Annalisa Di Santi | Università della Campania Luigi Vanvitelli (original) (raw)
Papers by Annalisa Di Santi
70° Congresso UZI - Rapallo (GE), 2009
Abstract: Lichens are valuable natural resources used for centuries throughout the world as medic... more Abstract: Lichens are valuable natural resources used for centuries throughout the world as medicine, food, fodder, perfume, spices and dyes, as well as for other miscellaneous purposes. This study investigates the antiproliferative, antibacterial and antifungal activity of the acetone extract of the lichen Xanthoria parietina (Linnaeus) Theodor Fries and its major secondary metabolite, parietin. The extract and parietin were tested for antimicrobial activity against nine American Type Culture Collection standard and clinically isolated OPEN ACCESS Int. J. Mol. Sci. 2015, 16 7862
Receptors & Clinical Investigation, 2015
The androgen receptor (AR) mediates differentiation, proliferation and transformation of target t... more The androgen receptor (AR) mediates differentiation, proliferation and transformation of target tissues. These processes require a crosstalk between epithelial and stromal cells. Prostate cancer (PCa) represents a major cause of cancer-related mortality in men, and is often associated with deregulation of androgen/AR axis. Clinical and molecular findings have highlighted the role of epithelial AR in PCa progression. In contrast, the functions of AR in mesenchymal cells are still unclear. We previously reported that low androgen concentration (1 pM) triggers interaction of AR with the Src tyrosine kinase and PI3-K, thus driving cell cycle progression in fibroblasts. In contrast, stimulation of fibroblasts and fibrosarcoma cells with physiological (10 nM) androgen concentration leads to interaction of AR with full-length filamin A (FLNa) and does not trigger DNA synthesis. On the basis of these findings, we reexamined the role of androgen/AR axis in fibroblasts and human fibrosarcoma HT1080 cells. Recently, we obtained two original and integrated findings on the decision of mesenchymal cells to undergo reversible quiescence and migrate upon stimulation with 10 nM androgens (Castoria et al. 2011 and 2014). This decision is dependent upon the interaction of AR with FLNa. Once assembled, the bipartite AR/FLNa complex recruits β1-integrin and triggers Rac1 activation, thereby enhancing on the one hand cell motility. On the other, Rac 1 activation triggers its downstream effector DYRK 1B, which phosphorylates Ser10 of p27. Stabilization of p27 and cell quiescence then follow. These results strengthen and extend our studies, adding a new and exciting piece to the complex puzzle of signaling networks activated by androgens in target cells. Our findings might have implications for current approaches to AR-related diseases.
International journal of molecular sciences, Jan 9, 2015
Lichens are valuable natural resources used for centuries throughout the world as medicine, food,... more Lichens are valuable natural resources used for centuries throughout the world as medicine, food, fodder, perfume, spices and dyes, as well as for other miscellaneous purposes. This study investigates the antiproliferative, antibacterial and antifungal activity of the acetone extract of the lichen Xanthoria parietina (Linnaeus) Theodor Fries and its major secondary metabolite, parietin. The extract and parietin were tested for antimicrobial activity against nine American Type Culture Collection standard and clinically isolated bacterial strains, and three fungal strains. Both showed strong antibacterial activity against all bacterial strains and matched clinical isolates, particularly against Staphylococcus aureus from standard and clinical sources. Among the fungi tested, Rhizoctonia solani was the most sensitive. The antiproliferative effects of the extract and parietin were also investigated in human breast cancer cells. The extract inhibited proliferation and induced apoptosis, ...
Journal of cellular physiology, 2015
Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that defends against oxidative d... more Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that defends against oxidative damage due to reactive oxygen species (ROS). A new isoform of MnSOD with cytotoxic activity was recently discovered in liposarcoma cells. Here, we tested the effectiveness of a recombinant form of this isoform (rMnSOD) on leukemic T cells, Jurkat cells, and lymphocytes. Our results confirm that leukemic T cells can internalize rMnSOD and that rMnSOD causes apoptosis of 99% of leukemic cells without showing toxic effects on healthy cells. Using light and electron microscopy, we determined that an rMnSOD concentration of 0.067 μM most effective on apoptosis induction. Western blot analysis showed that treatment with 0.067 μM rMnSOD resulted in high expression of the pro-apoptotic protein Bax and low expression of the anti-apoptotic protein Bcl-2 in leukemia cells. Concerning signal transduction pathway no influence was observed after treatment except for Jurkat cells showing a slightly decr...
Cell Death & Differentiation, 2014
Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 2011
The aim of this study was to clarify the role of the erythrocyte inclusions found during the hema... more The aim of this study was to clarify the role of the erythrocyte inclusions found during the hematological screening of loggerhead population of the Mediterranean Sea. We studied the erythrocyte inclusions in blood specimens collected from six juvenile and nine adult specimens of the loggerhead turtle, Caretta caretta, from the Adriatic and Tyrrhenian Seas. Our study indicates that the percentage of mature erythrocytes containing inclusions ranged from 3 to 82%. Each erythrocyte contained only one round inclusion body. Inclusion bodies stained with May Grünwald–Giemsa show that their cytochemical and ultrastructure characteristics are identical to those of human Heinz bodies. Because Heinz bodies originate from the precipitation of unstable hemoglobin (Hb) and cause globular osmotic resistance to increase, we analyzed loggerhead Hb using electrophoresis and high‐performance liquid chromatography to detect and quantitate Hb fractions. We also tested the resistance of Hb to alkaline p...
Comparative Clinical Pathology, 2012
The eosinophils and heterophils of loggerheads display a similar rounded shape, an eccentric and ... more The eosinophils and heterophils of loggerheads display a similar rounded shape, an eccentric and elongated or slightly lobulated nucleus and a cytoplasm containing acidophilic pleomorphic granules. The characterization of the different types of granulocytes from nine Mediterranean loggerheads (Caretta caretta), based on cytochemical and immunocytochemical reactions and ultrastructural studies, is reported. On the basis of their positivity after myeloperoxidase, alkaline phosphatase and chloroacetate esterase reactions (typical of mammalian neutrophils), the granulocytes were classified as heterophils, while those myeloperoxidase-negative, major basic protein-positive granulocytes (like mammalian eosinophils), displaying a weak expression of interleukin 5 (a growth and differentiation factor, activator and chemoattractant for eosinophils), were classified as eosinophils. The immunocytochemical and TEM studies allowed the identification of these two granulocyte types, both of which show an eosinophilic reaction with May-Grünwald-Giemsa, periodic acid-Schiff and neutral red staining positivity.
Comparative Clinical Pathology, 2011
In order to establish baseline hematological and biochemical values in loggerhead turtles from th... more In order to establish baseline hematological and biochemical values in loggerhead turtles from the Mediterranean Sea, 84 specimens were sampled, comprising 24 wild turtles in good health at the time of capture and 60 turtles tested after indoor rehabilitation at the Sea Turtle Rescue and Rehabilitation Centre of the Zoological Station Anton Dohrn in Naples, Italy. The following parameters were evaluated: red cell counts (RBC, 488-575×10 3 /μL), white cell counts (WBC, 17-24×10 3 /μL) and thrombocyte counts (TBC, 19-49×10 2 /μL), hemoglobin (Hb, 8-14 g/dL), hematocrit (Ht, 23-34%), mean corpuscular volume (MCV, 487-723 fL), mean corpuscular hemoglobin (MCH, 170-261 pg), mean corpuscular hemoglobin concentration (MCHC, 34-42%), white and red blood cell differential counts, and a panel of hematochemical tests, composed of glucose (97-164 mg/ dL), cholesterol (74-144 mg/dL), blood urea nitrogen (35-200 mg/dL), uric acid (1-2.7 mg/dL), total bilirubin (0.20-0.40 mg/dL), GOT (44-184 IU/L) and GPT (6 IU/L) transaminases, calcium (6.7-8.7 mg/dL), and magnesium (3.6-5.4 mEq/L). Comparisons of the statistically analyzed data from the turtles which were divided into groups on the basis of age and/or lifestyle (wild or captive) revealed that erythroid parameters attained higher values in captive turtles. This suggested a positive influence of the rich and complete diet fed in captivity upon the hemopoietic process of the turtle. On the other hand, data suggest a more intense and active hemopoiesis in young turtles, compared to adult specimens.
Comparative Clinical Pathology, 2010
Abstract Theileria is a small Apicomplexan parasite considered as the scourge of the cattle indus... more Abstract Theileria is a small Apicomplexan parasite considered as the scourge of the cattle industry, here detected for the first time in the blood cells of Caretta caretta from the Mediterranean Sea. In the early stage of infection by Theileria, rounded forms, defined schizonts, occur in ...
Cell Death & Differentiation, Jun 27, 2014
Nell’ambito della sperimentazione clinica, che viene svolta nei paesi industrializzati, si rende ... more Nell’ambito della sperimentazione clinica, che viene svolta nei paesi industrializzati, si rende necessario porre una specifica attenzione ai criteri etici di riferimento affinche siano salvaguardati i beni e i valori umani fondamentali. Il rischio e che gli interessi commerciali possano nascondersi dietro gli interessi scientifici, danneggiando il benessere psichico del malato e la sua dignita di essere umano. Nei pazienti oncologici la sofferenza fisica, che rappresenta soltanto un sintomo della malattia, determina necessariamente un’alterazione della qualita di vita e puo condizionare la vita di relazione. Tutto questo viene ovviamente amplificato quando il malato e un bambino che, talora, e incapace di esprimere, localizzare e quantificare la sua sofferenza. Gli oncologi pediatrici lamentano la mancanza di farmaci antitumorali studiati per i bambini, in considerazione del fatto che i chemioterapici utilizzati nei protocolli di cura degli adulti non sono spesso egualmente efficaci in eta pediatrica. Le aziende farmaceutiche hanno poco interesse a sviluppare farmaci per la popolazione pediatrica oncologica per motivi di natura economica, dipendenti dalla esiguita del numero di pazienti pediatrici rispetto alla popolazione adulta. In generale la produzione di farmaci antitumorali alternativi trova resistenza nell’ambito della industria farmaceutica coinvolta nei profitti relativi al mercato dei chemio e radioterapici. Obiettivo primario del mio progetto di ricerca e stato, pertanto, la messa a punto di una terapia non tossica, o a ridotta citotossicita, per la cura delle neoplasie ematologiche in eta pediatrica. In particolare l’obiettivo di tale progetto e stato, nella prima fase, testare l’efficacia d’azione, sulle cellule di Leucemia Linfoblastica Acuta (LLA), la patologia piu frequente in eta pediatrica, del peptide leader della rMnSOD (manganese superossido dismutasi ricombinante) quale carrier di citostatici (cisplatino ed epirubicina) allo scopo di ridurre la dose di citostatico necessaria per una efficace internalizzazione dello stesso nelle cellule tumorali e per ottenerne quindi la morte riducendo gli effetti collaterali. Nella seconda fase, e stata studiata l’azione, sulle cellule in coltura di LLA B e T ex vivo e su una linea stabilizzata, della r-MnSOD, una proteina isolata e sequenziata, per la prima volta, da cellule di liposarcoma umano (LSA) presso il laboratorio di Oncologia Sperimentale dell’Istituto dei Tumori di Napoli, G. Pascale, dal Dott. A. Mancini. Tale molecola ha dimostrato attivita antitumorale su altri tipi di tumore. I dati ottenuti in questo lavoro hanno dimostrato che il peptide leader e un buon carrier di citostatici all’interno delle cellule leucemiche, tuttavia i citostatici legati al suddetto peptide non si sono rivelati efficaci nell’indurre la morte delle cellule leucemiche in coltura. Sara necessario quindi, quale proseguimento della ricerca, testare l’azione di altri citostatici legati al peptide. L’utilizzo del peptide leader coniugato a dosi micro molari di un diverso citostatico, efficace nella LLA pediatrica, potra, se confermato dai risultati, ottenere una riduzione notevole degli effetti collaterali migliorando la qualita di vita e la prognosi dei pazienti leucemici pediatrici ad alto rischio. Diversamente, il trattamento con l’intera molecola rMnSOD ha dimostrato sia la sua capacita di internalizzazione (dovuta alla presenza del peptide leader) nelle cellule leucemiche di LLA B e T in coltura, che la sua potente azione antitumorale, avendo indotto la morte di un alto numero di cellule leucemiche trattate. E stato, cosi, dimostrato l’effetto citotossico di tale proteina nei confronti delle cellule neoplastiche trattate e la mancanza di effetti tossici sulle cellule sane di controllo, sulle quali svolge, invece, una benefica ossigenazione. L’entita delle alterazioni morfologiche osservate in microscopia ottica ed elettronica nelle cellule trattate con rMnSOD alla concentrazione di 2 µg/mL ha suggerito che questa sia la concentrazione piu efficace, tra quelle sperimentate. Tale dato e stato confermato dai risultati di biologia molecolare: e stata riscontrata, infatti, a seguito del suddetto trattamento, la massima espressione dei geni proapoptotici (Bax) e la minima espressione dei geni antiapoptotici (Bcl-2); cio a conferma dell’attivazione della via apoptotica nelle cellule leucemiche. Pertanto, la rMnSOD potrebbe rivelarsi un efficace farmaco antitumorale, non tossico come i chemioterapici, per il trattamento delle leucemie ad alto rischio.
The androgen receptor (AR) mediates differentiation, proliferation and transformation of target t... more The androgen receptor (AR) mediates differentiation, proliferation and transformation of target tissues. These processes require a crosstalk between epithelial and stromal cells. Prostate cancer (PCa) represents a major cause of cancer-related mortality in men, and is often associated with deregulation of androgen/AR axis. Clinical and molecular findings have highlighted the role of epithelial AR in PCa progression. In contrast, the functions of AR in mesenchymal cells are still unclear. We previously reported that low androgen concentration (1 pM) triggers interaction of AR with the Src tyrosine kinase and PI3-K, thus driving cell cycle progression in fibroblasts. In contrast, stimulation of fibroblasts and fibrosarcoma cells with physiological (10 nM) androgen concentration leads to interaction of AR with full-length filamin A (FLNa) and does not trigger DNA synthesis. On the basis of these findings, we re-examined the role of androgen/AR axis in fibroblasts and human fibrosarcoma...
Oncotarget, Jan 24, 2015
Prostate cancer is one of the most commonly diagnosed cancers in men, and androgen deprivation th... more Prostate cancer is one of the most commonly diagnosed cancers in men, and androgen deprivation therapy still represents the primary treatment for prostate cancer patients. This approach, however, frequently fails and patients develop castration-resistant prostate cancer, which is almost untreatable.Cancer cells are characterized by a hierarchical organization, and stem/progenitor cells are endowed with tumor-initiating activity. Accumulating evidence indicates that prostate cancer stem cells lack the androgen receptor and are, indeed, resistant to androgen deprivation therapy. In contrast, these cells express classical (α and/or β) and novel (GPR30) estrogen receptors, which may represent new putative targets in prostate cancer treatment.In the present review, we discuss the still-debated mechanisms, both genomic and non-genomic, by which androgen and estradiol receptors (classical and novel) mediate the hormonal control of prostate cell stemness, transformation, and the continued g...
Journal of cellular physiology, 2015
Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that defends against oxidative d... more Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that defends against oxidative damage due to reactive oxygen species (ROS). A new isoform of MnSOD with cytotoxic activity was recently discovered in liposarcoma cells. Here, we tested the effectiveness of a recombinant form of this isoform (rMnSOD) on leukemic T cells, Jurkat cells, and lymphocytes. Our results confirm that leukemic T cells can internalize rMnSOD and that rMnSOD causes apoptosis of 99% of leukemic cells without showing toxic effects on healthy cells. Using light and electron microscopy, we determined that an rMnSOD concentration of 0.067 μM most effective on apoptosis induction. Western blot analysis showed that treatment with 0.067 μM rMnSOD resulted in high expression of the pro-apoptotic protein Bax and low expression of the anti-apoptotic protein Bcl-2 in leukemia cells. Concerning signal transduction pathway no influence was observed after treatment except for Jurkat cells showing a slightly decr...
Methods in Molecular Biology, 2014
It is well established that histone posttranslational modifications mediate the control of gene e... more It is well established that histone posttranslational modifications mediate the control of gene expression played by chromatin. Such modifications are commonly reversible and many alternatives are open to drive transcription of inducible genes. Estrogens govern growth and survival of hormone-sensitive cells by inducing expression of genes important for cell cycle progression and apoptosis. Transcription of estrogen-responsive genes is triggered by the lysine-specific demethylase 1 (LSD1)-dependent demethylation of dimethylated lysine 9 in histone H3 (H3K9me2) that accompanies to local generation of oxygen reactive species (ROS). Production of ROS modifies guanines in neighbor DNA with consequent recruitment of base-excision repair (BER) enzymes and formation of breaks that support creation of bridges between sites that, although distant on linear DNA, establish strategic contacts useful for productive transcription.
Frontiers in Endocrinology, 2015
Prostate cancer (PCa) is the major cause of cancer-related death among the male population of Wes... more Prostate cancer (PCa) is the major cause of cancer-related death among the male population of Western society, and androgen-deprivation therapy (ADT) represents the first line in PCa treatment. However, although androgen receptor (AR) expression is maintained throughout the various stages of PCa, ADT frequently fails. Clinical studies have demonstrated that different androgen/AR signaling pathways operate in target tissues. AR stimulates growth and transformation of target cells, but under certain conditions slows down their proliferation. In this review, we discuss the role of AR in controlling different functions of mesenchymal and transformed mesenchymal cells. Findings here presented support the role of AR in suppressing proliferation and stimulating migration of stromal cells, with implications for current approaches to cancer therapy.
Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 2011
The aim of this study was to clarify the role of the erythrocyte inclusions found during the hema... more The aim of this study was to clarify the role of the erythrocyte inclusions found during the hematological screening of loggerhead population of the Mediterranean Sea. We studied the erythrocyte inclusions in blood specimens collected from six juvenile and nine adult specimens of the loggerhead turtle, Caretta caretta, from the Adriatic and Tyrrhenian Seas. Our study indicates that the percentage of mature erythrocytes containing inclusions ranged from 3 to 82%. Each erythrocyte contained only one round inclusion body. Inclusion bodies stained with May Grünwald-Giemsa show that their cytochemical and ultrastructure characteristics are identical to those of human Heinz bodies. Because Heinz bodies originate from the precipitation of unstable hemoglobin (Hb) and cause globular osmotic resistance to increase, we analyzed loggerhead Hb using electrophoresis and high-performance liquid chromatography to detect and quantitate Hb fractions. We also tested the resistance of Hb to alkaline pH, heat, isopropanol denaturation, and globular osmosis. Our hemogram results excluded the occurrence of any infection, which could be associated with an inclusion body, in all the specimens. Negative Feulgen staining indicated that the inclusion bodies are not derived from DNA fragmentation. We hypothesize that amino acid substitutions could explain why loggerhead Hb precipitates under normal physiologic conditions, forming Heinz bodies. The identification of inclusion bodies in loggerhead erythrocytes allow us to better understand the haematological characteristics and the physiology of these ancient reptiles, thus aiding efforts to conserve such an endangered species.
70° Congresso UZI - Rapallo (GE), 2009
Abstract: Lichens are valuable natural resources used for centuries throughout the world as medic... more Abstract: Lichens are valuable natural resources used for centuries throughout the world as medicine, food, fodder, perfume, spices and dyes, as well as for other miscellaneous purposes. This study investigates the antiproliferative, antibacterial and antifungal activity of the acetone extract of the lichen Xanthoria parietina (Linnaeus) Theodor Fries and its major secondary metabolite, parietin. The extract and parietin were tested for antimicrobial activity against nine American Type Culture Collection standard and clinically isolated OPEN ACCESS Int. J. Mol. Sci. 2015, 16 7862
Receptors & Clinical Investigation, 2015
The androgen receptor (AR) mediates differentiation, proliferation and transformation of target t... more The androgen receptor (AR) mediates differentiation, proliferation and transformation of target tissues. These processes require a crosstalk between epithelial and stromal cells. Prostate cancer (PCa) represents a major cause of cancer-related mortality in men, and is often associated with deregulation of androgen/AR axis. Clinical and molecular findings have highlighted the role of epithelial AR in PCa progression. In contrast, the functions of AR in mesenchymal cells are still unclear. We previously reported that low androgen concentration (1 pM) triggers interaction of AR with the Src tyrosine kinase and PI3-K, thus driving cell cycle progression in fibroblasts. In contrast, stimulation of fibroblasts and fibrosarcoma cells with physiological (10 nM) androgen concentration leads to interaction of AR with full-length filamin A (FLNa) and does not trigger DNA synthesis. On the basis of these findings, we reexamined the role of androgen/AR axis in fibroblasts and human fibrosarcoma HT1080 cells. Recently, we obtained two original and integrated findings on the decision of mesenchymal cells to undergo reversible quiescence and migrate upon stimulation with 10 nM androgens (Castoria et al. 2011 and 2014). This decision is dependent upon the interaction of AR with FLNa. Once assembled, the bipartite AR/FLNa complex recruits β1-integrin and triggers Rac1 activation, thereby enhancing on the one hand cell motility. On the other, Rac 1 activation triggers its downstream effector DYRK 1B, which phosphorylates Ser10 of p27. Stabilization of p27 and cell quiescence then follow. These results strengthen and extend our studies, adding a new and exciting piece to the complex puzzle of signaling networks activated by androgens in target cells. Our findings might have implications for current approaches to AR-related diseases.
International journal of molecular sciences, Jan 9, 2015
Lichens are valuable natural resources used for centuries throughout the world as medicine, food,... more Lichens are valuable natural resources used for centuries throughout the world as medicine, food, fodder, perfume, spices and dyes, as well as for other miscellaneous purposes. This study investigates the antiproliferative, antibacterial and antifungal activity of the acetone extract of the lichen Xanthoria parietina (Linnaeus) Theodor Fries and its major secondary metabolite, parietin. The extract and parietin were tested for antimicrobial activity against nine American Type Culture Collection standard and clinically isolated bacterial strains, and three fungal strains. Both showed strong antibacterial activity against all bacterial strains and matched clinical isolates, particularly against Staphylococcus aureus from standard and clinical sources. Among the fungi tested, Rhizoctonia solani was the most sensitive. The antiproliferative effects of the extract and parietin were also investigated in human breast cancer cells. The extract inhibited proliferation and induced apoptosis, ...
Journal of cellular physiology, 2015
Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that defends against oxidative d... more Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that defends against oxidative damage due to reactive oxygen species (ROS). A new isoform of MnSOD with cytotoxic activity was recently discovered in liposarcoma cells. Here, we tested the effectiveness of a recombinant form of this isoform (rMnSOD) on leukemic T cells, Jurkat cells, and lymphocytes. Our results confirm that leukemic T cells can internalize rMnSOD and that rMnSOD causes apoptosis of 99% of leukemic cells without showing toxic effects on healthy cells. Using light and electron microscopy, we determined that an rMnSOD concentration of 0.067 μM most effective on apoptosis induction. Western blot analysis showed that treatment with 0.067 μM rMnSOD resulted in high expression of the pro-apoptotic protein Bax and low expression of the anti-apoptotic protein Bcl-2 in leukemia cells. Concerning signal transduction pathway no influence was observed after treatment except for Jurkat cells showing a slightly decr...
Cell Death & Differentiation, 2014
Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 2011
The aim of this study was to clarify the role of the erythrocyte inclusions found during the hema... more The aim of this study was to clarify the role of the erythrocyte inclusions found during the hematological screening of loggerhead population of the Mediterranean Sea. We studied the erythrocyte inclusions in blood specimens collected from six juvenile and nine adult specimens of the loggerhead turtle, Caretta caretta, from the Adriatic and Tyrrhenian Seas. Our study indicates that the percentage of mature erythrocytes containing inclusions ranged from 3 to 82%. Each erythrocyte contained only one round inclusion body. Inclusion bodies stained with May Grünwald–Giemsa show that their cytochemical and ultrastructure characteristics are identical to those of human Heinz bodies. Because Heinz bodies originate from the precipitation of unstable hemoglobin (Hb) and cause globular osmotic resistance to increase, we analyzed loggerhead Hb using electrophoresis and high‐performance liquid chromatography to detect and quantitate Hb fractions. We also tested the resistance of Hb to alkaline p...
Comparative Clinical Pathology, 2012
The eosinophils and heterophils of loggerheads display a similar rounded shape, an eccentric and ... more The eosinophils and heterophils of loggerheads display a similar rounded shape, an eccentric and elongated or slightly lobulated nucleus and a cytoplasm containing acidophilic pleomorphic granules. The characterization of the different types of granulocytes from nine Mediterranean loggerheads (Caretta caretta), based on cytochemical and immunocytochemical reactions and ultrastructural studies, is reported. On the basis of their positivity after myeloperoxidase, alkaline phosphatase and chloroacetate esterase reactions (typical of mammalian neutrophils), the granulocytes were classified as heterophils, while those myeloperoxidase-negative, major basic protein-positive granulocytes (like mammalian eosinophils), displaying a weak expression of interleukin 5 (a growth and differentiation factor, activator and chemoattractant for eosinophils), were classified as eosinophils. The immunocytochemical and TEM studies allowed the identification of these two granulocyte types, both of which show an eosinophilic reaction with May-Grünwald-Giemsa, periodic acid-Schiff and neutral red staining positivity.
Comparative Clinical Pathology, 2011
In order to establish baseline hematological and biochemical values in loggerhead turtles from th... more In order to establish baseline hematological and biochemical values in loggerhead turtles from the Mediterranean Sea, 84 specimens were sampled, comprising 24 wild turtles in good health at the time of capture and 60 turtles tested after indoor rehabilitation at the Sea Turtle Rescue and Rehabilitation Centre of the Zoological Station Anton Dohrn in Naples, Italy. The following parameters were evaluated: red cell counts (RBC, 488-575×10 3 /μL), white cell counts (WBC, 17-24×10 3 /μL) and thrombocyte counts (TBC, 19-49×10 2 /μL), hemoglobin (Hb, 8-14 g/dL), hematocrit (Ht, 23-34%), mean corpuscular volume (MCV, 487-723 fL), mean corpuscular hemoglobin (MCH, 170-261 pg), mean corpuscular hemoglobin concentration (MCHC, 34-42%), white and red blood cell differential counts, and a panel of hematochemical tests, composed of glucose (97-164 mg/ dL), cholesterol (74-144 mg/dL), blood urea nitrogen (35-200 mg/dL), uric acid (1-2.7 mg/dL), total bilirubin (0.20-0.40 mg/dL), GOT (44-184 IU/L) and GPT (6 IU/L) transaminases, calcium (6.7-8.7 mg/dL), and magnesium (3.6-5.4 mEq/L). Comparisons of the statistically analyzed data from the turtles which were divided into groups on the basis of age and/or lifestyle (wild or captive) revealed that erythroid parameters attained higher values in captive turtles. This suggested a positive influence of the rich and complete diet fed in captivity upon the hemopoietic process of the turtle. On the other hand, data suggest a more intense and active hemopoiesis in young turtles, compared to adult specimens.
Comparative Clinical Pathology, 2010
Abstract Theileria is a small Apicomplexan parasite considered as the scourge of the cattle indus... more Abstract Theileria is a small Apicomplexan parasite considered as the scourge of the cattle industry, here detected for the first time in the blood cells of Caretta caretta from the Mediterranean Sea. In the early stage of infection by Theileria, rounded forms, defined schizonts, occur in ...
Cell Death & Differentiation, Jun 27, 2014
Nell’ambito della sperimentazione clinica, che viene svolta nei paesi industrializzati, si rende ... more Nell’ambito della sperimentazione clinica, che viene svolta nei paesi industrializzati, si rende necessario porre una specifica attenzione ai criteri etici di riferimento affinche siano salvaguardati i beni e i valori umani fondamentali. Il rischio e che gli interessi commerciali possano nascondersi dietro gli interessi scientifici, danneggiando il benessere psichico del malato e la sua dignita di essere umano. Nei pazienti oncologici la sofferenza fisica, che rappresenta soltanto un sintomo della malattia, determina necessariamente un’alterazione della qualita di vita e puo condizionare la vita di relazione. Tutto questo viene ovviamente amplificato quando il malato e un bambino che, talora, e incapace di esprimere, localizzare e quantificare la sua sofferenza. Gli oncologi pediatrici lamentano la mancanza di farmaci antitumorali studiati per i bambini, in considerazione del fatto che i chemioterapici utilizzati nei protocolli di cura degli adulti non sono spesso egualmente efficaci in eta pediatrica. Le aziende farmaceutiche hanno poco interesse a sviluppare farmaci per la popolazione pediatrica oncologica per motivi di natura economica, dipendenti dalla esiguita del numero di pazienti pediatrici rispetto alla popolazione adulta. In generale la produzione di farmaci antitumorali alternativi trova resistenza nell’ambito della industria farmaceutica coinvolta nei profitti relativi al mercato dei chemio e radioterapici. Obiettivo primario del mio progetto di ricerca e stato, pertanto, la messa a punto di una terapia non tossica, o a ridotta citotossicita, per la cura delle neoplasie ematologiche in eta pediatrica. In particolare l’obiettivo di tale progetto e stato, nella prima fase, testare l’efficacia d’azione, sulle cellule di Leucemia Linfoblastica Acuta (LLA), la patologia piu frequente in eta pediatrica, del peptide leader della rMnSOD (manganese superossido dismutasi ricombinante) quale carrier di citostatici (cisplatino ed epirubicina) allo scopo di ridurre la dose di citostatico necessaria per una efficace internalizzazione dello stesso nelle cellule tumorali e per ottenerne quindi la morte riducendo gli effetti collaterali. Nella seconda fase, e stata studiata l’azione, sulle cellule in coltura di LLA B e T ex vivo e su una linea stabilizzata, della r-MnSOD, una proteina isolata e sequenziata, per la prima volta, da cellule di liposarcoma umano (LSA) presso il laboratorio di Oncologia Sperimentale dell’Istituto dei Tumori di Napoli, G. Pascale, dal Dott. A. Mancini. Tale molecola ha dimostrato attivita antitumorale su altri tipi di tumore. I dati ottenuti in questo lavoro hanno dimostrato che il peptide leader e un buon carrier di citostatici all’interno delle cellule leucemiche, tuttavia i citostatici legati al suddetto peptide non si sono rivelati efficaci nell’indurre la morte delle cellule leucemiche in coltura. Sara necessario quindi, quale proseguimento della ricerca, testare l’azione di altri citostatici legati al peptide. L’utilizzo del peptide leader coniugato a dosi micro molari di un diverso citostatico, efficace nella LLA pediatrica, potra, se confermato dai risultati, ottenere una riduzione notevole degli effetti collaterali migliorando la qualita di vita e la prognosi dei pazienti leucemici pediatrici ad alto rischio. Diversamente, il trattamento con l’intera molecola rMnSOD ha dimostrato sia la sua capacita di internalizzazione (dovuta alla presenza del peptide leader) nelle cellule leucemiche di LLA B e T in coltura, che la sua potente azione antitumorale, avendo indotto la morte di un alto numero di cellule leucemiche trattate. E stato, cosi, dimostrato l’effetto citotossico di tale proteina nei confronti delle cellule neoplastiche trattate e la mancanza di effetti tossici sulle cellule sane di controllo, sulle quali svolge, invece, una benefica ossigenazione. L’entita delle alterazioni morfologiche osservate in microscopia ottica ed elettronica nelle cellule trattate con rMnSOD alla concentrazione di 2 µg/mL ha suggerito che questa sia la concentrazione piu efficace, tra quelle sperimentate. Tale dato e stato confermato dai risultati di biologia molecolare: e stata riscontrata, infatti, a seguito del suddetto trattamento, la massima espressione dei geni proapoptotici (Bax) e la minima espressione dei geni antiapoptotici (Bcl-2); cio a conferma dell’attivazione della via apoptotica nelle cellule leucemiche. Pertanto, la rMnSOD potrebbe rivelarsi un efficace farmaco antitumorale, non tossico come i chemioterapici, per il trattamento delle leucemie ad alto rischio.
The androgen receptor (AR) mediates differentiation, proliferation and transformation of target t... more The androgen receptor (AR) mediates differentiation, proliferation and transformation of target tissues. These processes require a crosstalk between epithelial and stromal cells. Prostate cancer (PCa) represents a major cause of cancer-related mortality in men, and is often associated with deregulation of androgen/AR axis. Clinical and molecular findings have highlighted the role of epithelial AR in PCa progression. In contrast, the functions of AR in mesenchymal cells are still unclear. We previously reported that low androgen concentration (1 pM) triggers interaction of AR with the Src tyrosine kinase and PI3-K, thus driving cell cycle progression in fibroblasts. In contrast, stimulation of fibroblasts and fibrosarcoma cells with physiological (10 nM) androgen concentration leads to interaction of AR with full-length filamin A (FLNa) and does not trigger DNA synthesis. On the basis of these findings, we re-examined the role of androgen/AR axis in fibroblasts and human fibrosarcoma...
Oncotarget, Jan 24, 2015
Prostate cancer is one of the most commonly diagnosed cancers in men, and androgen deprivation th... more Prostate cancer is one of the most commonly diagnosed cancers in men, and androgen deprivation therapy still represents the primary treatment for prostate cancer patients. This approach, however, frequently fails and patients develop castration-resistant prostate cancer, which is almost untreatable.Cancer cells are characterized by a hierarchical organization, and stem/progenitor cells are endowed with tumor-initiating activity. Accumulating evidence indicates that prostate cancer stem cells lack the androgen receptor and are, indeed, resistant to androgen deprivation therapy. In contrast, these cells express classical (α and/or β) and novel (GPR30) estrogen receptors, which may represent new putative targets in prostate cancer treatment.In the present review, we discuss the still-debated mechanisms, both genomic and non-genomic, by which androgen and estradiol receptors (classical and novel) mediate the hormonal control of prostate cell stemness, transformation, and the continued g...
Journal of cellular physiology, 2015
Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that defends against oxidative d... more Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that defends against oxidative damage due to reactive oxygen species (ROS). A new isoform of MnSOD with cytotoxic activity was recently discovered in liposarcoma cells. Here, we tested the effectiveness of a recombinant form of this isoform (rMnSOD) on leukemic T cells, Jurkat cells, and lymphocytes. Our results confirm that leukemic T cells can internalize rMnSOD and that rMnSOD causes apoptosis of 99% of leukemic cells without showing toxic effects on healthy cells. Using light and electron microscopy, we determined that an rMnSOD concentration of 0.067 μM most effective on apoptosis induction. Western blot analysis showed that treatment with 0.067 μM rMnSOD resulted in high expression of the pro-apoptotic protein Bax and low expression of the anti-apoptotic protein Bcl-2 in leukemia cells. Concerning signal transduction pathway no influence was observed after treatment except for Jurkat cells showing a slightly decr...
Methods in Molecular Biology, 2014
It is well established that histone posttranslational modifications mediate the control of gene e... more It is well established that histone posttranslational modifications mediate the control of gene expression played by chromatin. Such modifications are commonly reversible and many alternatives are open to drive transcription of inducible genes. Estrogens govern growth and survival of hormone-sensitive cells by inducing expression of genes important for cell cycle progression and apoptosis. Transcription of estrogen-responsive genes is triggered by the lysine-specific demethylase 1 (LSD1)-dependent demethylation of dimethylated lysine 9 in histone H3 (H3K9me2) that accompanies to local generation of oxygen reactive species (ROS). Production of ROS modifies guanines in neighbor DNA with consequent recruitment of base-excision repair (BER) enzymes and formation of breaks that support creation of bridges between sites that, although distant on linear DNA, establish strategic contacts useful for productive transcription.
Frontiers in Endocrinology, 2015
Prostate cancer (PCa) is the major cause of cancer-related death among the male population of Wes... more Prostate cancer (PCa) is the major cause of cancer-related death among the male population of Western society, and androgen-deprivation therapy (ADT) represents the first line in PCa treatment. However, although androgen receptor (AR) expression is maintained throughout the various stages of PCa, ADT frequently fails. Clinical studies have demonstrated that different androgen/AR signaling pathways operate in target tissues. AR stimulates growth and transformation of target cells, but under certain conditions slows down their proliferation. In this review, we discuss the role of AR in controlling different functions of mesenchymal and transformed mesenchymal cells. Findings here presented support the role of AR in suppressing proliferation and stimulating migration of stromal cells, with implications for current approaches to cancer therapy.
Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 2011
The aim of this study was to clarify the role of the erythrocyte inclusions found during the hema... more The aim of this study was to clarify the role of the erythrocyte inclusions found during the hematological screening of loggerhead population of the Mediterranean Sea. We studied the erythrocyte inclusions in blood specimens collected from six juvenile and nine adult specimens of the loggerhead turtle, Caretta caretta, from the Adriatic and Tyrrhenian Seas. Our study indicates that the percentage of mature erythrocytes containing inclusions ranged from 3 to 82%. Each erythrocyte contained only one round inclusion body. Inclusion bodies stained with May Grünwald-Giemsa show that their cytochemical and ultrastructure characteristics are identical to those of human Heinz bodies. Because Heinz bodies originate from the precipitation of unstable hemoglobin (Hb) and cause globular osmotic resistance to increase, we analyzed loggerhead Hb using electrophoresis and high-performance liquid chromatography to detect and quantitate Hb fractions. We also tested the resistance of Hb to alkaline pH, heat, isopropanol denaturation, and globular osmosis. Our hemogram results excluded the occurrence of any infection, which could be associated with an inclusion body, in all the specimens. Negative Feulgen staining indicated that the inclusion bodies are not derived from DNA fragmentation. We hypothesize that amino acid substitutions could explain why loggerhead Hb precipitates under normal physiologic conditions, forming Heinz bodies. The identification of inclusion bodies in loggerhead erythrocytes allow us to better understand the haematological characteristics and the physiology of these ancient reptiles, thus aiding efforts to conserve such an endangered species.