The high potential of micro-magnetic resonance imaging for the identification of archaeological reeds. The case study of Tutankhamun

This study explores the potential of micro-magnetic resonance imaging ( -MRI) for identifying archaeological reeds found in the tomb of Tutankhamun. Reed plants had various historical uses in the past, with ancient Egyptians extensively employing them for crafting a wide range of items. The distinct cross-sectional characteristics of Arundo donax (giant reed) and Phragmites australis (common reed) are observed and described via optical microscopy and  -MRI in this study. While optical microscopy offers higher resolution,  -MRI provides advantages for studying archaeobotanical specimens, as it eliminates the need for mechanical sectioning and potentially damaging fragile samples. The application of  -MRI on a selected archaeological reed allowed us to identify it as Phragmites australis, showing that  -MRI can yield clear images, maintaining the integrity of the sample. In contrast, diagnostic features appeared greatly deformed on the thin section observed via optical microscopy. Despite the limitations related to the sample size and the need for sample soaking,  -MRI presents a valuable tool for analyzing archaeological remains in the field of cultural heritage, with the potential for broader applications. Overall, this study contributes to expanding the toolkit available to researchers studying plant remains, providing insights into reed identification and preservation in archaeological contexts

A Unique Case of Bilateral Thalamic High-Grade Glioma in a Pediatric Patient with LI-Fraumeni Syndrome: Case Presentation and Review of the Literature

Li-Fraumeni syndrome (LFS) is a rare high-penetrance and autosomal-dominant pathological condition caused by the germline mutation of the TP53 gene, predisposing to the development of tumors from pediatric age. We conducted a qualitative systematic review following the ENTREQ (Enhancing Transparency in Reporting the Synthesis of Qualitative Research) framework. A search was made in MEDLINE/Pubmed and MeSH Database using the terms “Li-Fraumeni” AND “pediatric high-grade glioma (HGG)”, identifying six cases of HGGs in pediatric patients with LFS. We added a further case with peculiar features such as no familiar history of LFS, association of embryonal rhabdomyosarcoma and bithalamic HGG, whose immunohistochemical profile was accurately defined by Next Generation Sequencing. Knowledge synthesis and case analysis grounded the discussion about challenges in the management of this pathology in pediatric age

The Q-LAMP Method Represents a Valid and Rapid Alternative for the Detection of the BCR-ABL1 Rearrangement in Philadelphia-Positive Leukemias

Molecular detection of the BCR-ABL1 fusion transcripts is necessary for the genetic confirmation of a chronic myeloid leukemia diagnosis and for the risk classification of acute lymphoblastic leukemia. BCR-ABL1 mRNAs are usually identified using a conventional RT-PCR technique according to the BIOMED-1 method. In this study, we evaluated 122 BCR-ABL1-positive samples with the Q-LAMP assay to establish if this technology may represent a valid alternative to the qualitative BIOMED-1 PCR technique usually employed for the detection and the discrimination of the common BCR-ABL1 transcripts (p190 and p210 isoforms). We found a 100% concordance rate between the two methods. Specifically, the p190- and p210-positive samples were amplified by Q-LAMP with a median threshold time (Tt) of 26.70 min (range: 24.45-31.80 min) and 20.26 min (range: 15.25-34.57 min), respectively. A median time of 19.63 was observed in samples displaying both (e13a2/e14a2) p210 isoforms. Moreover, the Q-LAMP assay allowed recognition of the BCR-ABL1 e13a2 and e14a2 isoforms (median Tts 18.48 for e13a2 vs. 26.08 min for e14a2; p < 0.001). Finally, 20 samples harboring rare BCR-ABL1 isoforms (e1a3, e13a3, e14a3, and e19a2) were correctly identified by the Q-LAMP assay. We conclude that the Q-LAMP assay may represent a faster and valid alternative to the qualitative BIOMED-1 RT-PCR for the diagnosis at BCR-ABL1-positive leukemias, especially when samples are analyzed in centers with restricted resources and/or limited technical expertise

A multi-parametric investigation on waterlogged wood using a magnetic resonance imaging clinical scanner

In cultural heritage conservation science, moisture content (MC) is an essential factor to determine. At the same time, it is essential to choose non-destructive and non-invasive approaches for more sustainable investigations and make them safe for the environment and the sample. The question addressed in this work concerns the possibility and the opportunity to investigate waterlogged wood by using nuclear magnetic resonance imaging (MRI) clinical scanners to carry out non-destructive volumetric diagnostics. In this study, MRI, the most important non-invasive medical imaging technique for human tissue analysis, was applied to study archaeological waterlogged wood samples. This type of archaeological material has a very high moisture content (400%–800%), thus, it is an ideal investigative subject for MRI which detects water molecules inside matter. By following this methodology, it was possible to obtain information about water content and conservation status through a T1, T2, and T2* weighted image analysis, without any sampling or handling, and the samples were directly scanned in the water where they were stored. Furthermore, it permited processing 3D reconstruction, which could be an innovative tool for the digitalization of marine archaeological collections. In this work, 16 modern species of wood and a waterlogged archaeological wood sample were studied and investigated using a clinical NMR scanner operating at 3T. The results were compared with X-ray computed tomography (CT) images, as they had already been used for dendrochronology. The comparison highlights the similar, different, and complementary information about moisture content and conservation status in an all-in-one methodology obtainable from both MRI and CT techniques

Multiparametric and multimodal nuclear magnetic resonance diagnostics of wood for cultural heritage applications

La presente tesi di dottorato introduce un nuovo paradigma per la diagnostica non invasiva e non distruttiva della struttura e della fisiologia del legno, delle opere lignee e dei resti di legno archeologico basata sulla tecnologica di Risonanza Magnetica Nucleare protonica (1H-NMR). In questa tesi, è stata studiata l’organizzazione topologica della macro- e microstruttura del legno attraverso l’Imaging a Risonanza Magnetica (MRI) ad alta risoluzione e la diffusione RMN dell’acqua nel legno. La fisiologia e le proprietà chimico-fisiche dei componenti del legno (cellulosa, lignina) insieme alle sezioni trasversali dei vasi/tracheidi e delle fibre del legno sono stati investigati tramite la rilassometria RMN e la spettroscopia RMN di correlazioni 2D (COSY). Diversi modelli che descrivono il comportamento del segnale RMN sono stati testati ed applicati. Diversi protocolli RMN sono stati ottimizzati per la diagnostica di oggetti lignei, sia contemporanei che archeologici e di interesse nei Beni Culturali, con particolare riguardo agli aspetti di non invasività e non distruttività delle misure. A tale scopo, oltre all’utilizzo di uno spettrometro RMN ad alto campo magnetico (9.4 T), che consente misure non distruttive su piccoli campioni, sono stati utilizzati anche uno scanner clinico, con campo magnetico di 3 T per investigare campioni più grandi, e uno spettrometro RMN portatile a basso campo magnetico (0.35 T), che permette misure non invasive su oggetti di qualsiasi dimensione. L’aspetto che accomuna tutte queste tecniche è l’impiego di molecole d’acqua liquida o in forma di vapore per sondare le caratteristiche strutturali del legno. Il metodo di imaging in risonanza magnetica (MRI) è stato utilizzato per implementare un protocollo ad-hoc per l’osservazione dei caratteri diagnostici del legno e del suo stato di conservazione. Le immagini RMN sono state acquisite, mediante sezionamento virtuale, su legni moderni ed archeologici di età romana perfezionando le sequenze di acquisizione in modo tale da ottenere immagini ad alta risoluzione (fino a risoluzione lineare massima di 8 μm) per l’osservazione di tutti gli elementi anatomici del legno. Inoltre, pesando le immagini RMN sui tempi di rilassamenti (T1, T2, T2*) o sul coefficiente di diffusione (D) è stato possibile ottenere un contrasto ottimizzato in grado di fornire informazioni chimico-fisiche e dinamiche aggiuntive. L’istologia virtuale completa ottenuta nelle tre direzioni anatomiche del legno, trasversale, radiale e tangenziale, ha dimostrato che l’MRI è una valida tecnica non distruttiva per investigare i legni sommersi, soprattutto quelli hardwoods, e che fornisce informazioni complementari a quelle fornite dalla microscopia ottica ed elettronica. Essa permette anche di ricostruire in 3D l’intero campione in modo non-distruttivo a partire dalle immagini RMN 2D. Inoltre, per superare le restrizioni sulla dimensione del campione imposte dall’MRI realizzato con spettrometro ad alto campo magnetico (9.4 T), questa tesi riporta i risultati MRI ottenuti con scanner clinico (3 T) su legno archeologico sommerso. Sono state acquisite le prime immagini RM pesate in T1, T2 e T2* con risoluzione lineare di 250 micrometri di un palo di legno (diametro di 8.7 cm) dell’antico porto romano di Napoli e queste vengono confrontate con le immagini di tomografia computerizzata (TC). Questo studio ha suggerito che l’analisi MRI fornisce informazioni complementari utili per valutare lo stato di conservazione del legno archeologico e l’osservazione di alcuni caratteri diagnostici permettendo di riconoscere un softwood da un hardwood. Gli esperimenti di diffusione RMN sia su legni moderni imbibiti che archeologici sommersi hanno mostrato l’esistenza di diversi compartimenti diffusivi associati a diversi valori del coefficiente di diffusione molecolare (D) dell’acqua. Grazie all’impiego di una sequenza di acquisizione Pulsed Field Gradient (PFG) è stato possibile misurare il D a diversi tempi di osservazione Δ. Lo studio dell’andamento di D(Δ) ha consentito di ricavare informazioni circa la dimensione e l’interconnessione dei pori del legno e la permeabilità delle pareti cellulari. Inoltre, dai dati di diffusione è stato possibile estrarre informazioni circa il legno come mezzo poroso, ossia la tortuosità dell’acqua e la sua anisotropia. Le tecniche di rilassometria RMN sono state utilizzate per studiare la dinamica delle molecole d’acqua nelle varie strutture anatomiche del legno mediante la misura del tempo di rilassamento spin-reticolo (T1) e del tempo di rilassamento spin-spin (T2), i quali rappresentano un indice del grado di mobilità dell’acqua e delle sue interazioni con la matrice lignea. In particolare, nel legno archeologico sommerso di età romana la misura dei tempi di rilassamento analizzati con la trasformata di Laplace ha permesso di ricavare la distribuzione delle dimensioni dei pori, evidenziando un incremento del diametro medio delle strutture del legno dovuto al degrado della parete cellulare. Dalla misura del tempo di rilassamento spin-reticolo (T1) è stato anche possibile rivelare la presenza di sostanze paramagnetiche sotto forma di impurezze depositate all’interno dei reperti lignei. Nei legni a temperatura ambiente e umidità pari al 94%, la correlazione tra tempi di rilassamento T1 e T2 delle molecole d’acqua presenti nella parete cellulare e misurati su 15 specie lignee diverse ha permesso di evidenziare il diverso comportamento igroscopico della parete cellulare dei softwoods in confronto alla parete cellulare degli hardwoods. Questo ha permesso di distinguere, in modo non invasivo, i legni softwoods dagli hardwoods. Un’ipotesi per spiegare questo risultato potrebbe essere la diversa composizione della parete cellulare (e.g., contenuto di emicellulosa) dei softwoods rispetto agli hardwoods, ma ulteriori studi verranno effettuati. Inoltre, utilizzando l’RMN unilaterale portatile e sfruttando i metodi di Laplace RMN multidimensionale (2D), in combinazione con gli esperimenti monodimensionali (1D), sono state ottenute le proprietà strutturali e dinamiche dell’acqua nel legno archeologico sommerso di età romana in maniera non invasiva. Gli esperimenti di correlazione T1-T2 e D-T2 hanno permesso di sondare ed identificare un maggior numero di compartimenti di acqua. Unendo gli esperimenti 1D e 2D è stata possibile una caratterizzazione completa di ciascun compartimento di acqua, corrispondente ad una specifica struttura del legno, mediante i suoi valori di T1, T2 e D. I legni archeologici sono risultati privi della componente caratteristica di rilassamento corrispondente all’acqua legata nelle pareti cellulari (pari a circa 1 ms), che invece è presente nei legni contemporanei. Tale risultato ha suggerito l’abilità dell’RMN portatile nel diagnosticare la presenza di degrado. Mentre veniva portato avanti il lavoro di tesi, la tecnologica, i protocolli ed i modelli di RMN portatile usati per studiare il legno, sono stati anche estesi e adattati allo studio di materiali lapidei e della carta. Per i primi, le misure dei tempi di rilassamento T1 e T2 hanno permesso di valutare l’efficacia pulente di un idrogel a base di PVA-PEO e borace; per i secondi invece è stato monitorato l’effetto dell’esposizione alla luce UV

Decay of a Roman age pine wood studied by micro magnetic resonance imaging, diffusion nuclear magnetic resonance and portable nuclear magnetic resonance

Wood is a hygroscopic biodegradable porous material widely used by men in the past to create artworks. Its total preservation over time is quite rare and one of the best preservation modalities is waterlogging. Observing the anatomy of waterlogged archaeological wood could also be complicated because of its bacterial degradation. However, the characterization of wood morphology and conservation state is a fundamental step before starting any restoration intervention as it allows to extract information about past climatic conditions and human activities. In this work, a micro-invasive approach based on the combined use of high-resolution magnetic resonance imaging (MRI) and diffusion-nuclear magnetic resonance (NMR) was tested both on a modern and an ancient pine wood sample. Furthermore, a completely non-invasive analysis was performed by using portable NMR. This multi-analytical NMR approach allowed to highlight the effect of decay on the wood microstructure, through alterations in the pores size, tortuosity, and images contrast of the ancient pine compared to the modern one. This work pointed out the different but complementary multi-parametric information that can be obtained by using NMR and tested the potential of high-field MRI and low-field portable NMR in the detection of wood diagnostic features

Effect of age on Pine wood microstructure studied by micro-MRI and diffusion-NMR

Wood is a natural complex material widely used from men in the past to create artworks. One of its main anatomical elements is the annual ring that varies according to the species, the weather conditions under which the tree has grown and to possible adversities. To observe the anatomy of waterlogged archaeological wood could be complicated because of its degradation. However, knowing the state of conservation is very important for the future restoration. In this work a non-destructive approach based on the combined use of MRI and diffusion on the modern and ancient pine wood is presented. Micro-MR images allow to observe the diagnostic features. The molecular NMR diffusion analysis, with the estimation of the pores diameter and the tortuosity, provide important information about the effect of age on the wood microstructure. At the end of the analysis the unaltered sample can be repositioned in its original location on the artwork

NMR PROTOCOL FOR MULTISCALE CHARACTERIZATION OF ARCHAEOLOGICAL WATERLOGGED WOOD

Wood is a natural porous material with pores size between 0-400 μm and its structures [1,2] vary according to the species and to the anatomical direction observed. Wood is also a hygroscopic material in which water molecules diffuse and the diffusion coefficient is vehicle of information about pores dimension and distribution. Aim of this work was to characterize archaeological waterlogged wood by non-destructive nuclear magnetic resonance diffusion, relaxometry, imaging and cryoporometry in order to investigate the samples at different length scales (nanometer, micrometer and sub-millimeter scale). The work is divided in two steps. In the frst step, the molecular diffusion protocol was validated by studying fve soaked modern woods (four hardwoods and one softwood) and measuring the diffusion coefficient along x axis (i.e. perpendicular to the main axis of vessels/tracheids) with a 400 MHz Bruker-Avance spectrometer. By introducing data correction to remove the diffusion of water through semi-permeable membranes (cell walls) [3], pores diameter was estimated. In the second step, the characterization of the porosity and microstructure of a small and archaeological submerged wood excavated in Naples and belonging to the 5th century A.D. was carried out. Relaxation times T 1 and T 2 were quantifed and magnetic resonance images were acquired by using a 300 MHz or 400 MHz spectrometer while cryoporometry was performed at 500 MHz using spin-echo and CPMG experiments over the temperature range 180-294 K, increasing temperature in 76 steps of 1.5 K [4]. T 1 and T 2 provided preliminary information about water compartments, wood density and porosity. T 2 and T 1-weighted images with an in-plane resolution of 18 x 18 μm 2 allowed to observe anatomical and physiological elements under the micrometer and sub-millimeter scale while T 2 *-weighted images highlighted the presence of paramagnetic impurities due to the decay. Thanks to the magnetic resonance images it was possible to get a complete and non-destructive virtual histology in the three anatomical sections of wood (Fig. 1)

Single-Sided Portable NMR Investigation to Assess and Monitor Cleaning Action of PVA-Borax Hydrogel in Travertine and Lecce Stone

In this work, we investigated the potential of PVA-borax hydrogel for cleaning limestones and the dependence of the cleaning on the porosity of the rock and on the action time of the hydrogel treatment. Towards this goal, we used a nuclear magnetic resonance (NMR) spectrometer, developed for non-invasive and non-destructive applications on cultural heritage. T2-NMR parameters were quantified on different samples of Lecce stone and Travertine cut perpendicular (Pe) and parallel (Pa) to the bedding planes under different experimental conditions: untreated samples, treated with Paraloid B72 and cleaned with PVA-PEO-borax hydrogel applied for 4 min and 2 h. The T2 results suggest that the effectiveness of the cleaning strongly depended on the porosity of the stones. In Lecce stone, the hydrogel seemed to eliminate both the paramagnetic impurities (in equal measure with 4 min and 2 h treatment) and Paraloid B72. In Travertine Pe, characterized by a smaller pore size compared to Lecce stone, no significant effects were found regarding both the cleaning and the treatment with Paraloid B72. In Travertine Pa, characterized by a larger pore size than the other two samples, the hydrogel seemed to clean the paramagnetic agents (it worked better if applied for a longer time) but it did not appear to have any effect on Paraloid B72 removal

Testing portable NMR to monitor the effect of paper exposure to UV-light

Paper-based works of art can be considered the most important carrier of information about culture, science, business, politic and history. Therefore, it is highly important to preserve the integrity of the paper these objects are made of. Paper is an organic material mainly made of cellulose fibers, whose durability depends on pH, heat, humidity, oxygen, pollution, metal ions, lignin, and UV-visible light. Cellulose absorbs more in the near UV region, therefore radiation with wavelength of 300–550 nm produces most of the paper damage. The aim of this work was to test the potential of single-sided portable NMR to highlight the effect of UV light on paper. To this end, the longitudinal relaxation time (T1) and the transversal relaxation time (T2) of structural-known filter paper before and after the exposure to UV light were measured and supported by Raman spectroscopy. The decrease of both T1 and T2 parameters with the increase of the UV exposure time indicates a modification of the cellulose chains, which was confirmed by Raman spectra. Moreover, this study presents a preliminary non-invasive protocol for assessing the effect of artificial UV irradiation on paper by using a portable NMR sensor