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

Early occurrence of lung adenocarcinoma and breast cancer after radiotherapy of a chest wall sarcoma in a patient with a de novo germline mutation in TP53

We report a 26-year-old female patient who was diagnosed within 4years with chest sarcoma, lung adenocarcinoma, and breast cancer. While her family history was unremarkable, DNA sequencing of TP53 revealed a germline de novo non-sense mutation in exon 6 p.Arg213X. One year later, she further developed a contralateral ductal carcinoma in situ, and 18months later a jaw osteosarcoma. This case illustrates the therapeutic pitfalls in the care of a young cancer patient with TP53 de novo germline mutations and the complications related to her first-line therapy. Suggestion is made to use the less stringent Chompret criteria for germline TP53 mutation screening. Our observation underlines the possibly negative effect of radiotherapy in generating second tumors in patients with a TP53 mutation. We also present a review of six previously reported cases, comparing their cancer phenotypes with those generally produced by TP53 mutation

A new hexapeptide from the leader peptide of rMnSOD enters cells through the oestrogen receptor to deliver therapeutic molecules

A 24-amino acid leader peptide of a new human recombinant manganese superoxide dismutase can enter cells and carry molecules. Here, we demonstrated that six of the 24 amino acids penetrate cells through a particular gate represented by a specific amino acid sequence of the oestrogen receptor (ER). We analysed the internalization of the synthetic hexapeptide and the cytotoxic activity of the hexapeptide conjugated to cisplatin on a cell line panel. In most cell lines, the hexapeptide delivered an amount of cisplatin that was 2 to 8 times greater than that released by cisplatin when the drug was used alone. This increased delivery increases the therapeutic index of cisplatin and reduces side effects caused by a high dosage or long-term treatment times. We may consider this hexapeptide a new molecular carrier to deliver molecules with therapeutic activity into ER+ cells for diagnostic purposes and clinical or immune therapy

Evolutionary conservation of a regulative pathway of erythropoiesis in Poikilothermic vertebrates

Apoptosis, programmed cell death, plays a central role in haematopoiesis. Mature erythrocytes of non-mammalian vertebrates maintain a permanent nucleus; these cells can undergo apoptosis (eryptosis), as do other somatic cells of a given non-mammalian vertebrate. In this study, we have investigated the expression and subcellular distribution of Bcl-2, Bcl-XL and Bax proteins in the maturation phases and after X-ray irradiation of nucleated erythrocytes of Torpedo marmorata and Caretta caretta and the effect of X-ray irradiation on nucleated circulating erythrocytes of Torpedo marmorata. The cellular distribution of proteins was detected in erythrocytes by using immunocytochemistry at light microscopy and immunoelectron microscopy. The electrophoretic separation and immunoblotting of pro- and anti-apoptotic proteins of immature and mature erythroid cells was performed too, after X-ray irradiation of torpedoes. The results of the immunocytochemical analyses show an increase, in the expression level of Bax in mature as compared to young erythrocytes and a corresponding decrease of Bcl-2 and Bcl-XL. This maturation pattern of Bax, Bcl-2 and Bcl-XL was abrogated in X-ray irradiated torpedo erythrocytes. On the basis of these observations, Bax, Bcl-2 and Bcl-XL seems to play a role in the erythropoiesis of Torpedo marmorata Risso and in Caretta caretta. In conclusion, the same apoptotic proteins of somatic cells appear to be conserved in circulating nucleated erythrocytes thus suggesting to play a role in the maturation of these cells

Survey of Hand Hygiene, High-Touch Device Use, and Proper Habits of Health Care Workers for Infection Risk Prevention: Protocol for a Cross-Sectional Study.

Background: Care-related infections are infectious diseases that occur in a care setting. The most important prevention methods are hand hygiene and the proper use of gloves and gowns. Recent literature points out that hand contact with mobile devices or potentially contaminated environments can promote an increased occurrence of health care–associated infections (HAIs). Objective: This study aims to analyze the correlation between the microbial population present on the hands of health care professionals in the wards of the Tor Vergata Hospital in Rome and the microorganisms present on the surfaces of their smartphones and tablets by searching for the main agents responsible for HAIs. Methods: Sterile swabs will be used to collect samples from hands and smartphones, which will then be plated on nutrient agar and other selective media for microbial count. Colonies showing growth and morphological characteristics suggestive of potential pathogens will be isolated and subjected to further analysis for identification at the species level and for antimicrobial resistance profiling (using a proprietary automated analytical profile index system). Sampling will be conducted quarterly (first 2 weeks of each quarter) to assess any changes in microbial flora. In parallel, an Italian version of the World Health Organization questionnaire on health care workers’ knowledge of hand hygiene and a questionnaire on the use of high-touch devices will be administered to participants. Each quarter, 30 health workers will be selected, resulting in a total of 120 health workers and 240 samples collected by the end of the study. For each sample, the analysis will focus on quantifying the total bacterial load at 37 °C and 22 °C, along with detecting coliforms or Escherichia coli, Enterococci, Staphylococci, Acinetobacter, Klebsiella, Pseudomonas, and any associated antimicrobial resistance. Results: The study aims to begin sample collection by June 2025. The protocol was properly evaluated and approved by the territorial ethics committee “Lazio Area 2” on March 21, 2024, with the code 76.24 CET2 utv_ptv. Conclusions: The findings of this study will be crucial in highlighting the need for targeted education and training of health care practitioners involved in the study, with a focus on the prevention of HAIs

Mental Health of Long Term Survivors of Childhood and Young Adult Cancer

Background/Objectives Leukemic cells produce higher amounts of ROS than healthy cells and express low levels of antioxidant enzymes such as catalase (CAT) and superoxide dismutase (SOD). Recently, it was demonstrated that rMnSOD, a recombinant new isoform of MnSOD, recently isolated from human liposarcoma cell line, is able to trigger apoptosis of human T-ALL cells (99%) without toxic effects on healthy cells. The present study was aimed to verify if rMnSOD is able to induce apoptosis and to stop in G0/G1 stage of the cell cycle in pediatric patients with B-cell acute lymphoblastic leukaemia (ALL) and if it displays synergic action with daunorubicin. Design and Methods Cells were collected from three patients diagnosed for B-cell ALL at the Pediatric Oncology of University “Luigi Vanvitelli”. Cell viability, apoptosis and cell cycle of lymphoblasts and SUP-B15 cell line (ATCC) were analyzed through “ Muse Kit” after rMnSOD treatment. ROS analysis was detected with NAC. Apoptotic fragmentations of SUP-B15 were demonstrated by confocal imaging. Results We observed that low concentrations of rMnSOD are able to trigger apoptosis of SUP-B15 cell line and B-ALL cells from pediatric patients. In detail, we observed an apoptosis rate from 30% to 46% in B-ALL pediatric patient cells and of 45% in SUP-B15 cell line. Instead, cell cycle analysis showed a cell fraction decrease in G0/G1 phase. Moreover, the synergic activity of rMnSOD and Daunorubicin induces apoptosis in 92% of cells, by comparing the treatment with the single substances, rMnSOD (64,7%) and Daunorubicin (23,9%). Confocal microscopy analysis revealed internalization of rMnSOD in SUP-B15 cells and evident apoptotic alterations, such as nuclear fragmentations and apoptotic-bodies. In conclusion, rMnSOD exerts toxic activity only against cancer cells, by enhancing drug effect, thus allowing to utilize lower concentration of standard chemotherapy to increase the apoptosis level

Cardiac magnetic resonance predictors of left ventricular remodelling following acute ST elevation myocardial infarction: The VavirimS study

Left ventricular (LV) remodelling (REM) ensuing after ST-elevation myocardial infarction (STEMI), has typically been studied by echocardiography, which has limitations, or cardiac magnetic resonance (CMR) in early phase that may overestimate infarct size (IS) due to tissue edema and stunning. This prospective, multicenter study investigated LV-REM performing CMR in the subacute phase, and 6 months after STEMI

Clinical practice in European centres treating paediatric posterior fossa tumours with pencil beam scanning proton therapy

BACKGROUND AND PURPOSE: As no guidelines for pencil beam scanning (PBS) proton therapy (PT) of paediatric posterior fossa (PF) tumours exist to date, this study investigated planning techniques across European PT centres, with special considerations for brainstem and spinal cord sparing.MATERIALS AND METHODS: A survey and a treatment planning comparison were initiated across nineteen European PBS-PT centres treating paediatric patients. The survey assessed all aspects of the treatment chain, including but not limited to delineations, dose constraints and treatment planning. Each centre planned two PF tumour cases for focal irradiation, according to their own clinical practice but based on common delineations. The prescription dose was 54 Gy(RBE) for Case 1 and 59.4 Gy(RBE) for Case 2. For both cases, planning strategies and relevant dose metrics were compared.RESULTS: Seventeen (89 %) centres answered the survey, and sixteen (80 %) participated in the treatment planning comparison. In the survey, thirteen (68 %) centres reported using the European Particle Therapy Network definition for brainstem delineation. In the treatment planning study, while most centres used three beam directions, their configurations varied widely across centres. Large variations were also seen in brainstem doses, with a brainstem near maximum dose (D2%) ranging from 52.7 Gy(RBE) to 55.7 Gy(RBE) (Case 1), and from 56.8 Gy(RBE) to 60.9 Gy(RBE) (Case 2).CONCLUSION: This study assessed the European PBS-PT planning of paediatric PF tumours. Agreement was achieved in e.g. delineation-practice, while wider variations were observed in planning approach and consequently dose to organs at risk. Collaboration between centres is still ongoing, striving towards common guidelines.</p