HUMAN MESENCHYMAL STEM CELLS AS A MODEL OF STUDY FOR NEW BIOMATERIALS IN BONE TISSUE REGENERATION

The reconstruction of large bone segments remains a critical clinical problem in the case of extensive bone loss due to traumatic or pathological events. Due to its ideal biocompatibility and osteogenic properties, autologous bone is still considered the gold standard for bone replacement applications, however its use has limitations such as supply amount and unpredictable healing kinetics. These limitations and recent progress in biotechnology have driven the development of materials/scaffolds for bone repair. Biocompatibility issues and materials/scaffolds features need to be investigated and the usefulness of human cell cultures for biocompatibility testing is confirmed by experimental studies. In-vitro models employing human cells to study the interactions between the cell system and the biomaterial/device allow for a reasonable prediction of its performance in vivo. To study coral derived Hydroxyl-Apatite (HA) osteo-conductivity and cyto-toxicity we employed an engineered osteoblast-like cellular model named Saos-eGFP. Although the efficacy of this cellular model, the need of an in-vitro tool resembling a more physiological-like cellular response, led to the development of an adult human mesenchymal stem cell (hMSCs) model. This model was employed to assay porous stoichiometric HA and non-stoichiometric Mg-HA bone substitutes. Our studies showed how human cells may be employed to analyze osteo- conductivity/inductivity and cyto-toxicity, and specifically how hMSCs allow a more insightful understanding of the earliest phase of cell-biomaterial interaction, addressing the question of which type of biomaterials could be better for bone regeneration applications

Multiple or metastatic clear cell chondrosarcoma: a case report

We report multiple synchronous clear-cell chondrosarcomas in a 43-year-old patient. The patient had a lesion in the right proximal humerus and in the left femoral condyle. Bone scintigraphy revealed increased uptake in both foci. Pathological analysis confirmed the diagnosis in both locations. In the proximal humerus, wide resection of the tumour was performed with allograft reconstruction of the joint with osteosynthesis. The femoral condyle was treated with curettage, phenolization, and cementation. Over a follow-up of 10 years no recurrence or metastasis was observed

Neurological involvement in Ile68Leu (p.Ile88Leu) ATTR amyloidosis: not only a cardiogenic mutation

Ile68Leu transthyretin-related amyloidosis (ATTR) is known as a mainly or exclusively cardiogenic variant. We hypothesized that an accurate specialized neurological evaluation could reveal a consistent frequency of mixed phenotypes.Forty-six consecutive subjects with transthyretin (TTR) Ile68Leu (p.Ile88Leu) mutation (29 patients and 17 unaffected carriers) underwent an in-depth cardiac and neurologic evaluation at a single center.All 29 patients showed cardiac involvement. In 20 (69%) cases, it was associated with neurological abnormalities (i.e. a mixed phenotype): 10 (35% of the total) had signs and symptoms of neuropathy, 5 (17%) had abnormalities at the neurologic specialist examination but without symptoms, and 5 (17%) had abnormal nerve conduction study only. None of the asymptomatic carriers showed neurological abnormalities or cardiac involvement. The Neuropathy Impairment Score was5 in seven patients at baseline, and became5 in six more patients during follow-up. The probability of experiencing a major adverse cardiac event (MACE) during follow-up was higher in the mixed than cardiologic phenotype (At least two-thirds of patients with Ile68Leu ATTR and amyloidotic cardiomyopathy show an associated - definite or probable - neurologic impairment of variable degree if accurately evaluated in a neurologic setting. This proportion can rise during follow-up. The mixed phenotype carries a worse prognosis compared to the exclusively cardiologic one. These observations show that more patients could be eligible for treatment with gene silencers than currently indicated and highlight the need for an in-depth and continuous multidisciplinary evaluation of Ile68Leu ATTR patients

Minorenni vittime di omicidio a Milano (Italia): 1993-2017

This study aims to examine the phenomenon of homicide with victims under 18 years-old in a wide and multiethnic metropolitanarea in the north of Italy. We’re dealing with a type of crime that generates a widespread alarm and a common concern.The data analysis, approached with a multidisciplinary strategy, presented in this paper is based on forensic reports of the Instituteof Legal Medicine of the University of Milan, to provide a detailed framework of the homicides of minors that took place inthe territorial jurisdiction of Milan and Monza Prosecutor’s Office between 1993 and 2017. Quantitative and qualitative analysiswere conducted regarding victims’ socio-demographic characteristics (gender, age, nationality), weapons used, places and modalityof body discovery, murderers, motives of the killing and type of relationship between offenders and victims. Purpose of thispaper is to compare the present set of data with the ones provided by the international literature, as well as make some criminologicalobservations on this matter with a view to preventing the phenomenon. To this end, some representative case studieshave been added, their characteristics and also the relationships between offender and victim have been discussed.Questo studio esamina il fenomeno dell'omicidio con vittime di età inferiore ai 18 anni in una vasta e multietnica area metropolitanadel nord Italia, un tipo di crimine che genera allarme e preoccupazione. L'analisi dei dati, affrontata con una metodologia multidisciplinare,è basata sui verbali autoptici dell'Istituto di Medicina Legale dell'Università di Milano, al fine di fornire un quadro dettagliatodegli omicidi con vittima minorenne che si sono verificati nella giurisdizione territoriale della Procura di Milano e Monzatra il 1993 e il 2017. Sono state condotte analisi quantitative e qualitative sulle vittime, quali le caratteristiche socio-demografiche(genere, età, nazionalità), le armi utilizzate, i luoghi e le modalità di rinvenimento del corpo, le caratteristiche dell’omicida, i motividell'uccisione e il tipo di relazione tra criminale e vittima. Scopo di questo studio è confrontare l'attuale insieme di dati con quelliforniti dalla letteratura internazionale, nonché porre alcune considerazioni di carattere preventivo. A tal fine, sono stati presentatianche alcuni casi studio rappresentativi, discutendone le caratteristiche peculiari e le relazioni tra l'autore del reato e la vittima

Techniques and machines for conservation and organic agriculture: the S.M.O.C.A. project.

The combination of low environmental impact farming techniques and conservation agriculture techniques is considered not feasible due to some limitations, mainly including is the strong dependence of conservation cropping systems by chemical control of weeds and the use of mineral fertilizers, which are considered essential for supporting to acceptable levels of crop productions. In order to apply the techniques of reduced tillage also in organic and integrated agriculture, is therefore indispensable, on one hand, the availability of specific versatile and efficient machines for non-chemical cover-crop management, weed control and sod-seeding/planting, on the other hand, the optimization of cropping systems in function of improved nutrient cycling and preventive weed control. The SMOCA project (Smart Management Conservation of Organic Agriculture) aims to integrate organic cropping systems and conservation agriculture techniques in three different scenarios (arable open field, open field vegetables and orchards) thanks to development of machines and technical itineraries that allow to apply the reduced tillage techniques even in the absence of pesticides. Within this project, prototypes of machines were realized in order to implement conservation cropping systems with high energy efficiency. For each scenario, three technical itineraries were compared: - control (INT): based on integrated agriculture without use of conservation agriculture techniques. The use of agrochemicals is allowed in accordance with the disciplinary for integrated production; - organic system (ORG): based on the organic agriculture, involves the use of cover crops (managed as green manure) and soil tillage. The non chemical management of weed flora is achieved with preventive methods and direct methods (mechanical and thermal weed control); - organic and conservation system (ORG +): based on the deep integration between the principles of conservation agriculture and organic farming. The system aims to achieve an almost continuous coverage of the soil through the use of no-tillage techniques and cover crops managed as living or dead mulches. The study of the overall sustainability of the different systems in comparison is conducted by means of a detailed analysis of the following aspects: - agronomic; - energy and environmental; - economic. The environmental effect of the compared systems will be mainly assessed through the study of the following aspects: greenhouse gas emissions, nutrient balance, preservation of soil quality, biological nitrogen fixation, conservation of water resources in the soil and the life cycle assessment (LCA). The efficiency and the impact of different management techniques are also continuously monitored on the orchard and vegetable crops through the study of the fruits of growth dynamics, thanks to an innovative methodology that uses a network of growth sensors spatially distributed in the field. The final product quality is also evaluated through the analysis of their levels of nutraceutical compounds. The research project is still ongoing and the prototypes of machines realized are presented in this paper

Renin-Angiotensin-Aldosterone System Inhibitors, Statins, and Beta-Blockers in Diabetic Patients With Critical Limb Ischemia and Foot Lesions

Medical therapy for secondary prevention is known to be under-used in patients with peripheral artery disease (PAD). Few data are available on the subgroup with critical limb ischemia (CLI). Prescription of cardiovascular preventive therapies was recorded at discharge in a large, prospective cohort of patients admitted for treatment of CLI and foot lesions, stratified for coronary artery disease (CAD) diagnosis. All patients were followed up for at least 1 year. The primary endpoint was major adverse cardiovascular events (MACE). 618 patients were observed for a median follow-up of 981 days. Renin-angiotensin-aldosterone system (RAAS) inhibitors, statins, beta-blockers, and antithrombotic drugs were prescribed in 52%, 80%, 51%, and 99% of patients, respectively. However, only 43% of patients received optimal medical therapy (OMT), defined as the combination of RAAS inhibitor plus statin plus at least one antithrombotic drug. It was observed that the prescription of OMT was not affected by the presence of a CAD diagnosis. On the other hand, it was noticed that the renal function affected the prescription of OMT. OMT was independently associated with MACE (HR 0.688, 95%CI 0.475-0.995, P = .047) and, after propensity matching, also with all-cause mortality (HR 0.626, 95%CI 0.409-0.958, P = .031). Beta-blockers prescription was not associated with any outcome. In conclusion, patients with critical limb ischemia are under-treated with cardiovascular preventive therapies, irrespective of a CAD diagnosis. This has consequences on their prognosis

Serum IgG antibodies from pregnant women reacting to mimotopes of simian virus 40 large T antigen, the viral oncoprotein

Simian virus 40 (SV40) large T antigen (LT) coding sequences were revealed in different human samples, whereas SV40 antibodies (Ab) were detected in human sera of cancer patients and healthy individuals, although with a lower prevalence. Previous studies carried out by the neutralization assay gave a SV40 seroprevalence, in the general population, up to 8%, although higher rates, 12%, were detected in kidney transplant children, in a group of HIV-positive patients, and in healthy females. In this study, serum samples from pregnant women, together with those from non-pregnant women, were analyzed to check the prevalence of IgG Ab reacting to SV40 LT antigens. Serum samples were collected from pregnant and non-pregnant women, with the same mean age. Women were in the range of 15-48 years old. Samples were assayed by an indirect ELISA employing specific SV40 LT mimotopes as antigens, whereas functional analysis was performed by neutralization of the viral infectivity in cell cultures. As a control, sera were analyzed for Ab against BK polyomavirus (BKPyV), which is a human polyomavirus homologous to SV40. Statistical analyses employed chi-square with Yates' correction, and Student's t tests. Indirect ELISAs indicated that pregnant women tested SV40 LT-positive with a prevalence of 17% (23/134), whereas non-pregnant women had a prevalence of 20% (36/180) (P > 0.05). Ab against BKPyV were detected with a prevalence of 80% in pregnant women and with a prevalence of 78% in non-pregnant women. These data indicate that SV40 infects at a low prevalence pregnant women. We may speculate that SV40, or a close human polyomavirus still undetected, could be transmitted from mother to fetus

Optimized pipeline of MuTect and GATK tools to improve the detection of somatic single nucleotide polymorphisms in whole- exome sequencing data

Background: Detecting somatic mutations in whole exome sequencing data of cancer samples has become a popular approach for profiling cancer development, progression and chemotherapy resistance. Several studies have proposed software packages, filters and parametrizations. However, many research groups reported low concordance among different methods. We aimed to develop a pipeline which detects a wide range of single nucleotide mutations with high validation rates. We combined two standard tools – Genome Analysis Toolkit (GATK) and MuTect – to create the GATK-LODN method. As proof of principle, we applied our pipeline to exome sequencing data of hematological (Acute Myeloid and Acute Lymphoblastic Leukemias) and solid (Gastrointestinal Stromal Tumor and Lung Adenocarcinoma) tumors. We performed experiments on simulated data to test the sensitivity and specificity of our pipeline. Results: The software MuTect presented the highest validation rate (90 %) for mutation detection, but limited number of somatic mutations detected. The GATK detected a high number of mutations but with low specificity. The GATK-LODN increased the performance of the GATK variant detection (from 5 of 14 to 3 of 4 confirmed variants), while preserving mutations not detected by MuTect. However, GATK-LODN filtered more variants in the hematological samples than in the solid tumors. Experiments in simulated data demonstrated that GATK-LODN increased both specificity and sensitivity of GATK results. Conclusion: We presented a pipeline that detects a wide range of somatic single nucleotide variants, with good validation rates, from exome sequencing data of cancer samples. We also showed the advantage of combining standard algorithms to create the GATK-LODN method, that increased specificity and sensitivity of GATK results. This pipeline can be helpful in discovery studies aimed to profile the somatic mutational landscape of cancer genomes

Acute Prosthetic Joint Infections with Poor Outcome Caused by Staphylococcus Aureus Strains Producing the Panton-Valentine Leukocidin

The aim of this study was to investigate whether the presence of Staphylococcus aureus (SA) producing the Panton-Valentine leukocidin (PVL) affects the outcome of Prosthetic Joint Infection (PJI). Patients with acute and chronic PJI sustained by SA were prospectively enrolled at the orthopedic unit of "Casa di Cura Santa Maria Maddalena", from January 2019 to October 2021. PJI diagnosis was reached according to the diagnostic criteria of the International Consensus Meeting on PJI of Philadelphia. Synovial fluid obtained via joint aspirations was collected in order to isolate SA. The detection of PVL was performed via real-time quantitative PCR (RT-qPCR). The outcome assessment was performed using the criteria of the Delphi-based International Multidisciplinary Consensus. Twelve cases of PJI caused by SA were included. Nine (75%) cases were acute PJI treated using debridement, antibiotic and implant retention (DAIR); the remaining three (25%) were chronic PJI treated using two-stage (n = 2) and one-stage revision (n = 1), respectively. The SA strains that tested positive for PVL genes were 5/12 (41.6%,). Treatment failure was documented in three cases of acute PJI treated using DAIR, all supported by SA-PVL strains (p < 0.045). The remaining two cases were chronic PJI treated with a revision arthroplasty (one and two stage, respectively), with a 100% eradication rate in a medium follow-up of 24 months. Although a small case series, our study showed a 100% failure rate in acute PJI, probably caused by SA PVL-producing strains treated conservatively (p < 0.04). In this setting, toxin research should guide radical surgical treatment and targeted antibiotic therapy

Integrazione di tecniche di agricoltura biologica e conservativa in sistemi colturali con crescente intensità ecologica: il progetto F.I.R.B. SMOCA

Il progetto SMOCA (Smart Management of Organic Conservative Agriculture) (2014- 2017) mira ad incrementare la sostenibilità dei sistemi colturali integrati/biologici mediante l’introduzione di tecniche di agricoltura conservativa, finalizzate alla riduzione dei consumi energetici e al miglioramento della fertilità del terreno. In SMOCA saranno sviluppate macchine e strategie agronomiche innovative che permettano di applicare le tecniche di lavorazione ridotta anche in assenza di mezzi chimici di sintesi