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ANTO DE POL
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Dipartimento Chirurgico, Medico, Odontoiatrico e di Scienze Morfologiche con interesse Trapiantologico, Oncologico e di Medicina Rigenerativa
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Pubblicazioni
2020
- Is diet partly responsible for differences in COVID-19 death rates between and within countries?
[Articolo su rivista]
Bousquet, J.; Anto, J. M.; Iaccarino, G.; Czarlewski, W.; Haahtela, T.; DE POL, Anto; Akdis, C. A.; Blain, H.; Canonica, G. W.; Cardona, V.; Cruz, A. A.; Illario, M.; Ivancevich, J. C.; Jutel, M.; Klimek, L.; Kuna, P.; Laune, D.; Larenas-linnemann, D.; Mullol, J.; Papadopoulos, N. G.; Pfaar, O.; Samolinski, B.; Valiulis, A.; Yorgancioglu, A.; Zuberbier, T.; Abdul Latiff, A. H.; Abdullah, B.; Aberer, W.; Abusada, N.; Adcock, I.; Afani, A.; Agache, I.; Aggelidis, X.; Agustin, J.; Akdis, C.; Akdis, M.; Al-Ahmad, M.; Bassam, A. A. -Z.; Aldrey-Palacios, O.; Cuesta, E. A.; Alzaabi, A.; Amad, S.; Ambrocio, G.; Annesi-Maesano, I.; Ansotegui, I.; Anto, J.; Arshad, H.; Artesani, M. C.; Asayag, E.; Avolio, F.; Azhari, K.; Baiardini, I.; Bajrovic, N.; Bakakos, P.; Mongono, S. B.; Balotro-Torres, C.; Barba, S.; Barbara, C.; Barbosa, E.; Barreto, B.; Bartra, J.; Bateman, E. D.; Battur, L.; Bedbrook, A.; Barajas, M. B.; Beghe, B.; Bel, E.; Kheder, A. B.; Benson, M.; Berghea, C.; Bergmann, K. -C.; Bernstein, D.; Bewick, M.; Bialek, S.; Bialoszewski, A.; Bieber, T.; Billo, N.; Bilo, M. B.; Bindslev-Jensen, C.; Bjermer, L.; Blain, H.; Marciniak, M. B.; Bond, C.; Boner, A.; Bonini, M.; Bonini, S.; Bosnic-Anticevich, S.; Bosse, I.; Botskariova, S.; Bouchard, J.; Boulet, L. -P.; Bourret, R.; Bousquet, P.; Braido, F.; Briggs, A.; Brightling, C.; Brozek, J.; Buhl, R.; Bumbacea, R.; Cabanas, M. T. B.; Bush, A.; Busse, W. W.; Buters, J.; Caballero-Fonseca, F.; Calderon, M. A.; Calvo, M.; Camargos, P.; Camuzat, T.; Cano, A.; Canonica, G. W.; Capriles-Hulett, A.; Caraballo, L.; Cardona, V.; Carlsen, K. -H.; Caro, J.; Carr, W.; Carreon-Asun-cion, F.; Carriazo, A. M.; Casale, T.; Castor, M. A.; Castro, E.; Cecchi, L.; Sarabia, A. C.; Chandrasekharan, R.; Chang, Y. -S.; Chato-Andeza, V.; Chatzi, L.; Chatzidaki, C.; Chavannes, N. H.; Chen, Y.; Cheng, L.; Chivato, T.; Chkhartishvili, E.; Christoff, G.; Chrystyn, H.; Chu, D. K.; Chua, A.; Chuchalin, A.; Chung, K. F.; Ciceran, A.; Cingi, C.; Ciprandi, G.; Cirule, I.; Coelho, A. C.; Constantinidis, J.; de Sousa, J. C.; Costa, E.; Costa, D.; Dominguez, M. C. C.; Coste, A.; Cox, L.; Cruz, A. A.; Cullen, J.; Custovic, A.; Cvetkovski, B.; Czarlewski, W.; D'Amato, G.; Silva, J. D.; Dahl, R.; Dahlen, S. -E.; Daniilidis, V.; Nahhas, L. D.; Darsow, U.; Blay, F.; Guia, E. D.; Santos, C.; Keenoy, E. D. M.; Vries, G. D.; Deleanu, D.; Demoly, P.; Denburg, J.; Devillier, P.; Didier, A.; Dimou, M.; Dinh-Xuan, A. T.; Djukanovic, R.; Dokic, D.; Dominguez Silva, M. G.; Douagui, H.; Douladiris, N.; Doulaptsi, M.; Dray, G.; Dubakiene, R.; Durham, S.; Dykewicz, M.; Ebo, D.; Edelbaher, N.; Eklund, P.; El-Gamal, Y.; El-Sayed, Z. A.; El-Sayed, S. S.; El-Seify, M.; Emuzyte, R.; Enecilla, L.; Espinoza, H.; Guillermo, J.; Contreras, E.; Farrell, J.; Fernandez, L.; Wagner, A. F.; Fiocchi, A.; Fokkens, W. J.; Fontaine, J. -F.; Forastiere, F.; Perez, J. M. F.; Gaerlan-resureccion, E.; Gaga, M.; Romero, J. L. G.; Gamkrelidze, A.; Garcia, A.; Cobas, C. Y. G.; Garcia Cruz, M. L. L. H.; Gayraud, J.; Gemicioglu, B.; Genova, S.; Gereda, J.; Wijk, R. G.; Gomez, M.; Diaz, S. G.; Gotua, M.; Grigoreas, C.; Grisle, I.; Guidacci, M.; Guldemond, N.; Gutter, Z.; Guzman, A.; Haahtela, T.; Halloum, R.; Hamelmann, E.; Hammadi, S.; Harvey, R.; Heinrich, J.; Hejjaoui, A.; Hellquist-Dahl, B.; Velazquez, L. H.; Hew, M.; Hossny, E.; Howarth, P.; Hrubisko, M.; Villalobos, Y. R. H.; Humbert, M.; Hyland, M.; Ibrahim, M.; Illario, M.; Ilyina, N.; Irani, C.; Ispayeva, Z.; Ivancevich, J. C.; Jares, E.; Jarvis, D.; Jassem, E.; Jenko, K.; Uscanga, R. D. J.; Johnston, S.; Joos, G.; Jost, M.; Julge, K.; Jung, K. -S.; Just, J.; Jutel, M.; Kaidashev, I.; Kalayci, O.; Kalyoncu, F.; Kapsali, J.; Kardas, P.; Karjalainen, J.; Kasala, C. A.; Katotomichelakis, M.; Kazi, B.; Keil, T.; Keith, P.; Khaitov, M.; Khaltaev, N.; Kim, Y. -Y.; Kleine-Tebbe, J.; Klimek, L.; Koffi N'Goran, B.; Kompoti, E.; Kopac, P.; Koppelman, G.; Jeverica, A. K.; Kosnik, M.; Kostov, K. V.; Kowalski, M. L.
abstract
Reported COVID-19 deaths in Germany are relatively low as compared to many European countries. Among the several explanations proposed, an early and large testing of the population was put forward. Most current debates on COVID-19 focus on the differences among countries, but little attention has been given to regional differences and diet. The low-death rate European countries (e.g. Austria, Baltic States, Czech Republic, Finland, Norway, Poland, Slovakia) have used different quarantine and/or confinement times and methods and none have performed as many early tests as Germany. Among other factors that may be significant are the dietary habits. It seems that some foods largely used in these countries may reduce angiotensin-converting enzyme activity or are anti-oxidants. Among the many possible areas of research, it might be important to understand diet and angiotensin-converting enzyme-2 (ACE2) levels in populations with different COVID-19 death rates since dietary interventions may be of great benefit.
2020
- Modulation of Cell Death and Promotion of Chondrogenic Differentiation by Fas/FasL in Human Dental Pulp Stem Cells (hDPSCs)
[Articolo su rivista]
Pisciotta, Alessandra; Bertani, Giulia; Bertoni, Laura; Di Tinco, Rosanna; De Biasi, Sara; Vallarola, Antonio; Pignatti, Elisa; Tupler, Rossella; Salvarani, Carlo; de Pol, Anto; Carnevale, Gianluca
abstract
2019
- Evaluation of Biological Response of STRO-1/c-Kit Enriched Human Dental Pulp Stem Cells to Titanium Surfaces Treated with Two Different Cleaning Systems.
[Articolo su rivista]
Conserva, E; Pisciotta, A; Bertoni, L; Bertani, Giulia; Meto, A; Colombari, B; Blasi, E; Bellini, P; de Pol, A; Consolo, U; Carnevale, G.
abstract
Peri-implantitis-an infection caused by bacterial deposition of biofilm-is a common complication in dentistry which may lead to implant loss. Several decontamination procedures have been investigated to identify the optimal approach being capable to remove the bacterial biofilm without modifying the implant surface properties. Our study evaluated whether two different systems-Ni-Ti Brushes (Brush) and Air-Polishing with 40 µm bicarbonate powder (Bic40)-might alter the physical/chemical features of two different titanium surfaces-machined (MCH) and Ca++ nanostructured (NCA)-and whether these decontamination systems may affect the biological properties of human STRO-1+/c-Kit+ dental pulp stem cells (hDPSCs) as well as the bacterial ability to produce biofilm. Cell morphology, proliferation and stemness markers were analysed in hDPSCs grown on both surfaces, before and after the decontamination treatments. Our findings highlighted that Bic40 treatment either maintained the surface characteristics of both implants and allowed hDPSCs to proliferate and preserve their stemness properties. Moreover, Bic40 treatment proved effective in removing bacterial biofilm from both titanium surfaces and consistently limited the biofilm re-growth. In conclusion, our data suggest that Bic40 treatment may operatively clean smooth and rough surfaces without altering their properties and, consequently, offer favourable conditions for reparative cells to hold their biological properties.
2019
- Regenerative potential of human dental pulp stem cells in the treatment of stress urinary incontinence: In vitro and in vivo study
[Articolo su rivista]
Zordani, Alessio; Pisciotta, Alessandra; Bertoni, Laura; Bertani, Giulia; Vallarola, Antonio; Giuliani, Daniela; Puliatti, Stefano; Mecugni, Daniela; Bianchi, Giampaolo; De Pol, Anto; Carnevale, Gianluca
abstract
OBJECTIVES:
To evaluate the regenerative potential of human dental pulp stem cells (hDPSCs) in an animal model of stress urinary incontinence (SUI). SUI, an involuntary leakage of urine, is due to physical stress involving an increase in bladder pressure and a damage of external urethral sphincter affecting muscles and nerves. Conventional therapies can only relieve the symptoms. Human DPSCs are characterized by peculiar stemness and immunomodulatory properties and might provide an alternative tool for SUI therapy.
MATERIALS AND METHODS:
In vitro phase: hDPSCs were induced towards the myogenic commitment following a 24 hours pre-conditioning with 5-aza-2'-deoxycytidine (5-Aza), then differentiation was evaluated. In vivo phase: pudendal nerve was transected in female rats to induce stress urinary incontinence; then, pre-differentiated hDPSCs were injected in the striated urethral sphincter. Four weeks later, urethral sphincter regeneration was assayed through histological, functional and immunohistochemical analyses.
RESULTS:
Human DPSCs were able to commit towards myogenic lineage in vitro and, four weeks after cell injection, hDPSCs engrafted in the external urethral sphincter whose thickness was almost recovered, committed towards myogenic lineage in vivo, promoted vascularization and an appreciable recovery of the continence. Moreover, hDPSCs were detected within the nerve, suggesting their participation in repair of transected nerve.
CONCLUSIONS:
These promising data and further investigations on immunomodulatory abilities of hDPSCs would allow to make them a potential tool for alternative therapies of SUI.
2019
- Titanium Surface Properties Influence the Biological Activity and FasL Expression of Craniofacial Stromal Cells.
[Articolo su rivista]
Conserva, E; Pisciotta, A; Borghi, F; Nasi, M; Pecorini, Simone; Bertoni, L; de Pol, A; Consolo, U; Carnevale, G.
abstract
Mesenchymal stromal cells (MSCs) can be easily isolated form craniofacial bones during routine dentistry procedures. Due to their embryological origin from neural crest, they represent a suitable cell population to study cell-biomaterial interaction in the craniofacial field, including osteoinductive/osteointegrative processes. The biological and immunomodulatory properties of MSCs may be influenced by chemistry and topography of implant surfaces. We investigated if and how three different titanium surfaces, machined (MCH), sandblasted with resorbable blasting medium (RBM), and Ca++-nanostructured (NCA), may affect biological activity, osseointegration, and immunomodulatory properties of craniofacial MSCs. Cell proliferation, morphology, osteogenic markers, and FasL were evaluated on MSCs isolated from the mandibular bone after seeding on these three different surfaces. No statistically significant differences in cell proliferation were observed whereas different morphologies and growth patterns were detected for each type of surface. No difference in the expression of osteogenic markers was revealed. Interestingly, FasL expression, involved in the immunomodulatory activity of stem cells, was influenced by surface properties. Particularly, immunofluorescence analysis indicated that FasL expression increased on MCH surface compared to the others confirming the suggested role of FasL in promoting osteogenic differentiation. Titanium surface treatments and topography might reflect different biological behaviours of craniofacial MSCs and influence their osseointegration/immunomodulation properties.
2018
- Anterior Capsule of the Lens: Comparison of Morphological Properties and Apoptosis Induction following FLACS and Standard Phacoemulsification Surgery
[Articolo su rivista]
Pisciotta, Alessandra; De Maria, Michele; Verdina, Tommaso; Fornasari, Elisa; De Pol, Anto; Cavallini, Gian Maria
abstract
Purpose. Comparative evaluation of morphological features of anterior capsules and apoptosis induction in epithelial cells after femtosecond laser-assisted cataract surgery (FLACS) and standard phacoemulsification surgery. Methods. Group 1: 30 FLACS anterior capsulotomies and Group 2: 30 manual anterior continuous curvilinear capsulorhexes. All patients were operated on by the same experienced surgeon. Morphological features of the anterior capsules and apoptosis induction in epithelial cells were evaluated. Results. All patients revealed a significant mean best-corrected visual acuity (BCVA) improvement 3 months after surgery, and no major intraoperative nor postoperative complications occurred. The capsular epithelium appeared to be preserved in both groups. Scanning electron microscopy analysis revealed irregular saw-tooth shaped edges in capsules from Group 1 whereas capsules from Group 2 showed regular and smooth edges. A statistically significant higher expression of the downstream apoptotic effector cleaved caspase 3 was observed in Group 1. Conclusions. The saw-tooth appearance was likely due to the progressive sequence of laser pulses on the capsule. The low energy/high frequency properties of the laser pulse, combined with an overlapped pulse pattern, resulted in highly continuous morphology of capsule edges. The higher apoptosis induction in FLACS group might be due to photodisruption-dependent plasma generation and formation of cavitation bubbles.
2018
- Human dental pulp stem cells expressing STRO-1, c-kit and CD34 markers in peripheral nerve regeneration
[Articolo su rivista]
Carnevale, Gianluca; Pisciotta, Alessandra; Riccio, Massimo; Bertoni, Laura; DE BIASI, Sara; Gibellini, Lara; Zordani, Alessio; Cavallini, Gian Maria; LA SALA, Giovanni Battista; Bruzzesi, Giacomo; Ferrari, Adriano; Cossarizza, Andrea; DE POL, Anto
abstract
Peripheral nerve injuries are a commonly encountered clinical problem and often result in long-term functional defects. The application of stem cells able to differentiate in Schwann cell-like cells in vitro and in vivo, could represent an attractive therapeutic approach for the treatment of nerve injuries. Further, stem cells sources sharing the same embryological origin as Schwann cells might be considered a suitable tool. The aim of this study was to demonstrate the ability of a neuroectodermal subpopulation of human STRO-1(+) /c-Kit(+) /CD34(+) DPSCs, expressing P75(NTR) , nestin and SOX-10, to differentiate into Schwann cell-like cells in vitro and to promote axonal regeneration in vivo, which led to functional recovery as measured by sustained gait improvement, in animal rat model of peripheral nerve injury. Transplanted human dental pulp stem cells (hDPSCs) engrafted into sciatic nerve defect, as revealed by the positive staining against human nuclei, showed the expression of typical Schwann cells markers, S100b and, noteworthy, a significant number of myelinated axons was detected. Moreover, hDPSCs promoted axonal regeneration from proximal to distal stumps 1 month after transplantation. This study demonstrates that STRO-1(+) /c-Kit(+) /CD34(+) hDPSCs, associated with neural crest derivation, represent a promising source of stem cells for the treatment of demyelinating disorders and might provide a valid alternative tool for future clinical applications to achieve functional recovery after injury or peripheral neuropathies besides minimizing ethical issues. Copyright © 2016 John Wiley & Sons, Ltd.
2018
- Use of a 3D floating sphere culture system to maintain the neural crest-related properties of human dental pulp stem cells
[Articolo su rivista]
Pisciotta, Alessandra; Bertoni, Laura; Riccio, Massimo; Mapelli, Jonathan; Bigiani, Albertino; Noce, Marcella La; Orciani, Monia; de Pol, Anto; Carnevale, Gianluca
abstract
Human dental pulp is considered an interesting source of adult stem cells, due to the low-invasive isolation procedures, high content of stem cells and its peculiar embryological origin from neural crest. Based on our previous findings, a dental pulp stem cells sub-population, enriched for the expression of STRO-1, c-Kit, and CD34, showed a higher neural commitment. However, their biological properties were compromised when cells were cultured in adherent standard conditions. The aim of this study was to evaluate the ability of three dimensional floating spheres to preserve embryological and biological properties of this sub-population. In addition, the expression of the inwardly rectifying potassium channel Kir4.1, Fas and FasL was investigated in 3D-sphere derived hDPSCs. Our data showed that 3D sphere-derived hDPSCs maintained their fibroblast-like morphology, preserved stemness markers expression and proliferative capability. The expression of neural crest markers and Kir4.1 was observed in undifferentiated hDPSCs, furthermore this culture system also preserved hDPSCs differentiation potential. The expression of Fas and FasL was observed in undifferentiated hDPSCs derived from sphere culture and, noteworthy, FasL was maintained even after the neurogenic commitment was reached, with a significantly higher expression compared to osteogenic and myogenic commitments. These data demonstrate that 3D sphere culture provides a favorable micro-environment for neural crest-derived hDPSCs to preserve their biological properties.
2017
- Activation of Fas/FasL pathway and the role of c-FLIP in primary culture of human cholangiocarcinoma cells
[Articolo su rivista]
CARNEVALE, Gianluca; Carpino, Guido; Cardinale, Vincenzo; PISCIOTTA, ALESSANDRA; RICCIO, Massimo; Bertoni, Laura; GIBELLINI, Lara; DE BIASI, SARA; Nevi, Lorenzo; Costantini, Daniele; Overi, Diletta; COSSARIZZA, Andrea; DE POL, Anto; Gaudio, Eugenio; Alvaro, Domenico
abstract
Intrahepatic cholangiocarcinoma (iCCA) represents a heterogeneous group of malignancies emerging from the biliary tree, often in the context of chronic bile ducts inflammation. The immunological features of iCCA cells and their capability to control the lymphocytes response have not yet been investigated. The aims of the present study were to evaluate the interaction between iCCA cells and human peripheral blood mononuclear cells (PBMCs) and the role of Fas/FasL in modulating T-cells and NK-cells response after direct co-culture. iCCA cells express high levels of Fas and FasL that increase after co-culture with PBMCs inducing apoptosis in CD4(+), CD8(+) T-cells and in CD56(+) NK-cells. In vitro, c-FLIP is expressed in iCCA cells and the co-culture with PBMCs induces an increase of c-FLIP in both iCCA cells and biliary tree stem cells. This c-FLIP increase does not trigger the caspase cascade, thus hindering apoptotis of iCCA cells which, instead, underwent proliferation. The increased expression of Fas, FasL and c-FLIP is confirmed in situ, in human CCA and in primary sclerosing cholangitis. In conclusion our data indicated that iCCA cells have immune-modulatory properties by which they induce apoptosis of T and NK cells, via Fas/FasL pathway, and escape inflammatory response by up-regulating c-FLIP system.
2017
- Apoptosis and inflammatory response in human astrocytes are induced by a transmissible cytotoxic agent of neurological origin
[Articolo su rivista]
Beretti, Francesca; Ardizzoni, Andrea; Cermelli, Claudio; Guida, Marianna; Maraldi, Tullia; Pietrosemoli, P; Paulone, Simona; De Pol, Anto; Blasi, Elisabetta; Portolani, Marinella
abstract
We demonstrated the presence of an in vitro transmissible cytotoxic agent (TCA) in the cerebrospinal fluid (CSF) of patients with different acute neurological diseases. The nature of this agent is still a matter of study since repeated attempts have failed to identify it as a conventional infectious agent. Here, we describe the mechanisms through which TCA affects human astrocytes, demonstrating:a late apoptotic process, mediated by caspases 9 and 3 activation, involving the Bcl2-Bak-axis;an early and late p38 MAPK activation;an interference with the IL-8 and MCP-1 secretory response. These in vitro data provide initial evidence of TCA involvement as a pro-apoptotic and pro-inflammatory signal, directly affecting astrocytic behavior. The implications of these findings in certain neurological diseases will be discussed.
2017
- Corrigendum to: Osteogenic differentiation of hDPSCs on biogenic bone apatite thin films (Stem Cells International (2017) 2017 (3579283) DOI: 10.1155/2017/3579283)
[Articolo su rivista]
Bianchi, Michele; Pisciotta, Alessandra; Bertoni, Laura; Berni, Matteo; Gambardella, Alessandro; Visani, Andrea; Russo, Alessandro; DE POL, Anto; Carnevale, Gianluca
abstract
In the article titled "Osteogenic differentiation of hDPSCs on biogenic bone apatite thin films" [1], the second affiliation was incorrect. The corrected affiliations are shown above.
2017
- DEVELOPMENT OF A NOVEL METHOD FOR AMNIOTIC FLUID STEM CELL STORAGE
[Articolo su rivista]
Zavatti, Manuela; Beretti, Francesca; Casciaro, Francesca; Comitini, Giuseppina; Franchi, Fabrizia; Barbieri, Veronica; Bertoni, Laura; DE POL, Anto; LA SALA, Giovanni Battista; Maraldi, Tullia
abstract
Background - Current procedures for collection of human
Amniotic Fluid Stem Cells (hAFSCs) imply that amniotic fluid cells were
cultured in flask for two weeks, than can be devoted to research purpose.
However, hAFSCs could be retrieved directly from a small amount of
amniotic fluid that can be obtained at the time of diagnostic
amniocentesis. The aim of the study was to verify if a direct freezing of
amniotic fluid cells is able to maintain and / or improve the potential
of the sub-population of stem cells. Methods - We compared the potential
of the hAFSCs depending on the moment in which they are frozen, cells
obtained directly from amniotic fluid aspiration (D samples) and cells
cultured in flask before freezing (C samples). Colony-forming-unit
ability, proliferation, morphology, stemness-related marker expression,
senescence, apoptosis, and differentiation potential of C and D samples
were compared. Results - hAFSCs isolated from D samples expressed MSC
markers until later passages, had a good proliferation rate, and
exhibited differentiation capacity similar to hAFSCs of C samples.
Interestingly, the direct freezing induce a higher concentration of cells
positive for pluripotency stem cell markers, without teratoma formation
in vivo.
Conclusions - This study suggests that minimal processing may be adequate
for the banking of amniotic fluid cells, avoiding in vitro passages
before the storage and exposure to high oxygen concentration affecting
stem cell properties. This technique might be a reasonable approach in
terms of costs and for the process of accreditation in GMP for a stem
cell bank.
2017
- Human biliary tree stem/progenitor cells immunomodulation: Role of hepatocyte growth factor
[Articolo su rivista]
Maraldi, Tullia; Guida, Marianna; Beretti, Francesca; Resca, Elisa; Carpino, Guido; Cardinale, Vincenzo; Gentile, Raffaele; Ardizzoni, Andrea; Murgia, Alba; Alvaro, Domenico; Gaudio, Eugenio; DE POL, Anto
abstract
Aim: Human biliary tree stem/progenitor cells (hBTSC) are multipotent epithelial stem cells with the potential for allogenic transplant in liver, biliary tree, and pancreatic diseases. Human mesenchymal stem cells, but also epithelial stem cells, are able to modulate immune responses with different types of secretion molecules. Methods: The initial aim of the present study was to develop for the first time a culture protocol in order to expand hBTSC invitro through passages, allowing to maintain a similar stem cell and secretome profile. Furthermore, we investigated the secretome profile of the hBTSC to assess the production of molecules capable of affecting immune feedback. Results: We found that hepatocyte growth factor produced by hBTSC exerts its cytoprotective role inducing apoptosis in human immune cells, such as lymphocytes. Conclusions: The present study, therefore, supports the hypothesis that hBTSC can be useful for the purpose of regenerative medicine, as they can be banked and expanded, and they can secrete immunoregulatory factors.
2017
- Osteogenic Differentiation of hDPSCs on Biogenic Bone Apatite Thin Films
[Articolo su rivista]
Bianchi, Michele; Pisciotta, Alessandra; Bertoni, Laura; Berni, Matteo; Gambardella, Alessandro; Visani, Andrea; Russo, Alessandro; DE POL, Anto; Carnevale, Gianluca
abstract
A previous study reported the structural characterization of biogenic apatite (BAp) thin films realized by a pulsed electron deposition system by ablation of deproteinized bovine bone. Thin films annealed at 400 degrees C exhibited composition and crystallinity degree very close to those of biogenic apatite; this affinity is crucial for obtaining faster osseointegration compared to conventional, thick hydroxyapatite (HA) coatings, for both orthopedics and dentistry. Here, we investigated the adhesion, proliferation, and osteogenic differentiation of human dental pulp stem cells (hDPCS) on as-deposited and heat-treated BAp and stoichiometric HA. First, we showed that heat-treated BAp films can significantly promote hDPSC adhesion and proliferation. Moreover, hDPSCs, while initially maintaining the typical fibroblast-like morphology and stemness surface markers, later started expressing osteogenic markers such as Runx-2 and OSX. Noteworthy, when cultured in an osteogenic medium on annealed BAp films, hDPSCs were also able to reach a more mature and terminal commitment, with respect to HA and as-deposited films. Our findings suggest that annealed BAp films not only preserve the typical biological properties of stemness of, hDPSCs but also improve their ability of osteogenic commitment.
2016
- Estrogen receptor signaling in the ferutinin-induced osteoblastic differentiation of human amniotic fluid stem cells
[Articolo su rivista]
Zavatti, Manuela; Guida, M; Maraldi, Tullia; Beretti, Francesca; Bertoni, Laura; LA SALA, Giovanni Battista; DE POL, Anto
abstract
Ferutinin is a diaucane sesquiterpene with a high estrogenic activity. Since ferutinin is able to enhance osteoblastic differentiation of human amniotic fluid stem cells (hAFSCs), the aim of this study was to evaluate the role of the estrogen receptors α (ERα) and G-protein coupled receptor 30 (GPR30) in ferutinin-mediated osteoblastic differentiation. Moreover, it was investigated if MEK/ERK and PI3K/Akt signaling pathways are involved in ferutinin-induced effects.
2016
- NADPH oxidase-4 and MATER expressions in granulosa cells: Relationships with ovarian aging
[Articolo su rivista]
Maraldi, Tullia; Resca, Elisa; Nicoli, Alessia; Beretti, Francesca; Zavatti, Manuela; Capodanno, Francesco; Morini, Daria; Palomba, Stefano; LA SALA, Giovanni Battista; DE POL, Anto
abstract
Aims Relevant roles in follicular development and ovulation are played by maternal antigen that embryos require (MATER), product of a maternal effect gene, and by reactive oxygen species (ROS), indispensable for the induction of ovulatory genes. At the moment, the relationship between these two biological systems and their involvement in the ovarian aging have not been still clarified. The aim of the current experimental study was to analyse the age-related changes of the MATER and NOX proteins. Materials and methods MATER and ROS homeostasis was studied in granulosa cells (GCs) and cumulus cells (CCs) of infertile patients who undergone oocyte retrieval for in vitro fertilization cycles using Western blot and confocal immunofluorescence analysis. Samples were obtained from subjects with age ≥ 40 years (cases) and with age ≤ 37 years (controls). Key findings The expression pattern of MATER and NOX observed in GCs was not different from that observed in CCs. High levels of both proteins were detected in the control samples. A significant lower expression of both MATER and NOX4 was observed in the case versus control samples. Significance The expression of MATER and NOX4 proteins are closely related to the follicular development and ovulation with particular regard for ovarian aging.
2016
- Optimized Cryopreservation and Banking of Human Bone-Marrow Fragments and Stem Cells
[Articolo su rivista]
Carnevale, Gianluca; Pisciotta, Alessandra; Riccio, Massimo; De Biasi, Sara; Gibellini, Lara; Ferrari, Adriano; La Sala, Giovanni Battista; Bruzzesi, Giacomo; Cossarizza, Andrea; De Pol, Anto
abstract
Adult mesenchymal stem cells are a promising source for cell therapies and tissue engineering applications. Current procedures for banking of human bone-marrow mesenchymal stem cells (hBM-MSCs) require cell isolation and expansion, and thus the use of large amounts of animal sera. However, animal-derived culture supplements have the potential to trigger infections and severe immune reactions. The aim of this study was to investigate an optimized method for cryopreservation of human bone-marrow fragments for application in cell banking procedures where stem-cell expansion and use are not immediately needed. Whole trabecular fragments enclosing the bone marrow were stored in liquid nitrogen for 1 year in a cryoprotective solution containing a low concentration of dimethyl sulfoxide and a high concentration of human serum (HuS). After thawing, the isolation, colony-forming-unit ability, proliferation, morphology, stemness-related marker expression, cell senescence, apoptosis, and multi-lineage differentiation potential of hBM-MSCs were tested in media containing HuS compared with hBM-MSCs isolated from fresh fragments. Human BM-MSCs isolated from cryopreserved fragments expressed MSC markers until later passages, had a good proliferation rate, and exhibited the capacity to differentiate toward osteogenic, adipogenic, and myogenic lineages similar to hBM-MSCs isolated from fresh fragments. Moreover, the cryopreservation method did not induce cell senescence or cell death. These results imply that minimal processing may be adequate for the banking of tissue samples with no requirement for the immediate isolation and use of hBM-MSCs, thus limiting cost and the risk of contamination, and facilitating banking for clinical use. Furthermore, the use of HuS for cryopreservation and expansion/differentiation has the potential for clinical application in compliance with good manufacturing practice standards.
2016
- Reversal of the glycolytic phenotype of primary effusion lymphoma cells by combined targeting of cellular metabolism and PI3K/Akt/ mTOR signaling
[Articolo su rivista]
Mediani, Laura; Gibellini, Federica; Bertacchini, Jessika; Frasson, Chiara; Bosco, Raffaella; Accordi, Benedetta; Basso, Giuseppe; Bonora, Massimo; Calabrò, Maria Luisa; Mattiolo, Adriana; Sgarbi, Gianluca; Baracca, Alessandra; Pinton, Paolo; Riva, Giovanni; Rampazzo, Enrico; Petrizza, Luca; Prodi, Luca; Milani, Daniela; Luppi, Mario; Potenza, Leonardo; De Pol, Anto; Cocco, Lucio; Capitani, Silvano; Marmiroli, Sandra
abstract
PEL is a B-cell non-Hodgkin lymphoma, occurring predominantly as a lymphomatous effusion in body cavities, characterized by aggressive clinical course, with no standard therapy. Based on previous reports that PEL cells display a Warburg phenotype, we hypothesized that the highly hypoxic environment in which they grow in vivo makes them more reliant on glycolysis, and more vulnerable to drugs targeting this pathway. We established here that indeed PEL cells in hypoxia are more sensitive to glycolysis inhibition. Furthermore, since PI3K/Akt/mTOR has been proposed as a drug target in PEL, we ascertained that pathway-specific inhibitors, namely the dual PI3K and mTOR inhibitor, PF-04691502, and the Akt inhibitor, Akti 1/2, display improved cytotoxicity to PEL cells in hypoxic conditions. Unexpectedly, we found that these drugs reduce lactate production/extracellular acidification rate, and, in combination with the glycolysis inhibitor 2-deoxyglucose (2-DG), they shift PEL cells metabolism from aerobic glycolysis towards oxidative respiration. Moreover, the associations possess strong synergistic cytotoxicity towards PEL cells, and thus may reduce adverse reaction in vivo, while displaying very low toxicity to normal lymphocytes. Finally, we showed that the association of 2-DG and PF-04691502 maintains its cytotoxic and proapoptotic effect also in PEL cells co-cultured with human primary mesothelial cells, a condition known to mimic the in vivo environment and to exert a protective and pro-survival action. All together, these results provide a compelling rationale for the clinical development of new therapies for the treatment of PEL, based on combined targeting of glycolytic metabolism and constitutively activated signaling pathways.
2015
- Critical-size bone defect repair using amniotic fluid stem cell/collagen constructs: Effect of oral ferutinin treatment in rats
[Articolo su rivista]
Zavatti, Manuela; Bertoni, Laura; Maraldi, Tullia; Resca, Elisa; Beretti, Francesca; Guida, Marianna; La Sala, Giovanni Battista; De Pol, Anto
abstract
Aims: This study aims to evaluate the bone regeneration in a rat calvarias critical size bone defect treated with a construct consisting of collagen type I and human amniotic fluid stem cells (AFSCs) after oral administration of phytoestrogen ferutinin.
Main methods: In 12 week old male rats (n = 10), we performed two symmetric full-thickness cranial defects on each parietal region, and a scaffold was implanted into each cranial defect. The rats were divided into four groups: 1) collagen scaffold, 2) collagen scaffold + ferutinin at a dose of 2 mg/kg/5 mL, 3) collagen scaffold + AFSCs, and 4) collagen scaffold + AFSCs + ferutinin. The rats were sacrificed after 4 weeks, and the calvariae were removed, fixed, embedded in paraffin and cut into 7 pm thick sections. Histomorphometric measures, immunohistochemical and immunofluorescence analyses were performed on the paraffin sections.
Key findings: The histomorphometric analysis on H&E stained sections showed a significant increase in the regenerated area of the 4th group compared with the other groups. Immunohistochemistly performed with a human anti-mitochondrial antibody showed the presence of AFSCs 4 weeks after the transplant. Immunofluorescence analysis revealed the presence of osteocalcin and estrogen receptors (ER alpha. and GPR30) in all-groups, with a greater expression of all markers in samples where the scaffold was treated with AFSCs and the rats were orally administered ferutinin.
Significance: Our results demonstrated that the oral administration of ferutinin is able to improve the bone regeneration of critical-size bone defects in vivo that is obtained with collagen-AFSCs constructs.
2015
- Different origin of adipogenic stem cells influences the response to antiretroviral drugs
[Articolo su rivista]
Gibellini, Lara; DE BIASI, Sara; Nasi, Milena; Carnevale, Gianluca; Pisciotta, Alessandra; Bianchini, Elena; Bartolomeo, Regina; Polo, Miriam; DE POL, Anto; Pinti, Marcello; Cossarizza, Andrea
abstract
Lipodystrophy (LD) is a main side effect of antiretroviral therapy for HIV infection, and can be provoked by nucleoside reverse transcriptase inhibitors (NRTIs) and protease inhibitors (PIs). LD exists in different forms, characterized by fat loss, accumulation, or both, but its pathogenesis is still unclear. In particular, few data exist concerning the effects of antiretroviral drugs on adipocyte differentiation. Adipose tissue can arise either from mesenchymal stem cells (MSCs), that include bone marrow-derived MSCs (hBM-MSCs), or from ectodermal stem cells, that include dental pulp stem cells (hDPSCs). To analyze whether the embryonal origin of adipocytes might impact the occurrence of different phenotypes in LD, we quantified the effects of several antiretroviral drugs on the adipogenic differentiation of hBM-MSCs and hDPSCs. hBM-MSCs and hDPSCs were isolated from healthy donors. Cells were treated with 10 and 50μM stavudine (d4T), efavirenz (EFV), atazanavir (ATV), ritonavir (RTV), and ATV-boosted RTV. Viability and adipogenesis were evaluated by staining with propidium iodide, oil red, and adipoRed; mRNA levels of genes involved in adipocyte differentiation, i.e. CCAAT/enhancer-binding protein alpha (CEBPα) and peroxisome proliferator-activated receptor gamma (PPARγ), and in adipocyte functions, i.e. fatty acid synthase (FASN), fatty acid binding protein-4 (FABP4), perilipin-1 (PLIN1) and 1-acylglycerol-3-phosphate O-acyltransferase-2 (AGPAT2), were quantified by real time PCR. We found that ATV, RTV, EFV, and ATV-boosted RTV, but not d4T, caused massive cell death in both cell types. EFV and d4T affected the accumulation of lipid droplets and induced changes in mRNA levels of genes involved in adipocyte functions in hBM-MSCs, while RTV and ATV had little effects. All drugs stimulated the accumulation of lipid droplets in hDPSCs. Thus, the adipogenic differentiation of human stem cells can be influenced by antiretroviral drugs, and depends, at least in part, on their embryonal origin.
2015
- Enrichment in c-Kit improved differentiation potential of amniotic membrane progenitor/stem cells
[Articolo su rivista]
Resca, Elisa; Zavatti, Manuela; Maraldi, Tullia; Bertoni, Laura; Beretti, Francesca; Guida, Marianna; La Sala, Giovanni Battista; Guillot, P. V; David, A. L; Sebire, N. J; De Pol, Anto; De Coppi, P.
abstract
Introduction Human term placenta has attracted increasing attention as an alternative source of stem cells for regenerative medicine since it is accessible without ethical objections. The amniotic membrane (AM) contains at least two stem cell types from different embryological origins: ectodermal amniotic epithelial stem cells, and mesodermal mesenchymal stromal cells. Among the second group we studied the characteristics of amniotic mesenchymal cells (AMC) versus the ones enriched for the commonly used surface marker c-Kit (amniotic progenitor/stem cells-ASC), a stem cell factor receptor with crucial functions in a variety of biological systems and presents in early progenitors of different origin, as been already demonstrated in the enriched chorionic stem cells.
Methods After isolation, cells from the amniotic membranes (amniotic cells-AC) were selected for c-Kit (ASC) and compared these cells with c-Kit unselected (AMC), evaluating the expression of other stem cell markers (Oct-4, Tra-1-81, SSEA-4), CD271 and Slug.
Results Immunofluorescence analysis showed that ASC cells exhibited greater stem cell marker expression and included more CD271 and Slug positive cells. This was consistent with the interpretation that c-Kit enriched AC show greater stemness capacity compared to c-Kit unselected AMC.
Discussion AMC and ASC can both differentiate into various cell types including adipogenic, osteogenic, chondrogenic, neurogenic and hepatic lineages, but the enrichment in c-Kit improved stemness and differentiation potential of ASC.
2015
- Human dental pulp stem cells (hDPSCs): isolation, enrichment and comparative differentiation of two sub-populations
[Articolo su rivista]
Pisciotta, Alessandra; Carnevale, Gianluca; Meloni, Simona; Riccio, Massimo; De Biasi, Sara; Gibellini, Lara; Ferrari, Adriano; Bruzzesi, Giacomo; De Pol, Anto
abstract
Human dental pulp represents a suitable alternative source of stem cells for the purpose of cell-based therapies in regenerative medicine, because it is relatively easy to obtain it, using low invasive procedures. This study characterized and compared two subpopulations of adult stem cells derived from human dental pulp (hDPSCs). Human DPSCs, formerly immune-selected for STRO-1 and c-Kit, were separated for negativity and positivity to CD34 expression respectively, and evaluated for cell proliferation, stemness maintenance, cell senescence and multipotency.
2015
- Inhibition of Lon protease by triterpenoids alters mitochondria and is associated to cell death in human cancer cells
[Articolo su rivista]
Gibellini, Lara; Pinti, Marcello; Bartolomeo, Regina; De Biasi, Sara; Cormio, Antonella; Musicco, Clara; Carnevale, Gianluca; Pecorini, Simone; Nasi, Milena; De Pol, Anto; Cossarizza, Andrea
abstract
Mitochondrial Lon protease (Lon) regulates several mitochondrial functions, and is inhibited by the anticancer molecule triterpenoid 2-cyano-3, 12-dioxooleana-1,9(11)-dien-28-oic acid (CDDO), or by its C-28 methyl ester derivative (CDDO-Me). To analyze the mechanism of action of triterpenoids, we investigated intramitochondrial reactive oxygen species (ROS), mitochondrial membrane potential, mitochondrial mass, mitochondrial dynamics and morphology, and Lon proteolytic activity in RKO human colon cancer cells, in HepG2 hepatocarcinoma cells and in MCF7 breast carcinoma cells. We found that CDDO and CDDO-Me are potent stressors for mitochondria in cancer cells, rather than normal non-transformed cells. In particular, they: i) cause depolarization; ii) increase mitochondrial ROS, iii) alter mitochondrial morphology and proteins involved in mitochondrial dynamics; iv) affect the levels of Lon and those of aconitase and human transcription factor A, which are targets of Lon activity; v) increase level of protein carbonyls in mitochondria; vi) lead to intrinsic apoptosis. The overexpression of Lon can rescue cells from cell death, providing an additional evidence on the role of Lon in conditions of excessive stress load.
2015
- Le tavole di Giovanni Paolo Mascagni nella sala del Mascagni del Museo Anatomico di Modena
[Relazione in Atti di Convegno]
Corradini, Elena; Caneé, Valerio; DE POL, Anto
abstract
La realizzazione della sala del Mascagni nel Museo Anatomico di modena fu resa possibile nel 1852, allorché era stata demolita la scala realizzata dopo il 1815 per collegare il Teatro Anatomico con il piano superiore che era stato costruito per ospitare le Scuole Mediche e il Museo Anatomico: come collegamento del pianterreno con il piano superiore era infatti stata costruita la scala con gradini in marmo tuttora esistente .
Nella sala di Mascagni Gaddi aveva previsto un allestimento con le grandi tavole anatomiche a colori stampate da incisioni su rame realizzate da Antonio Serantoni che rappresentavano in forma naturale un uomo adulto alto cm. 175. Appartenevano alla monumentale opera di Paolo Mascagni Anatomiae Universae Icones, pubblicata postuma, dopo la sua morte avvenuta nel 1815, dallo stampatore pisano Niccolò Capurro in nove fascicoli, uno per anno, dal 1823 al 1832, per un totale di 88 tavole di cui 44 a colori e il loro equivalente a fronte in bianco e nero grazie a tre professori dello Studio pisano, A. Vacca Berlinghieri, G. Barzellotti e G. Rosini. Sei delle tavole, come ricorda lo stesso Gaddi, si trovavano già nella stessa sala a corredo di “ricche serie di finissime iniezioni delle arterie e vene capillari nei diversi tessuti organici” che erano collocate “entro dodici cassettoni a cristallo”. Quattordici tavole del Mascagni sono state di recente recuperate dai depositi dell’Ateneo e collocate nella sala riunioni e nel corridoio antistante del Dipartimento Chirurgico, Medico, Odontoiatrico e di Scienze Morfologiche con Interesse Trapiantologico, Oncologico e di Medicina Rigenerativa, in attesa di essere adeguatamente ricollocate nella sala del Mascagni del Museo Anatomico, così come è stato fatto all’Università di Pisa dove nell’ambito del Museo di Anatomia Umana “Filippo Civinini” è stata allestita la Galleria Mascagni.
2015
- Neural crest derived niche of human dental pulp stem cells promotes peripheral nerve regeneration and remyelination in animal model of critical sized sciatic nerve injury
[Articolo su rivista]
Carnevale, Gianluca; Pisciotta, Alessandra; DE BIASI, Sara; Gibellini, Lara; Cossarizza, Andrea; Bruzzesi, Giacomo; Ferrari, Adriano; DE POL, Anto
abstract
ABSTRACT
Peripheral nerve injuries are a commonly encountered clinical problem and often result in long-term functional defects. The use of stem cells, easily accessible, capable of rapid expansion in culture as well as fully integrate into the host tissue and capable to differentiate in myelinating cells of the peripheral nervous system, represent an attractive therapeutic approach for the treatment of nerve injuries. Farther, stem cells sources sharing the same embryological origin of Schwann cells, might be considered a suitable tool. The aim of this study was to demonstrate the ability of a neuroectodermal sub-population of STRO-1+/c-Kit+/CD34+ hDPSCs (1, 2), most of which being positive for neural crest (P75NTR) and neural progenitor cells (nestin) markers, to differentiate into Schwann cells-like cells in vitro and to promote axonal regeneration in vivo. As a matter of fact, following culture in appropriate induction medium, STRO-1+/c-Kit+/CD34+ hDPSCs were able to commit towards Schwann cells express- ing P75NTR, GFAP and S100b. After transplantation in animal model of sciatic nerve defect, hDPSCs promoted axonal regeneration from proximal to distal stumps, providing guidance to newly formed myelinated nerve fibers, which led to functional recovery as measured by sustained gait improvement. Particularly, transplanted hDP- SCs engrafted into critical sized sciatic nerve defect, as revealed by the positive stain- ing against human nuclei, showed the expression of typical Schwann cells markers, S100b and GFAP. In conclusion this study demonstrates that STRO-1+/c-Kit+/CD34+ hDPSCs, associated to neural crest derivation, represent a promising source of stem cells for the treatment of demyelinating disorders and might provide a valid alternative tool for future clinical applications to achieve functional recovery after injury or peripheral neuropathies besides minimizing ethical issues.
2015
- Nuclear Nox4 role in stemness power of human amniotic fluid stem cells
[Articolo su rivista]
Maraldi, Tullia; Guida, Marianna; Zavatti, Manuela; Resca, Elisa; Bertoni, Laura; La Sala, Giovanni Battista; De Pol, Anto
abstract
Human amniotic fluid stem cells (AFSC) are an attractive source for cell therapy due to their multilineage differentiation potential and accessibility advantages. However the clinical application of human stem cells largely depends on their capacity to expand in vitro, since there is an extensive donor-to-donor heterogeneity. Reactive oxygen species (ROS) and cellular oxidative stress are involved in many physiological and pathophysiological processes of stem cells, including pluripotency, proliferation, differentiation, and stress resistance. The mode of action of ROS is also dependent on the localization of their target molecules. Thus, the modifications induced by ROS can be separated depending on the cellular compartments they affect. NAD(P)H oxidase family, particularly Nox4, has been known to produce ROS in the nucleus. In the present study we show that Nox4 nuclear expression (nNox4) depends on the donor and it correlates with the expression of transcription factors involved in stemness regulation, such as Oct4, SSEA-4, and Sox2. Moreover nNox4 is linked with the nuclear localization of redox sensitive transcription factors, as Nrf2 and NF-κB, and with the differentiation potential. Taken together, these results suggest that nNox4 regulation may have important effects in stem cell capability through modulation of transcription factors and DNA damage.
2015
- Role of hepatocyte growth factor in the immunomodulation potential of amniotic fluid stem cells
[Articolo su rivista]
Maraldi, Tullia; Beretti, Francesca; Guida, Marianna; Zavatti, Manuela; De Pol, Anto
abstract
Human amniotic fluid stem cells (hAFSCs) may be useful for regenerative medicine because of their potential to differentiate into all three germ layers and to modulate immune response with different types of secretion molecules. This last issue has not been completely elucidated. The aim of this study was to investigate the secretome profile of the hAFSC, focusing on the role of hepatocyte growth factor (HGF) in immunoregulation through short and long cocultures with human peripheral blood mononuclear cells. We found that HGF produced by hAFSCs exerts a cytoprotective role, inducing an increase in caspase-dependent apoptosis in human immune cells. This study provides evidence supporting the hypothesis that amniotic fluid is an ideal source of stem cells for expansion and banking properties for therapeutic use. hAFSCs not only are less immunogenic but also can secrete immunoregulatory factors that may be useful in autoimmune diseases or allogenic implants.
SIGNIFICANCE:
New information about the secretome pattern is reported in this paper. Human amniotic fluid stem cells (hAFSCs) possess immunomodulatory properties involving hepatocyte growth factor production. hAFSCs could be used in immunotherapies and might be able to avoid allogenic rejection
2015
- Stem cells isolated from human dental pulp and amniotic fluid improve skeletal muscle histopathology in mdx/SCID mice
[Articolo su rivista]
Pisciotta, Alessandra; Riccio, Massimo; Carnevale, Gianluca; Lu, Aiping; DE BIASI, Sara; Gibellini, Lara; LA SALA, Giovanni Battista; Bruzzesi, Giacomo; Ferrari, Adriano; Huard, Johnny; DE POL, Anto
abstract
Introduction: Duchenne muscular dystrophy (DMD), caused by a lack of the functional structural protein dystrophin, leads to severe muscle degeneration where the patients are typically wheelchair-bound and die in their mid-twenties from cardiac or respiratory failure or both. The aim of this study was to investigate the potential of human dental pulp stem cells (hDPSCs) and human amniotic fluid stem cells (hAFSCs) to differentiate toward a skeletal myogenic lineage using several different protocols in order to determine the optimal conditions for achieving myogenic commitment and to subsequently evaluate their contribution in the improvement of the pathological features associated with dystrophic skeletal muscle when intramuscularly injected into mdx/SCID mice, an immune-compromised animal model of DMD. Methods: Human DPSCs and AFSCs were differentiated toward myogenic lineage in vitro through the direct co-culture with a myogenic cell line (C2C12 cells) and through a preliminary demethylation treatment with 5-Aza-2′-deoxycytidine (5-Aza), respectively. The commitment and differentiation of both hDPSCs and hAFSCs were evaluated by immunofluorescence and Western blot analysis. Subsequently, hDPSCs and hAFSCs, preliminarily demethylated and pre-differentiated toward a myogenic lineage for 2 weeks, were injected into the dystrophic gastrocnemius muscles of mdx/SCID mice. After 1, 2, and 4 weeks, the gastrocnemius muscles were taken for immunofluorescence and histological analyses. Results: Both populations of cells engrafted within the host muscle of mdx/SCID mice and through a paracrine effect promoted angiogenesis and reduced fibrosis, which eventually led to an improvement of the histopathology of the dystrophic muscle. Conclusion: This study shows that hAFSCs and hDPSCs represent potential sources of stem cells for translational strategies to improve the histopathology and potentially alleviate the muscle weakness in patients with DMD.
2014
- Feedbacks and adaptive capabilities of the PI3K/Akt/mTOR axis in acute myeloid leukemia revealed by pathway selective inhibition and phosphoproteome analysis
[Articolo su rivista]
Bertacchini, Jessika; Guida, M; Accordi, B; Mediani, Laura; Martelli, A. M; Barozzi, Patrizia; Petricoin, E; Liotta, L; Milani, G; Giordan, M; Luppi, Mario; Forghieri, Fabio; DE POL, Anto; Cocco, L; Basso, G; Marmiroli, Sandra
abstract
Acute myeloid leukemia (AML) primary cells express high levels of phosphorylated Akt, a master regulator of cellular functions regarded as a promising drug target. By means of reverse phase protein arrays, we examined the response of 80 samples of primary cells from AML patients to selective inhibitors of the phosphatidylinositol 3 kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) axis. We confirm that >60% of the samples analyzed are characterized by high pathway phosphorylation. Unexpectedly, however, we show here that targeting Akt and mTOR with the specific inhibitors Akti 1/2 and Torin1, alone or in combination, result in paradoxical Akt phosphorylation and activation of downstream signaling in 70% of the samples. Indeed, we demonstrate that cropping Akt or mTOR activity can stabilize the Akt/mTOR downstream effectors Forkhead box O and insulin receptor substrate-1, which in turn potentiate signaling through upregulation of the expression/phosphorylation of selected growth factor receptor tyrosine kinases (RTKs). Activation of RTKs in turn reactivates PI3K and downstream signaling, thus overruling the action of the drugs. We finally demonstrate that dual inhibition of Akt and RTKs displays strong synergistic cytotoxic effects in AML cells and downmodulates Akt signaling to a much greater extent than either drug alone, and should therefore be explored in AML clinical setting.
2014
- Human amniotic fluid stem cells: neural differentiation in vitro and in vivo
[Articolo su rivista]
Maraldi, Tullia; Bertoni, Laura; Riccio, Massimo; Zavatti, Manuela; Carnevale, Gianluca; Resca, Elisa; Guida, Marianna; Beretti, Francesca; LA SALA, Giovanni Battista; DE POL, Anto
abstract
The successful integration of stem cells after their implantation into the brain has become a central issue in modern neuroscience. In this study, we test the neural differentiation potential of c-Kit(+)/Oct-4(+) human amniotic fluid stem cells (hAFSCs) in vitro and their survival and integration in vivo. hAFSCs were induced towards neural differentiation and specific markers (GFAP, β-III tubulin, CNPase, MAP2, NeuN, synapsines, S100, PMP22) were detected by immunofluorescence and Western blot analysis. Glial proteins were expressed as early as 2 weeks after the initial differentiation stimulus, whereas neuronal markers started to appear from the third week of differentiation under culturing conditions of high cell density. This timeline suggested that glial cells possessed a promoting role in the differentiation of hAFSCs towards a neuronal fate. hAFSCs were then implanted into the lateral ventricle of the brain of 1-day-old rats, since neuronal development occurs up to 1 month after birth in this animal model. Our data showed that hAFSCs survived for up to 6 weeks post-implantation, were integrated into various areas of the central nervous system and migrated away from the graft giving rise to mature neurons and oligodendrocytes. We conclude that hAFSCs are able to differentiate and integrate into nervous tissue during development in vivo.
2014
- HUMAN BILIARY TREE STEM/PROGENITOR CELLS (hbTSCS) FROM PERIBILIARY GLANDS (PBGS) OF ADULT LIVER DISPLAY IMMUNOMODULATORY PROPERTIES THROUGH Fas/Fas LIGAND INDUCED T-CELL LYMPHOCYTE APOPTOSIS
[Abstract in Rivista]
Carnevale, G; Riccio, M; Cardinale, V; Gibelini, L; De Biasi, S; Pisciotta, A; Carpino, G; Gentile, R; Berloco, Pb; Brunelli, R; Bastianelli, C; Cossarizza, A; Gaudio, E; Alvaro, D; De Pol, A
abstract
Background and Aims: hBTSCs have the potential for regenerative
medicine in liver and pancreas diseases. T-cell control was recently
demonstrated for mesenchymal stem cells. The aims of this study
were to evaluate Fas-L expression within the stem cell niches of
adult biliary tree (PBGs), and to study the interaction between
hBTSCs and human lymphocytes.
Methods: HLA antigens, Fas and Fas-L expression were evaluated by
immunofluorescence and western blotting (WB) in cells of human
biliary tree in comparison with fibroblast cells, dental pulp stem
cells and bone marrow mesenchymal stem cells. The influence of
hBTSCs on lymphocytes’ activation and apoptosis were assessed by
co-culturing experiments.
Results: Adult hBTSCs expressed both class I and class
II HLA antigens, whereas fetal hBTSCs only class I HLA
antigens. 10 to 30% of the hBTSCs in PBGs were positive
for Fas-L. Fas-L+ cells were mostly located at the bottom
of PBGs and co-expressed EpCAM (Epithelial-Cell-AdhesionMolecule)
and proliferation marker (PCNA:Proliferating-CellNuclear-Antigen).
Mature cells at the bile duct surface epithelium
(mature cholangiocytes) were almost all negative for Fas-L. In
culture experiments confocal microscopy demonstrated that Fas-L
expression was restricted to EpCAM+/LGR5+ (a marker associated
with endodermal stem cells) hBTSCs. WB confirmed that hBTSCs
constitutively expressed high level of Fas-L which increased after
co-culture with T-cells. FACS analysis reveled that activated CD4+
and CD8+ T-cells co-cultured with hBTSCs underwent to a massiveinduction of apoptosis. Fas receptor appeared over-expressed in
T-cells co-cultured with hBTSCs respect to resting T-cells.
Conclusions: Our data demonstrated that hBTSCs can induce
“premature” apoptosis in T-cells trough the activation of Fas/Fas-L
pathway
2014
- Human biliary tree stem/progenitor cells (hBTSCs) from peribiliary glands (PBGs) of adult liver display immunomodulatory properties through Fas/Fas ligand induced T-cell lymphocyte apoptosis
[Abstract in Rivista]
Carnevale, G.; Riccio, M.; Cardinale, V.; Gibelini, L.; De Biasi, S.; Pisciotta, A.; Carpino, G.; Gentile, R.; Berloco, P. B.; Brunelli, R.; Bastianelli, C.; Cossarizza, A.; Gaudio, E.; Alvaro, D.; De Pol, A.
abstract
Background and aim: hBTSCs have been retrieved in peribiliary glands
(PBGs) of adult and fetal biliary tree, and have the potential for regenerative
medicine in liver, biliary tree, and pancreas diseases. The ability of stem cells
to control T-cells’ immune responses was recently demonstrated by human
mesenchymal stem cells. The aims of the present study were to evaluate Fas-L
expression within the stem cell niches of adult biliary tree, and to study the in
vitro interaction between hBTSCs and human lymphocytes.
Material and methods: HLA antigens, Fas and Fas-L expression were evaluated
in situ and in vitro by immunofluorescence and Western blots in cells
of the human biliary tree in comparison with fibroblast cells, dental pulp
stem cells and bone marrow mesechymal stem cells. Co-cultures of hBTSCs
with human leucocytes were used to analyze the influence of hBTSCs on
lymphocytes’ activation and apoptosis.
Results: Adult hBTSCs expressed both class I and class II HLA antigens,
whereas fetal hBTSCs had class I HLA antigens only. In PBG niche 10-30%
BTSCs were positive for Fas-L. Fas-L positive cells were mostly located at the
bottom of PBGs and co-expressed EpCAM (epithelial cell adhesion molecule)
and a marker of proliferation (PCNA: Proliferating Cell Nuclear Antigen).
Conversely, mature cells at the surface epithelium and cholangiocytes of large
intrahepatic ducts were almost all negative for Fas-L. In culture experiments
confocal microscopy demonstrated that Fas-L expression was restricted to
EpCAM+/LGR5+(a marker associated with endodermal stem cells) cells.
Western blot data confirmed that hBTSCs constitutively expressed high level
of Fas-L that increased after co-culture with T-cells. FACS analysis of T-cells
co-cultured with hBTSCs indicated that hBTSCs were able to induce apoptosis
in activated CD4+ and CD8+ T-cell populations. Moreover, Fas receptor
appears to be more expressed in T-cells co-cultured with hBTSCs than in
resting T-cells.
Conclusions: In conclusion our data suggest that hBTSCs could modulate
the T-cells response through the production of Fas-L, which influences the
lymphocyte Fas/Fas-L pathway by inducing “premature” apoptosis in CD4+
and CD8+ T-cells.
2014
- Nuclear Nox4-derived reactive oxygen species in myelodysplastic syndromes
[Articolo su rivista]
Guida, Marianna; Maraldi, Tullia; Beretti, Francesca; Follo, Matilde Y; Manzoli, Lucia; DE POL, Anto
abstract
A role for intracellular ROS production has been recently implicated in the pathogenesis and progression of a wide variety of neoplasias. ROS sources, such as NAD(P)H oxidase (Nox) complexes, are frequently activated in AML (acute myeloid leukemia) blasts and strongly contribute to their proliferation, survival, and drug resistance. Myelodysplastic syndromes (MDS) comprise a heterogeneous group of disorders characterized by ineffective hematopoiesis, with an increased propensity to develop AML. The molecular basis for MDS progression is unknown, but a key element in MDS disease progression is the genomic instability. NADPH oxidases are now recognized to have specific subcellular localizations, this targeting to specific compartments for localized ROS production. Local Nox-dependent ROS production in the nucleus may contribute to the regulation of redox-dependent cell growth, differentiation, senescence, DNA damage, and apoptosis. We observed that Nox1, 2, and 4 isoforms and p22phox and Rac1 subunits are expressed in MDS/AML cell lines and MDS samples, also in the nuclear fractions. Interestingly, Nox4 interacts with ERK and Akt1 within nuclear speckle domain, suggesting that Nox4 could be involved in regulating gene expression and splicing factor activity. These data contribute to the elucidation of the molecular mechanisms used by nuclear ROS to drive MDS evolution to AML.
2014
- Sirtuin 3 interacts with Lon protease and regulates its acetylation status.
[Articolo su rivista]
Gibellini, Lara; Pinti, Marcello; Beretti, Francesca; Pierri, Cl; Onofrio, A; Riccio, Massimo; Carnevale, Gianluca; DE BIASI, Sara; Nasi, Milena; Torelli, F; Boraldi, Federica; DE POL, Anto; Cossarizza, Andrea
abstract
Lon is a mitochondrial protease that degrades oxidized damaged proteins, assists protein folding and participates in maintaining mitochondrial DNA levels. Changes in Lon mRNA levels, protein levels and activity are not always directly correlated, suggesting that Lon could be regulated at post translational level. We found that Lon and SIRT3, the most important mitochondrial sirtuin, colocalize and coimmunoprecipitate in breast cancer cells, and silencing or inhibition of Lon did not alter SIRT3 levels. Silencing of SIRT3 increased the levels of Lon protein and of its acetylation, suggesting that Lon is a target of SIRT3, likely at K917.
2014
- Stem cell properties in cell cultures from different stage of melanoma progression.
[Articolo su rivista]
Magnoni, Cristina; S., Guidice; Pellacani, Giovanni; G., Bertazzoni; Longo, Caterina; E., Veratti; D., Morini; L., Benassi; S., Al Jalbout; C., Vaschieri; P., Azzoni; DE POL, Anto; S., Seidenari; Tomasi, Aldo; Ponti, Giovanni
abstract
Cutaneous Melanoma is an extremely heterogeneous human cancer. The most aggressive melanoma may contain deregulated cells with undifferentiated/stem cell-like phenotype. A critical mechanism by which melanoma cells enhance their invasive capacity is the dissolution of the intercellular adhesions and the acquisition of mesenchymal features as a part of an epithelial-to-mesenchimal transition (EMT). The aim of this study was to clarify the role of stem cell-like population in human melanomas through the in vitro analysis of melanocytic cell cultures, obtained from distinct histotypes of primary and metastatic malignant melanoma. Patients with advanced melanoma larger than 2cm diameter and/or wider than 300mm2 surface were enrolled. The melanoma cells were isolated from skin biopsies of lentigo melanoma (LM), superficial spreading (SS), nodular melanoma (NM) and metastatic melanoma (MM) and maintained in different media in order to evaluate the colony-forming unit assay, alkaline phosphatase and toluidine blue stain and the cell ability to differentiate into osteogenic and adipogenic lineages. Immunohistochemistry and flow cytometry analysis were performed in order to evaluate antigenic markers CD90, CD73, CD105, CD146, CD20, CD166 and nestin.
This study confirms that melanoma can include an heterogeneous cell population with both abilities to self-renew and to give rise to differentiated progeny. Melanoma cells displayed the intra-tumoral heterogeneity and dynamic antigen phenotypes. Histologically, the transitions from normal skin to DN to LM to NM to MM was associated with a gradual increase in the expression of CD146, CD20, CD133, Nestin and CD73 by melanoma cells. These molecular evidences could be a milestone for the development of novel biomolecular targeted-therapy approaches.
2014
- The Fas/Fas ligand apoptosis pathway underlies immunomodulatory properties of human biliary tree stem/progenitor cells
[Articolo su rivista]
Riccio, Massimo; Carnevale, Gianluca; Cardinale, Vincenzo; Gibellini, Lara; DE BIASI, Sara; Pisciotta, Alessandra; Carpino, Guido; Gentile, Raffaele; Berloco, Pasquale B; Brunelli, Roberto; Bastianelli, Carlo; Napoletano, Chiara; Cantafora, Alfredo; Cossarizza, Andrea; Gaudio, Eugenio; Alvaro, Domenico; DE POL, Anto
abstract
Human biliary tree stem/progenitor cells (hBTSCs) are multipotent epithelial stem cells, easily obtained from the biliary tree, with the potential for regenerative medicine in liver, biliary tree, and pancreas diseases. Recent reports indicate that human mesenchymal stem cells are able to modulate the T cell immune response. However, no information exists on the capabilities of hBTSCs to control the allogeneic response. The aims of this study were to evaluate FasL expression in hBTSCs, to study the in vitro interaction between hBTSCs and human lymphocytes, and the role of Fas/FasL modulation in inducing T cell apoptosis in hBTSCs/T cell co-cultures.
2013
- Enrichment in c-Kit(+) enhances mesodermal and neural differentiation of human chorionic placental cells
[Articolo su rivista]
Resca, Elisa; Zavatti, Manuela; Bertoni, Laura; Maraldi, Tullia; DE BIASI, Sara; Pisciotta, Alessandra; A., Nicoli; LA SALA, Giovanni Battista; P. V., Guillot; A. L., David; N. J., Sebire; P. D., Coppi; DE POL, Anto
abstract
OBJECTIVE: Human term placenta (HTP) has attracted increasing attention as an alternative source of stem cells for regenerative medicine since the amniochorionic membrane harbors stem cells populations that are easily accessible, abundantly available without ethical objections. In the chorionic side of HTP we found a progenitor perivascular "niche" in which rare cells co-express Oct-4 and c-Kit. We investigated the stem cell characteristics and differentiation potential of a chorionic derived population enriched in c-Kit(+) cells and compared this to the unenriched population. STUDY DESIGN: Cells, isolated from the chorion of HTP, were expanded and enriched in c-Kit(+) cells (Chorionic Stem Cells-CSC). Histological staining, immunofluorescence, Western blot and flow cytometry were used to verify the stem cells characteristics of the populations and to compare the differentiation capability towards mesodermal and neural lineages in vitro. RESULTS: The expression of the pluripotent marker Oct-4 was greater in the CSCs compared to the unselected cells (Chorionic Cell-CC) but both Oct-4 and c-Kit expression decreased during passages. After differentiation, CSC displayed stronger chondrogenic and osteogenic potential and a greater adipogenic forming capacity compared to unselected ones. CSC differentiated better into immature oligodendrocytes while CC showed a neuronal progenitor differentiation potential. Moreover, both populations were able to differentiate in hepatogenic lineage. CONCLUSION: CSC display improved Oct-4 expression and a high differentiation potential into mesodermal lineages and oligodendrocytes.
2013
- Ferutinin promotes proliferation and osteoblastic differentiation in human amniotic fluid and dental pulp stem cells.
[Articolo su rivista]
Zavatti, Manuela; Resca, Elisa; Bertoni, Laura; Maraldi, Tullia; Guida, Marianna; Carnevale, Gianluca; Ferrari, Adriano; DE POL, Anto
abstract
The phytoestrogen Ferutinin plays an important role in prevention of osteoporosis caused by ovariectomy-induced estrogen deficiency in rats, but there is no evidence of its effect on osteoblastic differentiation in vitro. In this study we investigated the effect of Ferutinin on proliferation and osteoblastic differentiation of two different human stem cells populations, one derived from the amniotic fluid (AFSCs) and the other from the dental pulp (DPSCs).AFSCs and DPSCs were cultured in a differentiation medium for 14 or 21days with or without the addition of Ferutinin at a concentration ranging from 10(-11) to 10(-4)M. 17β-Estradiol was used as a positive drug at 10(-8)M. Cell proliferation and expression of specific osteoblast phenotype markers were analyzed.MTT assay revealed that Ferutinin, at concentrations of 10(-8) and 10(-9)M, enhanced proliferation of both AFSCs and DPSCs after 72h of exposure. Moreover, in both stem cell populations, Ferutinin treatment induced greater expression of the osteoblast phenotype markers osteocalcin (OCN), osteopontin (OPN), collagen I, RUNX-2 and osterix (OSX), increased calcium deposition and osteocalcin secretion in the culture medium compared to controls. These effects were more pronounced after 14days of culture in both populations.The enhancing capabilities on proliferation and osteoblastic differentiation displayed by the phytoestrogen Ferutinin make this compound an interesting candidate to promote bone formation in vivo.
2013
- Human amniotic fluid-derived and dental pulp-derived stem cells seeded into collagen scaffold repair critical-size bone defects promoting vascularization.
[Articolo su rivista]
Maraldi, Tullia; Riccio, Massimo; Pisciotta, Alessandra; Zavatti, Manuela; Carnevale, Gianluca; Beretti, Francesca; LA SALA, Giovanni Battista; A., Motta; DE POL, Anto
abstract
INTRODUCTION: The main aim of this study is to evaluate potential human stem cells, such as dental pulp stem cells (DPSC) and amniotic fluid stem cells (AFSC), combined with collagen scaffold, to reconstruct critical size cranial bone defects in animal model. METHODS: We performed two symmetric full-thickness cranial defects on each parietal region of rats and we replenished them with collagen scaffolds with or without stem cells already seeded into and addressed towards osteogenic lineage in vitro. After 4 and 8 weeks cranial tissue samples were taken for histological and immunofluorescence analysis. RESULTS: We observed a new bone formation in all the samples but the most relevant difference in defect correction were shown by stem cell-collagen samples at 4 weeks after implant, suggesting a faster regeneration ability of the combined constructs. The presence of human cells in the newly-formed bone was confirmed by confocal analysis with an antibody directed to a human mitochondrial protein. Furthermore, human cells were found to be an essential part of new vessel formation in the scaffold. CONCLUSIONS: All these data confirmed the strong potential of bioengineered constructs of stem cell-collagen scaffold for correcting large cranial defects in animal model and highlighting the role of stem cells in neo vascularization during skeletal defect reconstruction.
2013
- In vitro differentiation into insulin-producing β-cells of stem cells isolated from human amniotic fluid and dental pulp.
[Articolo su rivista]
Carnevale, Gianluca; Riccio, Massimo; Pisciotta, Alessandra; Beretti, Francesca; Maraldi, Tullia; Zavatti, Manuela; Cavallini, Gian Maria; LA SALA, Giovanni Battista; Ferrari, Adriano; DE POL, Anto
abstract
AIM: To investigate the ability of human amniotic fluid stem cells and human dental pulp stem cells to differentiate into insulin-producing cells. METHODS: Human amniotic fluid stem cells and human dental pulp stem cells were induced to differentiate into pancreatic β-cells by a multistep protocol. Islet-like structures were assessed in differentiated human amniotic fluid stem cells and human dental pulp stem cells after 21 days of culture by dithizone staining. Pancreatic and duodenal homebox-1, insulin and Glut-2 expression were detected by immunofluorescence and confocal microscopy. Insulin secreted from differentiated cells was tested with SELDI-TOF MS and by enzyme-linked immunosorbent assay. RESULTS: Human amniotic fluid stem cells and human dental pulp stem cells, after 7 days of differentiation started to form islet-like structures that became evident after 14 days of induction. SELDI-TOF MS analysis, revealed the presence of insulin in the media of differentiated cells at day 14, further confirmed by enzyme-linked immunosorbent assay after 7, 14 and 21 days. Both stem cell types expressed, after differentiation, pancreatic and duodenal homebox-1, insulin and Glut-2 and were positively stained by dithizone. Either the cytosol to nucleus translocation of pancreatic and duodenal homebox-1, either the expression of insulin, are regulated by glucose concentration changes. Day 21 islet-like structures derived from both human amniotic fluid stem cells and human dental pulp stem cell release insulin in a glucose-dependent manner. CONCLUSION: The present study demonstrates the ability of human amniotic fluid stem cells and human dental pulp stem cell to differentiate into insulin-producing cells, offering a non-pancreatic, low-invasive source of cells for islet regeneration.
2013
- Inhibition of nuclear nox4 activity by plumbagin: effect on proliferative capacity in human amniotic stem cells.
[Articolo su rivista]
Guida, Marianna; Maraldi, Tullia; Resca, Elisa; Beretti, Francesca; Zavatti, Manuela; Bertoni, Laura; LA SALA, Giovanni Battista; DE POL, Anto
abstract
Human amniotic fluid stem cells (AFSC) with multilineage differentiation potential are novel source for cell therapy. However, in vitro expansion leads to senescence affecting differentiation and proliferative capacities. Reactive oxygen species (ROS) have been involved in the regulation of stem cell pluripotency, proliferation, and differentiation. Redox-regulated signal transduction is coordinated by spatially controlled production of ROS within subcellular compartments. NAD(P)H oxidase family, in particular Nox4, has been known to produce ROS in the nucleus; however, the mechanisms and the meaning of this function remain largely unknown. In the present study, we show that Nox4 nuclear expression (nNox4) increases during culture passages up to cell cycle arrest and the serum starvation causes the same effect. With the decrease of Nox4 activity, obtained with plumbagin, a decline of nuclear ROS production and of DNA damage occurs. Moreover, plumbagin exposure reduces the binding between nNox4 and nucleoskeleton components, as Matrin 3. The same effect was observed also for the binding with phospho-ERK, although nuclear ERK and P-ERK are unchanged. Taken together, we suggest that nNox4 regulation may have important pathophysiologic effects in stem cell proliferation through modulation of nuclear signaling and DNA damage.
2013
- The protein kinase Akt/PKB regulates both prelamin A degradation and Lmna gene expression
[Articolo su rivista]
Bertacchini, Jessika; Beretti, Francesca; Vittoria, Cenni; Guida, Marianna; Federica, Gibellini; Mediani, Laura; Oriano, Marin; Nadir M., Maraldi; DE POL, Anto; Giovanna, Lattanzi; Lucio, Cocco; Marmiroli, Sandra
abstract
The serine/threonine kinase Akt/PKB is a major signaling hub integrating metabolic, survival, growth and cell cycle regulatory signals. The definition of the phospho-motif cipher driving phosphorylation by Akt led to the identification of hundreds of putative substrates, and it is therefore pivotal to name those whose phosphorylation by Akt is of consequence to biological processes. The Lmna gene products lamin A/C and their precursor prelamin A (collectively called A-type lamins) are type V intermediate filaments proteins forming a filamentous meshwork, the lamina, underneath the inner nuclear membrane, for nuclear envelope structures organization and interphase chromatin anchoring. In our previous work we reported that A-type lamins are phosphorylated by Akt at S301 and S404 in physiological conditions, and are therefore bona fide substrates of Akt. We describe here that Akt phosphorylation at S404 targets the precursor prelamin A for degradation. We further demonstrate that Akt regulates also Lmna transcription. All together, our study unveils a previously unknown function of Akt in the control of prelamin A stability and expression. Moreover, given the large number of diseases related to prelamin A, our findings represent a further important step bridging basic A-type lamins physiology to therapeutic approaches for lamin A-linked disorders.
2012
- Fibroin scaffold repairs critical-size bone defects in vivo supported by human amniotic fluid and dental pulp stem cells.
[Articolo su rivista]
Riccio, Massimo; Maraldi, Tullia; Pisciotta, Alessandra; LA SALA, Giovanni Battista; Ferrari, Adriano; G., Bruzzesi; A., Motta; C., Migliaresi; DE POL, Anto
abstract
The main aim of this study was the comparative evaluation of fibroin scaffolds combined with human stem cells, such as dental pulp stem cells (hDPSCs) and amniotic fluid stem cells (hAFSCs), used to repair critical-size cranial bone defects in immunocompromised rats. Two symmetric full-thickness cranial defects on each parietal region of rats have been replenished with silk fibroin scaffolds with or without preseeded stem cells addressed toward osteogenic lineage in vitro. Animals were euthanized after 4 weeks postoperatively and cranial tissue samples were taken for histological analysis. The presence of human cells in the new-formed bone was confirmed by confocal analysis with an antibody directed to a human mitochondrial protein. Fibroin scaffolds induced mature bone formation and defect correction, with higher bone amount produced by hAFSC-seeded scaffolds. Our findings demonstrated the strong potential of stem cells/fibroin bioengineered constructs for correcting large cranial defects in animal model and is likely a promising approach for the reconstruction of human large skeletal defects in craniofacial surgery.
2012
- Human serum promotes osteogenic differentiation of Human Dental Pulp Stem Cells in vitro and in vivo.
[Articolo su rivista]
Pisciotta, Alessandra; Riccio, Massimo; Carnevale, Gianluca; Beretti, Francesca; Gibellini, Lara; Maraldi, Tullia; Cavallini, Gian Maria; Ferrari, Adriano; Bruzzesi, G; DE POL, Anto
abstract
Human dental pulp is a promising alternative source of stem cells for cell-based tissue engineering in regenerative medicine, for the easily recruitment with low invasivity for the patient and for the self-renewal and differentiation potential of cells. So far, in vitro culture of mesenchymal stem cells is usually based on supplementing culture and differentiation media with foetal calf serum (FCS). FCS is known to contain a great quantity of growth factors, and thus to promote cell attachment on plastic surface as well as expansion and differentiation. Nevertheless, FCS as an animal origin supplement may represent a potential means for disease transmission besides leading to a xenogenic immune response. Therefore, a significant interest is focused on investigating alternative supplements, in order to obtain a sufficient cell number for clinical application, avoiding the inconvenients of FCS use. In our study we have demonstrated that human serum (HS) is a suitable alternative to FCS, indeed its addition to culture medium induces a high hDPSCs proliferation rate and improves the in vitro osteogenic differentiation. Furthermore, hDPSCs-collagen constructs, pre-differentiated with HS-medium in vitro for 10 days, when implanted in immunocompromised rats, are able to restore critical size parietal bone defects. Therefore these data indicate that HS is a valid substitute for FCS to culture and differentiate in vitro hDPSCs in order to obtain a successful bone regeneration in vivo.
2012
- The protease inhibitor atazanavir triggers autophagy and mitophagy in human preadipocytes
[Articolo su rivista]
Gibellini, Lara; DE BIASI, Sara; Pinti, Marcello; Nasi, Milena; Riccio, Massimo; Carnevale, Gianluca; Cavallini, Gian Maria; F. J., Sala De Oyanguren; J. E., O’Connor; Mussini, Cristina; DE POL, Anto; Cossarizza, Andrea
abstract
Background: The association betweenHAARTand lipodystrophy iswell established, but lipodystrophy pathogenesis is still poorly understood. Drugs, and in particular protease inhibitors, accumulate in adipose tissue affecting adipocyte physiology and gene expression by several mechanisms. Recent studies have identified autophagy as another process affected by these classes of drugs, but no studies have been performed in adipose cells. Methods: SW872 preadipocytic human cell line was used to evaluate changes induced by amprenavir (APV), ritonavir (RTV), or atazanavir (ATV), all used at 10–200mmol/l. A subline was stably transfected with murine stem cell virus (pMSCV)-enhanced green fluorescent protein (EGFP)-LC3 plasmid (to obtain a fluorescent LC3 protein) and treated with ATV at different doses. The distribution of LC3 and the colocalization of mitochondria, lysosome, and autophagosome were assessed by confocal microscopy. Transmission electron microscopy of ATV-treated cells was also performed. The cellular content of lysosomes was assessed using Lysotracker Green; apoptosis was evaluated by annexin V/propidium iodide staining, and mitochondrial superoxide anion (mtO2-) was analyzed by mitoSOX red. Lysosomes, apoptosis, and mtO2 - were studied by flow cytometry and multispectral imaging flow cytometry. Results: In SW872 cells, RTV caused massive apoptosis, more than autophagy, whereas APV was almost ineffective. ATV induced both apoptosis (high doses) and autophagy (low doses). ATV-treated cells displayed LC3-specific punctae, suggesting the formation of autophagosomes that enclosed mitochondria, as revealed by electron microscopy. At low doses, ATV promoted mitochondrial superoxide generation, whereas at high doses, it induced mitochondrial membrane depolarization. Conclusion: Autophagy/mitophagy can be considered a mechanism triggered by ATV in SW872 preadipocytes.
2011
- A novel biomarker harvesting nanotechnology identifies Bak as a candidate melanoma biomarker in serum.
[Articolo su rivista]
Longo, Caterina; G., Gambara; V., Espina; A., Luchini; B., Bishop; A. S., Patanarut; E. F., Petricoin; Beretti, Francesca; B., Ferrari; E., Garaci; DE POL, Anto; Pellacani, Giovanni; L. A., Liotta
abstract
Melanoma represents only 4\% of all skin cancers, but nearly 80\% of skin cancer deaths. This manuscript applies several new measurement technologies with the purpose of elucidating molecular signatures of melanoma aggressiveness.We sought to determine whether low-abundant serum proteins related to apoptotic pathways could be measured and correlated with defined melanoma subtypes. Hydrogel core shell nanoparticles, a new technology capable of selectively entrapping low molecular weight proteins and protecting them from enzymatic degradation, were used to capture candidate serum biomarkers. Biomarker levels were correlated with confocal microscopy, thereby representing a combination of new technologies for in vivo histologic documentation.Among a panel of analyzed serum proteins, Bak was differentially expressed between nevi and melanomas. Melanomas with higher Bak serum levels exhibited more pronounced junctional activity on confocal imaging, whereas lesions with 'sparse' dermal nests had weak Bak expression.Our study links serum proteome analysis with confocal microscopic clinical in vivo histologic classification of melanomas. Bak has not been previously measured in serum. Bak differential expression among melanoma subtypes confirms the importance of the apoptotic pathway as a contributor to melanoma aggressiveness.
2011
- Embriologia umana
[Monografia/Trattato scientifico]
Armato, Bani; Baroni, Lumare; Bressan, Buscemi; Ceccoli, Ciccarelli; Cimini, ; Dal, Pra; DE POL, Anto; Galdieri, Gerbino; Marotta, Nerducci; Ricci, Romagnoli; Sica, Vannucchi; Zani,
abstract
2011
- Human amniotic fluid stem cells seeded in fibroin scaffold produce in vivo mineralized matrix.
[Articolo su rivista]
Maraldi, Tullia; Riccio, Massimo; Resca, Elisa; Pisciotta, Alessandra; LA SALA, Giovanni Battista; Ferrari, Adriano; Bruzzesi, G; Motta, A; Migliaresi, C; Marzona, Laura; DE POL, Anto
abstract
This study investigated the potential of amniotic fluid stem cells (AFSCs) to synthesize mineralized extracellular matrix (ECM) within different porous scaffolds of collagen, poly-D,L-lactic acid (PDLLA), and silk fibroin. The AFSCs were initially differentiated by using an osteogenic medium in two-dimensional culture, and expression of specific bone proteins and the physiologic mineral production by the AFSCs were analyzed. In particular, during differentiation process, AFSCs expressed proteins like Runt-related transcription factor 2 (Runx2), Osterix, Osteopontin, and Osteocalcin with a sequential expression, analogous to those occurring during osteoblast differentiation, and produced extracellular calcium stores. AFSCs were then cultured on three-dimensional (3D) scaffolds and evaluated for their ability to differentiate into osteoblastic cells in vivo. Stem cells were cultured in vitro for 1 week in collagen, fibroin, and PDLLA scaffolds. The effect of predifferentiation of the stem cells in scaffolds on the subsequent bone formation in vivo was determined in a rat subcutaneous model. With the addition of a third dimension, osteogenic differentiation and mineralized ECM production by AFSCs were significantly higher. This study demonstrated the strong potential of AFSCs to produce 3D mineralized bioengineered constructs in vivo and suggests that fibroin may be an effective scaffold material for functional repair of critical size bone defects.
2011
- Istologia umana
[Monografia/Trattato scientifico]
Bani, Baroni; Becchetti, Lumare; Bressan, Buscemi; Caramelli, Caruso; Cimini, ; DE POL, Anto; Gerbino, Luca; Marotta, Nico; Nicolin, Pugnaloni; Romagnoli, Sica; Vannucchi,
abstract
2011
- Low levels of selenium compounds are selectively toxic for a human neuron cell line through ROS/RNS increase and apoptotic process activation
[Articolo su rivista]
Maraldi, Tullia; Riccio, Massimo; L., Zambonin; Vinceti, Marco; DE POL, Anto; G., Hakimb
abstract
Organic and inorganic selenium compounds were used to examinewhether low selenium concentrationis able to trigger apoptotic degeneration in a human neuron cell line in vitro and to explore changes inreactive oxygen and nitrogen species and antioxidant protein content during the apoptotic processes.The results indicated that: (1) SKNBE neuroblastoma cells treated with sodium selenite, sodium selenateand seleno-methionine (0.1, 0.5 and 0.5 mM, respectively) for 24 h exhibited a viability decrease, unlikekidney or prostatic cells; (2) the PARP (poly-ADP-ribose-polymerase) degradation and caspase activationdetected by Western blot and flow cytometry fluorimetric examination showed induction of apoptosis;(3) during selenium treatment, a ROS/RNS increase occurred despite the GSH increment, as revealed byfluorimetric analysis; (4) the RNS production could be blocked by a peroxynitrite scavenger; (5) afterexposure to selenium compounds, the concentration of nitric oxide synthase, manganese superoxidedismutase (SOD2), P-NF-kB (phospho nuclear factor kB), glutathione reductase and glutathioneperoxidase increased, whereas that of P-ERK (phospho extracellular signal-regulated kinase) decreased;(6) selenium presence induced copper/zinc superoxide dismutase (SOD1) translocation into mitochondria,in a way similar to what is observed in amyotrophic lateral sclerosis (ALS). This study supportsepidemiologic studies showing the possibility that excess environmental exposure to Se represents a riskfactor for a devastating human neurodegenerative disease.
2011
- Reverse-phase protein microarrays (RPPA) as a diagnosticand therapeutic guide in multidrug resistant leukemia
[Articolo su rivista]
Maraldi, Tullia; Bertacchini, Jessika; Benincasa, Marta; M., Guida; DE POL, Anto; L., Liotta LA; E., Petricoin; L., Cocco; Marmiroli, Sandra
abstract
Abstract. Reverse-phase microarray assays using phosphospecificantibodies (RPPA) can directly measure levels ofphosphorylated protein isoforms. In the current study, lysatesfrom parental and multidrug resistant (MDR) CEM leukemiacells were spotted onto reverse-phase protein microarraysand probed with a panel of phospho-antibodies to ERK, PCKand Akt pathways. In particular, the Akt pathway is consideredto play significant roles in leukemia and Akt inhibitor therapyhas been proposed as a potential tool in the treatment of thisdisease. The RPPA data prompted us to investigate deeperthis pathway. Here, we found that whereas total Akt1 proteinlevel is higher in parental CEM cells, the activated isoformcontent, p-Akt1, increases in doxorubicin-selected CEM cells(MDR-CEM). This was backed up by Western blot analysis,confirming that Akt1 activity/phosphorylation may be upregulatedin MDR-CEM cells. Further exploration of inhibitorytherapy in this system was evaluated. The TNF-relatedapoptosis-inducing ligand, TRAIL, has been shown toselectively kill tumor cells. Herein, we describe that in MDRCEMcells TRAIL responsiveness correlates with a reducedexpression of endogenous Akt1, suggesting that the MDRphenotype associated to P-gp sensitizes cells to TRAIL therapy.
2011
- RGB method in immunofluorescence investigations on stem cells
[Articolo su rivista]
Riccio, Massimo; E., Resca; Bertoni, Laura; Cavani, Francesco; Sena, Paola; Ferretti, Marzia; Baldini, Andrea; Palumbo, Carla; DE POL, Anto
abstract
Colour is not related to a particular discipline, but it is transversely present in many circles and inalmost all the aspects of life. It has a special value in art, but also as far as other disciplines areconcerned, like the sciences, the colour is at the basis of some of their intrinsic significances and it oftenneeded to allow the interpretation of some of their phenomena as well. As regards the development ofcell biology knowledge, colour acquired more and more importance in revealing the observations of theresearchers. A field in which the methods based on the colours are particularly employed is theimmunofluorescence, used to identify specific proteins in cells and tissues. These techniques combinethe fluorochrome properties with specific molecules, i.e. antibodies, directed against particularsubstances to investigate, for example a specific protein. In single immunofluorescence analysis, thesignal from an excited fluorochrome corresponds to a particular protein. In multiple immunofluorescenceanalysis, two or more signals are simultaneously detected to show the localization of differentproteins on the same sample. The three primary colours red, green and blue were currently assigned tothe signals from immunofluorescence-processed samples and visualized by the RGB method. In thepresent work, different examples of RGB applications in immunocytochemical investigations areshowed: the first concerns the multiple analysis of three markers, localized in different loci of the cellplasma membrane; the second is related to the co-localization of two signals in the same site of specificsubcellular structures. In this case the secondary colours, obtained by overlapping the primary ones,demonstrate the specific co-presence of two proteins in the same site. With the present paper, theauthors wish to underline the relevant role of colours also in those areas in which colours are the meansnot the end.
2010
- Altered Expression of Apoptosis Biomarkers in Human Colorectal Microadenomas
[Articolo su rivista]
Sena, Paola; Roncucci, Luca; Marzona, Laura; Mariani, Francesco; Maffei, Stefania; Manenti, Antonio; DE POL, Anto
abstract
Human colorectal microadenomas are considered the earliest detectable premalignant lesions in the colon. They can be identified as aggregates of enlarged crypts with thicker epithelial linings and elongated luminal openings on the colonic mucosal surface after methylene blue staining and observation under a dissecting microscope. Multiple lines of evidence suggest that a central role in neoplastic development is played by the inhibition of apoptosis, followed by disruption of DNA repair. Understanding the early mechanisms of colorectal carcinogenesis may help develop new approaches of colorectal cancer prevention and treatment. The aim of the present study was to quantify poly-ADP ribose polymerase 1 (PARP-1)-positive cells and to evaluate apoptotic control mechanisms through Caspase-3 active and Bcl-2 protein expression in human microadenomas and in normal colorectal mucosa using immunofluorescence techniques coupled with confocal microscopy and immunoblot experiments. The mean percentage of PARP-1-positive epithelial cells was 3.0 +/- 0.37% (SD) and 15.67 +/- 0.40% in microadenoma and in normal mucosa, respectively. Proteins involved in programmed cell death were differently expressed in microadenoma and in normal mucosa. Indeed, by semiquantitative immunoflourescence analysis, confirmed byWestern blot, microadenoma showed high levels of Caspase-3 active and low levels of Bcl-2 expression, whereas the opposite was true for normal colorectal mucosa. In the stroma of normal colorectal mucosa, fibroblast-like cells and neutrophils were the cells that underwent apoptosis to a greater extent. In conclusion, malfunction of the control mechanisms of programmed cell death seems present in the early stages of colorectal cancer development. Cancer Epidemiol Biomarkers Prev; 19(2); 351-7. (C) 2010 AACR.
2010
- Altered expression of apoptosis biomarkers in human colorectal microadenomas (Cancer Epidemiology, Biomarkers & Prevention (2010) 19, (351-357))
[Articolo su rivista]
Sena, P.; Roncucci, L.; Marzona, L.; Mariani, F.; Maffei, S.; Manenti, A.; De Pol, A.
abstract
2010
- Fondamenti di Anatomia, Lineamenti di Istologia e Fisiologia
[Monografia/Trattato scientifico]
Palumbo, Carla; Rezzani, R.; DE POL, Anto; Bigiani, Albertino
abstract
Il volume è rivolto agli Studenti delle Professioni Sanitarie e contiene la trattazione omnicomprensiva dell'Anatomia umana normale, dell'Istologia (ad essa propedeutica) e della Fisiologia (ad essa conseguente). E' un testo di Anatomia e Istologia funzionale adatto allo studio di tre discipline fondamentali che offre, nell'ambito della formazione nelle Professioni Sanitarie, un quadro complessivo dei livelli organizzativi e funzionali del corpo umano, nella spiegazione dell'espletamento delle complesse e sofisticate funzioni dell'organismo.
2010
- Human dental pulp stem cells produce mineralized matrix in 2D and 3D cultures
[Articolo su rivista]
Riccio, Massimo; Resca, Elisa; Maraldi, Tullia; Pisciotta, Alessandra; Ferrari, Adriano; Bruzzesi, G; DE POL, Anto
abstract
The aim of this study was to characterize the in vitro osteogenic differentiation of dental pulp stem cells (DPSCs) in 2D cultures and 3D biomaterials. DPSCs, separated from dental pulp by enzymatic digestion, and isolated by magnetic cell sorting were differentiated toward osteogenic lineage on 2D surface by using an osteogenic medium. During the differentiation process, DPSCs express specific bone proteins like Runx-2, Osx, OPN and OCN with a sequential expression, analogous to those occurring during osteoblast differentiation, and produce extracellular calcium deposits. In order to differentiate cells in a 3D space that mimes the physiological environment, DPSCs were cultured in two distinct bioscaffolds, MatrigelTM and Collagen sponge. With the addition of a third dimension, osteogenic differentiation and mineralized extracellular matrix production significantly improved. In particular, in MatrigelTM DPSCs differentiated with osteoblast/osteocyte characteristics and connected by gap junction, and therefore formed calcified nodules with a 3D intercellular network. Furthermore, DPSCs differentiated in collagen sponge actively secrete human type I collagen micro-fibrils and form calcified matrix containing trabecular-like structures. These neo-formed DPSCs-scaffold devices may be used in regenerative surgical applications in order to resolve pathologies and traumas characterized by critical size bone defects.
2009
- A-type lamins and signaling: the PI 3-kinase/Akt pathway moves forward
[Articolo su rivista]
Marmiroli, Sandra; Bertacchini, Jessika; Beretti, Francesca; V., Cenni; M., Guida; DE POL, Anto; N. M., Maraldi; G., Lattanzi
abstract
Lamin A/C is a nuclear lamina constituent mutated in a number of human inherited disorders collectively referred to as laminopathies. The occurrence and significance of lamin A/C interplay with signaling molecules is an old question, suggested by pioneer studies performed in vitro. However, this relevant question has remained substantially unanswered, until data obtained in cellular and organismal models of laminopathies have indicated two main aspects of lamin A function. The first aspect is that lamins establish functional interactions with different protein platforms, the second aspect is that lamin A/C activity and altered function may elicit different effects in different cells and tissue types and even in different districts of the same tissue. Both these observations strongly suggest that signaling mechanisms targeting lamin A/C or its binding partners may regulate such a plastic behavior. A number of very recent data show involvement of kinases, as Akt and Erk, or phosphatases, as PP1 and PP2, in lamin A-linked cellular mechanisms. Moreover, altered activation of signaling in laminopathies and rescue of the pathological phenotype in animal models by inhibitors of signaling pathways, strongly suggest that signaling effectors related to lamin A/C may be implicated in the pathogenesis of laminopathies and may represent targets of therapeutic intervention. In face of such an open perspective of basic and applied research, we review current evidence of lamin A/C interplay with signaling molecules, with particular emphasis on the lamin A-Akt interaction and on the biological significance of their relationship.
2009
- Cyclooxygenase-2 and Hypoxia-Inducible Factor-1 alpha protein expression is related to inflammation, and up-regulated since the early steps of colorectal carcinogenesis
[Articolo su rivista]
Mariani, Francesco; Sena, Paola; Marzona, Laura; Riccio, Massimo; Fano, Rita Adriana; Manni, Paola; C., Di Gregorio; A., Pezzi; PONZ DE LEON, Maurizio; Monni, Sebastiano Graziano; DE POL, Anto; Roncucci, Luca
abstract
Chronic mucosal inflammation is considered a risk factor for colorectal cancer. Neutrophils are a major source of oxidants, whereas cyclooxygenase 2 (COX-2) and Hypoxia Inducible Factor-1 alpha (HIF-1 alpha) protein expression levels are increased in inflammatory and malignant lesions. The main purpose of the present study was to evaluate myeloperoxidase (MPO) positive cell infiltration, COX-2 and HIF-1 alpha protein expression in colorectal carcinogenesis, especially in its early phases, using immunohistochemistry and immunofluorescence confocal microscopy techniques. MPO, COX-2 and HIF-1 alpha proteins were expressed at higher rates in the normal colorectal mucosa of patients with inflammatory bowel diseases and colorectal tumours than in patients with normal colonoscopy. A gradual increase in COX-2 and HIF-1 alpha protein expression was observed in dysplastic aberrant crypt foci, adenomas and carcinomas, showing a strong relation to dysplasia. In conclusion, the present study supports the hypothesis of a key role of inflammation in malignant transformation of colorectal mucosa. The evaluation of some early markers related to inflammation in the mucosa of the large bowel may serve as potential tool for prognosis and therapeutic strategies. (C) 2009 Elsevier Ireland Ltd. All rights reserved.
2009
- Human MATER localization in specific cell domains of oocytes and follicular cells
[Articolo su rivista]
Sena, Paola; Riccio, Massimo; Marzona, Laura; A., Nicoli; T., Marsella; Marmiroli, Sandra; Bertacchini, Jessika; Fano, Rita Adriana; LA SALA, Giovanni Battista; DE POL, Anto
abstract
MATER (Maternal Antigen That Embryos Require) is an oocyte-specific protein dependent on the maternal genome and required for early embryonic development. The gene products expressed in oocytes play important roles in folliculogenesis, fertilization and pre-implantation development. The aim of this study was to characterize the localization and distribution pattern of the human MATER protein during follicular development and after ovulation, to determine its functional role. Immunocytochemistry experiments coupled with confocal and electron microscopy analysis were carried out to determine the ultrastructural localization of MATER in human ovarian tissue and in isolated oocytes, obtained during IVF procedures. Human cumulus cells were cultured, with or without cycloheximide, to confirm endogenous biosynthesis of the protein. Human MATER is detectable at the onset of the follicular maturation process, suggesting this protein has a role at earlier stages in the human compared with other mammalian species. The presence of MATER is specific to the oocyte and follicular cells that, during maturation, are spatially and functionally associated with the oocyte. The nuclear, nucleolar and mitochondrial localization hints at a possible role in RNA processing and the metabolic activity of the cell.
2009
- MATER protein as substrate of PKCepsilon in human cumulus cells.
[Articolo su rivista]
Maraldi, Tullia; Riccio, Massimo; Sena, Paola; Marzona, Laura; A., Nicoli; Marca, A. L.; Marmiroli, Sandra; Bertacchini, Jessika; LA SALA, Giovanni Battista; DE POL, Anto
abstract
High activity of the phosphoinositide 3-kinase/Akt pathway in cumulus cells plays an important role in FSH regulation of cell function and Protein Kinase C epsilon (PKCepsilon) collaborates with these signalling pathways to regulate cell proliferation. Relevant roles in follicular development are played by Maternal Antigen That Embryos Require (MATER) that is a cumulus cell- and oocyte-specific protein dependent on the maternal genome. We recently demonstrated that human MATER localizes at specific domains of oocytes and, for the first time, also in cumulus cells. MATER contains a carboxy-terminal leucine-rich repeat domain involved in protein-protein interactions regulating different cellular functions. Here we investigated the functional role of MATER. Thus, we performed coimmunoprecipitation experiments using HEK293T cells expressing human MATER; a similar approach was then followed in human cumulus/follicular cells. In MATER(+)HEK293T cells, we observed that this protein acts as a phosphorylation substrate of PKCepsilon. Western blot experiments indicate that, unlike oocytes, human cumulus cells express PKCepsilon. Immunoprecipitation and confocal analysis suggest for the first time that MATER protein interacts with this protein kinase in cumulus cells under physiological conditions. Since PKCepsilon is known to collaborate with antiapoptotic signalling pathways, this suggests a novel mechanism for the function of MATER in follicular maturation.
2009
- Multipotent stem cells in vitro differentiation on 3-D scaffolds.
[Abstract in Rivista]
Palumbo, Carla; Riccio, M.; Resca, E.; Maraldi, Tullia; Bertoni, Laura; Sena, Paola; DE POL, Anto
abstract
Goal of the study is to obtain cell/scaffold complexes to use in regenerative medicine.
2009
- OSTEOGENIC DIFFERENTIATION OF DENTAL PULP STEM CELLS (DPSC) IN 3D-MATRICES TO USE IN REGENERATIVE MEDICINE
[Abstract in Rivista]
Resca, Elisa; Riccio, Massimo; Bertoni, Laura; Maraldi, Tullia; Palumbo, Carla; DE POL, Anto
abstract
The investigation concerns the selection of mesenchymal stem cells, derived from adult human dental pulp (DPSC), to commit towards osteogenic differentiation in order to perform new strategies for the regenerative medicine. The final aim of this study is to use DPSC in regenerative medicine approaches for recovering wide gaps of bone tissue due to post-traumatic locomotor apparatus damages.
2008
- Biomechanical aspects in dental replacements
[Capitolo/Saggio]
Baldini, Andrea; Bruzzesi, G.; Zaffe, Davide; Giacopini, Matteo; Strozzi, Antonio; DE POL, Anto
abstract
This chapter deals with biomechanical aspects in dental replacements. The state of the art is critically reviewed taking into account the body of the literature results. The initial section is devoted to the mechanical properties of bone and to a description of the jaw geometry and of its loading and constraining. The following section presents a classifi cation of the various tooth replacement confi gurations and of the various materials adopted, where single and multiple replacements are considered. A specifi c section is devoted to the solid modelling of the jaw as input to numerical analyses, where the aid offered by reverse engineering and tomography
is underlined. The fi nite element method as well as alternative numerical and experimental approaches are reviewed in a dedicated section. The stress analysis forecasts and measurements are biomechanically interpreted in the light of the current literature results. The chapter ends with a section devoted to biological aspects.
2008
- Human dental pulp stem cells (HDPSC) versus osteoblast-like cells: comparison for capability of adhesion, growth and bone matrix formation on differently-shaped surfaces of biomaterials.
[Abstract in Rivista]
Palumbo, Carla; Riccio, M.; Resca, E.; Bretoni, L.; Ferretti, Marzia; Cavani, Francesco; Bruzzesi, G.; Baldini, Andrea; DE POL, Anto
abstract
Comparisons between HDPSC and osteoblast-like cells are made in regards to bone matrix production as well as distribution, density and adhesion to the biomaterial surfaces.
2008
- Human ovarian tissue cryopreservation: effect of sucrose concentration on morphological features after thawing
[Articolo su rivista]
Marsella, Tiziana; Sena, Paola; Xella, Susanna; LA MARCA, Antonio; Giulini, Simone; DE POL, Anto; Volpe, Annibale; Marzona, Laura
abstract
Recent improvements in techniques in clinical assisted reproduction have led to an increased interest in the cryopreservation of human ovarian tissue as a way of preserving fertility and ovarian steroidogenic activity in young cancer patients. Acceptable follicular survival in frozen-thawed human ovarian tissue has generally been reported. Since a 0.3 mol/l sucrose concentration in cryopreservation solutions evidently increases human oocyte survival after cryopreservation, the aim of this study was to observe the effect of sucrose concentrations of 0.2 mol/l and 0.3 mol/l on human ovarian tissue survival after thawing. Ovarian cortical slices from 10 patients, 22-36 years of age, were cryopreserved slowly using 0.2 mol/l or 0.3 mol/l sucrose with 1,2-propanediol (1.5 mol/l) as the cryoprotectants. Light and electron microscopy were used for the histological analyses. Results showed that both treatments produced an increase in damaged cells; however, the use of 0.3 mol/l sucrose showed a smaller percentage of damaged germ cells than 0.2 mol/l sucrose, and therefore was less detrimental to the thawed ovarian tissue. However as the damage occurred principally in the stroma and follicular cells rather than in the oocytes, the suitability of these cryopreservation protocols must be further evaluated prior to considering the use of stored ovarian cortex for autografting after thawing.
2008
- Lamin A Ser404 Is a Nuclear Target of Akt Phosphorylation in C2C12 Cells
[Articolo su rivista]
Vittoria, Cenni; Bertacchini, Jessika; Beretti, Francesca; Giovanna, Lattanzi; Alberto, Bavelloni; Riccio, Massimo; Maria, Ruzzene; Oriano, Marin; Giorgio, Arrigoni; Veena, Parnaik; Manfred, Wehnert; Nadir M., Maraldi; DE POL, Anto; Lucio, Cocco; Marmiroli, Sandra
abstract
Akt/PKB is a central activator of multiple signaling pathways coupled with a large number of stimuli. Although both localization and activity of Akt in the nuclear compartment are well-documented, most Akt substrates identified so far are located in the cytoplasm, while nuclear substrates have remained elusive. A proteomic-based search for nuclear substrates of Akt was undertaken, exploiting 2D-electrophoresis/MS in combination with an anti-Akt phosphosubstrate antibody. This analysis indicated lamin A/C as a putative substrate of Akt in C2C12 cells. In vitro phosphorylation of endogenous lamin A/C by recombinant Akt further validated this result. Moreover, by phosphopeptide analysis and point mutation, we established that lamin A/C is phosphorylated by Akt at Ser404, in an evolutionary conserved Akt motif. To delve deeper into this, we raised an antibody against the lamin A Ser404 phosphopeptide which allowed us to determine that phosphorylation of lamin A Ser404 is triggered by the well-known Akt activator insulin, and is therefore to be regarded as a physiological response. Remarkably, expression of S404A lamin A in primary cells from healthy tissue caused the nuclear abnormalities that are a hallmark of Emery-Dreifuss muscular dystrophy (EDMD) cells. Indeed, it is known that mutations at several sites in lamin A/C cause autosomal dominant EDMD. Very importantly, we show here that Akt failed to phosphorylate lamin A/C in primary cells from an EDMD-2 patient with lamin A/C mutated in the Akt consensus motif. Together, our data demonstrate that lamin A/C is a novel signaling target of Akt, and implicate Akt phosphorylation of lamin A/C in the correct function of the nuclear lamina.
2008
- Propagation of a transmissible cytotoxic activity on cultures of human peripheral blood lymphocytes
[Articolo su rivista]
Marinella, Portolani; Beretti, Francesca; Anna M., Bartoletti; Paola, Pietrosemoli; Stefano, Salvioli; Claudio, Franceschi; Sena, Paola; DE POL, Anto
abstract
Positive results were attained when human peripheral blood lymphocytes (PBLs) were investigated for their ability to propagate a transmissible cytotoxic activity (TCA) isolated on VERO cell cultures from a sample of cerebrospinal fluid (CSF) drawn from a woman with ischemic brain injury. In consideration of this finding it can be assumed that "in vivo" blood lymphocytes contributed to give rise to the TCA detected "in vitro" in the CSF inoculum.
2008
- The oral protein-kinase C beta inhibitor enzastaurin (LY317615) suppresses signalling through the AKT pathway, inhibits proliferation and induces apoptosis in multiple myeloma cell lines
[Articolo su rivista]
Antonino, Neri; Marmiroli, Sandra; Pierfrancesco, Tassone; Luigia, Lombardi; Lucia, Nobili; Donata, Verdelli; Monica, Civallero; Maria, Cosenza; Bertacchini, Jessika; Federico, Massimo; DE POL, Anto; Giorgio Lambertenghi, Deliliers; Sacchi, Stefano
abstract
Deregulation of the protein kinase C (PKC) signalling pathway has been implicated in tumor progression. Here we investigated the PKC inhibitor enzastaurin for its activity against multiple myeloma (MM) cells. Enzastaurin suppresses cell proliferation in a large panel of human myeloma cell lines (HMCLs), with IC50 values ranging from 1.3 to 12.5 mu M and induces apoptosis, which is prevented by the ZVAD-fmk broad caspase inhibitor. These results are consistent with decreased phosphorylation of AKT and GSK3-beta, a downstream target of the AKT pathway and a pharmacodynamic marker for enzastaurin. Furthermore, enzastaurin cytotoxicity is retained when HMCLs were cocultured with multipotent mesenchymal stromal cells. Enzastaurin has additive or synergistic cytotoxic effects with bortezomib or thalidomide. Considering the strong anti-myeloma activity of enzastaurin in vitro and in animal models and its safe toxicity profile, phase II studies in MM patients of enzastaurin alone or in combination with other drugs are warranted.
2007
- Adult human dental pulp stem cells (DPSC): preliminary observations for selecting and conditioning DPSC according osteogenic aims.
[Abstract in Rivista]
Palumbo, Carla; Riccio, Massimo; Resca, Elisa; Bertoni, Laura; Baldini, Andrea; Strozzi, Antonio; G., Bruzzesi; DE POL, Anto
abstract
The previlinary observations have shown the possibility to obtain in vitro bone formation to apply in regenerative medicine.
2006
- Subcellular localization of beta-catenin and APC proteins in colorectal preneoplastic and neoplastic lesions RID B-2583-2012
[Articolo su rivista]
Sena, Paola; Saviano, Massimo; Monni, Sebastiano Graziano; Losi, Lorena; Roncucci, Luca; Marzona, Laura; DE POL, Anto
abstract
Adenomatous polyposis coli (APC) is a tumor suppressor gene whose main function is the destabilization of beta-catenin, a key effector of the Wnt signaling pathway. This gene is defective in familial adenomatous polyposis (FAP), a dominantly inherited disease, but inactivation of APC has been reported also in most sporadic colorectal tumors and it is considered an early event in colorectal tumorigenesis. The aim of the present study was to evaluate the intracellular ultrastructural distribution of beta-catenin and APC proteins in epithelial cells of normal colorectal mucosa, aberrant crypt foci (ACF, an early premalignant lesion) and cancer. We used the immunogold electron microscopic method to identify both proteins. Normal colonic epithelial cells showed a strong membranous expression of beta-catenin and lacked cytoplasmic and nuclear expression. Normal cells showed APC localization pattern characterized by diffuse nuclear expression and along the plasma membrane. In ACF and in carcinoma an absent or reduced membranous expression of beta-catenin was associated with an increased nuclear and cytoplasmatic expression. In aberrant crypt foci and carcinoma, APC was evident inside the nucleus and at the level of cell-cell junctions, but it was decreased in the cytoplasm. This method allowed the accurate localization of proteins of the Writ signaling pathway in the early steps of colorectal carcinogenesis. The similar pattern of subcellular distribution of APC and beta-catenin in dysplastic ACF and colorectal cancer suggests that ACF are precursor lesions of sporadic and FAP-associated colorectal carcinoma. (c) 2005 Elsevier Ireland Ltd. All rights reserved.
2006
- The Oral PKC-ß Inhibitor Enzastaurin (LY317615) Suppress Phosphorylation and Induces Apoptosis in Multiple Myeloma Cell Lines by Inhibition of AKT Pathway.
[Abstract in Rivista]
Antonino, Neri; Marmiroli, Sandra; Pierfrancesco, Tassone; Luigia, Lombardi; Lucia, Nobili; Donata, Verdelli; Civallero, Monica; Cosenza, Maria; Jessika, Bertacchini; Federico, Massimo; DE POL, Anto; Giorgio Lambertenghi, Deliliers; Sacchi, Stefano
abstract
The PKC pathway has been shown to play a role in the regulation of cell proliferation in several hematologic malignancies. In this study we tested the oral PKC-ß inhibitor, Enzastaurin (LY317615 - Eli Lilly) for its therapeutic efficacy in Multiple Myeloma (MM). We first analyzed PKC-ß I and II expression by Western blot in a panel of 19 human MM cell lines, showing that 9 cell lines express either 1 or both isoforms. We next examined the growth inhibition effect of Enzastaurin in the same panel of MM cell lines using either WST-1 or MTT assay and cell viability assessment by Tripan Blue exclusion. Eighteen cell lines have IC50 value ranging from 1,2 µM to 12,5 µM. To examine molecular mechanisms whereby Enzastaurin induces cytotoxicity, we performed cell cycle profiling using PI and observed a significant increase of the percentage of cells in the sub G0–G1 fraction. To determine whether Enzastaurin-induced cell death is mediated by apoptosis, we studied by ELISA and Western blot caspase 3 and PARP cleavage. We observed induction of caspase 3 and PARP cleavage in a dose and time dependent fashion. Notably, the broad caspase (Z-VAD-FMK) inhibitor reduced Enzastaurin-induced cytotoxicity. We next determined whether Enzastaurin could inhibit AKT phosphorylation in MM cell lines with constitutive phosphorylation of AKT. Enzastaurin decreased AKT phosphorylation in a dose and time dependent fashion. Phosphorylation of GSK3ß, a downstream target protein of AKT, was also markedly inhibited. Phosphorylation of PDK-1, a known upstream activator of AKT, was not affected by Enzastaurin. In conclusion, our results indicate that Enzastaurin-induced cytotoxicity is mediated via activation of caspase. This effect is associated with significant inhibition of AKT activity and its downstream target GSK3 ß. Enzastaurin does not alter the phosphorylation of the upstream AKT activator PDK-1. These data suggest that Enzastaurin inhibit AKT signalling pathway and support its evaluation in a murine model of human MM.
2005
- Cell culture isolation of a transmissible cytotoxicity from a human sample of cerebrospinal fluid
[Articolo su rivista]
Portolani, Marinella; Beretti, Francesca; Cermelli, Claudio; Am, Bartoletti; P., Pietrosemoli; Bargellini, Annalisa; DE POL, Anto; Rossini, Gian Paolo
abstract
We investigated a transmissible cytotoxicity isolated in VERO cell cultures from a sample of cerebrospinal fluid (CSF) drawn from a woman with ischemic brain injury. Amorphous aggregates formed by subunities of similar to 11nm of diameter were detected in ultracentrifugates from partially purified cytotoxic cell preparations in the absence of virion-like particles which might justify the trasmissibility of this cytotoxic activity. Results of chemico-physical studies provided indications on the presence in the CSF of two protease-resistant acidic glycoproteins of about 39 and 27 kDa, respectively. The conformational change of a proteinic molecule may associate with particular properties such as tendency to aggregation, resistance to proteolysis, cytotoxicity. Considering that these same properties are shared by proteins present in the CSF sample under study, a hypothesis to pursue is that the CSF inoculum we isolated contained misfolded proteins formed in vivo following the ischemic injury of brain tissue. As far as the in vitro transmissibility of the cytotoxic activity, this could take place following the reproduction of the alterations of those proteins, independently of the original cause(s) which have fostered their formation in vivo.
2005
- Citologia e istologia funzionale
[Monografia/Trattato scientifico]
Calligaro, Colombo; DE POL, Anto; Guidotti, Maraldi; Millo, Narducci; Rana, Roncali; Sica, Tessitore; Urzì,
abstract
2005
- Preservation of inosine on renal tissue during shockwave application in rat model
[Poster]
De Stefani, S; De Carne, C; Di Pietro, C; Micali, Salvatore; Marzona, L; DE POL, Anto; Volpi, Nicola; Bianchi, G.
abstract
Preservation of inosine on renal tissue during shockwave application in rat model
2005
- Preservation of Inosine on renal tissue durino shockwave application in rat model.
[Poster]
Micali, Salvatore; DE STEFANI, Stefano; C., De Carne; C., Di Pietro; L., Marzola; DE POL, Anto; Volpi, Nicola; M. C., Sighinolfi; A., Celia; Bianchi, Giampaolo
abstract
Preservation of Inosine on renal tissue durino shockwave application in rat model.
2004
- Developmental expression and subcellular localization of mouse MATER, an oocyte-specific protein essential for early development
[Articolo su rivista]
Zb, Tong; L., Gold; DE POL, Anto; K., Vanevski; H., Dorward; Sena, Paola; Palumbo, Carla; Ca, Bondy; Lm, Nelson
abstract
We reported previously that Mater is a maternal effect gene that is required for early embryonic development beyond the two-cell stage in mice. Here we show the expressional profile of Mater and its protein during oogenesis and embryogenesis as well as its subcellular localization in oocytes. Mater mRNA was detectable earliest in oocytes of type 2 follicles, whereas MATER protein appeared earliest in oocytes of type 3a primary follicles. Both mRNA and protein accumulated during oocyte growth. In situ hybridization showed that Mater mRNA appeared progressively less abundant in oocytes beyond type 5a primary follicles. By ribonuclease protection assay, Mater mRNA was abundant in germinal vesicle oocytes, but was undetectable in all stages of preimplantation embryos. In contrast, the protein persisted throughout preimplantation development. Immunogold electron microscopic analysis revealed that MATER was located in oocyte mitochondria and nucleoli, and close to nuclear pores. Taken together, our data indicate that Mater gene transcription and protein translation are active during oogenesis, but appear inactive during early embryogenesis. Thus, Mater and its protein are expressed in a manner typical of maternal effect genes. The presence of MATER protein in mitochondria and nucleoli suggests that it may participate in both cytoplasmic and nuclear events during early development.
2004
- Inducible nitric oxide synthase (iNOS) in immune-mediated demyelination and Wallerian degeneration of the rat peripheral nervous system
[Articolo su rivista]
G., Conti; A., Rostami; E., Scarpini; P., Baron; D., Galimberti; N., Bresolin; Contri, Miranda; Palumbo, Carla; DE POL, Anto
abstract
The inducible isoform of nitric oxide synthase (iNOS), produces nitric oxide (NO) from L-arginine in response to inflammatory stimuli. NO sub-serves different functions from cytotoxicity to neuroprotection and triggers either necrosis or apoptosis. This study shows by Northern blot analysis that during experimental allergic neuritis (EAN), at the beginning of clinical signs, there is a transient extensive iNOS mRNA induction in nerve roots, in which morphology is mainly characterized by severe demyelination, but not in sciatic nerve, where scattered axonal degeneration is evident. Immunocytochemistry performed on teased nerve fibers and ultrastructural analysis showed that iNOS was localized in both inflammatory and Schwann cells, and the study of cell membrane permeability detected with fluorescent dyes showed a diffuse necrotic phenotype in the whole peripheral nervous system (PNS). With EAN clinical progression toward spontaneous recovery, endoneurial iNOS was rapidly down-regulated and in nerve roots almost all cells shifted their membrane permeability to an apoptotic phenotype, while necrosis persisted in sciatic nerve, until complete clinical recovery, when both root and nerve returned to normal. During wallerian degeneration following sciatic nerve transection, iNOS was undetectable in PNS, while endoneurial cell membrane had a diffuse necrotic phenotype. These data support the hypothesis that, during cell-mediated demyelination, iNOS may influence Schwann cell-axon relationship causing axonal damage and regulating endoneurial cell life and death.
2004
- Sensitization of multidrug resistant human ostesarcoma cells to Apo2 Ligand/TRAIL-induced apoptosis by inhibition of the Akt/PKB kinase
[Articolo su rivista]
V., Cenni; Nm, Maraldi; A., Ruggeri; P., Secchiero; R., Del Coco; DE POL, Anto; L., Cocco; Marmiroli, Sandra
abstract
Chemotherapeutic agents have been used for the treatment of patients with osteosarcoma (OS). However, inherent or acquired resistance to these agents is a serious problem in the management of OS patients. The emergence of the multidrug resistance (MDR) phenotype in cancer cells is often associated with the overexpression of P-glycoprotein, encoded by the multidrug resistance gene MDR-1. The administration of some of the most common chemotherapeutic agents to these cells becomes ineffective because of their P-gp-driven efflux from the cell. Apo2L/TRAIL is a member of the tumor necrosis factor (TNF) family of cytokines that is considered to induce death of cancer cells but not normal cells. Its powerful apoptotic activity is mediated through its cell surface death domain-containing receptors, TRAIL-R1/DR4 and TRAIL-R2/DR5, which in turn spread the signal in the cytosol through the activation of the caspase cascade. The Akt/PKB kinase is an important cell survival protein which is regulated by D3-phosphoinositides. High Akt expression and activity levels are well documented in many types of tumors., which very often show an altered PI3-K/Akt/PTEN pathway. In this study the U2OS human osteosarcoma cell line and its multidrug resistant (MDR) subline that overexpresses MDR-1 gene, MDR-U2OS. have been analyzed for their responsiveness to TRAIL. In conflict with the presence of active DR4 and DR5 receptors in both clones, U2OS cells exhibited only a low responsiveness to TRAIL, while the MDR-U2OS subline did exhibit a marked TRAIL sensitivity. An analysis of the post-receptor events showed that TRAIL responsiveness correlates with a reduced expression of endogenous Akt. In fact, expression in MDR-U2OS cells of a constitutively active Akt strongly decreased their sensitivity to TRAIL. The identification of Akt as a key modulator of TRAIL responsiveness could help to design TRAIL-based combinations for treatment of osteosarcoma. Moreover, the discovery that multidrug resistant osteosarcomas are highly sensitive to TRAIL-induced apoptosis indicates TRAIL as a new candidate for the treatment of multidrug resistant bone malignancies.
2003
- Apoptosis during intramernbranous ossification
[Articolo su rivista]
Palumbo, Carla; Ferretti, Marzia; DE POL, Anto
abstract
This paper concerns the role of apoptosis during the onset of bone histogenesis. Previous investigations by us performed on intramembranous ossification revealed the existence of two types of osteogenesis: static (SBF) and dynamic bone formation (DBF). During SBF, the first to occur, stationary osteoblasts transform into osteocytes in the same location where they differentiated, forming the primary spongiosa. DBF takes place later, when movable osteoblastic laminae differentiate along the surface of the primary trabeculae. The main distinctive feature between SBF and DBF is that the latter involves the invasion of pre-existing adjacent tissue, whereas the former does not. To ascertain whether programmed cell death during the invasive DBF process determines the fate of surrounding pre-existing mesenchyme differently from that occurring during the non-invasive SBF process, we studied apoptosis in ossification centres of tibial diaphysis in chick embryos and newborn rabbits with TUNEL and TEM. It emerged that, in both SBF and DBF, apoptosis affects mesenchymal cells located between the forming trabeculae and capillaries. However, apoptotic cells were observed more frequently during DBF than during SBF. This suggests that, during bone histogenesis, apoptosis, which is mostly associated with the invasive process of DBF, is probably dedicated to making space for advancing bone growth.
2003
- Apoptosis of striate muscle fibres after tendon lesions: preliminary observations.
[Abstract in Rivista]
Palumbo, Carla; Rovesta, Claudio; Leigheb, M.; Ferretti, Marzia; DE POL, Anto
abstract
Tendon lesions imply the degeneration of the injuried tendon that, in its turn, induces the partial disuse of the muscle mass that undergoes atrophy by apoptosis.
2003
- Interleukin-1-receptor-associated kinase 2 (IRAK2)-mediated interleukin-1-dependent nuclear factor kappa B transactivation in Saos2 cells requires the Akt/protein kinase B kinase
[Articolo su rivista]
V., Cenni; A., Sirri; DE POL, Anto; N. M., Maraldi; Marmiroli, Sandra
abstract
The post-receptor pathway that leads to nuclear factor kappaB (NF-kappaB) activation begins with the assembly of a membrane-proximal complex among the interleukin 1 (IL-1) receptors and the adaptor molecules, myeloid differentiation protein 88 (MyD88), IL-1-receptor-associated kinases (IRAKs) and tumour-necrosis-factor-receptor-associated factor 6. Eventually, phosphorylation of the inhibitor of NF-kappaB (IkappaB) by the IkappaB kinases releases NF-kappaB, which translocates to the nucleus and modulates gene expression. In this paper, we report that IRAK2 and MyD88, but not IRAK1, interact physically with Akt, as demonstrated by coimmunoprecipitation and pull-down experiments. Interestingly, the association of Akt with recombinant IRAK2 is decreased by stimulation with IL-1, and is favoured by pre-treatment with phosphatase. Likewise, Akt association with IRAK2 is increased considerably by overexpression of PTEN (phosphatase and tensin homologue deleted on chromosome 10), while it is completely abrogated by overexpression of phosphoinositide-dependent protein kinase 1. These data indicate that Akt takes part in the formation of the signalling complex that conveys the signal from the IL-1 receptors to NF-kappaB, a step that is much more membrane-proximal than was reported previously. We also demonstrate that Akt activity is necessary for IL-1-dependent NF-kappaB transactivation, since a kinase-defective mutant of Akt impairs IRAK2- and MyD88-dependent, but not IRAK1-dependent, NF-kappaB activity, as monitored by a gene reporter assay. Accordingly, IRAK2 failed to trigger inducible nitric oxide synthase and IL-1beta production in cells expressing dominant-negative Akt. However, NF-kappaB binding to DNA was not affected by inhibition of Akt, indicating that Akt regulates NF-kappaB at a level distinct from the dissociation of p65 from IkappaBalpha and its translocation to the nucleus, possibly involving phosphorylation of the p65 transactivation domain.
2003
- Targeting of the Akt/PKB kinase to the actin skeleton
[Articolo su rivista]
V., Cenni; A., Sirri; Riccio, Massimo; G., Lattanzi; S., Santi; DE POL, Anto; Nm, Maraldi; Marmiroli, Sandra
abstract
Serine/threonine kinase Akt/PKB intracellular distribution undergoes rapid changes in response to agonists such as Platelet-derived growth factor (PDGF) or Insulin-like growth factor (IGF). The concept has recently emerged that Akt subcellular movements are facilitated by interaction with nonsubstrate ligands. Here we show that Akt is bound to the actin skeleton in in situ cytoskeletal matrix preparations from PDGF-treated Saos2 cells, suggesting an interaction between the two proteins. Indeed, by immunoprecipitation and subcellular fractioning, we demonstrate that endogenous Akt and actin physically interact. Using recombinant proteins in in vitro binding and overlay assays, we further demonstrate that Akt interacts with actin directly. Expression of Akt mutants strongly indicates that the N-terminal PH domain of Akt mediates this interaction. More important, we show that the partition between actin bound and unbound Akt is not constant, but is modulated by growth factor stimulation. In fact, PDGF treatment of serum-starved cells triggers an increase in the amount of Akt associated with the actin skeleton, concomitant with an increase in Akt phosphorylation. Conversely, expression of an Akt mutant in which both Ser473 and Thr308 have been mutated to alanine completely abrogates PDGF-induced binding. The small GTPases Rac1 and Cdc42 seem to facilitate actin binding, possibly increasing Akt phosphorylation.
2002
- Apoptosis during static and dynamic bone formation.
[Abstract in Rivista]
Palumbo, Carla; Ferretti, Marzia; DE POL, Anto; Marotti, Gastone
abstract
The study shows that apoptotic phenomena occur in cells of fibroblastic type, located between the newly-forming bony trabeculae and blood capillaries; however, the number of apoptotic cells is incomparably higher during dynamic bone formation than static bone formation.
2002
- Avian scleral bones and human auditory ossicles, two different models to study osteocyte apoptosis in relation with bone remodeling.
[Abstract in Rivista]
Palumbo, Carla; Ferretti, Marzia; DE POL, Anto
abstract
Osteocyte death could represent a programmed phenomenon (apoptosis) in those skeletal segments that should not undergo bone remodeling, because are submitted to steriotyped mechanical stresses for the whole life.
2002
- Interleukin-1 beta and interferon-gamma induce proliferation and apoptosis in cultured Schwann cells
[Articolo su rivista]
G., Conti; DE POL, Anto; E., Scarpini; F., Vaccina; M., De Riz; P., Baron; M., Tiriticco; G., Scarlato
abstract
This study reports that in Schwann cell tissue culture the administration of the two pro-inflammatory cytokines, interleukin-1 beta (IL-1beta) and interferon-gamma (IFN-gamma), at different dosages, singly or in combination, can induce apoptosis and/or mitosis. Schwann cell apoptosis was maximal within 24 h of stimulation with 50 U/ml of IFN-gamma, while proliferation was at its peak within 24 h with 10 U/ml IL-1beta, and both processes decreased progressively by 48 and 72 h. Moreover, the combination of the two cytokines did not show any synergistic effect. These data can be interpreted as a possible involvement of pro-inflammatory cytokines not only in myelin disruption but also in promoting remyelination. (C) 2002 Elsevier Science B.V. All rights reserved.
2002
- Osteocyte dendrogenesis in static bone formation.
[Abstract in Rivista]
Palumbo, Carla; Ferretti, Marzia; DE POL, Anto; Marotti, Gastone
abstract
All osteocytes, independently of their SBF or DBF origin, take part in the formation on the continuous osteocytic network, connected by electric synapsis (gap junctions), potentially capable of modulating by wiring transmission the cells covering the bone surfaces.
2002
- TUNEL REVEALS THE ONSET OF APOPTOSIS ON MESENCHYMAL CELLS IN STATIC AND DYNAMIC BONE FORMATION.
[Abstract in Rivista]
Ferretti, Marzia; Palumbo, Carla; Benincasa, Marta; DE POL, Anto
abstract
Apoptosis is mostly associated with DBF with respect SBF during intramembranous ossification.
2001
- Apoptosis during intramembranous ossification: ultrastructural observations.
[Abstract in Rivista]
Palumbo, Carla; Ferretti, Marzia; Marzona, Laura; DE POL, Anto
abstract
During intramembranous ossification cell apoptosis occurs in the periosteal fibrous tissue in parallel to bone growth and in association with blood vessels.
2001
- Effects of estrogens and oxytocin on the development of neonatal mammalian ovary
[Articolo su rivista]
Marzona, Laura; R., Arletti; Benelli, Augusta; Sena, Paola; DE POL, Anto
abstract
The preservation and death of germ cells in the neonatal mammalian ovary are linked with the presence of hormones. Estrogens and oxytocin are present at birth in all mammalian vertebrates. The aim of this study was to examine their role in the development of the neonatal ovary and also in the preservation and death of germ cells in the neonatal period: apoptotic phenomena play a fundamental role in the control of their number. Female neonatal mice were treated at birth with estradiol monobenzoate or oxytocin and sacrificed after 5 days. The ovaries were sectioned in toto into semi-thin sections, in order to calculate their volume. Thin sections were also carried out to verify, under the transmission electron microscope (T.E.M.), the cells in apoptosis. The ovaries treated with the greater concentration of estradiol monobenzoate showed a volume that was significantly greater than that of the controls and a reduction of germ cells in apoptosis. The ovaries treated with oxytocin at all degrees of concentration had a volume significantly less than the controls and they also had a higher number of germ cells in apoptosis.
2001
- Genetic alterations at the nuclear localization signal of the RB2/p130 gene occur in lymphoid tumor but not in osteosarcoma cell lines
[Articolo su rivista]
Maraldi, Nm; Giordano, A; Manzoli, L; Falconi, M; DE POL, Anto; Cinti, C.
abstract
2001
- Increased activity and nuclear localisation of inositol lipid signal transduction enzymes in rat hepatoma cells
[Articolo su rivista]
P., Santi; N., Zini; S., Santi; Riccio, Massimo; G. G., Piccari; De Pol, Anto; N. M., Maraldi
abstract
To elucidate the relationship between inositol lipid signal transduction and oncogenic transformation, the activity and subcellular distribution of phospholipase C isoforms and of phosphatidylinositol 3-kinase were analysed in Morris hepatoma cells, MH1C1, with respect to normal rat liver cells. The results provide evidence of a gain of: function of the enzymes involved in inositide signal transduction, the amount of which increased mainly at the nuclear level. Phospholipase C and phosphatidylinositol 3-kinase activities are significantly higher in rat hepatoma than in rat liver cells. Moreover, some phospholipase C isoforms are expressed at higher levels at the nuclear level; this is particularly evident in the case of the delta (1) isoform which is not expressed at the nuclear level in rat liver cells. Therefore, the autonomous nuclear signal transduction system, formerly reported as involved in the modulation of cell proliferation and differentiation, appears also affected in oncogenic transformation.
2001
- Influence of estrogens and oxytocin on germ cells death in the neonatal mammalian ovary
[Articolo su rivista]
DE POL, Anto; Benelli, Augusta; Arletti, R.; Cavazzuti, E.; Sena, Paola; Vaccina, F.; Marzona, Laura
abstract
During mammalian oogenesis, some processes involve proliferation and others drastic reduction of germ cells. This study reports on the role played by two hormones, estradiol monobenzoate and oxytocin, in the control of the number of germ cells in the neonatal mouse ovary. Female neonatal mice were treated with doses ranging between 0.1 and 1 microg/mouse of estradiol monobenzoate or oxytocin and sacrificed at 5 days of postnatal age. The results showed that in the animals treated with estrogen, follicular development was more advanced than that of controls. Further the number of germ cells in apoptosis was drastically reduced. In the animals treated with oxytocin, the follicular development was arrested at the stage of primary follicles. In addition, the number of apoptotic germ cells increased if compared with that of the controls.
2001
- Microsatellite instability and mismatch-repair protein expression in hereditary and sporadic colorectal carcinogenesis
[Articolo su rivista]
Pedroni, Monica; Sala, E; Scarselli, A; Borghi, F; Menigatti, M; Benatti, Piero; Percesepe, Antonio; Rossi, Giorgio; Foroni, M; Losi, Lorena; Di Gregorio, C; DE POL, Anto; Nascimbeni, R; Di Betta, E; Salerni, B; PONZ DE LEON, Maurizio; Roncucci, Luca
abstract
Aberrant crypt foci (ACF) are microscopic clusters of altered colonic crypts considered premalignant lesions in the large bowel. Genomic instability at short tandem repeats in the DNA, referred to as microsatellite instability (MSI) is the hallmark of hereditary nonpolyposis colorectal carcinoma (HNPCC) caused by mutations in DNA mismatch-repair genes, mostly hMLH1 and LMSH2. In this study, we evaluated for MSI ACF (n = 16), adenomas (n = 18), carcinomas (n = 22), and lymph node metastases (n = 3) from 17 patients with colorectal cancer positive for MSI, Ten patients were members of HNPCC families; 7 patients had no family history of cancer. MSI was found in 7 of 7 (100%) ACF and 11 of 12 (91%) adenomas from patients with HNPCC, MSI was not related to histology and size of ACF. A progressive increase in instability as estimated by the number of shifted bands was observed along the ACF-adenoma-carcinoma sequence, In contrast, two of nine (228) ACF and none of six adenomas from patients with MSI sporadic carcinoma were unstable at microsatellite loci. hMLH1 or hMSH2 protein expression was altered only in MSI-positive premalignant lesions (ACF and/or adenomas), but not in all MSI-positive lesions in patients with HNPCC. These observations provide evidence of the premalignant nature of ACF in HNPCC and suggest that MSI is a very early event both in HNPCC and in sporadic colorectal carcinogenesis, although in the latter it seems infrequent.
2000
- Aberrant crypt foci in colorectal carcinogenesis. Cell and crypt dynamics
[Articolo su rivista]
Roncucci, Luca; Pedroni, Monica; F., Vaccina; Benatti, Piero; Marzona, Laura; DE POL, Anto
abstract
Aberrant crypt foci (ACF) have been identified on the colonic mucosal surface of rodents treated with colon carcinogens and of humans after methylene-blue staining and observation under a light microscope. Several lines of evidence strongly suggest that ACF with certain morphological, histological, cell kinetics, and genetic features are precursor lesions of colon cancer both in rodents and in humans. Thus, ACF represent the earliest step in colorectal carcinogenesis. This paper has the main purpose of reviewing the evidence supporting this view, with particular emphasis on cell and crypt dynamics in ACF. ACF have been used as intermediate biomarkers of cancer development in animal studies aimed at the identification of colon carcinogens and chemopreventive agents. Recently, evidence has also shown that ACF can be effectively employed in chemopreventive studies also in humans.
2000
- [In vitro study of periodontal ligament cells].
[Articolo su rivista]
R., Galetti; Benelli, Augusta; G., Bruzzesi; Consolo, Ugo; Filaferro, Monica; Genedani, Susanna; S., Palazzini; G., Rasio; DE POL, Anto
abstract
The aim of this research is to outline a procedure able to promote specific cellular differentiation and proliferation with consequent periodontal regeneration. To achieve this goal, use was made of various compounds supposed to have the capacity of aiding periodontal regeneration.The cells utilised for this study were obtained from explants of human periodontal ligaments. Their proliferation and differentiation capacity was examined in the presence of: coral granules (350, 500 mu), collagene type 1, growth factors (Platelet derived growth factor, PDGF and Transforming growth factor beta 1, TGF beta 1), both on their own and in different combination with one another. The differentiation activity was evaluated by ultrastructural morphological method (Transmission electron microscope-TEM) and by spectrophotometric investigation of the alkaline phosphatasis (ALP).The data show that the coral granules and among the growth factors used only TGF beta 1 stimulate the differentiation activity of the periodontal ligament cells valued on the basis of their capacity of producing ALP. These data are supported by the observation with TEM.From these results it is suggested that there may be therapeutic efficiency in the periodontal field of substances promoting cellular proliferation and differentiation.
2000
- Insulin selectively stimulates nuclear phosphoinositide-specific phospholipase C (PI-PLC) beta 1 activity through a mitogen-activated protein (MAP) kinase-dependent serine phosphorylation
[Articolo su rivista]
Martelli, A. M.; Billi, A. M.; Manzoli, L.; Faenza, I.; Aluigi, M.; Falconi, M.; DE POL, Anto; Gilmour, R. S.; Cocco, L.
abstract
Using NIH 3T3 cells, me have investigated nuclear phosphoinositide metabolism in response to insulin, a molecule, which acts as a proliferating factor for this cell line and which is known as a powerful activator of the mitogen-activated protein (MAP) kinase pathway. Insulin stimulated inositol lipid metabolism in the nucleus, as demonstrated by measurement of the diacylglycerol mass produced in vivo and by in vitro nuclear phosphoinositide-specific phospholipase C (PI-PLC) activity assay. Despite the fact that nuclei of NIH 3T3 cells contained all of the four isozymes of the beta family of PI-PLC (i.e. beta1, beta2, beta3, and beta4), insulin only activated the beta1 isoform. Insulin also induced nuclear translocation of MAP kinase, as demonstrated by Western blotting analysis, enzyme activity assays, and immunofluorescence staining, and this translocation was blocked by the specific MAP kinase kinase inhibitor PD98059. By means of both a monoclonal antibody recognizing phosphoserine and in vivo labeling,vith [P-32]orthophosphate, we ascertained that nuclear PI-PLC-beta1 (and in particular the b subtype) was phosphorylated on serine residues in response to insulin. Both phosphorylation and activation of nuclear PI-PLC-beta1 mere substantially reduced by PD98059. Our results conclusively demonstrate that activation of nuclear PI-PLC-beta1 strictly depends on its phosphorylation which is mediated through the MAP kinase pathway. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
2000
- L-sulpiride, at antidepressant dosage, prevents conditioned-fear stress-induced gastric lesions in rats
[Articolo su rivista]
Benelli, Augusta; DE POL, Anto; R., Poggioli; E., Cavazzuti; R., Arletti; A., Bertolini; A. V., Vergoni
abstract
It has been previously shown that long-term treatment with low doses of L-sulpiride is highly effective in rat models of depression and of anticipatory anxiety/panic behavior. The present study was aimed at investigating whether the same treatment can prevent the ulcerogenic effect of repeated inescapable stresses. In adult rats, the repeated (7 consecutive days) exposure to an uncontrollable stressful condition (inescapable 2.5 mA scrambled shock for 60 s) produced the development of gastric lesions (multiple punctiform telangiectasias in all rats, with superficial erosions or more severe ulcerations in 10 out 13 rats; score 4.67 +/- 0.44). L-sulpiride, intraperitoneally injected once a day at an antidepressant dose level (4 mg kg(-1) per day), starting 21 days before the beginning of the 7-day sequence of inescapable punishments (= 28 daily treatments), almost completely prevented the stress-induced gastric injury (score 1.67 +/- 0.29; P < 0.001 vs saline-treated rats, Mann-Whitney U test). These results show that, in rats, a long-term treatment with low doses of L-sulpiride prevents the development of gastric lesions induced by chronic exposure to uncontrollable stress. (C) 2000 Academic Press.
1999
- Human Schwann cell proliferation and IL-6 production following TNF-alpha stimulation in vitro
[Articolo su rivista]
Scarpini, E; Conti, Gc; Bussini, S; Clerici, R; Siglienti, I; Piccio, L; DE POL, Anto; Baron, Pl; Scarlato, G.
abstract
1998
- Apoptosis in different stages of human oogenesis.
[Articolo su rivista]
De Pol, Anto; Marzona, Laura; Vaccina, F.; Negro, R.; Sena, Paola; Forabosco, A.
abstract
Apoptosis is an active form of cell death characterized by a series of morphological changes that become particularly evident at the ultrastructural level. The majority of ovarian germ cells undergo degeneration during prenatal and reproductive life and only in recent studies has it been demonstred that this drop is due to an apoptotic process. We evaluated this process during human oogenesis in prenatal life and we studied the ultrastructural changes that occur in apoptosis in various phases of the meiotic process. From our observations it is clear that apoptosis involves two main phases of the meiotic process: an earlier one concerning the oogonia and oocytes in the preleptotene stage, and a later one that mainly concerns the oocytes in the pachytene stage.
1998
- Scanning electron microscopy of aberrant crypt foci in human colorectal mucosa
[Articolo su rivista]
F., Vaccina; F., Scorcioni; Pedroni, Monica; Mg, Tamassia; PONZ DE LEON, Maurizio; DE POL, Anto; Marzona, Laura; Roncucci, Luca
abstract
Background: Aberrant crypt foci (ACF) are clusters of morphologically altered crypts which can be observed by light or stereomicroscopy on the mucosal surface of the colon after staining with methylene-blue. They probably represent one of the earliest events in human colorectal carcinogenesis. The main purpose of the present study was to observe the surface features of aberrant and normal colonic crypts in humans using scanning electron microscopy (SEM) in order to find and measure differences between aberrant and normal. Materials and Methods: Fifteen mucosal specimens containing ACF and 8 with normal mucosa taken from patients operated on for colon cancer were observed under a scanning electron microscope. Results: By SEM ACF were easily observed on the mucosal surface because they showed a well defined border and were elevated on the mucosal surface. Under higher magnification luminal openings of aberrant crypts had a larger overall average diameter than normal (37.6 mu m +/- 13.5, mean a SD, vs 15.9 mu m +/- 4.9, P=0.001), though when crypt multiplicity of ACF (number of crypts per ACF) was higher, the diameter of luminal openings tended to be smaller and similar to those of normal crypts, with weak negative correlation between crypt multiplicity of ACF and mean diameter of aberrant luminal openings (r=-0.27). Finally, the mucosal surface among aberrant crypts was flattened because of a loss of microvilli. In conclusion, scanning electron microscopy allows a better definition of the topological features of aberrant crypt foci than light or stereomicroscopy.
1998
- Ultrastructural localization of gelsolin in lattice corneal dystrophy type I
[Articolo su rivista]
L., Longanesi; Cavallini, Gian Maria; A., Iammarino; F., Vaccina; Guerra, Roberto; DE POL, Anto
abstract
In the light of recent studies into lattice corneal dystrophies, with particular reference to gelsolin immunoreactivity, the authors set out to determine the ultrastructural localization of gelsolin molecules in lattice corneal dystrophy type I. Immunoelectron microscopy with a monoclonal antibody against the COOH-tenminal of the native gelsolin molecule (clone GS-2C4) was used to compare antigelsolin reactivity in normal and dystrophic corneas. A gelsolin-like protein was observed at the level of the rough endoplasmic reticulum in both epithelial and endothelial cells, together with mild positive staining in stromal keratocytes of normal corneas, increased keratocytic immunoreactivity with positive staining within and/or around corneal amyloid deposits was revealed in dystrophic corneas. Observed intra- and extracellular immunoreactivity suggests that amyloid deposition may induce gelsolin synthesis; this actin-related protein could be involved in the rearrangement of corneal stroma in lattice corneal dystrophy.
1996
- Quantitative study of definitive histogenesis in normal and trisomy 21 ovaries.
[Articolo su rivista]
Forabosco, Antonino; C., Sforza; Marzona, Laura; DE POL, Anto; V. F., Ferrario
abstract
1993
- Developmental Pathways Of Vertebral Centra And Neural Arches In Human Embryos And Fetuses
[Articolo su rivista]
R., Bareggi; V., Grill; M. A., Sandrucci; G., Baldini; DE POL, Anto; Forabosco, Antonino; P., Narducci
abstract
The ossification pathways of both vertebral centra (i.e., vertebral bodies) and neural arches were studied in human embryos and fetuses (CR-length between 38 and 116 mm). A clearing and double-staining method for whole embryo or fetus, using alcian blue and alizarin red S, allowed an easy and precise detection of the morphology of the whole vertebral column and every single vertebra. Both cartilaginous and bony components were clearly visible. Different temporal and topographical patterns of ossification were shown for the centra and arches; the latter were respectively proximal-distal (i.e., bidirectional from a defined starting tract in T10-L1) and cranial-caudal (i.e., monodirectional). The patterns could be related to the morphogenetic processes of other structures (i.e., muscles and nerves). Moreover, the numerical survey of ossification centers provided a possible parameter for the determination of the fetal developmental age. This could be useful in the study of pathological conditions.
1993
- Human neonatal ovary: proposal of a tree- dimensional model.
[Articolo su rivista]
V. F., Ferrario; Marzona, Laura; C., Sforza; DE POL, Anto; A., Miani; A. E. A., Bertelli; Forabosco, Antonino
abstract
A model of human neonatal ovary is presented, derived from morphometric, evaluations carried out on left ovaries removed from five full-term neonates with a 46, XX karyotype, free from malformations of the genital apparatus. According to this model, the gonad can be represented by a triaxial ellipsoid with a central medullary core surrounded by a cortical stratum of constant thickness. The germinal population, consisting of follicles and primitive cortical tissue, occupies the cortex, intermingled with the interstitium or stroma. In the cortex it is then possible to describe an outer layer formed by primitive cortical tissue, and an inner portion occupied by follicles. The primary and secondary follicles fill the portion near the medulla and the primordial ones are contained in the middle and outer zones. Since the variability observed among ovaries is slight, we can propose a mean model of neonatal ovary in which the spatial relationships among the different components, the total number of follicles and their position in the cortex can be calculated.
1993
- Morphometric Study Of The Human Ovary During Compartmentalization
[Articolo su rivista]
Sforza, C; Ferrario, Vf; DE POL, Anto; Marzona, Laura; Forni, M; Forabosco, Antonino
abstract
The volumetric composition of the human ovary during the compartmentalization stage has been investigated using current stereological methods. Eight left ovaries removed from three fetuses (developmental age 20-25 weeks), four neonates, and one 8-month-old child all with a 46,XX karyotype, free from malformations of the genital apparatus, were completely cut obtaining serial sections and one 1 mum-thick section every 1,000 mum was examined. Ovarian volume was 30 mm3 at the 20th week of development, 36 mm3 at the 25th week, 129 mm3 at birth, and 287 mm3 at the eighth postnatal month. The primitive cortical tissue was the largest component of the fetal ovaries (17 mm3, corresponding to 60.2% of the organ). The second component was the interstitium (21% of the organ), followed by the medulla (11.8% of the organ). The primordial follicles occupied a small part of the organs: 1.8 mm3 at 20 weeks and 3.4 mm3 at 25 weeks (respectively 6.7% and 5.4% of the volumes of the relevant ovaries). At birth, most of the organ was composed of interstitial tissue (57 mm3. 44.2% of the volume) followed by the medulla (25 mm3 20.3% of the volume). The germinal tissue occupied 46 mm3 mainly primitive cortical tissue (14.9% of the ovary) and primordial follicles (16.3% of the ovary), with a minor contribution from the antral follicles (about 3% of the ovary). At 8 months, the somatic tissue formed the majority of the organ (143 MM3 of Stroma, corresponding to about 50% of the volume, and 43 mm3 of medulla, about 15% of the volume); the germinal tissue occupied about 101 mm3; MoSt of this volume was given by the antral follicles (28.6% of the ovarian volume).
1992
- Epikeratophakia - Histopathological And Cultural-Study
[Articolo su rivista]
Cavallini, Gian Maria; L., Longanesi; DE POL, Anto; E. C., Campos; R., Guerra
abstract
Three epikeratoplasty buttons (one aphakic and two myopic) prepared by the freeze technique were removed two to nineteen months after surgery. A complete morphological and immunohistochemical study was performed on these buttons in order to gain insight into the reasons for epikeratophakia failure. Histopathological studies with light microscopy and scanning and transmission electron microscopy were performed. All three cases showed an anomalous process of re-epithelialization: in the first, the epithelium over the donor cap was almost completely absent, with many abnormalities; in the second, the epithelium was irregular, with a varying number of cell layers and poor adhesion between cells; in the third, the development of an epithelial cyst between the tissue lens and the cornea of the host caused the failure of the epikeratoplasty. The specimens were cultured 'in vitro' and the cells grown typed for HLA antigens. The antigen panel was the same as that of the host. Immunohistochemistry showed CD3- and CD8-positive cells in the stroma, proving the activity of T-suppressor lymphocytes in a cell-mediated immunoreaction.
1991
- Morphometric study of human neonatal ovary.
[Articolo su rivista]
Forabosco, Antonino; C., Sforza; DE POL, Anto; L., Vizzotto; Marzona, Laura; V. F., Ferrario
abstract
A morphometric analysis, based on mathematical evaluations and stereological methods, has been used to study five left neonatal ovaries, removed from full-term neonates with a 46,XX karyotype free from malformations of the genital apparatus. Each ovary was completely cut obtaining serial sections and one 1-mu-m-thick section every 1,000-mu-m was examined. Ovarian length ranged from 9 to 17 mm (mean 13 mm), width from 3.5 to 7 mm (mean 5.7 mm), thickness from 2.5 to 5 mm (mean 4 mm), and volume from 82.23 to 198.3 mm3 (mean 125.88 mm3). In the ovarian cortex, primitive cortical tissue accounted for 10-20% of the total volume, follicles for 10-25% and interstitium for 35-45%; 10-30% of the organ consisted of inner medulla. The total follicle number ranged from 130,000 to 385,000 per ovary, with an average of 266,000 with 95% being represented by primordial follicles. In all ovaries examined follicular growth was still in process, with follicles at different stages of development.
1983
- Surveillance of constitutional chromosome abnormalities in western Emilia
[Articolo su rivista]
Forabosco, A; Croci, G; Manini, M; Temperani, P; Caselli, L; Marzona, L; DE POL, Anto
abstract
1980
- Ittiosi lamellare in età neonatale
[Relazione in Atti di Convegno]
Malmusi, L; DE POL, Anto; Grosoli, Mv; Ferrari, Fabrizio
abstract
NO ABSTRACT
1979
- 46,XX21p-/46XX,i(21q) chromosomal pattern: a rare formula in Down's syndrome
[Articolo su rivista]
DE POL, Anto; Salsi, M; Gavioli, G; Temperani, P.
abstract
1977
- Male with 45,X karyotype.
[Articolo su rivista]
A., Forabosco; A., Carratu; M., Assuma; DE POL, Anto; B., Dutrillaux; E., Cheli
abstract
1977
- Sull'impiego del Bismarck Brown quale reagente “tipo-Schiff”
[Articolo su rivista]
Zaffe, Davide; DE POL, Anto
abstract
Le divergenze tra i diversi ricercatori hanno stimolato l'approfondimento dello studio sul comportamento del Bismarck Brown quale sostituente della Pararosanilina nel reattivo di Schiff. I risultati ottenuti utilizzando il reattivo nelle reazioni Feulgen e PAS, portano alla conclusione che non è possibile considerare questo colorante come un vero "tipo-Schiff".