81 results on '"Pisano I"'
Search Results
2. In Vitro Effects of Low-energy Ultrasound Treatment on Healthy CD3/CD8+ Lymphocytes, Red blood cells, Acute Myeloid leukemia cells, and Jurkat cell line
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De Leonardis, F., Barile, S. N., Cianci, C., Pisano, I., Merla, G., Pappalettera, G., Casavola, C., Pappalettere, C., De Leonardis, Francesco, Barile, Simona Nicole, Cianci, Claudia, Pisano, Isabella, Merla, Giuseppe, Pappalettera, Giovanni, Casavola, Caterina, and Pappalettere, Carmine
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CD3/CD8 lymphocytes proliferation/activation ,Oncology ,cytotoxic functions ,Lymphocytes ,Red blood cells ,Lymphocyte ,CD3/CD8 lymphocytes proliferation/activation and cytotoxic function - Abstract
The study of the biological effects of low-energy ultrasound and its applications is a rapidly expanding research area. Low-energy ultrasound could be used as anti-tumoral therapy with or without the pharmacological combination even if the second situation has been scarcely investigated up to now. Very little information is available about the ultrasound effects on healthy red blood cells, CD3, and mainly CD8 subset lymphocytes which is the main subset cell having cytotoxic function towards cancer cells. In this study, we investigated in vitro the bioeffects of low energy ultrasound on red blood cells and PBMCs isolated from healthy donors as well as on two myeloid leukemia cell lines (OCI- AML-3 MOLM-13) and lymphoblastic Jurkat cell line. Using low-energy ultrasound (US), a study was conducted to determine how it affects CD3/CD8 lymphocytes and leukemia cells, as well as its potential role in treating blood cancers, by analyzing changes in mitochondrial membrane potential, phosphatidylserine asymmetry, morphological changes for myeloid AML cell lines, proliferation and cytotoxic activation of healthy lymphocytes, and apoptosis for RBCs after US exposure. Overall, we demonstrated that CD3/CD8 lymphocytes proliferation/activation and cytotoxic functions are fully preserved after ultrasound treatments, whereas leukemia cell lines undergo apoptosis and stop proliferating suggesting a potential method of treating blood cancer.
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- 2023
3. Cheap and environmentally sustainable stereoselective arylketones reduction by Lactobacillus reuteri whole cells
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Perna, F.M., Ricci, M.A., Scilimati, A., Mena, M.C., Pisano, I., Palmieri, L., Agrimi, G., and Vitale, P.
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- 2016
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4. P.0502 Sex-dependent effects of a chronic treatment with lurasidone on the vulnerability to chronic stress exposure
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Begni, V., primary, Creutzberg, K.C., additional, Marchisella, F., additional, Pisano, I., additional, De Rosa, F., additional, Papp, M., additional, and Riva, M.A., additional
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- 2021
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5. P.0552 The early therapeutic efficacy of lurasidone in a rodent model of depression: a behavioural and biomolecular study
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Pisano, I., primary, Begni, V., additional, Creutzberg, K.C., additional, Marchisella, F., additional, De Rosa, F., additional, Papp, M., additional, and Riva, M.A., additional
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- 2021
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6. P.0497 Acute stress differentially regulates BDNF expression in prefrontal excitatory and inhibitory neurons of chronically stressed rats: modulation by lurasidone treatment
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Marchisella, F., primary, Pisano, I., additional, De Rosa, F., additional, Creutzberg, K.C., additional, Begni, V., additional, Papp, M., additional, and Riva, M.A., additional
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- 2021
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7. Long-chain polyphosphates impair SARS-CoV-2 infection and replication: a route for therapy in man
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Giuseppe Castaldo, Borriello G, Hyeri Kim, Fusco G, Ferrucci, Siciliano R, Angelo Boccia, Tiberio C, Kong D, Pierri Bm, Quarantelli F, Yun K, Marika Comegna, Monica Rd, Martina Bianchi, G. Paolella, Pisano I, Zollo M, Atripaldi L, Viscardi M, Barbara Izzo, Criscuolo G, Cerino P, Brandi S, Asadzadeh F, Marrone L, Jae Ho Cheong, Stefano Pascarella, and Chiariotti L
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chemistry.chemical_classification ,viruses ,RNA ,Biology ,Virology ,In vitro ,Virus ,Amino acid ,chemistry.chemical_compound ,chemistry ,Viral replication ,Transcription (biology) ,RNA polymerase ,Subgenomic mRNA - Abstract
Anti-viral activities of long-chain inorganic polyphosphates (PolyPs) against severe acute respiratory syndrome coronavirus (SARS-CoV)-2 infection were investigated. In molecular docking analyses, PolyPs interacted with several conserved angiotensin-converting enzyme (ACE)2 and RNA-dependent RNA polymerase (RdRp) amino acids. We thus tested PolyPs for functional interactionsin vitroin SARS-CoV-2–infected Vero E6, Caco2 and human primary nasal epithelial cells. Immunofluorescence, qPCR, direct RNA sequencing, FISH and Immunoblotting were used to determine virus loads and transcription levels of genomic(g)RNAs and sub-genomic(sg)RNAs. We show that PolyP120 binds to ACE2 and enhances its proteasomal degradation. PolyP120 shows steric hindrance of the genomic Sars-CoV-2-RNA/RdRP complex, to impair synthesis of positive-sense gRNAs, viral subgenomic transcripts and structural proteins needed for viral replication. Thus, PolyP120 impairs infection and replication of Korean and European (containing non-synonymous variants) SARS-CoV-2 strains. As PolyPs have no toxic activities, we envision their use as a nebulised formula for oropharyngeal delivery to prevent infections of SARS-CoV-2 and during early phases of antiviral therapy.
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- 2020
8. P.223 Time course of antidepressant actions of lurasidone treatment towards the identification of early biomarkers of response
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Begni, V., primary, Creutzberg, K.C., additional, Marchisella, F., additional, Pisano, I., additional, Papp, M., additional, and Riva, M.A., additional
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- 2021
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9. CLINICAL OUTCOME OF CHRONIC MYELOID LEUKEMIA (CML) PATIENTS WITH DELETION AND INSERTION EVENTS (DI) IN THE TYROSINE KINASE DOMAIN OF BCR-ABL
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Peluso AL, Seneca E, Cosenza MR, Musella F, Cacciapuoti V, Esposito N, Muccioli Casadei G, Pisano I, De Angelis B, Annunziata M, Sessa U, Nunziata G, Palmieri F, Palmieri R, Danise P, Pezzullo L, Serio B, Esposito MR, Villa MR, Spiezia M, Vallone R, De Falco G, Sica A, Camera A, Iovine M, Svanera L, Soverini S, Martinelli G, Luciano L, QUINTARELLI, CONCETTA, IZZO, BARBARA, PANE, FABRIZIO, Peluso AL, Peluso, Al, Quintarelli, Concetta, Seneca, E, Cosenza, Mr, Musella, F, Cacciapuoti, V, Esposito, N, Izzo, Barbara, Muccioli Casadei, G, Pisano, I, De Angelis, B, Annunziata, M, Sessa, U, Nunziata, G, Palmieri, F, Palmieri, R, Danise, P, Pezzullo, L, Serio, B, Esposito, Mr, Villa, Mr, Spiezia, M, Vallone, R, De Falco, G, Sica, A, Camera, A, Iovine, M, Svanera, L, Soverini, S, Martinelli, G, Luciano, L, and Pane, Fabrizio
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- 2013
10. MOLECULAR EVALUATION OF ZNF224 MRNA EXPRESSION IN CML PATIENTS AS A NOVEL DETERMINANT OF TKI RESPONSIVENESS
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Errichiello, S., Caruso, S., BIAGIO DE ANGELIS, Quintarelli, C., Pisano, I., Izzo, B., Muccioli, G., Musella, F., Del Prete, C., Visconti, R., Galdiero, A., Cacciapuoti, V., Siciliano, M., Pugliese, N., Della Pepa, R., Pane, F., Errichiello, Santa, Caruso, Simona, DE ANGELIS, Biagio, Quintarelli, Concetta, Pisano, Ida, Izzo, Barbara, G., Muccioli, F., Musella, C. Del, Prete, R., Visconti, A., Galdiero, V., Cacciapuoti, M., Siciliano, Pugliese, Novella, DELLA PEPA, Roberta, and Pane, Fabrizio
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The transcription factor Wilms’ tumor gene 1, WT1, is implicated both in normal developmental processes and in the generation of a variety of solid tumors and hematological malignancies. WT1 is highly expressed in leukemia cells and its overexpression is associated with a poor response to therapy. Recently the Krüppel-like zinc-finger protein, ZNF224 was identified as a novel WT1-interacting factor involved in WT1 transcriptional regulation. ZNF224 itself could be modulated by cytosine arabinoside (ara-C), a drug widely used in the treatment of myeloid leukemia and that ZNF224 overexpression increases susceptibility to apoptosis of Ph+ K562 cell lines. In our retrospective analysis we evaluated the relative expression of ZNF224 mRNA in 30 adult patients with BCR-ABL–positive chronic phase chronic myeloid leukaemia (CP-CML) as a determinant of imatinib sensitivity. Methods: Response to tyrosine kinase inhibitor (TKI) imatinib is assessed with standardized real quantitative polymerase chain reaction and/or cytogenetics at 3, 6, and 12 months. Response to the therapy was classified as optimal, warning, and failure, according to the recent ELN criteria. We compared the ZNF224 expression at diagnosis with molecular response over the first 12 month of imatinib therapy. Sample have been selected, for retrospective analysis, for them interim molecular results a 12 month, showing 15 patients in optimal response (OR), 10 patients in a warning response (WR) and 5 patients in failure response (FR). 5 healthy donors (HDs) were included to the study. All patients signed informed consent in accordance with the Declaration of Helsinki. RT-qPCR results were normalized by the expression of ABL mRNA (Normalized mRNA copy Number: NCN).Results:ZNF224 mRNA were significantly up-regulated in PB samples at diagnosis of patients with OR compared to patients with WR/FR, (1.13±0.76 vs 0.62±0.25 NCN,respectively; p=0.05). Interesting the ZNF224 mRNA expression in HDs was significantly higher (2.11±0.98 NCN vs OR patients, p=0.05 and WR/FR patients; p=0.0005). The treatment for 12 month with imatinib increase the ZNF224 expression in both CML categories (2.91±1.72 NCN in OR and1.77±1.52 NCN in WR/FR; p=0.05). Conclusions:We observed that the OR patients express a significantly higher number of copies of the ZNF224 transcript than WR/FR. Furthermore, in both groups of patients at diagnosis, ZNF224 protein levels are lower than those after therapy with TKI at 12 months
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- 2015
11. Identification of a novel Sp1 splice variant as a strong transcriptional activator
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Infantino V, Convertini P, Iacobazzi F, Pisano I, Scarcia P, and Iacobazzi V.
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- 2011
12. Studio caso-controllo sull’epidemiologia di stipiti antibiotico resistenti di escherichia coli e enterococcus spp. in pecore e ungulati selvatici nel parco nazionale dei monti sibillini
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Soncini, G., Sechi, Paola, Pisano, I., Monticelli, C., Nicchiarelli, I., and CENCI GOGA, Beniamino Terzo
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- 2009
13. Imipenem/cilastatin (1.5 g daily) versus meropenem (3.0 g daily) in patients with intra-abdominal infections: Results of a prospective, randomized, multicentre trial
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Basoli, Antonio, Chirletti, Piero, Paolo, Mazzocchi, Vincenzo, Speranza, Lezoche, E., Guerrieri, M., Marrano, D., Minni, F., Giulini, S. M., Nodari, F., Brotzu, G., Loddo, P., Latteri, F., Scuderi, Gianluca, Rodolico, G., Cavallaro, L., Donini, I., Sortini, A., Tonelli, F., Spini, S., Natale, C., Musto, V., Vio, A., Verdecchia, G., Morgagni, D., Mariani, L., Montefusco, A., Gerosa, E., Tiberio, G., Nardone, A., Mazzeo, F., Benassai, G., D'Amico, D., Tropea, A., Piervittori, M., Becelli, S., Cazzaniga, M., Stagnitti, F., Crucitti, F., Pacelli, F., Gargiulo, A., Panichi, Giovanni, DI ROSA, Roberta, Porzio, R., Lombardi, U., Stipa, V., Chirletti, P., De Anna, D., Pisano, I., Armenio, S., Salvestrini, E., Baglioni, A., Iafrate, G., Donadio, F., Paron, L., Saccia, A., Di Girolamo, P., A., Basoli, E. Z., Meli, P., Mazzocchi, V., Speranza, E., Lezoche, M., Guerrieri, D., Marrano, F., Minni, S. M., Giulini, F., Nodari, G., Brotzu, P., Loddo, F., Latteri, G., Scuderi, G., Rodolico, L., Cavallaro, I., Donini, A., Sortini, F., Tonelli, S., Spini, C., Natale, V., Musto, A., Vio, G., Verdecchia, D., Morgagni, L., Mariani, A., Montefusco, E., Gerosa, G., Tiberio, Nardone, GERARDO ANTONIO PIO, F., Mazzeo, Benassai, Giacomo, D., Damico, A., Tropea, M., Piervittori, S., Becelli, M., Cazzaniga, F., Stagnitti, F., Crucitti, F., Pacelli, A., Gargiulo, G., Panichi, R., Dirosa, R., Porzio, U., Lombardi, V., Stipa, P., Chirletti, D., Deanna, I., Pisano, S., Armenio, E., Salvestrini, A., Baglioni, G., Iafrate, F., Donadio, L., Paron, A., Saccia, and P., Digirolamo
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Adult ,Male ,Microbiology (medical) ,Imipenem ,Meropenem ,law.invention ,Randomized controlled trial ,law ,Abdomen ,polycyclic compounds ,medicine ,Humans ,Protease Inhibitors ,Prospective Studies ,Infusions, Intravenous ,Prospective cohort study ,APACHE ,Aged ,General Immunology and Microbiology ,Cilastatin ,business.industry ,Imipenem/cilastatin ,Bacterial Infections ,General Medicine ,Middle Aged ,biochemical phenomena, metabolism, and nutrition ,bacterial infections and mycoses ,Anti-Bacterial Agents ,Clinical trial ,Treatment Outcome ,Infectious Diseases ,Tolerability ,Anesthesia ,Female ,Thienamycins ,business ,medicine.drug - Abstract
30 citazioni su Scopus. 46 citazioni su GoogleScholar. ------ http://www.scopus.com/record/display.url?eid=2-s2.0-0347771343&origin=resultslist&sort=plf-f&src=s&sid=1BklTwBDkRxWaxt9FLi7YAz%3a70&sot=aut&sdt=a&sl=37&s=AU-ID%28%22Benassai%2c+Giacomo%22+6602920646%29&relpos=11&relpos=11&searchTerm=AU-ID(\"Benassai, Giacomo\" 6602920646) ------ An open-label prospective, randomized, parallel multicentre study was undertaken to compare the efficacy and tolerability of 1.5 g/day intravenous imipenem/cilastatin with 3 g/day intravenous meropenem in the treatment of intra-abdominal infections, A total of 287 patients mere enrolled; 201 patients, divided between the 2 treatment groups, were evaluable. Clinical outcome, bacteriological outcome, untoward microbiological effects, and clinical and laboratory adverse experiences mere evaluated, 98% of patients receiving imipenem/cilastatin therapy mere cured, with 96% showing eradication of infection, 95% of those on meropenem mere cured, with 98% showing eradication. These differences in clinical and bacteriological outcome between the 2 treatments were not statistically significant, Two patients receiving imipenem/cilastatin and 5 receiving meropenem had untoward microbiological effects. There mas a 0.7% frequency (1/139 patients) of possibly or probably drug-related clinical or laboratory adverse experiences with imipenem/cilastatin and a 2.7% frequency (4/148) with meropenem, The mean time to defervescence was significantly less for patients in the imipenem/cilastatin treatment group than for those receiving meropenem. This study shows that 1.5 g/day of imipenem/cilastatin is equivalent to 3.0 g/day meropenem in clinical and bacteriological outcome, as well as in incidence of side effects.
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- 1997
14. CONSERVAZIONE DI VENE PER ETEROTRAPIANTO: STUDIO SPERIMENTALE
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Pisano, I., Deiana, A., Bresadola, Vittorio, Virdis, A., Sanna, E., Trignano, M., and De Anna, D.
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- 1994
15. PREVALENCE OF ANTIBIOTIC RESISTANT STRAINS OF ESCHERICHIA COLI AND ENTEROCOCCUS SPP. IN ROE DEER (CAPREOLUS CAPREOLUS) AND RED DEER (CERVUS ELAPHUS) AT THE PARCO NAZIONALE DEI MONTI SIBILLINI, ITALY
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Cenci Goga, B., primary, Vizzani, A., additional, Monticelli, C., additional, Nicchiarelli, I., additional, Sechi, P., additional, and Pisano, I., additional
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- 2009
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16. 34 P Management and outcomes of patients with early colorectal cancer
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Scognamillo, F., primary, Carboni, G., additional, Cavallo, G., additional, Pala, C., additional, Mastino, G., additional, Pisano, I., additional, Soro, P., additional, and Trignano, M., additional
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- 2002
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17. Cystic pancreatic tumor treated by distal splenopancreasectomy with occasional diagnosis of neuroendocrine tumor | Tumore cistico pancreatico trattato mediante splenopancreasectomia distale con riscontro occasionale di neoplasia neuroendocrina
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Pala, C., Serventi, F., Scognamillo, F., Federico Attene, Pisano, I. P., Cugia, L., Meloni, M., and Trignano, M.
18. NANOG: ITS ROLE IN THE TKI RESISTANCE OBSERVED IN PATIENTS WITH CHRONIC MYELOID LEUKEMIA
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Caruso, S., Errichiello, S., Pisano, I., Quintarelli, C., BIAGIO DE ANGELIS, Izzo, B., Muccioli, G., Pugliese, M., Della Pepe, R., Musella, F., Del Prete, C., Visconti, R., Galdiero, A., Cacciapuoti, V., Siciliano, M., Pane, F., FONDAZIONE FERRARA STORTI, Caruso, Simona, Errichiello, Santa, Pisano, Ida, Quintarelli, Concetta, B. De, Angeli, Izzo, Barbara, G., Muccioli, M., Pugliese, R. Della, Pepe, F., Musella, C. Del, Prete, R., Visconti, A., Galdiero, V., Cacciapuoti, M., Siciliano, and Pane, Fabrizio
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Treatment of patients with Chronic Myeloid Leukemia in chronic phase (CML-CP) with tyrosine kinase inhibitors (TKIs) showed a substantially improving of patient life expectancy. However,it is becoming evident that persistent leukemic stem cells, which are in-sensitive to TKIs in their quiescent state, can lead to CML recurring.Nanog is a pluripotency gene associated to a vital role in neoplasia, correlating with cell proliferation, clonogenic growth, tumorigenicity, and therapeutic resistance. Our group carried out microarray experiments on Ph+ KCL22 cell line with a sensible (Kcl22-S) or resistant (Kcl22-R) phenotype to Imatinib (Ima). The gene expression of Nanog was significantly increased in the Kcl22-R. Thus, we sought to investigate the role of Nanog in the TKI resistance observed in patients with CML-CP. Methods: Real Time RT-PCR (RT-qPCR) for the expression of Nanog was con- ducted on Ph+ K562 cell line treated with increasing doses of Ima. Western blotting (WB) analysis was conducted for the protein expression of Nanog on K562 cells treated with 5uM Ima. RNA was purified from mononuclear cells of 27 CML patients at diagnosis and after 3 months of TKI treatment. Patients were monitored by RT-qPCR for the expres- sion of the fusion BCR-ABL mRNA. RT-qPCR for the expression of Nanog, was conducted. RT-qPCR results were normalized by the ex- pression of Gus mRNA (Normalized mRNA copy Number: NCN). Re- sults: We observed a significant increase of Nanog mRNA expression in K562 cells treated with 0.5 uM of Imatinib. Moreover, we were also able to observe a significant increase of Nanog protein expression in K562 cells treated with 1-5uM Imatinib by WB. In peripheral blood samples of CML patients at diagnosis, we observed a significant higher mRNA expression of Nanog in No-Optimal Responder compared to Optimal Responder patients (NANOg mRNA: 0.3±0.25 NCN by GUS mRNA vs 0.6±0.7 NCN by GUS mRNA) Conclusions and Summary: These data sug- gest that the expression analysis of Nanog at CML patient baseline, may assist in the early prediction of molecular response in patients treated with TKI. Further studies are ongoing to functionally evaluate whether Nanog is regulated by endogenous or exogenous signals in leukemic cells and evaluate the role of Nanog stemness power in induction of trans- formation of hematopoietic stem cell.
19. Prophylactic central neck lymphadenectomy in high risk patients with T1 or T2 papillary thyroid carcinoma: Is it useful?
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Delogu, D., Pisano, I. P., Pala, C., Pulighe, F., Denti, S., Antonio Giuseppe Maria COSSU, and Trignano, M.
20. Neutrophil to lymphocyte ratio as a predictor of thyroid papillary carcinoma
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Paliogiannis, P., Scognamillo, F., Bellomo, M., Pittalis, M. L., Pisano, I. P., Karligkiotis, A., Bozzo, C., Sotgiu, G., and Federico Attene
21. Mucosa associated lymphoid tissue lymphoma of the thyroid gland: A case report and literature review | MALT linfoma della tiroide: Caso clinico e revisione della letteratura
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Pulighe, F., Paliogiannis, P., Pisano, I. P., Federico Attene, Scognamillo, F., and Trignano, M.
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immune system diseases ,hemic and lymphatic diseases ,MED/18 Chirurgia generale - Abstract
Mucosa associated lymphoid tissue (MALT) lymphomas are low-grade, non-Hodgkin’s B cell lymphomas, mainly occurring in the gastrointestinal tract, but also in other tissues. We describe the management of a patient with hypothyroidism, tracheoesophageal compressive symptoms and chest tightness affected by a thyroid MALT lymphoma. The patient underwent debulking thyroidectomy and temporary tracheostomy in order to reduce dysphonia and dysphagia, followed by adjuvant chemotherapy and subsequently radiation therapy. A CT scan performed at the end of radiotherapy 6 months after surgery revealed remnants of residual tissue from the thyroidectomy without any pathological findings. I linfomi MALT sono dei linfomi non-Hodgkin a cellule B a basso grado che in genere insorgono a livello del tratto gastrointestinale, ma anche in altri tessuti. Descriviamo in questo articolo il management clinico-chirurgico di un paziente con ipotiroidismo, sintomi da compressione tracheo-esofagea e senso di oppressione toracica, affetto da linfoma MALT della tiroide. Il paziente è stato sottoposto a parziale asportazione della massa tiroidea e tracheostomia allo scopo di ridurre i sintomi compressivi ed in seguito a trattamento chemioterapico e radioterapico. L’esame TC effettuato una volta conclusa la radioterapia, circa 6 mesi dopo l’intervento, ha evidenziato gli esiti della tiroidectomia parziale in assenza di altri reperti patologici.
22. Accidental parathyroidectomy as a risk factor for postoperative hypocalcemia in thyroid surgery
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Panagiotis Paliogiannis, Pisano, I. P., Marrosu, A., Pulighe, F., Fara, P., Biddau, C., Sotgiu, G., and Trignano, M.
23. Industrial Production of Proteins with Pichia pastoris-Komagataella phaffii
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Giovanni Davide Barone, Anita Emmerstorfer-Augustin, Antonino Biundo, Isabella Pisano, Paola Coccetti, Valeria Mapelli, Andrea Camattari, Barone, G, Emmerstorfer-Augustin, A, Biundo, A, Pisano, I, Coccetti, P, Mapelli, V, and Camattari, A
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methylotrophic yeast ,applied biotechnology ,bioreactor-based approache ,industrial biotechnology ,Pichia pastori ,protein production ,Molecular Biology ,Biochemistry ,BIO/10 - BIOCHIMICA ,biotechnology ,Komagataella phaffii - Abstract
Since the mid-1960s, methylotrophic yeast Komagataella phaffii (previously described as Pichia pastoris) has received increasing scientific attention. The interest for the industrial production of proteins for different applications (e.g., feed, food additives, detergent, waste treatment processes, and textile) is a well-consolidated scientific topic, and the importance for this approach is rising in the current era of environmental transition in human societies. This review aims to summarize fundamental and specific information in this scientific field. Additionally, an updated description of the relevant products produced with K. phaffii at industrial levels by a variety of companies—describing how the industry has leveraged its key features, from products for the ingredients of meat-free burgers (e.g., IMPOSSIBLE™ FOODS, USA) to diabetes therapeutics (e.g., Biocon, India)—is provided. Furthermore, active patents and the typical workflow for industrial protein production with this strain are reported.
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- 2023
24. Loss of Detection of sgN Precedes Viral Abridged Replication in COVID-19-Affected Patients—A Target for SARS-CoV-2 Propagation
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Veronica Ferrucci, Pasqualino de Antonellis, Fabrizio Quarantelli, Fatemeh Asadzadeh, Francesca Bibbò, Roberto Siciliano, Carmen Sorice, Ida Pisano, Barbara Izzo, Carmela Di Domenico, Angelo Boccia, Maria Vargas, Biancamaria Pierri, Maurizio Viscardi, Sergio Brandi, Giovanna Fusco, Pellegrino Cerino, Livia De Pietro, Ciro Furfaro, Leonardo Antonio Napolitano, Giovanni Paolella, Lidia Festa, Stefania Marzinotto, Maria Concetta Conte, Ivan Gentile, Giuseppe Servillo, Francesco Curcio, Tiziana de Cristofaro, Francesco Broccolo, Ettore Capoluongo, Massimo Zollo, Ferrucci, V., de Antonellis, P., Quarantelli, F., Asadzadeh, F., Bibbo, F., Siciliano, R., Sorice, C., Pisano, I., Izzo, B., Di Domenico, C., Boccia, A., Vargas, M., Pierri, B., Viscardi, M., Brandi, S., Fusco, G., Cerino, P., De Pietro, L., Furfaro, C., Napolitano, L. A., Paolella, G., Festa, L., Marzinotto, S., Conte, M. C., Gentile, I., Servillo, G., Curcio, F., de Cristofaro, T., Broccolo, F., Capoluongo, E., and Zollo, M.
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SgE ,Virus Replication ,Giant Cells ,Sensitivity and Specificity ,Catalysis ,Ribonuclease P ,Viroporin Proteins ,Inorganic Chemistry ,QPCR methods ,Limit of Detection ,SgN ,Nasopharynx ,QPCR method ,SARS-CoV-2 virus particle ,Coronavirus Nucleocapsid Proteins ,Humans ,RNA, Antisense ,Viral ,Physical and Theoretical Chemistry ,Antisense ,Molecular Biology ,Spectroscopy ,SARS-CoV-2 virus particles ,qPCR methods ,2′-O-methyl antisense RNA ,sgN ,sgE ,SARS-CoV-2 ,Organic Chemistry ,COVID-19 ,General Medicine ,Viral Load ,Phosphoproteins ,Computer Science Applications ,HEK293 Cells ,RNA, Viral ,Social Isolation ,RNA - Abstract
The development of prophylactic agents against the SARS-CoV-2 virus is a public health priority in the search for new surrogate markers of active virus replication. Early detection markers are needed to follow disease progression and foresee patient negativization. Subgenomic RNA transcripts (with a focus on sgN) were evaluated in oro/nasopharyngeal swabs from COVID-19-affected patients with an analysis of 315 positive samples using qPCR technology. Cut-off Cq values for sgN (Cq < 33.15) and sgE (Cq < 34.06) showed correlations to high viral loads. The specific loss of sgN in home-isolated and hospitalized COVID-19-positive patients indicated negativization of patient condition, 3–7 days from the first swab, respectively. A new detection kit for sgN, gene E, gene ORF1ab, and gene RNAse P was developed recently. In addition, in vitro studies have shown that 2’-O-methyl antisense RNA (related to the sgN sequence) can impair SARS-CoV-2 N protein synthesis, viral replication, and syncytia formation in human cells (i.e., HEK-293T cells overexpressing ACE2) upon infection with VOC Alpha (B.1.1.7)-SARS-CoV-2 variant, defining the use that this procedure might have for future therapeutic actions against SARS-CoV-2.
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- 2022
25. Long-chain polyphosphates impair SARS-CoV-2 infection and replication
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Sergio Brandi, Bianca Maria Pierri, Giorgia Borriello, Ettore Capoluongo, Barbara Izzo, Giuseppe Castaldo, Angelo Boccia, Hong-Yeoul Kim, Lorenzo Chiariotti, Giovanna Fusco, Rosa Della Monica, Dae-Young Kong, Ilaria Iacobucci, Maurizio Viscardi, Margherita Passariello, Roberto Siciliano, Stefano Pascarella, Claudia Tiberio, Camilla Anastasio, Giovanni Paolella, Fatemeh Asadzadeh, Jae-Ho Cheong, Pellegrino Cerino, Luigi Atripaldi, Marika Comegna, Martina Bianchi, Maria Chiara Monti, Fabrizio Quarantelli, Laura Marrone, Kyong-Seop Yun, Ida Pisano, Massimo Zollo, Giuseppina Criscuolo, Claudia De Lorenzo, Veronica Ferrucci, Ferrucci, V., Kong, D. -Y., Asadzadeh, F., Marrone, L., Boccia, A., Siciliano, R., Criscuolo, G., Anastasio, C., Quarantelli, F., Comegna, M., Pisano, I., Passariello, M., Iacobucci, I., della Monica, R., Izzo, B., Cerino, P., Fusco, G., Viscardi, M., Brandi, S., Pierri, B. M., Borriello, G., Tiberio, C., Atripaldi, L., Bianchi, M., Paolella, G., Capoluongo, E., Castaldo, G., Chiariotti, L., Monti, M., de Lorenzo, C., Yun, K. -S., Pascarella, S., Cheong, J. -H., Kim, H. -Y., and Zollo, M.
- Subjects
0301 basic medicine ,viruses ,Virus Replication ,Biochemistry ,chemistry.chemical_compound ,Host Microbial Interaction ,0302 clinical medicine ,HEK293 Cell ,Polyphosphates ,RNA polymerase ,Chlorocebus aethiops ,Proteolysi ,skin and connective tissue diseases ,Peptide sequence ,Protein Interaction Domains and Motif ,Research Articles ,Subgenomic mRNA ,Caco-2 Cell ,Coronavirus RNA-Dependent RNA Polymerase ,Chemistry ,Molecular Docking Simulation ,030220 oncology & carcinogenesis ,Cytokines ,RNA, Viral ,Angiotensin-Converting Enzyme 2 ,inorganic polyphosphate ,Human ,Signal Transduction ,Research Article ,Proteasome Endopeptidase Complex ,In Vitro Techniques ,Chlorocebus aethiop ,Antiviral Agents ,Models, Biological ,Virus ,Microbiology ,03 medical and health sciences ,Viral entry ,Virology ,Polyphosphate ,Administration, Inhalation ,Animals ,Humans ,Protein Interaction Domains and Motifs ,Amino Acid Sequence ,Cytokine ,Molecular Biology ,Vero Cells ,Antiviral Agent ,Host Microbial Interactions ,Sequence Homology, Amino Acid ,Animal ,In Vitro Technique ,SARS-CoV-2 ,Nebulizers and Vaporizers ,fungi ,RNA ,COVID-19 ,Cell Biology ,STKE Research Articles ,respiratory tract diseases ,COVID-19 Drug Treatment ,body regions ,Coronavirus ,030104 developmental biology ,HEK293 Cells ,Viral replication ,Proteolysis ,Vero Cell ,Vero cell ,Sars-CoV-2 ,Caco-2 Cells ,Nebulizers and Vaporizer - Abstract
Long-chain polyphosphates inhibit SARS-CoV-2 infection by targeting a host receptor and a viral RNA polymerase., Polyphosphates versus SARS-CoV-2 Long-chain, inorganic polyphosphates (polyPs), which are found in many cells in the blood, have cytoprotective and antiviral activities, particularly against HIV-1 infection. Ferrucci et al. tested the effects of polyPs of various lengths on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in vitro. Molecular docking and binding analyses showed that polyPs bound to the host receptor ACE2, which facilitates viral entry, and a viral RNA polymerase required for replication. Both proteins underwent proteasomal degradation in cells incubated with polyP120, the optimal species tested, resulting in inhibition of SARS-CoV-2 replication and a reduced inflammatory response. Given that polyPs have low toxicity, these results suggest that their potential therapeutic use should be further explored., Inorganic polyphosphates (polyPs) are linear polymers composed of repeated phosphate (PO43−) units linked together by multiple high-energy phosphoanhydride bonds. In addition to being a source of energy, polyPs have cytoprotective and antiviral activities. Here, we investigated the antiviral activities of long-chain polyPs against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In molecular docking analyses, polyPs interacted with several conserved amino acid residues in angiotensin-converting enzyme 2 (ACE2), the host receptor that facilitates virus entry, and in viral RNA-dependent RNA polymerase (RdRp). ELISA and limited proteolysis assays using nano– LC-MS/MS mapped polyP120 binding to ACE2, and site-directed mutagenesis confirmed interactions between ACE2 and SARS-CoV-2 RdRp and identified the specific amino acid residues involved. PolyP120 enhanced the proteasomal degradation of both ACE2 and RdRp, thus impairing replication of the British B.1.1.7 SARS-CoV-2 variant. We thus tested polyPs for functional interactions with the virus in SARS-CoV-2–infected Vero E6 and Caco2 cells and in primary human nasal epithelial cells. Delivery of a nebulized form of polyP120 reduced the amounts of viral positive-sense genomic and subgenomic RNAs, of RNA transcripts encoding proinflammatory cytokines, and of viral structural proteins, thereby presenting SARS-CoV-2 infection in cells in vitro.
- Published
- 2021
26. KRAS-regulated glutamine metabolism requires UCP2-mediated aspartate transport to support pancreatic cancer growth
- Author
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Vittoria Rago, Rocco Malivindi, Giuseppe E. De Benedetto, Isabella Pisano, Giuseppe Fiermonte, Francesco M. Lasorsa, Carmela Piazzolla, Christopher L. Riley, Angelo Vozza, Stephan J. Reshkin, Wolfgang Sommergruber, Gennaro Agrimi, Francesca Pezzuto, Rosa Angela Cardone, Simona N. Barile, Yuan Li, Pasquale Scarcia, Carlo M.T. Marobbio, Maria C. Vegliante, Ruggiero Gorgoglione, Edward M. Mills, Luigi Palmieri, Loredana Capobianco, Deborah Fratantonio, Susanna Raho, Maria Raffaella Greco, Francesco De Leonardis, Vincenza Dolce, Raho, Susanna, Capobianco, Loredana, Malivindi, Rocco, Vozza, Angelo, Piazzolla, Carmela, De Leonardis, Francesco, Gorgoglione, Ruggiero, Scarcia, Pasquale, Pezzuto, Francesca, Agrimi, Gennaro, Barile, Simona N., Pisano, Isabella, Reshkin, Stephan J., Greco, Maria R., Cardone, Rosa A., Rago, Vittoria, Li, Yuan, Marobbio, Carlo M. T., Sommergruber, Wolfgang, Riley, Christopher L., Lasorsa, Francesco M., Mills, Edward, Vegliante, Maria C., De Benedetto, Giuseppe E., Fratantonio, Deborah, Palmieri, Luigi, Dolce &, Vincenza, Fiermonte, Giuseppe, Raho, S., Capobianco, L., Malivindi, R., Vozza, A., Piazzolla, C., De Leonardis, F., Gorgoglione, R., Scarcia, P., Pezzuto, F., Agrimi, G., Barile, S. N., Pisano, I., Reshkin, S. J., Greco, M. R., Cardone, R. A., Rago, V., Li, Y., Marobbio, C. M. T., Sommergruber, W., Riley, C. L., Lasorsa, F. M., Mills, E., Vegliante, M. C., De Benedetto, G. E., Fratantonio, D., Palmieri, L., Dolce, V., and Fiermonte, G.
- Subjects
endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,Glutamine ,Biological Transport, Active ,Mice, SCID ,Mitochondrion ,Proto-Oncogene Proteins p21(ras) ,chemistry.chemical_compound ,Mice ,Cytosol ,Physiology (medical) ,Cell Line, Tumor ,Internal Medicine ,Animals ,Humans ,Uncoupling Protein 2 ,oncogenic Kras, mitochondrial carrier, UCP2, human pancreatic ductal adenocarcinoma (PDAC) ,chemistry.chemical_classification ,Reactive oxygen species ,Aspartic Acid ,Glutaminolysis ,Cell growth ,Animal ,Pancreatic Neoplasm ,Cell Biology ,Xenograft Model Antitumor Assays ,Cell biology ,Mitochondria ,Pancreatic Neoplasms ,chemistry ,Glutathione disulfide ,Female ,Aspartate transport ,Reactive Oxygen Species ,Reactive Oxygen Specie ,Oxidation-Reduction ,NADP ,Carcinoma, Pancreatic Ductal ,Human - Abstract
The oncogenic KRAS mutation has a critical role in the initiation of human pancreatic ductal adenocarcinoma (PDAC) since it rewires glutamine metabolism to increase reduced nicotinamide adenine dinucleotide phosphate (NADPH) production, balancing cellular redox homeostasis with macromolecular synthesis1,2. Mitochondrial glutamine-derived aspartate must be transported into the cytosol to generate metabolic precursors for NADPH production2. The mitochondrial transporter responsible for this aspartate efflux has remained elusive. Here, we show that mitochondrial uncoupling protein 2 (UCP2) catalyses this transport and promotes tumour growth. UCP2-silenced KRASmut cell lines display decreased glutaminolysis, lower NADPH/NADP+ and glutathione/glutathione disulfide ratios and higher reactive oxygen species levels compared to wild-type counterparts. UCP2 silencing reduces glutaminolysis also in KRASWT PDAC cells but does not affect their redox homeostasis or proliferation rates. In vitro and in vivo, UCP2 silencing strongly suppresses KRASmut PDAC cell growth. Collectively, these results demonstrate that UCP2 plays a vital role in PDAC, since its aspartate transport activity connects the mitochondrial and cytosolic reactions necessary for KRASmut rewired glutamine metabolism2, and thus it should be considered a key metabolic target for the treatment of this refractory tumour. UCP2 is shown in yeast and mammalian cells to transport aspartate out of mitochondria, thus enabling KRAS-mutated pancreatic ductal adenocarcinoma cells to perform glutaminolysis to support cancer growth.
- Published
- 2020
27. Syngas Derived from Lignocellulosic Biomass Gasification as an Alternative Resource for Innovative Bioprocesses
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Isabella Pisano, Giacobbe Braccio, Roberto Albergo, Cosetta Ciliberti, Gennaro Agrimi, Antonino Biundo, Isabella De Bari, Ciliberti, C., Biundo, A., Albergo, R., Agrimi, G., Braccio, G., de Bari, I., and Pisano, I.
- Subjects
Commodity chemicals ,Bioconversion ,020209 energy ,gasification ,Lignocellulosic biomass ,Bioengineering ,02 engineering and technology ,010501 environmental sciences ,lcsh:Chemical technology ,01 natural sciences ,lcsh:Chemistry ,chemistry.chemical_compound ,Syngasfermentation ,0202 electrical engineering, electronic engineering, information engineering ,Chemical Engineering (miscellaneous) ,lcsh:TP1-1185 ,Lignocellulosicbiomass ,lignocellulosic biomass ,0105 earth and related environmental sciences ,biorefinery ,syngas fermentation ,Process Chemistry and Technology ,Butanol ,Wood–Ljungdahlpathway ,Pulp and paper industry ,Biorefinery ,lcsh:QD1-999 ,chemistry ,Syngas fermentation ,Wood–Ljungdahl pathway ,Environmental science ,Syngas - Abstract
A hybrid system based on lignocellulosic biomass gasification and syngas fermentation represents a second-generation biorefinery approach that is currently in the development phase. Lignocellulosic biomass can be gasified to produce syngas, which is a gas mixture consisting mainly of H2, CO, and CO2. The major challenge of biomass gasification is the syngas’s final quality. Consequently, the development of effective syngas clean-up technologies has gained increased interest in recent years. Furthermore, the bioconversion of syngas components has been intensively studied using acetogenic bacteria and their Wood–Ljungdahl pathway to produce, among others, acetate, ethanol, butyrate, butanol, caproate, hexanol, 2,3-butanediol, and lactate. Nowadays, syngas fermentation appears to be a promising alternative for producing commodity chemicals in comparison to fossil-based processes. Research studies on syngas fermentation have been focused on process design and optimization, investigating the medium composition, operating parameters, and bioreactor design. Moreover, metabolic engineering efforts have been made to develop genetically modified strains with improved production. In 2018, for the first time, a syngas fermentation pilot plant from biomass gasification was built by LanzaTech Inc. in cooperation with Aemetis, Inc. Future research will focus on coupling syngas fermentation with additional bioprocesses and/or on identifying new non-acetogenic microorganisms to produce high-value chemicals beyond acetate and ethanol.
- Published
- 2020
28. Metastatic group 3 medulloblastoma is driven by PRUNE1 targeting NME1-TGF-β-OTX2-SNAIL via PTEN inhibition
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Roberto Fattorusso, Sara Gargiulo, Francesco Salvatore, Donatella Diana, Iolanda Boffa, Matteo Gramanzini, Antonella Virgilio, Maria Elena Errico, William A. Weiss, Aldo Galeone, Louis Chesler, Valeria D'Argenio, Valentina Del Monaco, Angela Mastronuzzi, Livia Garzia, Iolanda Scognamiglio, Felice Tirone, Pasqualino De Antonellis, Emilia Pedone, Daniel Picard, Arturo Brunetti, Marianeve Carotenuto, Michael D. Taylor, Olivier Delattre, Laura Danielson, Antonio Verrico, Fatemeh Asadzadeh, Marc Remke, Fredrik J. Swartling, Donatella Montanaro, Luigi Navas, Craig Daniels, Veronica Ferrucci, Lucia Quaglietta, Ida Pisano, Massimo Zollo, Lucia Liguori, Felice Giangaspero, Francesco Paolo Pennino, Giuseppe Cinalli, Vittoria Donofrio, Ferrucci, V, de Antonellis, P, Pennino, FRANCESCO PAOLO, Asadzadeh, F, Virgilio, A, Montanaro, D, Galeone, A, Boffa, I, Pisano, I, Scognamiglio, I, Navas, L, Diana, D, Pedone, E, Gargiulo, S, Gramanzini, M, Brunetti, A, Danielson, L, Carotenuto, M, Liguori, L, Verrico, A, Quaglietta, L, Errico, Me, Del Monaco, V, D'Argenio, V, Tirone, F, Mastronuzzi, A, Donofrio, V, Giangaspero, F, Picard, D, Remke, M, Garzia, L, Daniels, C, Delattre, O, Swartling, Fj, Weiss, Wa, Salvatore, F, Fattorusso, R, Chesler, L, Taylor, Md, Cinalli, G, Zollo, M., Ferrucci, Veronica, de Antonellis, Pasqualino, Pennino, Francesco Paolo, Asadzadeh, Fatemeh, Virgilio, Antonella, Montanaro, Donatella, Galeone, Aldo, Boffa, Iolanda, Pisano, Ida, Scognamiglio, Iolanda, Navas, Luigi, Diana, Donatella, Pedone, Emilia, Gargiulo, Sara, Gramanzini, Matteo, Brunetti, Arturo, Danielson, Laura, Carotenuto, Marianeve, Liguori, Lucia, Verrico, Antonio, Quaglietta, Lucia, Errico, Maria Elena, Del Monaco, Valentina, D'Argenio, Valeria, Tirone, Felice, Mastronuzzi, Angela, Donofrio, Vittoria, Giangaspero, Felice, Picard, Daniel, Remke, Marc, Garzia, Livia, Daniels, Craig, Delattre, Olivier, Swartling, Fredrik J, Weiss, William A, Salvatore, Francesco, Fattorusso, Roberto, Chesler, Loui, Taylor, Michael D, Cinalli, Giuseppe, and Zollo, Massimo
- Subjects
0301 basic medicine ,Male ,Models, Molecular ,Mice ,Cell Movement ,Transforming Growth Factor beta ,molecular genetic ,Gene Regulatory Networks ,Neoplasm Metastasis ,Child ,Regulation of gene expression ,metastatic CNS tumour ,Mice, Inbred BALB C ,biology ,Prune ,Hedgehog signaling pathway ,Gene Expression Regulation, Neoplastic ,Child, Preschool ,oncology ,Female ,Signal transduction ,Signal Transduction ,cerebellum ,Adolescent ,Pyrimidinones ,medulloblastoma ,03 medical and health sciences ,Downregulation and upregulation ,Cell Line, Tumor ,medicine ,metastasis ,PTEN ,Animals ,Humans ,groups 3 and 4 medulloblastoma ,paediatric ,PRUNE1 ,NME1-TGF-β-OTX2-SNAIL ,PTEN inhibition ,Cerebellar Neoplasms ,Cell Proliferation ,Medulloblastoma ,Cancer och onkologi ,genetic network ,PTEN Phosphohydrolase ,Infant ,medicine.disease ,Phosphoric Monoester Hydrolases ,030104 developmental biology ,Cancer and Oncology ,SNAI1 ,molecular genetics ,Cancer research ,biology.protein ,Neurology (clinical) ,Snail Family Transcription Factors ,Carrier Proteins ,Transforming growth factor - Abstract
Genetic modifications during development of paediatric groups 3 and 4 medulloblastoma are responsible for their highly metastatic properties and poor patient survival rates. PRUNE1 is highly expressed in metastatic medulloblastoma group 3, which is characterized by TGF-β signalling activation, c-MYC amplification, and OTX2 expression. We describe the process of activation of the PRUNE1 signalling pathway that includes its binding to NME1, TGF-β activation, OTX2 upregulation, SNAIL (SNAI1) upregulation, and PTEN inhibition. The newly identified small molecule pyrimido-pyrimidine derivative AA7.1 enhances PRUNE1 degradation, inhibits this activation network, and augments PTEN expression. Both AA7.1 and a competitive permeable peptide that impairs PRUNE1/NME1 complex formation, impair tumour growth and metastatic dissemination in orthotopic xenograft models with a metastatic medulloblastoma group 3 cell line (D425-Med cells). Using whole exome sequencing technology in metastatic medulloblastoma primary tumour cells, we also define 23 common 'non-synonymous homozygous' deleterious gene variants as part of the protein molecular network of relevance for metastatic processes. This PRUNE1/TGF-β/OTX2/PTEN axis, together with the medulloblastoma-driver mutations, is of relevance for future rational and targeted therapies for metastatic medulloblastoma group 3. Genetic modifications during development of paediatric groups 3 and 4 medulloblastoma are responsible for their highly metastatic properties and poor patient survival rates. PRUNE1 is highly expressed in metastatic medulloblastoma group 3, which is characterized by TGF-β signalling activation, c-MYC amplification, and OTX2 expression. We describe the process of activation of the PRUNE1 signalling pathway that includes its binding to NME1, TGF-β activation, OTX2 upregulation, SNAIL (SNAI1) upregulation, and PTEN inhibition. The newly identified small molecule pyrimido-pyrimidine derivative AA7.1 enhances PRUNE1 degradation, inhibits this activation network, and augments PTEN expression. Both AA7.1 and a competitive permeable peptide that impairs PRUNE1/NME1 complex formation, impair tumour growth and metastatic dissemination in orthotopic xenograft models with a metastatic medulloblastoma group 3 cell line (D425-Med cells). Using whole exome sequencing technology in metastatic medulloblastoma primary tumour cells, we also define 23 common 'non-synonymous homozygous' deleterious gene variants as part of the protein molecular network of relevance for metastatic processes. This PRUNE1/TGF-β/OTX2/PTEN axis, together with the medulloblastoma-driver mutations, is of relevance for future rational and targeted therapies for metastatic medulloblastoma group 3. 10.1093/brain/awy039-video1 awy039media1 5742053534001
- Published
- 2017
29. Role of lenalidomide in the management of myelodysplastic syndromes with del(5q) associated with pure red cell aplasia (PRCA)
- Author
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Lucio Catalano, Marco Picardi, Antonio M. Risitano, Fabrizio Pane, Claudio Cerchione, Fiorella Alfinito, Ida Pisano, Simona Avilia, Giuseppe Cerciello, Cerchione, C, Catalano, L, Cerciello, G, Avilia, S, Picardi, Marco, Risitano, Am, Pisano, I, Alfinito, F, and Pane, Fabrizio
- Subjects
Oncology ,medicine.medical_specialty ,Hematology ,business.industry ,Myelodysplastic syndromes ,lenalidomide ,Pure red cell aplasia ,General Medicine ,medicine.disease ,myelodysplastic syndromes ,Internal medicine ,medicine ,business ,Lenalidomide ,medicine.drug - Published
- 2015
30. Deletion or Overexpression of Mitochondrial NAD(+) Carriers in Saccharomyces cerevisiae Alters Cellular NAD and ATP Contents and Affects Mitochondrial Metabolism and the Rate of Glycolysis
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Gianni Frascotti, Luigi Palmieri, Isabella Pisano, Luca Brambilla, Gennaro Agrimi, Marina Vai, Danilo Porro, Agrimi, G, Brambilla, L, Frascotti, G, Pisano, I, Porro, D, Vai, M, and Palmieri, L
- Subjects
Saccharomyces cerevisiae Proteins ,Bioenergetics ,Physiology ,Saccharomyces cerevisiae ,Oxidative phosphorylation ,Mitochondrion ,Biology ,Applied Microbiology and Biotechnology ,Mitochondrial Proteins ,chemistry.chemical_compound ,Adenosine Triphosphate ,Glycolysis ,Sequence Deletion ,Ecology ,NAD ,CHIM/11 - CHIMICA E BIOTECNOLOGIA DELLE FERMENTAZIONI ,Culture Media ,Glucose ,Glycerol-3-phosphate dehydrogenase ,Biochemistry ,chemistry ,Fermentation ,Yeast, NAD transporters, mitochondria, NAD homeostasis ,Crabtree effect ,NAD+ kinase ,Carrier Proteins ,Oxidation-Reduction ,Adenosine triphosphate ,Food Science ,Biotechnology - Abstract
The modification of enzyme cofactor concentrations can be used as a method for both studying and engineering metabolism. We varied Saccharomyces cerevisiae mitochondrial NAD levels by altering expression of its specific mitochondrial carriers. Changes in mitochondrial NAD levels affected the overall cellular concentration of this coenzyme and the cellular metabolism. In batch culture, a strain with a severe NAD depletion in mitochondria succeeded in growing, albeit at a low rate, on fully respiratory media. Although the strain increased the efficiency of its oxidative phosphorylation, the ATP concentration was low. Under the same growth conditions, a strain with a mitochondrial NAD concentration higher than that of the wild type similarly displayed a low cellular ATP level, but its growth rate was not affected. In chemostat cultures, when cellular metabolism was fully respiratory, both mutants showed low biomass yields, indicative of impaired energetic efficiency. The two mutants increased their glycolytic fluxes, and as a consequence, the Crabtree effect was triggered at lower dilution rates. Strikingly, the mutants switched from a fully respiratory metabolism to a respirofermentative one at the same specific glucose flux as that of the wild type. This result seems to indicate that the specific glucose uptake rate and/or glycolytic flux should be considered one of the most important independent variables for establishing the long-term Crabtree effect. In cells growing under oxidative conditions, bioenergetic efficiency was affected by both low and high mitochondrial NAD availability, which suggests the existence of a critical mitochondrial NAD concentration in order to achieve optimal mitochondrial functionality.
- Published
- 2011
31. Systematic screening for the biocatalytic hydration of fatty acids from different oily substrates by Elizabethkingia meningoseptica oleate hydratase through a Design-of-experiments approach.
- Author
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Biundo A, Lima S, Ciaccia M, Ciliberti C, Serpico A, Agrimi G, Scargiali F, and Pisano I
- Subjects
- Oleic Acid metabolism, Flavobacteriaceae metabolism, Flavobacteriaceae enzymology, Hydro-Lyases metabolism, Fatty Acids metabolism, Olive Oil metabolism, Olive Oil chemistry, Lipase metabolism, Sunflower Oil metabolism, Triglycerides metabolism, Wastewater chemistry, Wastewater microbiology, Saccharomycetales, Escherichia coli metabolism, Escherichia coli genetics, Plant Oils metabolism, Biocatalysis
- Abstract
The edible plant oils production is associated with the release of different types of by-products. The latter represent cheap and available substrates to produce valuable compounds, such as flavours and fragrances, biologically active compounds and bio-based polymers. Elizabethkingia meningoseptica Oleate hydratases (Em_OhyA) can selectively catalyze the conversion of unsaturated fatty acids, specifically oleic acid, into hydroxy fatty acids, which find different industrial applications. In this study, Design-of-experiment (DoE) strategy was used to screen and identify conditions for reaching high yields in the reaction carried out by Escherichia coli whole-cell carrying the recombinant enzyme Em_OhyA using Waste Cooking Oils (WCO)-derived free fatty acids (FFA) as substrate. The identified reaction conditions for high oleic acid conversion were also tested on untreated triglycerides-containing substrates, such as pomace oil, sunflower oil, olive oil and oil mill wastewater (OMW), combining the triglyceride hydrolysis by the lipase from Candida rugosa and the E. coli whole-cell containing Em_OhyA for the production of hydroxy fatty acids. When WCO, sunflower oil and OMW were used as substrate, the one-pot bioconversion led to an increase of oleic acid conversion compared to the standard reaction. This work highlights the efficiency of the DoE approach to screen and identify conditions for an enzymatic reaction for the production of industrially-relevant products., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2024
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32. Keratinous and corneous-based products towards circular bioeconomy: A research review.
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Barone GD, Tagliaro I, Oliver-Simancas R, Radice M, Kalossaka LM, Mattei M, Biundo A, Pisano I, and Jiménez-Quero A
- Abstract
Keratins and corneous proteins are key components of biomaterials used in a wide range of applications and are potential substitutes for petrochemical-based products. Horns, hooves, feathers, claws, and similar animal tissues are abundant sources of α-keratin and corneous β-proteins, which are by-products of the food industry. Their close association with the meat industry raises environmental and ethical concerns regarding their disposal. To promote an eco-friendly and circular use of these materials in novel applications, efforts have focused on recovering these residues to develop sustainable, non-animal-related, affordable, and scalable procedures. Here, we review and examine biotechnological methods for extracting and expressing α-keratins and corneous β-proteins in microorganisms. This review highlights consolidated research trends in biomaterials, medical devices, food supplements, and packaging, demonstrating the keratin industry's potential to create innovative value-added products. Additionally, it analyzes the state of the art of related intellectual property and market size to underscore the potential within a circular bioeconomic model., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)
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- 2024
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33. Lack of Mitochondrial DNA Provides Metabolic Advantage in Yeast Osmoadaptation.
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Di Noia MA, Ocheja OB, Scarcia P, Pisano I, Messina E, Agrimi G, Palmieri L, and Guaragnella N
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- Mitochondria metabolism, Mitochondria genetics, Adaptation, Physiological genetics, Oxidative Stress genetics, Glycerol metabolism, Ethidium metabolism, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism, DNA, Mitochondrial genetics, DNA, Mitochondrial metabolism, Saccharomyces cerevisiae Proteins metabolism, Saccharomyces cerevisiae Proteins genetics
- Abstract
Alterations in mitochondrial function have been linked to a variety of cellular and organismal stress responses including apoptosis, aging, neurodegeneration and tumorigenesis. However, adaptation to mitochondrial dysfunction can occur through the activation of survival pathways, whose mechanisms are still poorly understood. The yeast Saccharomyces cerevisiae is an invaluable model organism for studying how mitochondrial dysfunction can affect stress response and adaptation processes. In this study, we analyzed and compared in the absence and in the presence of osmostress wild-type cells with two models of cells lacking mitochondrial DNA: ethidium bromide-treated cells (ρ
0 ) and cells lacking the mitochondrial pyrimidine nucleotide transporter RIM2 (Δ RIM2 ). Our results revealed that the lack of mitochondrial DNA provides an advantage in the kinetics of stress response. Additionally, wild-type cells exhibited higher osmosensitivity in the presence of respiratory metabolism. Mitochondrial mutants showed increased glycerol levels, required in the short-term response of yeast osmoadaptation, and prolonged oxidative stress. The involvement of the mitochondrial retrograde signaling in osmoadaptation has been previously demonstrated. The expression of CIT2 , encoding the peroxisomal isoform of citrate synthase and whose up-regulation is prototypical of RTG pathway activation, appeared to be increased in the mutants. Interestingly, selected TCA cycle genes, CIT1 and ACO1 , whose expression depends on RTG signaling upon stress, showed a different regulation in ρ0 and Δ RIM2 cells. These data suggest that osmoadaptation can occur through different mechanisms in the presence of mitochondrial defects and will allow us to gain insight into the relationships among metabolism, mitochondria-mediated stress response, and cell adaptation., Competing Interests: The authors declare no conflicts of interest.- Published
- 2024
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34. Hydrothermal carbonization of milk/dairy processing sludge: Fate of plant nutrients.
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Kwapinska M, Pisano I, and Leahy JJ
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- Animals, Nutrients, Phosphorus, Milk, Sewage
- Abstract
Dairy processing sludge (DPS) is a byproduct generated in wastewater treatment plants located in dairy (milk) processing companies (waste activated sludge). DPS presents challenges in terms of its management (as biosolids) due to its high moisture content, prolonged storage required, uncontrolled nutrient loss and accumulation of certain substances in soil in the proximity of dairy companies. This study investigates the potential of hydrothermal carbonization (HTC) for recovery of nutrients in the form of solid hydrochar (biochar) produced from DPS originating from four different dairy processing companies. The HTC tests were carried out at 160 °C, 180 °C, 200 °C and 220 °C, and a residence time of 1h. The elemental properties of hydrochars (biochars), the content of primary and secondary nutrients, as well as contaminants were examined. The transformation of phosphorus in DPS during HTC was investigated. The fraction of plant available phosphorus was determined. The properties of hydrochar (biochar) were compared against the European Union Fertilizing Products Regulation. The findings of this study demonstrate that the content of nutrient in hydrochars (biochars) meet the requirements for organo-mineral fertilizer with nitrogen and phosphorus as the declared nutrients (13.9-26.7%). Further research on plant growth and field tests are needed to fully assess the agronomic potential of HTC hydrochar (biochar)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2023
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35. Genetic inactivation of the Carnitine/Acetyl-Carnitine mitochondrial carrier of Yarrowia lipolytica leads to enhanced odd-chain fatty acid production.
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Messina E, de Souza CP, Cappella C, Barile SN, Scarcia P, Pisano I, Palmieri L, Nicaud JM, and Agrimi G
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- Acetyl Coenzyme A metabolism, Acetylcarnitine metabolism, Fatty Acids metabolism, Propionates metabolism, Mitochondria metabolism, Metabolic Engineering, Carnitine metabolism, Yarrowia genetics, Yarrowia metabolism
- Abstract
Background: Mitochondrial carriers (MCs) can deeply affect the intracellular flux distribution of metabolic pathways. The manipulation of their expression level, to redirect the flux toward the production of a molecule of interest, is an attractive target for the metabolic engineering of eukaryotic microorganisms. The non-conventional yeast Yarrowia lipolytica is able to use a wide range of substrates. As oleaginous yeast, it directs most of the acetyl-CoA therefrom generated towards the synthesis of lipids, which occurs in the cytoplasm. Among them, the odd-chain fatty acids (OCFAs) are promising microbial-based compounds with several applications in the medical, cosmetic, chemical and agricultural industries., Results: In this study, we have identified the MC involved in the Carnitine/Acetyl-Carnitine shuttle in Y. lipolytica, YlCrc1. The Y. lipolytica Ylcrc1 knock-out strain failed to grow on ethanol, acetate and oleic acid, demonstrating the fundamental role of this MC in the transport of acetyl-CoA from peroxisomes and cytoplasm into mitochondria. A metabolic engineering strategy involving the deletion of YlCRC1, and the recombinant expression of propionyl-CoA transferase from Ralstonia eutropha (RePCT), improved propionate utilization and its conversion into OCFAs. These genetic modifications and a lipogenic medium supplemented with glucose and propionate as the sole carbon sources, led to enhanced accumulation of OCFAs in Y. lipolytica., Conclusions: The Carnitine/Acetyl-Carnitine shuttle of Y. lipolytica involving YlCrc1, is the sole pathway for transporting peroxisomal or cytosolic acetyl-CoA to mitochondria. Manipulation of this carrier can be a promising target for metabolic engineering approaches involving cytosolic acetyl-CoA, as demonstrated by the effect of YlCRC1 deletion on OCFAs synthesis., (© 2023. The Author(s).)
- Published
- 2023
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36. Inactivation of HAP4 Accelerates RTG -Dependent Osmoadaptation in Saccharomyces cerevisiae .
- Author
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Di Noia MA, Scarcia P, Agrimi G, Ocheja OB, Wahid E, Pisano I, Paradies E, Palmieri L, Guaragnella C, and Guaragnella N
- Subjects
- Citric Acid Cycle genetics, Citrate (si)-Synthase metabolism, Signal Transduction, Gene Expression Regulation, Fungal, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism, Saccharomyces cerevisiae Proteins genetics, Saccharomyces cerevisiae Proteins metabolism
- Abstract
Mitochondrial RTG (an acronym for ReTroGrade) signaling plays a cytoprotective role under various intracellular or environmental stresses. We have previously shown its contribution to osmoadaptation and capacity to sustain mitochondrial respiration in yeast. Here, we studied the interplay between RTG2 , the main positive regulator of the RTG pathway, and HAP4 , encoding the catalytic subunit of the Hap2-5 complex required for the expression of many mitochondrial proteins that function in the tricarboxylic acid (TCA) cycle and electron transport, upon osmotic stress. Cell growth features, mitochondrial respiratory competence, retrograde signaling activation, and TCA cycle gene expression were comparatively evaluated in wild type and mutant cells in the presence and in the absence of salt stress. We showed that the inactivation of HAP4 improved the kinetics of osmoadaptation by eliciting both the activation of retrograde signaling and the upregulation of three TCA cycle genes: citrate synthase 1 ( CIT1 ), aconitase 1 ( ACO1 ), and isocitrate dehydrogenase 1 ( IDH1 ). Interestingly, their increased expression was mostly dependent on RTG2 . Impaired respiratory competence in the HAP4 mutant does not affect its faster adaptive response to stress. These findings indicate that the involvement of the RTG pathway in osmostress is fostered in a cellular context of constitutively reduced respiratory capacity. Moreover, it is evident that the RTG pathway mediates peroxisomes-mitochondria communication by modulating the metabolic function of mitochondria in osmoadaptation.
- Published
- 2023
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37. Industrial Production of Proteins with Pichia pastoris - Komagataella phaffii .
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Barone GD, Emmerstorfer-Augustin A, Biundo A, Pisano I, Coccetti P, Mapelli V, and Camattari A
- Subjects
- Humans, Yeasts, Recombinant Proteins genetics, Recombinant Proteins metabolism, Pichia genetics, Pichia metabolism, Saccharomycetales
- Abstract
Since the mid-1960s, methylotrophic yeast Komagataella phaffii (previously described as Pichia pastoris ) has received increasing scientific attention. The interest for the industrial production of proteins for different applications (e.g., feed, food additives, detergent, waste treatment processes, and textile) is a well-consolidated scientific topic, and the importance for this approach is rising in the current era of environmental transition in human societies. This review aims to summarize fundamental and specific information in this scientific field. Additionally, an updated description of the relevant products produced with K . phaffii at industrial levels by a variety of companies-describing how the industry has leveraged its key features, from products for the ingredients of meat-free burgers (e.g., IMPOSSIBLE™ FOODS, USA) to diabetes therapeutics (e.g., Biocon, India)-is provided. Furthermore, active patents and the typical workflow for industrial protein production with this strain are reported.
- Published
- 2023
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38. Regio- and stereoselective biocatalytic hydration of fatty acids from waste cooking oils en route to hydroxy fatty acids and bio-based polyesters.
- Author
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Biundo A, Stamm A, Gorgoglione R, Syrén PO, Curia S, Hauer B, Capriati V, Vitale P, Perna F, Agrimi G, and Pisano I
- Subjects
- Oils, Biofuels, Fatty Acids, Unsaturated, Cooking, Esters, Fatty Acids, Polyesters
- Abstract
The development of biorefinery approaches is of great relevance for the sustainable production of valuable compounds. In accordance with circular economy principles, waste cooking oils (WCOs) are renewable resources and biorefinery feedstocks, which contribute to a reduced impact on the environment. Frequently, this waste is wrongly disposed of into municipal sewage systems, thereby creating problems for the environment and increasing treatment costs in wastewater treatment plants. In this study, regenerated WCOs, which were intended for the production of biofuels, were transformed through a chemo-enzymatic approach to produce hydroxy fatty acids, which were further used in polycondensation reaction for polyester production. Escherichia coli whole cell biocatalyst containing the recombinantly produced Elizabethkingia meningoseptica Oleate hydratase (Em_OhyA) was used for the biocatalytic hydration of crude WCOs-derived unsaturated free fatty acids for the production of hydroxy fatty acids. Further hydrogenation reaction and methylation of the crude mixture allowed the production of (R)- 10-hydroxystearic acid methyl ester that was further purified with a high purity (> 90%), at gram scale. The purified (R)- 10-hydroxystearic acid methyl ester was polymerized through a polycondensation reaction to produce the corresponding polyester. This work highlights the potential of waste products to obtain bio-based hydroxy fatty acids and polyesters through a biorefinery approach., Competing Interests: Declaration of Competing Interest All contributing authors declare that they have no conflict of interest., (Copyright © 2022. Published by Elsevier Inc.)
- Published
- 2023
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39. A comparative screening of laccase-mediator systems by white-rot fungi laccases for biocatalytic benzyl alcohol oxidation.
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Marino I, Pignataro E, Danzi D, Cellini F, Cardellicchio C, Biundo A, Pisano I, and Capozzi MAM
- Subjects
- Benzyl Alcohol, Renal Dialysis, Oxidation-Reduction, Solvents, Laccase metabolism, Trametes metabolism
- Abstract
Production of value-added compounds from waste materials is of utmost importance for the development of a sustainable society especially regarding their use as catalysts in industrially relevant synthetic reactions. Herein, we show the production of laccases from four white-rot fungi, which were grown on agricultural residues, specifically Trametes versicolor 11269, Pleurotus ostreatus 1020, Panus tigrinus 707 and Lentinula edodes SC-495. The produced laccases were tested on a laccase-mediator system (LMS) for the biocatalytic oxidation of the model substrate benzyl alcohol into benzaldehyde. The LMS was carried out in the presence both of tetrahydrofuran as co-solvent and of the mediator 2,2,6,6-tetramethyl-1-piperidinyloxyl (TEMPO) due to its high redox potential and its ability to perform the oxidation. Tolerance studies showed that the dialyzed solutions were able to tolerate 1% (99:1 v/v) of co-solvent, whereas a concentration of 10% v/v had a detrimental activity. Performances in the biocatalytic oxidation of laccase solutions from different purification steps were compared. Similar conversion was observed for laccase in dialysis (raw) and gel filtration (GF) product versus commercial T. versicolor laccase. The latter oxidized almost 99% of substrate while the other laccase solutions were able to reach a conversion from 91% for the laccase solution from P. tigrinus 707 after dialysis, to 50% for the laccase solution from P. ostreatus 1020 after gel filtration. This work highlights the potential of unpurified laccase solutions to be used as catalysts in synthetic reactions., (© 2022. The Author(s).)
- Published
- 2022
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40. Mitochondrial Role in Intrinsic Apoptosis Induced by a New Synthesized Chalcone in Hepatocellular Carcinoma Cells.
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Santarsiero A, Pappalardo I, Rosa GM, Pisano I, Superchi S, Convertini P, Todisco S, Scafato P, and Infantino V
- Abstract
Hepatocellular carcinoma (HCC) is the most common type of liver cancer and the fourth cause of cancer-related deaths worldwide. Presently, a few drugs are available for HCC treatment and prevention, including both natural and synthetic compounds. In this study, a new chalcone, ( E )-1-(2,4,6-triethoxyphenyl)-3-(3,4,5-trimethoxyphenyl)prop-2-en-1-one (ETTC), was synthesized and its effects and mechanisms of action over human hepatoma cells were investigated. Cytotoxic activity was revealed in HCC cells, while no effects were observed in normal hepatocytes. In HCC cells, ETTC caused subG1 cell cycle arrest and apoptosis, characterized by nuclear fragmentation. The activation of caspases 3/7 and 9, the increase in pro-apoptotic BAX, and the decrease in anti-apoptotic BCL-2 suggest the activation of the intrinsic pathway of apoptosis. ETTC mitochondrial targeting is confirmed by the reduction in mitochondrial membrane potential and Complex I activity together with levels of superoxide anion increasing. Our outcomes prove the potential mitochondria-mediated antitumor effect of newly synthesized chalcone ETTC in HCC.
- Published
- 2022
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41. Corrigendum: Bioisosteric Modification of To042: Synthesis and Evaluation of Promising Use-Dependent Inhibitors of Voltage-Gated Sodium Channels.
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Milani G, Cavalluzzi MM, Altamura C, Santoro A, Perrone M, Muraglia M, Colabufo NA, Corbo F, Casalino E, Franchini C, Pisano I, Desaphy JF, Carrieri A, Carocci A, and Lentini G
- Published
- 2022
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42. Loss of Detection of sgN Precedes Viral Abridged Replication in COVID-19-Affected Patients-A Target for SARS-CoV-2 Propagation.
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Ferrucci V, de Antonellis P, Quarantelli F, Asadzadeh F, Bibbò F, Siciliano R, Sorice C, Pisano I, Izzo B, Di Domenico C, Boccia A, Vargas M, Pierri B, Viscardi M, Brandi S, Fusco G, Cerino P, De Pietro L, Furfaro C, Napolitano LA, Paolella G, Festa L, Marzinotto S, Conte MC, Gentile I, Servillo G, Curcio F, de Cristofaro T, Broccolo F, Capoluongo E, and Zollo M
- Subjects
- Coronavirus Nucleocapsid Proteins analysis, Giant Cells drug effects, Giant Cells virology, HEK293 Cells, Humans, Limit of Detection, Nasopharynx virology, Phosphoproteins analysis, Phosphoproteins genetics, RNA, Antisense pharmacology, RNA, Viral, Ribonuclease P genetics, SARS-CoV-2 drug effects, SARS-CoV-2 genetics, Sensitivity and Specificity, Social Isolation, Viral Load, Viroporin Proteins genetics, Virus Replication drug effects, COVID-19 virology, Coronavirus Nucleocapsid Proteins genetics, SARS-CoV-2 physiology, Virus Replication physiology
- Abstract
The development of prophylactic agents against the SARS-CoV-2 virus is a public health priority in the search for new surrogate markers of active virus replication. Early detection markers are needed to follow disease progression and foresee patient negativization. Subgenomic RNA transcripts (with a focus on sgN) were evaluated in oro/nasopharyngeal swabs from COVID-19-affected patients with an analysis of 315 positive samples using qPCR technology. Cut-off Cq values for sgN (Cq < 33.15) and sgE (Cq < 34.06) showed correlations to high viral loads. The specific loss of sgN in home-isolated and hospitalized COVID-19-positive patients indicated negativization of patient condition, 3-7 days from the first swab, respectively. A new detection kit for sgN, gene E, gene ORF1ab, and gene RNAse P was developed recently. In addition, in vitro studies have shown that 2'- O -methyl antisense RNA (related to the sgN sequence) can impair SARS-CoV-2 N protein synthesis, viral replication, and syncytia formation in human cells (i.e., HEK-293T cells overexpressing ACE2) upon infection with VOC Alpha (B.1.1.7)-SARS-CoV-2 variant, defining the use that this procedure might have for future therapeutic actions against SARS-CoV-2.
- Published
- 2022
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43. Bioisosteric Modification of To042: Synthesis and Evaluation of Promising Use-Dependent Inhibitors of Voltage-Gated Sodium Channels.
- Author
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Milani G, Cavalluzzi MM, Altamura C, Santoro A, Perrone M, Muraglia M, Colabufo NA, Corbo F, Casalino E, Franchini C, Pisano I, Desaphy JF, Carrieri A, Carocci A, and Lentini G
- Subjects
- Antioxidants chemical synthesis, Antioxidants metabolism, Antioxidants pharmacology, Antioxidants toxicity, Butylamines chemical synthesis, Butylamines metabolism, Butylamines toxicity, HEK293 Cells, HeLa Cells, Humans, Mexiletine pharmacology, Molecular Docking Simulation, Phenyl Ethers chemical synthesis, Phenyl Ethers metabolism, Phenyl Ethers toxicity, Protein Binding, Reactive Oxygen Species metabolism, Voltage-Gated Sodium Channel Blockers chemical synthesis, Voltage-Gated Sodium Channel Blockers metabolism, Voltage-Gated Sodium Channel Blockers toxicity, Butylamines pharmacology, NAV1.4 Voltage-Gated Sodium Channel metabolism, Phenyl Ethers pharmacology, Voltage-Gated Sodium Channel Blockers pharmacology
- Abstract
Three analogues of To042, a tocainide-related lead compound recently reported for the treatment of myotonia, were synthesized and evaluated in vitro as skeletal muscle sodium channel blockers possibly endowed with enhanced use-dependent behavior. Patch-clamp experiments on hNav1.4 expressed in HEK293 cells showed that N-[(naphthalen-1-yl)methyl]-4-[(2,6-dimethyl)phenoxy]butan-2-amine, the aryloxyalkyl bioisostere of To042, exerted a higher use-dependent block than To042 thus being able to preferentially block the channels in over-excited membranes while preserving healthy tissue function. It also showed the lowest active transport across BBB according to the results of P-glycoprotein (P-gp) interacting activity evaluation and the highest cytoprotective effect on HeLa cells. Quantum mechanical calculations and dockings gave insights on the most probable conformation of the aryloxyalkyl bioisostere of To042 in solution and the target residues involved in the binding, respectively. Both approaches indicated the conformations that might be adopted in both the unbound and bound state of the ligand. Overall, N-[(naphthalen-1-yl)methyl]-4-[(2,6-dimethyl)phenoxy]butan-2-amine exhibits an interesting toxico-pharmacological profile and deserves further investigation., (© 2021 The Authors. ChemMedChem published by Wiley-VCH GmbH.)
- Published
- 2021
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44. RTG Signaling Sustains Mitochondrial Respiratory Capacity in HOG1 -Dependent Osmoadaptation.
- Author
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Guaragnella N, Agrimi G, Scarcia P, Suriano C, Pisano I, Bobba A, Mazzoni C, Palmieri L, and Giannattasio S
- Abstract
Mitochondrial RTG -dependent retrograde signaling, whose regulators have been characterized in Saccharomyces cerevisiae , plays a recognized role under various environmental stresses. Of special significance, the activity of the transcriptional complex Rtg1/3 has been shown to be modulated by Hog1, the master regulator of the high osmolarity glycerol pathway, in response to osmotic stress. The present work focuses on the role of RTG signaling in salt-induced osmotic stress and its interaction with HOG1 . Wild-type and mutant cells, lacking HOG1 and/or RTG genes, are compared with respect to cell growth features, retrograde signaling activation and mitochondrial function in the presence and in the absence of high osmostress. We show that RTG2, the main upstream regulator of the RTG pathway, contributes to osmoadaptation in an HOG1 -dependent manner and that, with RTG3 , it is notably involved in a late phase of growth. Our data demonstrate that impairment of RTG signaling causes a decrease in mitochondrial respiratory capacity exclusively under osmostress. Overall, these results suggest that HOG1 and the RTG pathway may interact sequentially in the stress signaling cascade and that the RTG pathway may play a role in inter-organellar metabolic communication for osmoadaptation.
- Published
- 2021
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45. Long-chain polyphosphates impair SARS-CoV-2 infection and replication.
- Author
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Ferrucci V, Kong DY, Asadzadeh F, Marrone L, Boccia A, Siciliano R, Criscuolo G, Anastasio C, Quarantelli F, Comegna M, Pisano I, Passariello M, Iacobucci I, Monica RD, Izzo B, Cerino P, Fusco G, Viscardi M, Brandi S, Pierri BM, Borriello G, Tiberio C, Atripaldi L, Bianchi M, Paolella G, Capoluongo E, Castaldo G, Chiariotti L, Monti M, De Lorenzo C, Yun KS, Pascarella S, Cheong JH, Kim HY, and Zollo M
- Subjects
- Administration, Inhalation, Amino Acid Sequence, Angiotensin-Converting Enzyme 2 chemistry, Angiotensin-Converting Enzyme 2 metabolism, Animals, Antiviral Agents administration & dosage, Antiviral Agents chemistry, COVID-19 metabolism, COVID-19 virology, Caco-2 Cells, Chlorocebus aethiops, Coronavirus RNA-Dependent RNA Polymerase chemistry, Coronavirus RNA-Dependent RNA Polymerase genetics, Coronavirus RNA-Dependent RNA Polymerase metabolism, Cytokines metabolism, HEK293 Cells, Host Microbial Interactions drug effects, Host Microbial Interactions genetics, Host Microbial Interactions physiology, Humans, In Vitro Techniques, Models, Biological, Molecular Docking Simulation, Nebulizers and Vaporizers, Polyphosphates administration & dosage, Polyphosphates chemistry, Proteasome Endopeptidase Complex metabolism, Protein Interaction Domains and Motifs, Proteolysis drug effects, RNA, Viral genetics, RNA, Viral metabolism, SARS-CoV-2 genetics, SARS-CoV-2 physiology, Sequence Homology, Amino Acid, Signal Transduction drug effects, Vero Cells, Virus Replication drug effects, Antiviral Agents pharmacology, Polyphosphates pharmacology, SARS-CoV-2 drug effects, COVID-19 Drug Treatment
- Abstract
Inorganic polyphosphates (polyPs) are linear polymers composed of repeated phosphate (PO
4 3- ) units linked together by multiple high-energy phosphoanhydride bonds. In addition to being a source of energy, polyPs have cytoprotective and antiviral activities. Here, we investigated the antiviral activities of long-chain polyPs against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In molecular docking analyses, polyPs interacted with several conserved amino acid residues in angiotensin-converting enzyme 2 (ACE2), the host receptor that facilitates virus entry, and in viral RNA-dependent RNA polymerase (RdRp). ELISA and limited proteolysis assays using nano- LC-MS/MS mapped polyP120 binding to ACE2, and site-directed mutagenesis confirmed interactions between ACE2 and SARS-CoV-2 RdRp and identified the specific amino acid residues involved. PolyP120 enhanced the proteasomal degradation of both ACE2 and RdRp, thus impairing replication of the British B.1.1.7 SARS-CoV-2 variant. We thus tested polyPs for functional interactions with the virus in SARS-CoV-2-infected Vero E6 and Caco2 cells and in primary human nasal epithelial cells. Delivery of a nebulized form of polyP120 reduced the amounts of viral positive-sense genomic and subgenomic RNAs, of RNA transcripts encoding proinflammatory cytokines, and of viral structural proteins, thereby presenting SARS-CoV-2 infection in cells in vitro., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)- Published
- 2021
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46. KRAS-regulated glutamine metabolism requires UCP2-mediated aspartate transport to support pancreatic cancer growth.
- Author
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Raho S, Capobianco L, Malivindi R, Vozza A, Piazzolla C, De Leonardis F, Gorgoglione R, Scarcia P, Pezzuto F, Agrimi G, Barile SN, Pisano I, Reshkin SJ, Greco MR, Cardone RA, Rago V, Li Y, Marobbio CMT, Sommergruber W, Riley CL, Lasorsa FM, Mills E, Vegliante MC, De Benedetto GE, Fratantonio D, Palmieri L, Dolce V, and Fiermonte G
- Subjects
- Animals, Biological Transport, Active, Cell Line, Tumor, Cytosol metabolism, Female, Humans, Mice, Mice, SCID, Mitochondria metabolism, NADP metabolism, Oxidation-Reduction, Reactive Oxygen Species metabolism, Xenograft Model Antitumor Assays, Aspartic Acid metabolism, Carcinoma, Pancreatic Ductal metabolism, Glutamine metabolism, Pancreatic Neoplasms metabolism, Proto-Oncogene Proteins p21(ras) metabolism, Uncoupling Protein 2 metabolism
- Abstract
The oncogenic KRAS mutation has a critical role in the initiation of human pancreatic ductal adenocarcinoma (PDAC) since it rewires glutamine metabolism to increase reduced nicotinamide adenine dinucleotide phosphate (NADPH) production, balancing cellular redox homeostasis with macromolecular synthesis
1,2 . Mitochondrial glutamine-derived aspartate must be transported into the cytosol to generate metabolic precursors for NADPH production2 . The mitochondrial transporter responsible for this aspartate efflux has remained elusive. Here, we show that mitochondrial uncoupling protein 2 (UCP2) catalyses this transport and promotes tumour growth. UCP2-silenced KRASmut cell lines display decreased glutaminolysis, lower NADPH/NADP+ and glutathione/glutathione disulfide ratios and higher reactive oxygen species levels compared to wild-type counterparts. UCP2 silencing reduces glutaminolysis also in KRASWT PDAC cells but does not affect their redox homeostasis or proliferation rates. In vitro and in vivo, UCP2 silencing strongly suppresses KRASmut PDAC cell growth. Collectively, these results demonstrate that UCP2 plays a vital role in PDAC, since its aspartate transport activity connects the mitochondrial and cytosolic reactions necessary for KRASmut rewired glutamine metabolism2 , and thus it should be considered a key metabolic target for the treatment of this refractory tumour.- Published
- 2020
- Full Text
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47. Optimization of Microwave-Assisted Extraction of Antioxidants from Bamboo Shoots of Phyllostachys pubescens .
- Author
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Milani G, Curci F, Cavalluzzi MM, Crupi P, Pisano I, Lentini G, Clodoveo ML, Franchini C, and Corbo F
- Subjects
- Polyphenols chemistry, Antioxidants chemistry, Microwaves, Plant Extracts chemistry, Poaceae chemistry
- Abstract
Bamboo is a well-known medicinal plant in Southeast Asia that recently has attracted attention for its high polyphenol content and its medical and nutraceutical applications. In this work, polyphenols have been recovered for the first time by microwave-assisted extraction (MAE) from an unusual Italian cultivar of Phyllostachys pubescens bamboo shoots. The effects of three independent variables, such as extraction time, temperature, and solid/liquid ratio, on polyphenol recovery yield were investigated and successfully optimized through the response surface methodology. We demonstrated that MAE is an excellent polyphenols extraction technique from bamboo shoots because the total phenolic content obtained under microwave irradiation optimal conditions (4 min at 105 °C with 6.25 mg/mL ratio) was about eight-fold higher than that obtained with the conventional extraction method. Furthermore, higher total flavonoid content was also obtained under MAE. Consistent with these results, MAE enhanced the extract antioxidant properties with significant improved DPPH, ABTS, and FRAP scavenging ability. Therefore, this innovative extraction process enhances the recovery of biologically active compounds from Phyllostachys pubescens bamboo shoots with a dramatic reduction of time and energy consumption, which paves the way for its industrial application in functional food production.
- Published
- 2020
- Full Text
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48. CaCO 3 as an Environmentally Friendly Renewable Material for Drug Delivery Systems: Uptake of HSA-CaCO 3 Nanocrystals Conjugates in Cancer Cell Lines.
- Author
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Vergaro V, Pisano I, Grisorio R, Baldassarre F, Mallamaci R, Santoro A, Suranna GP, Papadia P, Fanizzi FP, and Ciccarella G
- Abstract
Chemical and biochemical functionalization of nanoparticles (NPs) can lead to an active cellular uptake enhancing their efficacy thanks to the targeted localization in tumors. In the present study calcium carbonate nano-crystals (CCNs), stabilized by an alcohol dehydration method, were successfully modified by grafting human serum albumin (HSA) on the surface to obtain a pure protein corona. Two types of CCNs were used: naked CaCO
3 and the (3-aminopropyl)triethoxysilane (APTES) modified CaCO3 -NH2 . The HSA conjugation with naked CCN and amino-functionalized CCN (CCN-NH2 ) was established through the investigation of modification in size, zeta potential, and morphology by Transmission Electron Microscopy (TEM). The amount of HSA coating on the CCNs surface was assessed by spectrophotometry. Thermogravimetric analysis (TGA) and Differential scanning calorimetry (DSC) confirmed the grafting of APTES to the surface and successive adsorption of HSA. Furthermore, to evaluate the effect of protein complexation of CCNs on cellular behavior, bioavailability, and biological responses, three human model cancer cell lines, breast cancer (MCF7), cervical cancer (HeLa), and colon carcinoma (Caco-2) were selected to characterize the internalization kinetics, localization, and bio-interaction of the protein-enclosed CCNs. To monitor internalization of the various conjugates, chemical modification with fluorescein-isothiocyanate (FITC) was performed, and their stability over time was measured. Confocal microscopy was used to probe the uptake and confirm localization in the perinuclear region of the cancer cells. Flow cytometry assays confirmed that the bio-functionalization influence cellular uptake and the CCNs behavior depends on both cell line and surface features., Competing Interests: The authors declare no conflict of interest.- Published
- 2019
- Full Text
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49. Metastatic group 3 medulloblastoma is driven by PRUNE1 targeting NME1-TGF-β-OTX2-SNAIL via PTEN inhibition.
- Author
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Ferrucci V, de Antonellis P, Pennino FP, Asadzadeh F, Virgilio A, Montanaro D, Galeone A, Boffa I, Pisano I, Scognamiglio I, Navas L, Diana D, Pedone E, Gargiulo S, Gramanzini M, Brunetti A, Danielson L, Carotenuto M, Liguori L, Verrico A, Quaglietta L, Errico ME, Del Monaco V, D'Argenio V, Tirone F, Mastronuzzi A, Donofrio V, Giangaspero F, Picard D, Remke M, Garzia L, Daniels C, Delattre O, Swartling FJ, Weiss WA, Salvatore F, Fattorusso R, Chesler L, Taylor MD, Cinalli G, and Zollo M
- Subjects
- Adolescent, Animals, Carrier Proteins genetics, Cell Line, Tumor, Cell Movement drug effects, Cell Proliferation drug effects, Cerebellar Neoplasms pathology, Child, Child, Preschool, Female, Gene Expression Regulation, Neoplastic drug effects, Gene Regulatory Networks, Humans, Infant, Male, Medulloblastoma pathology, Mice, Mice, Inbred BALB C, Models, Molecular, Neoplasm Metastasis genetics, PTEN Phosphohydrolase genetics, Phosphoric Monoester Hydrolases, Pyrimidinones chemistry, Pyrimidinones pharmacology, Signal Transduction drug effects, Signal Transduction genetics, Snail Family Transcription Factors metabolism, Transforming Growth Factor beta metabolism, Carrier Proteins metabolism, Cerebellar Neoplasms metabolism, Gene Expression Regulation, Neoplastic physiology, Medulloblastoma metabolism, Neoplasm Metastasis physiopathology, PTEN Phosphohydrolase metabolism
- Abstract
Genetic modifications during development of paediatric groups 3 and 4 medulloblastoma are responsible for their highly metastatic properties and poor patient survival rates. PRUNE1 is highly expressed in metastatic medulloblastoma group 3, which is characterized by TGF-β signalling activation, c-MYC amplification, and OTX2 expression. We describe the process of activation of the PRUNE1 signalling pathway that includes its binding to NME1, TGF-β activation, OTX2 upregulation, SNAIL (SNAI1) upregulation, and PTEN inhibition. The newly identified small molecule pyrimido-pyrimidine derivative AA7.1 enhances PRUNE1 degradation, inhibits this activation network, and augments PTEN expression. Both AA7.1 and a competitive permeable peptide that impairs PRUNE1/NME1 complex formation, impair tumour growth and metastatic dissemination in orthotopic xenograft models with a metastatic medulloblastoma group 3 cell line (D425-Med cells). Using whole exome sequencing technology in metastatic medulloblastoma primary tumour cells, we also define 23 common 'non-synonymous homozygous' deleterious gene variants as part of the protein molecular network of relevance for metastatic processes. This PRUNE1/TGF-β/OTX2/PTEN axis, together with the medulloblastoma-driver mutations, is of relevance for future rational and targeted therapies for metastatic medulloblastoma group 3.10.1093/brain/awy039_video1awy039media15742053534001.
- Published
- 2018
- Full Text
- View/download PDF
50. SLC25A10 biallelic mutations in intractable epileptic encephalopathy with complex I deficiency.
- Author
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Punzi G, Porcelli V, Ruggiu M, Hossain MF, Menga A, Scarcia P, Castegna A, Gorgoglione R, Pierri CL, Laera L, Lasorsa FM, Paradies E, Pisano I, Marobbio CMT, Lamantea E, Ghezzi D, Tiranti V, Giannattasio S, Donati MA, Guerrini R, Palmieri L, Palmieri F, and De Grassi A
- Subjects
- Antioxidants metabolism, Child, DNA, Mitochondrial genetics, Heterozygote, Humans, Male, Metabolism, Inborn Errors genetics, Metabolism, Inborn Errors metabolism, Mitochondria metabolism, Oxidative Phosphorylation, Oxidative Stress genetics, Pedigree, RNA Splicing genetics, Brain Diseases genetics, Brain Diseases metabolism, Dicarboxylic Acid Transporters genetics, Dicarboxylic Acid Transporters metabolism, Mitochondrial Diseases genetics, Mitochondrial Diseases metabolism, Mutation genetics
- Abstract
Mitochondrial diseases are a plethora of inherited neuromuscular disorders sharing defects in mitochondrial respiration, but largely different from one another for genetic basis and pathogenic mechanism. Whole exome sequencing was performed in a familiar trio (trio-WES) with a child affected by severe epileptic encephalopathy associated with respiratory complex I deficiency and mitochondrial DNA depletion in skeletal muscle. By trio-WES we identified biallelic mutations in SLC25A10, a nuclear gene encoding a member of the mitochondrial carrier family. Genetic and functional analyses conducted on patient fibroblasts showed that SLC25A10 mutations are associated with reduction in RNA quantity and aberrant RNA splicing, and to absence of SLC25A10 protein and its transporting function. The yeast SLC25A10 ortholog knockout strain showed defects in mitochondrial respiration and mitochondrial DNA content, similarly to what observed in the patient skeletal muscle, and growth susceptibility to oxidative stress. Albeit patient fibroblasts were depleted in the main antioxidant molecules NADPH and glutathione, transport assays demonstrated that SLC25A10 is unable to transport glutathione. Here, we report the first recessive mutations of SLC25A10 associated to an inherited severe mitochondrial neurodegenerative disorder. We propose that SLC25A10 loss-of-function causes pathological disarrangements in respiratory-demanding conditions and oxidative stress vulnerability., (© The Author(s) 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Published
- 2018
- Full Text
- View/download PDF
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