defence mechanisms, including the synthesis of HSPs and the onset of apoptosis and/or autophagy. In this paper we investigated the functional relationship between autophagy and apoptosis, evaluating apoptosis signals in cadmium-exposed Paracentrotus lividus embryos with inhibited autophagy. We found that the inhibition of autophagy produced the concurrent reduction of apoptosis, suggesting that the two p…

Proteolytic activity and MMP-14-like protein levels are affected by Vanadium in Paracentrotus lividus Embryo

2023

The increasing industrial use of vanadium (V), as well as its recent medical use in various pathologies has intensified its environmental release, making it an emerging pollutant [1]. The sea urchin embryo has long been used to study the effects induced by metals, including V. In this study we used an integrated approach that correlates the biological effects on embryo development with proteolytic activities of gelatinases that could better reflect any metal induced imbalances. V-exposure caused morphological/morphometric aberrations, mainly concerning the correct dis-tribution of embryonic cells, the development of the skeleton and the embryo volume [2]. Moreover, V induced a concentration…

Sea urchin embryos cadmium exposed as an experimental model system for studying the relationship between autophagy and apoptosis

2012

Gadolinium toxicity on sea urchin embryos of geographically and phylogenetically distant species

2016

Pharmaceuticals are a class of emerging environmental contaminants. Gadolinium (Gd) is a metal of the lanthanide series of the elements whose chelates are commonly employed as contrast agents for magnetic resonance imaging, and subsequently released into the aquatic environment. We investigated the consequences of sea urchin embryo exposure to sublethal Gd concentrations, comparing the effects on the development of four phylogenetically and geographically distant species: two Mediterranean species, Paracentrotus lividus and Arbacia lixula, and two species living in the East coast of Australia, Heliocidaris tuberculata and Centrostephanus rodgersii. Measures of the Gd and Ca content inside e…

Toxicity of manganese on embryos of the sea urchin, Paracentrotus lividus.

2008

Vanadium perturbs the fertilization outcome and the metalloproteinase activity in sea urchin embryos

2021

Metal toxicology represents a current major topic due to the disper- sion of these elements in the environment. Metals are released from both natural sources and industrial activities. Some of them have also a clinical interest due to their application as metallodrugs (i.e., Pt, Cu, Au, Ru, and Y) or in medical diagnosis (Gd).1,2 Recently, V derivatives are considered as potential therapeutic factors in some diseases (e.g., obesity, diabetes, cancer, neurodegenerative and heart disorders). As a consequence, pharmaceutical residues could repre- sent emerging pollutants of aquatic environments, as wastewater treatment plants do not sufficiently remove these compounds3. Embryonic models repres…

JA47, a new histone deacetylase inhibitor that induces cytotoxic effects on triple-negative MDA-MB231 breast cancer cells in vitro

2012

The toxic effect of gadolinium ions on sea urchin embryos: comparison among phylogenetically distant species and focus on mechanisms regulating stres…

2016

Gadolinium (Gd) is a metal of the lanthanide series of the elements whose chelates are commonly employed as contrast agents for magnetic resonance imaging, and subsequently released into the aquatic environment. We investigated the consequences of sea urchin embryo exposure to sublethal Gd concentrations, comparing the effects on the development of four phylogenetically and geographically distant species: two Mediterranean species, Paracentrotus lividus and Arbacia lixula, and two species living in the East coast of Australia, Heliocidaris tuberculata and Centrostephanus rodgersii. Measures of the Gd content inside embryos by ICP-MS showed a time- and dose-dependent increase. In all these s…

Gadolinium-induced stress response causes a time-dependent miss-expression of regulative and structural genes involved in the development of the sea …

2015

Autophagy is related to apoptosis in sea urchin embryos exposed to cadmium

2014

The sea urchin embryo is a suitable model system that offers an excellent opportunity to investigate different defense strategies activated in stress conditions. We previously showed that cadmium treatment provokes the accumulation of metal in dose-time dependent manner in embryonic cells and the activation of defense systems, such as the synthesis of HSPs and/or the initiation of apoptosis. Analyzing autophagy, by neutral red, acridine orange and LC3-detection, we demonstrated that Cd-exposed embryos adopt this process as an additional stratagem to safeguard the developmental program. We observed that embryos treated with subletal Cd concentration activate massive autophagic response after…

Apoptosis rate in human germinal cells: quality assay.

2009

Apoptosis is a process that eliminates superfluous or genetically compromised cells. Cell death is one of the most important aspects of organization of the developing, as alteration in timing, level, or pattern of cell death can lead to developmental anomalies. In our studies, we investigated apoptosis in follicular somatic and germ cells of patients with infertility problems. We found that cell death occur in human germinal vesicle, in some mature oocytes and in the oocytes that failed fertilization after ICSI (Intracytoplasmic Sperm Injection), this failure was strictly correlated with apoptosis. Still, after the Italian law number 40 of 2004, it was not available oocytes to research, and…

Autophagy is related to apoptosis in Paracentrotus lividus embryos cadmium exposed

2012

P. lividus embryo offers an excellent opportunity to investigate the adaptive response of cells exposed to different stress. We previously demonstrated that cadmium treatment triggers the accumulation of metal in embryonic cells and the activation of defense system depending on concentration and exposure time, through the synthesis of HSPs and/or the initiation of apoptosis. Analysing autophagy, by neutral red, acridine orange and LC3-detection, we demonstrated that Cd-exposed embryos adopt this process as an additional stratagem to safeguard the developmental program. We observed that embryos treated at subletal Cd concentration activate a massive autophagic response after 18h, which decre…

Asimmetric skeleton patterns induced by gadolinium ions in sea urchin embryos: focus on mechanisms regulating skeletogenesis and comparison among phy…

2014

Gadolinium induces autophagy in sea urchin embryos

2014

Apoptosis rate in cumulus cells can be considerate as an indicator for the selection of embryos to improve ongoing pregnancy and implantation rate

2014

AN INNOVATIVE STRATEGY IN SELECTING OOCYTES WITH HIGH IMPLANTATION POTENTIALITY FOR INTRACYTOPLASMIC SPERM INJECTION PROCEDURE

2015

The aim of the research was to investigate the apoptosis rate of individual cumulus cell-oocyte complexes (COC), associated to the level of pAKT, to verify the difference between oocytes who produce embryos able to reach the blastocyst stage compared with embryos arrested during the in vitro culture. It was demonstrated that DNA fragmentation in cumulus cells was remarkably lower in patients who achieved a pregnancy after ICSI cycles, related to the quality of oocytes and embryos1,2. AKT pathway plays a critical role in the regulation of cell survival, and most growth factors activate this pathway3. The study focused on 53 patients, involved after informed consent. In this prospective and r…

Methylation decrease of BECN1 gene induced by phytochemical Indicaxantin in Caco2 cells: an epigenetic hypothesis of autophagy

2014

Autophagy is a highly conserved catabolic process that degrades and recycles intracellular components through the lysosomes [1]. The role of this process in tumorigenesis and tumor progression is controversial: in the early stages, it can block tumor growth and conversely it can promote its progression in the later stages [2]. The tumor suppressor BECN1 gene, encodes the protein Beclin 1, a marker of autophagy down-regulated in several types of cancer, such as colorectal cancer [3]. There are a lot of both genetic and environmental risk factors for colorectal cancer, including diet: for this reason, in accordance with epidemiological studies, consumption of foods rich in phytochemicals is w…

L’autofagia durante l’oogenesi e i primi stadi di sviluppo embrionale in Paracentrotus lividus

2013

Autophagy during development of sea urchin embryos

2009

Autophagy is a highly regulated mechanism that enhances cell eukaryotic survival under various environmental and cellular stresses, by breakdown and recycling of macromolecules and organelles. Here we report that in Paracentrotus lividus embryos autophagic process occur, at a lesser extend during physiological development and at greater levels after a cadmium treatment. By Acridine Orange staining, we found that embryonic cells exposed to cadmium display green fluorescence in cytoplasm and nucleus, and show considerable red fluorescent dots in cytoplasm. This evidence suggests formation of acidic auto-phagolysosomal vacuoles. By Neutral Red vital staining, specific for lysosomes and cellula…

The autophagic demand during oogenesis and early development of sea urchin

2015

The autophagic pathway is an evolutionarily conserved homeostatic process, responsible for degradation and recycling of long-term proteins and cytoplasmic organelles in eukaryotic cells. This process constitutively occurs at basal levels and is involved in cell survival. Increased autophagy is induced by environmental cues, such as starvation and many stress agents, while excessive levels of autophagy can lead to autophagic Programmed Cell Death, with features that differ from those of the apoptotic process. We recently demonstrated massive activation of autophagy in P. lividus embryos, in cadmium stress conditions, and the existence of a temporal relationship between induced autophagy end …

Sea urchin embryo as a model system for studing autophagy induced by cadmium stress

2010

L'embrione di riccio di mare come modello di studio dell'autofagia indotta da stress

2011

Gli embrioni di riccio di mare, Paracentrotus lividus, sono in grado di attivare differenti strategie di difesa come risposta a stress chimico/fisici. Recentemente abbiamo dimostrato che il cadmio, metallo pesante altamente embriotossico, induce la sintesi di specifiche hsps e/o l’innesco di processi apoptotici, via via che si accumula nelle cellule embrionali. Nel presente lavoro, mostriamo che gli embrioni di P. lividus sono in grado di attivare l’autofagia come un aggiuntivo meccanismo atto a salvaguardare il programma di sviluppo, in seguito a esposizione a dosi citotossiche di CdCl2. L’autofagia è un meccanismo molecolare che può stimolare la sopravvivenza, attraverso la degradazione e…

Cadmium induces autophagy during development of Paracentrotus lividus embryos.

2010

Evaluation of DNA damage in human spermatozoa: a sperm quality assay.

2008

Several studies have demonstrated the relationship between gonadotrophine serum levels and the apoptosis rate in germinal cells. In human, lower FSH serum levels are associated with reduced Sperm Standard Parameters in term of concentration, motility and morphology and higher sperm DNA fragmentation. Hypogonadotrope hypogonadism is a clinical condition associated with male infertility, characterized by low serum levels of gonadotrophins. In this pilot study we investigated the spermatozoa quality of 11 patients with development of hypogonadotrope hypogonadism urdergoing in vitro fertilization programs. Recombinant-FSH administered to improve the sperm parameters, apoptosis rate, aiming to i…

Apoptosis rate in human spermatozoa: a sperm quality assay.

2009

Several studies have demonstrated the relationship between gonadotrophine serum levels and the apoptosis rate in germinal cells (Bosco et al 2005). In human, lower FSH serum levels are associated with reduced Sperm Standard Parameters in term of concentration, motility and morphology and higher sperm DNA fragmentation. Hypogonadotrope hypogonadism is a clinical condition associated with male infertility, characterized by low serum levels of gonadotrophins. In this pilot study we investigated the spermatozoa quality of 11 patients with development of hypogonadotrope hypogonadism urdergoing in vitro fertilization programs. Recombinant-FSH administered to improve the sperm parameters, apoptosi…

Strategie di difesa attivate in risposta a stress, in embrioni di Paracentrotus lividus

2012

Sea urchin embryos are able to activate different defense strategies against stress. This model system allows to investigate numerous phenomena in multipotent cells, which interact among themselves, in their natural position, bypassing the disadvantages of isolated cells, deprived of their normal network. Cadmium (Cd) treatment triggers the accumulation of metal in embryonic cells and the activation of defense systems depending on concentration and exposure time, through the synthesis of heat shock proteins and/or the initiation of apoptosis. Here we show that Paracentrotus lividus embryos exposed to subacute/sublethal concentrations of Cd adopt autophagy as an additional stratagem to safeg…

A possible role of autophagy for the execution of apoptosis in cadmium-exposed sea urchin embryos

2012

The sea urchin embryo is a suitable model system that offers an excellent opportunity to investigate different defence strategies activated in stress conditions. We previously showed that cadmium treatment provokes the accumulation of metal in dose- and time-dependent manner in embryonic cells and the activation of defence systems, such as the synthesis of HSPs and/or the initiation of apoptosis. Analysing autophagy, by neutral red, acridine orange and LC3-detection, we demonstrated that Cd-exposed embryos adopt this process as an additional stratagem to safeguard the developmental program. We observed that embryos treated with subletal Cd concentration activate a massive autophagic respons…

Differences beetween physiological and induced apoptosis in sea urchin embryos.

2008

Autophagy induction in sea urchin embryos exposed to gadolinium ions

2014

Sea urchin embryos are highly sensitive to several kinds of stressors, and able to activate different defense strategies. Gadolinium (Gd) is a metal of the lanthanide series of the elements: its chelates are employed as contrast agents for magnetic resonance imaging since the 1980s. Gd complexes are released in the aquatic environment, making Gd an emergent environmental pollutant. In this study we focused on the effects of Gd ions on sea urchin embryos development. The study was conducted looking at three different processes: general development, apoptosis and autophagy. At the whole morphological level, Paracentrotus lividus sea urchin embryos continuously exposed to Gd ions displayed mor…

P. lividus oogenesis and early development require autophagy

2014

It is well known that autophagy is a major intracellular pathway for the degradation and recycling of cytosolic components, in both basal and stress conditions. We have recently demonstrated the activation of autophagy in P. lividus embryos: at high levels, after a stress induced by cadmium, and at basal levels, during the physiological development (1) (2). Here we report ours recent data about autophagy during oogenesis and segmentation. In order to detect autophagolysosomes (AVOs) and autolysosomes, we respectively performed incubation in vivo with acridine orange (AO), and in situ immunofluorescence (IF) with the anti-LC3 antibody (autophagy marker). All observations were carried out by …

JAHA, a novel histone deacetylase inhibitor: cytotoxic effect on triple-negative breast cancer cells

2013

The effects of manganese on embryos of the sea urchin, Paracentrotus lividus: stress or not?.

2008

Autophagy as defence strategy against cadmium stress in Paracentrotus lividus embryos.

2009

Sea urchin embryo is a developmental model that offers an excellent opportunity to investigate the possible adaptive response of cells exposed to different stress during differentiation. These embryos are able to respond to many stress by synthesizing a set of highly conserved proteins, the hsps and/or by activation of apoptosis. The exposure to cadmium trigger the accumulation of metal in embryo cells and the activation of both defence mechanisms depending on concentration and exposure time [1-4]. Recent experimental evidences demonstrate that by autophagy, a highly regulated mechanism that enhances cell survival under various environmental and cellular stress, the breakdown and recycling …

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition) 1

2021

Contains fulltext : 232759.pdf (Publisher’s version ) (Closed access) In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to…

Relationship between autophagy and apoptosis in Paracentrotus lividus embryos cadmium exposed

2011

Cadmium is a strong toxicant for living organism since it does not have biological roles and can cause several cellular damages comprising genotoxicity, oxidative stress, and other biochemical dysfunctions. Marine invertebrate embryos represent a suitable model system where to investigate the effects of many stressors on development and cell viability. Here we investigated the toxic effect of cadmium on sea urchin Paracentrotus lividus embryos focusing our attention on metal-inducead autophagy and the possible temporal and functional relationship with apoptosis. Using several techniques to detect autophagy (neutral red, acridine orange and LC3-detection) we demonstrated that Cd-exposed P. l…

A new strategy in selecting oocytes using cumulus cells analysis of specific molecules of the apoptotic pathway, according to the ability to reach bl…

2015

Study question: The aim of the research was to investigate the apoptosis rate of individual cumulus cell–oocyte complexes (COC), associated to the level of p-Akt, to verify the difference between oocytes who produce embryos able to reach the blastocyst stage compared with embryos arrested during the in vitro colture. Summary answer: It was demonstrated that DNA fragmentation in cumulus cells was remarkably lower in patients who achieved a pregnancy after ICSI cycles, related to the quality of oocytes and embryos. Akt pathway plays a critical role in the regulation of cell survival, and most growth factors activate this pathway. What is known already: Studies on oocyte maturation have shown …

Autophagy

2021

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide…

Ruolo dell’autofagia nell’oogenesi di riccio di mare Paracentrotus lividus

2013

Relazione tra autofagia e apoptosi in embrioni di riccio di mare esposti a stress

2012

L’embrione di riccio di mare Paracentrotus lividus è stato utilizzato come organismo modello della biologia dello sviluppo per molti anni ed è considerato il deuterostoma più primitivo, con scheletro calcificato, correlato a protocordati e vertebrati. Tale sistema offre un'eccellente opportunità per studiare le numerose strategie di difesa che gli embrioni mettono in atto contro diverse condizioni di stress. In precedenza, abbiamo riportato che l’esposizione di embrioni a dosi citotossiche di cadmio, provoca l'accumulo intracellulare del metallo e l'attivazione del sistema difensivo, in modo dose-tempo dipendente, attraverso la sintesi di specifiche HSPs e/o l’innesco di apoptosi. Mediante …

Effetto citotossico di un nuovo inibitore delle deacetilasi istoniche, JAHA, su cellule di tumore mammario umano triplo negativo

2013

Toxicity induced by Gadolinium ions on sea urchin embryos: comparison among phylogenetically distant species and focus on stress response and skeleto…

2016

Pharmaceuticals are a class of emerging environmental contaminants. Gadolinium (Gd) is a lanthanide metal whose chelates are employed as contrast agents for magnetic resonance imaging, and subsequently released into the aquatic environment. We investigated the effects of exposure to sublethal Gd concentrations on the development of four phylogenetically and geographically distant sea urchin species: two Mediterranean, Paracentrotus lividus and Arbacia lixula, and two from Australia, Heliocidaris tuberculata and Centrostephanus rodgersii. Sensitivity to Gd greatly varied, with EC50 ranging from 56 nM to 132 µM across the four species. Measures of the Gd and Ca content inside embryos showed a…

Developmental abnormalities induced by Gadolinium causes a time-dependent miss-expression of regulative and structural genes in P. lividus sea urchin…

2015

Gadolinium (Gd) is a metal of the lanthanide series of the elements whose chelates are commonly used as contrast agents for magnetic resonance imaging. Its release into the aquatic milieu has posed serious concerns regarding its noxious effects, and therefore Gd is now considered an emerging environmental pollutant. The sea urchin embryo is an excellent model used in both toxicological and developmental research. We analysed the consequences of embryo exposure to sublethal concentrations of Gd on embryo development, focusing on skeletogenesis and developmental symmetry. We observed a strong inhibition of skeleton growth, frequently displayed by an asymmetrical pattern. Continuous exposure t…