Biophysical and biochemical characterization of a liposarcoma-derived recombinant MnSOD protein acting as an anticancer agent

A recombinant MnSOD (rMnSOD) synthesized by specific cDNA clones derived from a liposarcoma cell line was shown to have the same sequence as the wild-type MnSOD expressed in the human myeloid leukaemia cell line U937, except for the presence of the leader peptide at the N-terminus. These results were fully confirmed by the molecular mass of rMnSOD as evaluated by ES/MS analysis (26662.7 Da) and the nucleotide sequence of the MnSOD cDNA. The role of the leader peptide in rMnSOD was investigated using a fluorescent and/or 68Gallium-labelled synthetic peptide. The labelled peptide permeated MCF-7 cells and uptake could be inhibited in the presence of an excess of oestrogen. In vivo it was taken up by the tumour, suggesting that the molecule can be used for both therapy and diagnosis. The in vitro and in vivo pharmacology tests confirmed that rMnSOD is only oncotoxic for tumour cells expressing oestrogen receptors. Pharmacokinetic studies in animals performed with 125I- and 131I-labelled proteins confirmed that, when administered systemically, rMnSOD selectively reached the tumour, where its presence was unambiguously demonstrated by scintigraphic and PET scans. PCR analysis revealed that Bax gene expression was increased and the Bcl2 gene was down regulated in MCF7 cells treated with rMnSOD, which suggests that the protein induces a pro-apoptotic mechanism

Ultrastructural immunolocalization of transferrin receptor in circulating erythroid cells: its role as biomarker for X ray damage in Elasmobranchs

Transferrin receptor (CD71), a membrane glycoprotein involved in iron absorption, that plays a key role in metabolism and cell homeostasis, is a biomarker of X-ray damage in man and rat(1). Following previous observations by FACS and immunocytochemical detection at L.M., that suggested an increased CD71 expression in the circulating erythroid cells (RBCs) of Torpedoes after X-ray exposure, the aim of this study was the ultrastructural localization of this marker before and after X-ray treatment of benthonic Elasmobranchs (Torpedo marmorata and Torpedo ocellata) in order to evaluate whether CD71 can be used as a biomarker of sea X-ray pollution. Ultrathin sections of RBCs, withdrawn from 3 specimens before and after irradiation (45, 80 and 90 Gy), were first incubated with polyclonal primary antibody anti-CD71, then with secondary antibody conjugated with colloidal gold and lastly contrasted and observed by T.E.M.. Before irradiation, only few RBCs showed low CD71-immunoreactivity, mainly in plasma membrane but also cytoplasm. After irradiation, morphological apoptotic-like changes in the nucleus and cytoplasm vacuolization were observed and a more intense CD71-ir was found mainly into cytoplasmic vesicles, demonstrating an increased receptor endocytosis and occasionally in mitochondria, injured by irradiation. These preliminary data, showing an increased CD71 expression and internalization in circulating RBCs of Torpedoes after X-ray exposure, may suggest, if confirmed by further experiments, the role of CD71 as biomarker of X-ray sea pollution

Active Internalization of a Mn-SOD in living tumor cells

MnSOD deserve not only to be considered for the treatment cancer but also to prevent or to restore the degeneration of normal cells, having atherapeutic potential against other pathological conditions known to be associated with oxidative stresses, such as neurodegenerative diseases and agin

Skin necrosis in sea turtle cold stunning: Regeneration following rMnSOD topic treatment in a specimen of Caretta caretta

A cell line derived from a human liposarcoma, named LSA, was previously reported to secrete a protein with a unique ability to penetrate solely cancer cells. The protein was revealed to be a variant isoform of Manganese SuperOxide Dismutase which was also obtained in recombinant form and so called rMnSOD (Mancini et al. 2006). The remarkable enzymatic capacity of this rMnSOD to neutralize free radicals occurring in damaged tissues was used to cure a diffuse skin necrosis of the neck, head and fins of a specimen of sea turtle Caretta caretta. The present work describes the total recovery of lesions in a specimen of C. caretta affected by hypothermia. Treatment with rMnSOD appears to have enzymatically transformed all the free radicals occurring in the lesion, thus avoiding necrosis and disruption of the tissue and generating new tissu

Screening ematologico delle tartarughe marine del Mediterraneo: determinazione degli intervalli di normalità dei principali parametri ematochimici in Caretta caretta

Presso il Turtle Point della Stazione Zoologica di Napoli, perviene annualmente un gran numero di esemplari di Caretta caretta, alcuni dei quali in stato di debilitazione, causato da traumi di varia origine. Sono stati scelti 15 esemplari, in apparente stato di buona salute, per la determinazione dei parametri ematochimici comunemente utilizzati in diagnostica, allo scopo di stabilire gli intervalli di normalità per la popolazione di tartarughe del mar Mediterraneo. I campioni di sangue sono stati prelevati dal seno venoso cervicale da esemplari di peso compreso tra 2.50 e 13.50 kg, per l’esecuzione di: glicemia, colesterolo, urea, acido urico, proteine totali, bilirubina totale, transaminasi GOT e GPT, Ca2+ e Mg2+; tali determinazioni sono state ripetute per 3 volte in ciascun esemplare, ad intervalli di almeno 2 mesi, nell’arco di 8 mesi di stabulazione. I risultati ottenuti, espressi come valore medio con relativa deviazione standard, sono riportati di seguito: glicemia 117 ± 19 mg/dL, colesterolo 102 ± 31 mg/dL, urea 137 ± 45 mg/dL, acido urico 1.2 ± 0.5 mg/dL, proteine totali 3.1 ± 0.5 g/dL, bilirubina totale 0.23 ± 0.05 mg/dL, GOT 92 ± 52 U/L GPT 6 ± 0 U/L, Ca2+ 7.3 ± 0.8 mg/dL e Mg2+ 4.4 ± 0.6 mEq/L. Tale rilevazione, in fase di progressivo ampliamento, sarà utilizzata per creare un database di valori normali di riferimento per la popolazione del Mediterraneo, indispensabile per la valutazione dello stato di salute di animali catturati in stato di debilitazione

Rigenerazione cutanea dopo necrosi a seguito di trattamento con rMnSOD nella tartaruga marina Caretta caretta

Una isoforma di Manganese superossido dismutasi ricombinante derivata da cellule di liposarcoma è stata adoperata in forma topica per la cura di una estesa e profonda necrosi con esposizione dell'osso negli arti, di una tartaruga marina caretta caretta in grave stato di debilitazione per esposizione al freddo. Il trattamento ha prodotto la completa rigenerazione cutanea a seguito di 120 giorni di cura

A MUTATED FORM OF MANGANESE SUPER OXIDE DISMUTASE INJECTABLE

rMnSOD is proposed as therapeutical or preventive agent for the treatment of neurological disorders in which it became crucial to assure a constant amount of molecular oxygen, thus protecting the nerve cells and the entire organism from free radicals, from which all disorders take their origi

Hbv/hdv co‐infection: Epidemiological and clinical changes, recent knowledge and future challenges

Several investigations have been published on Hepatitis Delta Virus (HDV) infection in recent years, from which we have drawn the salient data to provide readers with useful information to improve their knowledge on the subject. HDV genotypes 5–8 have been recently imported to Western countries from central Africa, whose clinical relevance deserves further investigation. On-going HDV replication has been identified as an independent predictor of progression to cirrhosis and HCC for patients with HDV chronic hepatitis (HDV‐CH). Long‐term treatments of HDV‐CH with standard or pegylated interferon alfa (peg‐IFN‐α) have all been unsatisfactory, leading to a sustained virological response (SVR) only in 20–30% of patients treated, faced with a poor tolerabil-ity and frequent serious adverse reactions; the addition of HBV nucleo(s)tide analogues to peg‐IFN-α did not improve the rate of SVR. The improved knowledge of the HDV life cycle has allowed the development of direct acting agents towards key‐points of the HDV life cycle, namely bulevirtide, lonafarnib and nucleic acid polymers. Preliminary data have shown that these drugs are more effective than interferon‐based therapies, but adverse reactions are also common, which however seem toned down in combination therapy with other antivirals