Publications by authors named "Francesco Salvatore"

188 Publications

A novel smaller β-defensin-derived peptide is active against multidrug-resistant bacterial strains.

FASEB J 2021 12;35(12):e22026

Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, Naples, Italy.

Antibiotic resistance is becoming a severe obstacle in the fight against acute and chronic infectious diseases that accompany most degenerative illnesses from neoplasia to osteo-arthritis and obesity. Currently, the race is on to identify pharmaceutical molecules or combinations of molecules able to prevent or reduce the insurgence and/or progression of infectivity. Attempts to substitute antibiotics with antimicrobial peptides have, thus far, met with little success against multidrug-resistant (MDR) bacterial strains. During the last decade, we designed and studied the activity and features of human β-defensin analogs, which are salt-resistant, and hence active also under high salt concentrations as, for instance, in cystic fibrosis. Herein, we describe the design, synthesis, and major features of a new 21 aa long molecule, peptide γ2. The latter derives from the γ-core of the β-defensin natural molecules, a small fragment of these molecules still bearing high antibacterial activity. We found that peptide γ2, which contains only one disulphide bond, recapitulates most of the biological properties of natural human β-defensins and can also counteract both Gram-positive and Gram-negative MDR bacterial strains and biofilm formation. Moreover, it has great stability in human serum thereby enhancing its antibacterial presence and activity without cytotoxicity in human cells. In conclusion, peptide γ2 is a promising new weapon also in the battle against intractable infectious diseases.
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http://dx.doi.org/10.1096/fj.202002330RRDOI Listing
December 2021

Comprehensive Molecular Analysis of Gene Increases the Diagnostic Value of Dystrophinopathies: A Pilot Study in a Southern Italy Cohort of Patients.

Diagnostics (Basel) 2021 Oct 15;11(10). Epub 2021 Oct 15.

CEINGE-Biotecnologie Avanzate s.c.ar.l., 80145 Naples, Italy.

Duchenne/Becker muscular dystrophy (DMD/BMD) is an X-linked neuromuscular disease due to pathogenic sequence variations in the dystrophin ( gene, one of the largest human genes. More than 70% of gene defects result from genomic rearrangements principally leading to large deletions, while the remaining are small nucleotide variants, including nonsense and missense variants, small insertions/deletions or splicing alterations. Considering the large size of the gene and the wide mutational spectrum, the comprehensive molecular diagnosis of DMD/BMD is complex and may require several laboratory methods, thus increasing the time and costs of the analysis. In an attempt to simplify DMD/BMD molecular diagnosis workflow, we tested an NGS method suitable for the detection of all the different types of genomic variations that may affect the gene. Forty previously analyzed patients were enrolled in this study and re-analyzed using the next generation sequencing (NGS)-based single-step procedure. The NGS results were compared with those from multiplex ligation-dependent probe amplification (MLPA)/multiplex PCR and/or Sanger sequencing. Most of the previously identified deleted/duplicated exons and point mutations were confirmed by NGS and 1 more pathogenic point mutation (a nonsense variant) was identified. Our results show that this NGS-based strategy overcomes limitations of traditionally used methods and is easily transferable to routine diagnostic procedures, thereby increasing the diagnostic power of molecular analysis.
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http://dx.doi.org/10.3390/diagnostics11101910DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8534830PMC
October 2021

16S rRNA of Mucosal Colon Microbiome and CCL2 Circulating Levels Are Potential Biomarkers in Colorectal Cancer.

Int J Mol Sci 2021 Oct 4;22(19). Epub 2021 Oct 4.

CEINGE Biotecnologie Avanzate S.C.a R.L., 80131 Naples, Italy.

Colorectal cancer (CRC) is one of the most common malignancies in the Western world and intestinal dysbiosis might contribute to its pathogenesis. The mucosal colon microbiome and C-C motif chemokine 2 (CCL2) were investigated in 20 healthy controls (HC) and 20 CRC patients using 16S rRNA sequencing and immunoluminescent assay, respectively. A total of 10 HC subjects were classified as overweight/obese (OW/OB_HC) and 10 subjects were normal weight (NW_HC); 15 CRC patients were classified as OW/OB_CRC and 5 patients were NW_CRC. Results: and were more abundant in OW/OB_HC than in NW_HC microbiomes. Globally, , , , and were significantly increased in CRC patient tumor/lesioned tissue (CRC_LT) and CRC patient unlesioned tissue (CRC_ULT) microbiomes compared to HC microbiomes. CCL2 circulating levels were associated with tumor presence and with the abundance of , and . Our data suggest that mucosal colon dysbiosis might contribute to CRC pathogenesis by inducing inflammation. Notably, , which was more abundant in the OW/OB_HC than in the NW_HC microbiomes, might represent a putative link between obesity and increased CRC risk.
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http://dx.doi.org/10.3390/ijms221910747DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8509685PMC
October 2021

Mitidjospirone, a new spirodioxynaphthalene and GC-MS screening of secondary metabolites produced by strains of associated to dieback.

Nat Prod Res 2021 Sep 13:1-10. Epub 2021 Sep 13.

Department of Chemical Sciences, University of Naples Federico II, Naples, Italy.

Mitidjospirone, a new spiridioxynaphthalene, was isolated from the mycelial extract of a strain of , a recently described species belonging to the family Botryosphaeriaceae. Its structure was elucidated by extensive spectroscopic analysis and the absolute configuration was determined by electronic circular dichroism (ECD) experiment. Furthermore, several known compounds were identified during the screening of secondary metabolites produced by four strains of .
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http://dx.doi.org/10.1080/14786419.2021.1977299DOI Listing
September 2021

An Integrated Analysis of Intracellular Metabolites and Virulence Gene Expression during Biofilm Development of a Clinical Isolate of on Distinct Surfaces.

Int J Mol Sci 2021 Aug 21;22(16). Epub 2021 Aug 21.

Department of Biology, University of Naples 'Federico II', Via Cinthia, 80126 Naples, Italy.

Emergence of , which causes potential life-threatening invasive candidiasis, is often associated with colonization of medical devices as biofilm. Biofilm plays an important role in the virulence of the pathogen because of its complex structure, which provides resistance to conventional antimicrobials. In this study, the metabolic response of a clinical strain of colonizing three distinct surfaces (polytetrafluoroethylene (PTFE), polystyrene, and polycarbonate) as well as the expression of virulence and stress related genes (, , , , and ), were explored. Our results showed that lesser biofilm was developed on PTFE compared to polystyrene and polycarbonate. GS-MS metabolic analysis identified a total of 36 metabolites in the intracellular extract of cells grown on polystyrene, polycarbonate, and PTFE, essentially belonging to central carbon metabolism, amino acids, and lipids metabolism. The metabolic analysis showed that saturated and unsaturated fatty acids are preferentially produced during biofilm development on polycarbonate, whereas trehalose and vitamin B6, known as cellular protectors against a variety of stressors, were characteristic of biofilm on PTFE. The results of the transcriptomic analysis consider the different degrees of colonization of the three substrates, being , which encodes the component of signaling pathway of hyphal formation-cAMP-PKA, downregulated in PTFE biofilm compared to polycarbonate or polystyrene biofilms, while was upregulated in concomitance with the potential unfavorable conditions for biofilm formation on PTFE. Overall, this work provides new insights into the knowledge of biofilm development on surfaces of medical relevance in the perspective of improving the management of infections.
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http://dx.doi.org/10.3390/ijms22169038DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8396647PMC
August 2021

Yield and clinical significance of genetic screening in elite and amateur athletes.

Eur J Prev Cardiol 2021 Aug;28(10):1081-1090

CEINGE - Biotecnologie Avanzate, Italy.

Aims: The purpose of this study was to assess the value of genetic testing in addition to a comprehensive clinical evaluation, as part of the diagnostic work-up of elite and/or amateur Italian athletes referred for suspicion of inherited cardiac disease, following a pre-participation screening programme.

Methods: Between January 2009-December 2018, of 5892 consecutive participants, 61 athletes were investigated: 30 elite and 31 amateur athletes. Elite and amateur athletes were selected, on the basis of clinical suspicion for inherited cardiac disease, from two experienced centres for a comprehensive cardiovascular evaluation. Furthermore, the elite and amateur athletes were investigated for variants at DNA level up to 138 genes suspected to bear predisposition for possible cardiac arrest or even sudden cardiac death.

Results: Of these 61 selected subjects, six (10%) had diagnosis made possible by a deeper clinical evaluation, while genetic testing allowed a definite diagnosis in eight (13%). The presence of >3 clinical markers (i.e. family history, electrocardiogram and/or echocardiographic abnormalities, exercise-induced ventricular arrhythmias) was associated with a higher probability of positive genetic diagnosis (75%), compared with the presence of two or one clinical markers (14.2%, 8.1%, respectively, p-value = 0.004).

Conclusion: A combined clinical and genetic evaluation, based on the subtle evidence of clinical markers for inherited disease, was able to identify an inherited cardiac disease in about one-quarter of the examined athletes.
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http://dx.doi.org/10.1177/2047487320934265DOI Listing
August 2021

Genetic evaluation in athletes and cascade family screening: reply.

Eur J Prev Cardiol 2021 Jul 22. Epub 2021 Jul 22.

CEINGE - Biotecnologie Avanzate, Via Gaetano Salvatore, 486 - 80145 Naples, Italy.

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http://dx.doi.org/10.1093/eurjpc/zwaa150DOI Listing
July 2021

Nutritional Controlled Preparation and Administration of Different Tomato Purées Indicate Increase of β-Carotene and Lycopene Isoforms, and of Antioxidant Potential in Human Blood Bioavailability: A Pilot Study.

Nutrients 2021 Apr 17;13(4). Epub 2021 Apr 17.

CEINGE-Biotecnologie Avanzate, Via G. Salvatore, 486, 80145 Naples, Italy.

The isoforms of lycopene, carotenoids, and their derivatives including precursors of vitamin A are compounds relevant for preventing chronic degenerative diseases such as cardiovascular diseases and cancer. Tomatoes are a major source of these compounds. However, cooking and successive metabolic processes determine the bioavailability of tomatoes in human nutrition. To evaluate the effect of acute/chronic cooking procedures on the bioavailability of lycopene and carotene isoforms in human plasma, we measured the blood levels of these compounds and of the serum antioxidant potential in volunteers after a meal containing two different types of tomato sauce ( or ). Using a randomized cross-over administration design, healthy volunteers were studied, and the above indicated compounds were determined by HPLC. The results indicate an increased bioavailability of the estimated compounds and of the serum antioxidant potential with both types of tomato purée and the subsequently derived sauces (the increase was greater with purée). This study sheds light on the content of nutrient precursors of vitamin A and other antioxidant compounds derived from tomatoes cooked with different strategies. Lastly, our study indicates that strained purée should be preferred over rustic purée.
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http://dx.doi.org/10.3390/nu13041336DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8073136PMC
April 2021

GC-MS-Based Metabolomics Study of Single- and Dual-Species Biofilms of and .

Int J Mol Sci 2021 Mar 28;22(7). Epub 2021 Mar 28.

Department of Biology, University of Naples 'Federico II', via Cinthia, 80126 Naples, Italy.

and frequently co-exist within the human host as a complex biofilm community. These pathogens are of interest because their association is also related to significantly increased morbidity and mortality in hospitalized patients. With the aim of highlighting metabolic shifts occurring in the dual-species biofilm, an untargeted GC-MS-based metabolomics approach was applied to single and mixed biofilms of and . Metabolomic results showed that among the extracellular metabolites identified, approximately 40 compounds had significantly changed relative abundance, mainly involving central carbon, amino acid, vitamin, and secondary metabolisms, such as serine, leucine, arabitol, phosphate, vitamin B6, -(Phe-Pro), trehalose, and nicotinic acid. The results were related to the strict interactions between the two species and the different microbial composition in the early and mature biofilms.
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http://dx.doi.org/10.3390/ijms22073496DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8037927PMC
March 2021

Impact of the Peptide WMR-K on Dual-Species Biofilm and on the Untargeted Metabolomic Profile.

Pathogens 2021 Feb 16;10(2). Epub 2021 Feb 16.

Department of Biology, University of Naples 'Federico II', via Cinthia, 80126 Naples, Italy.

In recent years, the scientific community has focused on the development of new antibiotics to address the difficulties linked to biofilm-forming microorganisms and drug-resistant infections. In this respect, synthetic antimicrobial peptides (AMPs) are particularly regarded for their therapeutic potential against a broad spectrum of pathogens. In this work, the antimicrobial and antibiofilm activities of the peptide WMR-K towards single and dual species cultures of and were investigated. We found minimum inhibitory concentration (MIC) values for WMR-K of 10 µM for and of 200 µM for . Furthermore, sub-MIC concentrations of peptide showed an in vitro inhibition of biofilm formation of mono and polymicrobial systems and also a good biofilm eradication even if higher concentrations of it are needed. In order to provide additional evidence for the effect of the examined peptide, a study of changes in extracellular metabolites excreted and/or uptaken from the culture medium (metabolomic footprinting) in the poly-microbial association of and in presence and absence of WMR-K was performed. Comparing to the untreated dual species biofilm culture, the metabolomic profile of the WMR-K treated culture appears significantly altered. The differentially expressed compounds are mainly related to the primary metabolic pathways, including amino acids, trehalose, pyruvic acid, glycerol and vitamin B6.
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http://dx.doi.org/10.3390/pathogens10020214DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7920046PMC
February 2021

Nano-bio interface between human plasma and niosomes with different formulations indicates protein corona patterns for nanoparticle cell targeting and uptake.

Nanoscale 2021 Mar;13(10):5251-5269

Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli "Federico II", Napoli, Italy. and CEINGE-Biotecnologie Avanzate S.c.a r.l., Napoli, Italy.

Unraveling the proteins interacting with nanoparticles (NPs) in biological fluids, such as blood, is pivotal to rationally design NPs for drug delivery. The protein corona (PrC), formed on the NP surface, represents an interface between biological components and NPs, dictating their pharmacokinetics and biodistribution. PrC composition depends on biological environments around NPs and on their intrinsic physicochemical properties. We generated different formulations of non-ionic surfactant/non-phospholipid vesicles, called niosomes (NIOs), using polysorbates which are biologically safe, cheap, non-toxic and scarcely immunogenic. PrC composition and relative protein abundance for all designed NIOs were evaluated ex vivo in human plasma (HP) by quantitative label-free proteomics. We studied the correlation of the relative protein abundance in the corona with cellular uptake of the PrC-NIOs in healthy and cancer human cell lines. Our results highlight the effects of polysorbates on nano-bio interactions to identify a protein pattern most properly aimed to drive the NIO targeting in vivo, and assess the best conditions of PrC-NIO NP uptake into the cells. This study dissected the biological identity in HP of polysorbate-NIOs, thus contributing to shorten their passage from preclinical to clinical studies and to lay the foundations for a personalized PrC.
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http://dx.doi.org/10.1039/d0nr07229jDOI Listing
March 2021

Coordination Properties of the Fungal Metabolite Harzianic Acid toward Toxic Heavy Metals.

Toxics 2021 Jan 20;9(2). Epub 2021 Jan 20.

Department of Chemical Sciences, University of Naples Federico II, 80126 Naples, Italy.

Some Trichoderma strains are known for their capacity to produce harzianic acid, a metabolite belonging to the tetramic acid derivatives. Harzianic acid has interesting biological properties, such as antimicrobial activities against phytopathogenic fungi and promotion of plant growth. It also possesses remarkable chemical properties, including the chelating properties toward essential transition metals, which might be related to the biological activities. Increasing knowledge on chelating properties might be relevant for understanding the various beneficial effects of harzianic acid in the interaction between the producer fungi and plants. In this work, the coordination capacity of harzianic acid was studied to evaluate the formation and stability of complexes formed with toxic heavy metals (i.e., Cd, Co, Ni, and Pb), which might have a crucial role in the tolerance of plants growing in metal-contaminated soils and in abiotic stress.
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http://dx.doi.org/10.3390/toxics9020019DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7909447PMC
January 2021

Potential role of imaging markers in predicting future disease expression of arrhythmogenic cardiomyopathy.

Future Cardiol 2021 07 21;17(4):647-654. Epub 2020 Oct 21.

Department of Translational Medical Sciences, Inherited & Rare Cardiovascular Diseases, University of Campania 'Luigi Vanvitelli', Monaldi Hospital, Naples, Italy.

To evaluate the predictive accuracy of trabecular hypertrophy/derangement and late gadolinium enhancement in predicting diagnosis of arrhythmogenic cardiomyopathy (AC). Fifty-nine suspected AC patients were evaluated. To evaluate the ability of these markers to detect patients fulfilling definite diagnosis at 4-year follow-up, sensitivity (Se), specificity (Sp), positive and negative predictive value (PPV and NPV) and predictive accuracy (PA) were calculated. Presence of trabecular hypertrophy/derangement showed high NPV, while late gadolinium enhancement at cardiac MRI high Sp, NPV and PA. Their combination with symptoms and/or revised Task Force Criteria showed a high Sp, NPV and PA for definitive diagnosis during follow-up. In suspected AC patients, the absence of these markers allows to identify those with lower risk of disease progression.
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http://dx.doi.org/10.2217/fca-2020-0107DOI Listing
July 2021

The abundance of the long intergenic non-coding RNA 01087 differentiates between luminal and triple-negative breast cancers and predicts patient outcome.

Pharmacol Res 2020 11 14;161:105249. Epub 2020 Oct 14.

CEINGE-Biotecnologie Avanzate s.c.ar.l, Naples 80145, Italy; Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, Naples 80131, Italy; Inter-University Center for multifactorial and multi genetic chronic human diseases, "Federico II"- Naples, Tor Vergata- Roma II, and Chieti-Pescara Universities, Italy. Electronic address:

The molecular complexity of human breast cancer (BC) renders the clinical management of the disease challenging. Long non-coding RNAs (lncRNAs) are promising biomarkers for BC patient stratification, early detection, and disease monitoring. Here, we identified the involvement of the long intergenic non-coding RNA 01087 (LINC01087) in breast oncogenesis. LINC01087 appeared significantly downregulated in triple-negative BCs (TNBCs) and upregulated in the luminal BC subtypes in comparison to mammary samples from cancer-free women and matched normal cancer pairs. Interestingly, deregulation of LINC01087 allowed to accurately distinguish between luminal and TNBC specimens, independently of the clinicopathological parameters, and of the histological and TP53 or BRCA1/2 mutational status. Moreover, increased expression of LINC01087 predicted a better prognosis in luminal BCs, while TNBC tumors that harbored lower levels of LINC01087 were associated with reduced relapse-free survival. Furthermore, bioinformatics analyses were performed on TNBC and luminal BC samples and suggested that the putative tumor suppressor activity of LINC01087 may rely on interferences with pathways involved in cell survival, proliferation, adhesion, invasion, inflammation and drug sensitivity. Altogether, these data suggest that the assessment of LINC01087 deregulation could represent a novel, specific and promising biomarker not only for the diagnosis and prognosis of luminal BC subtypes and TNBCs, but also as a predictive biomarker of pharmacological interventions.
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http://dx.doi.org/10.1016/j.phrs.2020.105249DOI Listing
November 2020

Effect of γ-Aminobutyric Acid (GABA) on the Metabolome of Two Strains of Isolated from Grapevine.

Molecules 2020 Aug 23;25(17). Epub 2020 Aug 23.

Department of Chemical Sciences, University of Naples 'Federico II', 80126 Naples, Italy.

The effect of γ-aminobutyric acid (GABA) on the metabolome of two strains of isolated from grapevine that hold a different degree of virulence to the host plant (LA-SOL3 (more virulent), LA-SV1 (less virulent)) was investigated. The culture filtrates and crude extracts from the two strains grown in the presence and absence of 10 mM of GABA were tested for phytotoxicity on tomato plant cuttings and leaves, respectively. Considering the opportunistic nature of this fungus for humans, crude extracts were also tested for cytotoxicity on mammalian cell lines. We found that culture filtrates and crude extracts have a decreased toxicity in the presence of GABA. Metabolomic analysis, conducted on both strains at both growth conditions, revealed the production of several compounds, such as indole-3-carboxylic acid (ICA, which is the main compound produced by ), 3-indolecarboxyaldehyde, (3,4)-botryodiplodin, ()-mellein. Finally, data demonstrate that GABA both induces a decrease in the amount of ICA, and a diversification of the metabolites produced by .
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http://dx.doi.org/10.3390/molecules25173833DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7503889PMC
August 2020

Yield and clinical significance of genetic screening in elite and amateur athletes.

Eur J Prev Cardiol 2020 Jul 2:2047487320934265. Epub 2020 Jul 2.

CEINGE - Biotecnologie Avanzate, Italy.

Aims: The purpose of this study was to assess the value of genetic testing in addition to a comprehensive clinical evaluation, as part of the diagnostic work-up of elite and/or amateur Italian athletes referred for suspicion of inherited cardiac disease, following a pre-participation screening programme.

Methods: Between January 2009-December 2018, of 5892 consecutive participants, 61 athletes were investigated: 30 elite and 31 amateur athletes. Elite and amateur athletes were selected, on the basis of clinical suspicion for inherited cardiac disease, from two experienced centres for a comprehensive cardiovascular evaluation. Furthermore, the elite and amateur athletes were investigated for variants at DNA level up to 138 genes suspected to bear predisposition for possible cardiac arrest or even sudden cardiac death.

Results: Of these 61 selected subjects, six (10%) had diagnosis made possible by a deeper clinical evaluation, while genetic testing allowed a definite diagnosis in eight (13%). The presence of 3 clinical markers (i.e. family history, electrocardiogram and/or echocardiographic abnormalities, exercise-induced ventricular arrhythmias) was associated with a higher probability of positive genetic diagnosis (75%), compared with the presence of two or one clinical markers (14.2%, 8.1%, respectively, -value = 0.004).

Conclusion: A combined clinical and genetic evaluation, based on the subtle evidence of clinical markers for inherited disease, was able to identify an inherited cardiac disease in about one-quarter of the examined athletes.
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http://dx.doi.org/10.1177/2047487320934265DOI Listing
July 2020

Liposome-Embedding Silicon Microparticle for Oxaliplatin Delivery in Tumor Chemotherapy.

Pharmaceutics 2020 Jun 17;12(6). Epub 2020 Jun 17.

Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli "Federico II", 80131 Napoli, Italy.

Mesoporous silicon microparticles (MSMPs) can incorporate drug-carrying nanoparticles (NPs) into their pores. An NP-loaded MSMP is a multistage vector (MSV) that forms a Matryoshka-like structure that protects the therapeutic cargo from degradation and prevents its dilution in the circulation during delivery to tumor cells. We developed an MSV constituted by 1 µm discoidal MSMPs embedded with PEGylated liposomes containing oxaliplatin (oxa) which is a therapeutic agent for colorectal cancer (CRC). To obtain extra-small liposomes able to fit the 60 nm pores of MSMP, we tested several liposomal formulations, and identified two optimal compositions, with a prevalence of the rigid lipid 1,2-distearoyl-sn-glycero-3-phosphocholine and of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000]. To improve the MSV assembly, we optimized the liposome-loading inside the MSMP and achieved a five-fold increase of the payload using an innovative lyophilization approach. This procedure also increased the load and limited dimensional changes of the liposomes released from the MSV in vitro. Lastly, we found that the cytotoxic efficacy of oxa-loaded liposomes and-oxa-liposome-MSV in CRC cell culture was similar to that of free oxa. This study increases knowledge about extra-small liposomes and their loading into porous materials and provides useful hints about alternative strategies for designing drug-encapsulating NPs.
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http://dx.doi.org/10.3390/pharmaceutics12060559DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7355455PMC
June 2020

Adapted recreational football small-sided games improve cardiac capacity, body composition and muscular fitness in patients with type 2 diabetes.

J Sports Med Phys Fitness 2020 Sep 12;60(9):1261-1268. Epub 2020 Jun 12.

Department of Human Movement Sciences and Wellbeing, Parthenope University, Naples, Italy -

Background: The usefulness of adapted small-sided games (SSGs) in improving cardiac function in subjects with T2DM is still debated. Here we evaluated the effects of 18 weeks indoor muscular activation training (6 weeks; IMA) followed by adapted SSGs football training (12 weeks) on cardiac function, muscular fitness, body composition and adiponectin expression in sedentary T2DM volunteers.

Methods: Six T2DM patients underwent IMA protocol of 6 weeks, twice a week followed by 12 weeks SSGs (5-a-side, once a week) training. Glucose, lipid profile and serum homocysteine concentration, body composition (BC), bone mineral density (DEXA), were determined at baseline and after 18 weeks (IMA+SSGs). VO2max and muscular fitness were recorded at baseline and after IMA (6 weeks) and SSGs (12 weeks), respectively.

Results: No significant differences were found for VO2max and muscular fitness after 6weeks of IMA. After 18 weeks (6 weeks IMA + 12 weeks SSGs) of training, significant improvements were found in the following parameters: work capacity, VO2peak, Ventilation (VEpeak), breathing reserve consumption and oxygen uptake efficiency slope (P<0.05); leg fitness (P<0.05), BC (P<0.05), vertebral column T-score (P<0.01) and adiponectin (total and high-molecular-weight; P<0.05). Compared to baseline, a reduction in serum homocysteine occurred after 18 weeks of training (P<0.05).

Conclusions: We evidenced that weekly adapted SSGs friendly football matches for 12 weeks improve cardiorespiratory capacity and the expression of independent markers associated with cardiovascular risk in T2DM patients, suggesting an overall reduced CVD-risk in these patients. These preliminary data encourage us to test the efficacy of this type of exercise in a larger population.
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http://dx.doi.org/10.23736/S0022-4707.20.10498-5DOI Listing
September 2020

Genotype-Phenotype Correlation: A Triple DNA Mutational Event in a Boy Entering Sport Conveys an Additional Pathogenicity Risk.

Genes (Basel) 2020 05 8;11(5). Epub 2020 May 8.

Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, Via Sergio Pansini, 5, 80131 Naples, Italy.

The purpose of this paper is to present a clinical and laboratory study of a family, in which a 12-year-old boy was examined to assess his health status before starting competitive sports. A variety of clinical and instrumental tests were used to evaluate the status of the heart and its functions. Using Sanger sequencing (SS), we sequenced six related genes to verify suspected arrhythmogenic right ventricular cardiomyopathy (ARVC) hypothesized at the cardiac assessment and, subsequently, by a next-generation sequencing (NGS)-based multi-gene panel for more paramount genetic risk of sudden cardiac death (SCD) assessment. SS revealed two variants in the gene, one was inherited from the father and the other from the mother. The analysis on a large panel of genes (n = 138), putatively associated with sudden cardiac death, revealed, in the proband, a third variant in a different gene that encodes the protein desmin. Our results indicate that: i) NGS revealed a mutational event in a gene not conventionally screened as a first-line test in the presence of clinical suspicion of the arrhythmic disease; ii) a plurality of variants in different genes in the same subject (the proband) may increase the risk of heart disease; iii) in silico analysis with various methodological software and bioinformatic prediction tools indicates that the cumulative effects of the three variants in the same subject constitute an additional risk factor. This case report indicates that more pathogenic variants or likely pathogenic variants can contribute to the clinical phenotype of an individual, thereby contributing to the diagnosis and prognosis of inherited heart diseases.
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http://dx.doi.org/10.3390/genes11050524DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7288460PMC
May 2020

Bivalent Metal-Chelating Properties of Harzianic Acid Produced by Associated to the Gastropod .

Molecules 2020 May 4;25(9). Epub 2020 May 4.

Department of Chemical Sciences, University of Naples Federico II, 80126 Naples, Italy.

Harzianic acid is a secondary metabolite of , structurally belonging to the dienyltetramic acid subgroup of the tetramic acids. Biological activities of harzianic acid are of great interest for its antimicrobial and plant growth-promoting activities, which might be related to its chelating properties. In the present work harzianic acid, isolated from cultures of a strain of associated to the gastropod , was studied as a complexant agent of a number of biologically relevant transition metals (i.e., Zn, Fe, Cu, and Mn), using UV-VIS, potentiometry, MS and NMR techniques. Our findings show the coordination capacity of harzianic acid toward the above cations through the formation of neutral or charged complexes in a variable ratio depending on the metal and pH conditions.
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http://dx.doi.org/10.3390/molecules25092147DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7248884PMC
May 2020

The shift of the paradigm between ageing and diseases.

Clin Chem Lab Med 2020 09;58(10):1635-1644

Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, Naples, Italy.

In the area of the Medical Sciences, the chronological age has always been, and still is, an indicator by which we try to understand the health status of an individual. However, besides considering people born with an already expressed disease, each human genome has sequence alterations called predisposing mutations; carriers of such genetic alterations have an increased risk of contracting diseases during their life. In addition, the exposome, i.e. the totality of environmental noxae ("hits") to which our body is exposed throughout life (through ingestion, breathing, body surface hits, and psychosociological stress agents, etc.) contributes to increase gradually but inexorably the frailty of an organism, and this process is usually referred to as "physiological ageing". This position paper proposes that we invert our visual angle and view the passage-of-time not as the cause of diseases, but consider the genome alterations present at birth and the noxae received during our life as the real major causes of ageing. The Biomedical Sciences are now increasingly unraveling the etiopathogenesis of most chronic degenerative diseases; thus, it will be possible to monitor and treat those that most contribute to the increased frailty of each person, which is now referred to with the misnomer "physiological ageing". These concepts are not banal; indeed, they imply that we must try to avoid the causes of alterations that result later in chronic degenerative diseases. Thus, we should shift our attention from the cure to the prevention of alterations/diseases also to improve both the length and quality of our life. Moreover, this approach involves real personalized or individualized medicine, thus conferring a more direct benefit to each of us by finalizing either the cure or the monitoring of diseases.
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http://dx.doi.org/10.1515/cclm-2020-0125DOI Listing
September 2020

Identification of the Main Metabolites of a Marine-Derived Strain of Using LC and GC MS Techniques.

Metabolites 2020 Jan 30;10(2). Epub 2020 Jan 30.

Department of Chemical Sciences, University of Naples Federico II, 80126 Naples, Italy.

Marine-derived fungi are an important source of many valuable compounds with original structures and diverse physico-chemical properties. In this work, the metabolomic profile of a strain of , recovered from a snakelocks sea anemone (), was investigated through the parallel application of LC-ESI-HRMS, GC-MS, and NMR. Our strategy allowed the identification of mycophenolic acid, brevianamide A, and several compounds belonging to the thiosilvatins. Among the latter, five products are reported for the first time in this species. The main product of this series, -bis(methylthio)silvatin, was also tested for antiproliferative activity on both cancer and non-tumoral colon cell lines.
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http://dx.doi.org/10.3390/metabo10020055DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7074196PMC
January 2020

Hypermethioninemia in Campania: Results from 10 years of newborn screening.

Mol Genet Metab Rep 2019 Dec 11;21:100520. Epub 2019 Oct 11.

Department of Molecular Medicine and Medical Biotechnologies, "Federico II" University, Naples, Italy.

In the last years tandem mass spectrometry (MS/MS) has become a leading technology used for neonatal screening purposes. Newborn screening by MS/MS on dried blood spot samples (DBS) has one of its items in methionine levels: the knowledge of this parameter allows the identification of infant affected by homocystinuria (cystathionine β-synthase, CBS, deficiency) but can also lead, as side effect, to identify cases of methionine adenosyltransferase (MAT) type I/III deficiency. We started an expanded newborn screening for inborn errors of metabolism in Campania region in 2007. Here we report our ten years experience on expanded newborn screening in identifying patients affected by hypermethioninemia. During this period we screened approximately 77,000 infants and identified two cases: one case of classical homocystinuria and one patient affected by defect of MAT I/III. In this paper we describe these patients and their biochemical follow-up and review the literature concerning worldwide newborn screening reports on incidence of CBS and MAT deficiency.
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http://dx.doi.org/10.1016/j.ymgmr.2019.100520DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6796781PMC
December 2019

A Functional Analysis of the Unclassified Pro2767Ser BRCA2 Variant Reveals Its Potential Pathogenicity that Acts by Hampering DNA Binding and Homology-Mediated DNA Repair.

Cancers (Basel) 2019 Sep 28;11(10). Epub 2019 Sep 28.

CEINGE-Biotecnologie Avanzate, 8014 Naples, Italy.

and are the genes most frequently associated with hereditary breast and ovarian cancer (HBOC). They are crucial for the maintenance of genome stability, particularly in the homologous recombination-mediated repair pathway of DNA double-strand breaks (HR-DSBR). Widespread next-generation sequencing (NGS) screening has revealed numerous variants of uncertain significance. Assessing the clinical significance of these variants is challenging, particularly regarding the clinical management of patients. Here, we report the functional characterization of the unclassified c.8299C > T variant, identified in a young breast cancer patient during NGS screening. This variant causes the change of Proline 2767 to Serine in the DNA binding domain (DBD) of the BRCA2 protein, necessary for the loading of RAD51 on ssDNA during the HR-DSBR. Our in silico analysis and 3D-structure modeling predicted that the p.Pro2767Ser substitution is likely to alter the BRCA2 DBD structure and function. Therefore, to evaluate the functional impact of the p.Pro2767Ser variant, we used a minigene encoding a truncated protein that contains the BRCA2 DBD and the nearby nuclear localization sequence. We found that the ectopically expressed truncated protein carrying the normal DBD, which retains the DNA binding function and lacks the central RAD51 binding domain, interferes with endogenous wild-type BRCA2 mediator functions in the HR-DSBR. We also demonstrated that the BRCA2 Pro2767Ser DBD is unable to compete with endogenous BRCA2 DNA binding, thereby suggesting that the p.Pro2767Ser substitution in the full-length protein causes the functional loss of BRCA2. Consequently, our data suggest that the p.Pro2767Ser variant should be considered pathogenic, thus supporting a revision of the ClinVar interpretation. Moreover, our experimental strategy could be a valid method with which to preliminarily evaluate the pathogenicity of the unclassified germline variants in the DBD and their risk of predisposing to HBOC.
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http://dx.doi.org/10.3390/cancers11101454DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6826418PMC
September 2019

Crosstalk between 14-3-3θ and AF4 enhances MLL-AF4 activity and promotes leukemia cell proliferation.

Cell Oncol (Dordr) 2019 Dec 6;42(6):829-845. Epub 2019 Sep 6.

IRCCS SDN, 80143, Naples, Italy.

Purpose: The t(4;11)(q21;q23) translocation characterizes a form of acute lymphoblastic leukemia with a poor prognosis. It results in a fusion gene encoding a chimeric transcription factor, MLL-AF4, that deregulates gene expression through a variety of still controversial mechanisms. To provide new insights into these mechanisms, we examined the interaction between AF4, the most common MLL fusion partner, and the scaffold protein 14-3-3θ, in the context of t(4;11)-positive leukemia.

Methods: Protein-protein interactions were analyzed using immunoprecipitation and in vitro binding assays, and by fluorescence microscopy in t(4;11)-positive RS4;11 and MV4-11 leukemia cells and in HEK293 cells. Protein and mRNA expression levels were determined by Western blotting and RT-qPCR, respectively. A 5-bromo-2'-deoxyuridine assay and an annexin V/propidium iodide assay were used to assess proliferation and apoptosis rates, respectively, in t(4;11)-positive and control cells. Chromatin immunoprecipitation was performed to assess binding of 14-3-3θ and AF4 to a specific promoter element.

Results: We found that AF4 and 14-3-3θ are nuclear interactors, that 14-3-3θ binds Ser of AF4 and that 14-3-3θ forms a complex with MLL-AF4. In addition, we found that in t(4;11)-positive cells, 14-3-3θ knockdown decreased the expression of MLL-AF4 target genes, induced apoptosis and hampered cell proliferation. Moreover, we found that 14-3-3θ knockdown impaired the recruitment of AF4, but not of MLL-AF4, to target chromatin. Overall, our data indicate that the activity of the chimeric transcription factor MLL-AF4 depends on the cellular availability of 14-3-3θ, which triggers the transactivating function and subsequent degradation of AF4.

Conclusions: From our data we conclude that the scaffold protein 14-3-3θ enhances the aberrant activity of the chimeric transcription factor MLL-AF4 and, therefore, represents a new player in the molecular pathogenesis of t(4;11)-positive leukemia and a new promising therapeutic target.
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http://dx.doi.org/10.1007/s13402-019-00468-6DOI Listing
December 2019

The Molecular Hallmarks of the Serrated Pathway in Colorectal Cancer.

Cancers (Basel) 2019 Jul 20;11(7). Epub 2019 Jul 20.

CEINGE-Biotecnologie Avanzate s.c.ar.l., 80145 Naples, Italy.

Colorectal cancer (CRC) is a leading cause of cancer death worldwide. It includes different subtypes that differ in their clinical and prognostic features. In the past decade, in addition to the conventional adenoma-carcinoma model, an alternative multistep mechanism of carcinogenesis, namely the "serrated pathway", has been described. Approximately, 15 to 30% of all CRCs arise from neoplastic serrated polyps, a heterogeneous group of lesions that are histologically classified into three morphologic categories: hyperplastic polyps, sessile serrated adenomas/polyps, and the traditional serrated adenomas/polyps. Serrated polyps are characterized by genetic ( or mutations) and epigenetic (CpG island methylator phenotype (CIMP)) alterations that cooperate to initiate and drive malignant transformation from normal colon mucosa to polyps, and then to CRC. The high heterogeneity of the serrated lesions renders their diagnostic and pathological interpretation difficult. Hence, novel genetic and epigenetic biomarkers are required for better classification and management of CRCs. To date, several molecular alterations have been associated with the serrated polyp-CRC sequence. In addition, the gut microbiota is emerging as a contributor to/modulator of the serrated pathway. This review summarizes the state of the art of the genetic, epigenetic and microbiota signatures associated with serrated CRCs, together with their clinical implications.
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http://dx.doi.org/10.3390/cancers11071017DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6678087PMC
July 2019

Publisher Correction: Randomized controlled trial on the influence of dietary intervention on epigenetic mechanisms in children with cow's milk allergy: the EPICMA study.

Sci Rep 2019 Jun 26;9(1):9504. Epub 2019 Jun 26.

Department of Translational Medical Science, University Federico II, Naples, Italy.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.
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http://dx.doi.org/10.1038/s41598-019-45226-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6593099PMC
June 2019

Sequence Variants in Myopathies: Expression and Functional Studies in Two Families.

Biomed Res Int 2019 21;2019:7638946. Epub 2019 Apr 21.

CEINGE-Advanced Biotechnologies, Via Gaetano Salvatore 486, 80145 Naples, Italy.

The skeletal muscle ryanodine receptor (RyR1), i.e., the Ca channel of the sarco/endoplasmic reticulum (S/ER), and the voltage-dependent calcium channel Cav1.1 are the principal channels involved in excitation-contraction coupling in skeletal muscle. gene variants are linked to distinct skeletal muscle disorders, including malignant hyperthermia susceptibility and central core disease (CCD), mainly with autosomal dominant inheritance, and autosomal recessive myopathies with a broad phenotypic and histopathological spectrum. The age at onset of -related myopathies varies from infancy to adulthood. We report the identification of four variants in two Italian families: one with myopathy and variants c.4003C>T (p.R1335C) and c.7035C>A (p.S2345R), and another with CCD and variants c.9293G>T (p.S3098I) and c.14771_14772insTAGACAGGGTGTTGCTCTGTTGCCCTTCTT (p.F4924_V4925insRQGVALLPFF). We demonstrate that, in patient-specific lymphoblastoid cells, the c.4003C>T (p.R1335C) variant is not expressed and the in-frame 30-nucleotide insertion variant is expressed at a low level. Moreover, Ca release in response to the RyR1 agonist 4-chloro-m-cresol and to thapsigargin showed that the c.7035C>A (p.S2345R) variant causes depletion of S/ER Ca stores and that the compound heterozygosity for variant c.9293G>T (p.S3098I) and the 30-nucleotide insertion increases RyR1-dependent Ca release without affecting ER Ca stores. In conclusion, we detected and functionally characterized disease-causing variants of the RyR1 channel in patient-specific lymphoblastoid cells. This paper is dedicated to the memory and contribution of Luigi Del Vecchio.
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http://dx.doi.org/10.1155/2019/7638946DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6500691PMC
December 2019
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