MRI characterization of rat brain aging at structural and functional level: Clues for translational applications.

Magnetic resonance imaging (MRI) paradigms, using non-invasive approaches, can provide relevant findings about brain aging. The attention has been primarily focused on neurodegenerative diseases, while little or nothing has been done to differentiate physiology from pathology. The present study aimed to test diffusion tensor imaging (DTI) and functional MRI (fMRI) metrics to analyze physiological age-related changes in rats at myelin structure and activation level; findings were validated by ex vivo histology. The purpose is to find comparable biomarkers in rodents and humans to allow a reliable translation from pre-clinical to clinical settings. Data evidenced: i) a significantly higher cerebrospinal fluid volume in middle-aged and aged vs. young rats; ii) a progressive alteration of white matter; iii) a significant reduction of evoked activity in aged animals. These results partially mirror the age-related changes in humans and may represent a preliminary step to find reliable tools for a lifelong monitoring with a value for the clinical practice (e.g., to provide support to the early diagnosis of dementia in asymptomatic subjects).

EGFR-Targeted Magnetic Nanovectors Recognize, in Vivo, Head and Neck Squamous Cells Carcinoma-Derived Tumors.

Head and neck squamous cell carcinomas (HNSCC) are a diverse group of tumors with high morbidity and mortality that have remained mostly unchanged over the past decades. The epidermal growth factor receptor (EGFR) is often overexpressed and activated in these tumors and strongly contributes to their pathogenesis. Still, EGFR-targeted therapies such as monoclonal antibodies and kinase inhibitors have demonstrated only limited improvements in the clinical outcome of this disease. Here, we take advantage of the extraordinary affinity of EGF for its cognate receptor to specifically target magnetite-containing nanoparticles to HNSCC cells and mediate, in vitro, their cellular upload. On the basis of this, we show efficient accumulation, in vivo, of such nanoparticles in subcutaneous xenograft tumor tissues in sufficient amounts to be able to mediate visualization by magnetic resonance imaging. Overall, our EGF-coated nanosystem may warrant, in the near future, novel and very efficient theranostic approaches to HNSCC.

Cryopreserved Subcutaneous Adipose Tissue for Fat Graft.

UNLABELLED: Cryopreservation of subcutaneous white adipose tissue (sWAT) avoids multiple surgeries in patients subjected to reconstructive procedure. Fat grafts were performed subcutaneously on 26 mice treated with fresh (13 mice) or cryopreserved (13 mice) human sWAT. Cytofluorometry for CD marker expression of stem cells, differentiation capability, and in vivo survival of fat grafts were evaluated. In vitro analysis evidenced that cryopreservation did not affect the stem potential of samples. In vivo MRI showed that grafts were well preserved in 13 mice treated with fresh sWAT, whereas in 13 animals treated with thawed fat, graft volumes were strongly reduced after 1 week. Ultrastructural studies performed both on fresh and thawed specimens demonstrated that grafts performed with thawed sWAT are able to store lipids more slowly with respect to grafts performed with fresh sWAT and adipocytes maintained a multilocular appearance. Collected data demonstrated that the protocol of cryopreservation could maintain the regenerative capability of the sWAT, but the rate of reabsorption after fat grafting is higher using cryopreserved sWAT. Maintaining the stem potential of sWAT after cryopreservation is a very important aspect for reconstructive and regenerative medicine. The employment of cryopreserved sWAT represents an interesting goal for surgeons. Surely there is the necessity to improve the protocol of cryopreservation. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors http://www.springer.com/00266 .

(1) H-MR spectroscopy characterization of the adipose tissue associated with colorectal tumor.

PURPOSE: To investigate colorectal tumor by looking at the lipid tissue around the lesion. Adipose tissue is not only an inert storage system for excess calories, but is involved in several pathological processes linked to tumor development. MATERIALS AND METHODS: We obtained 24 colorectal specimens after surgical excision from patients affected by colorectal cancer at different tumor stages. (1) H-MR spectoscopy (MRS) spectra were collected from two voxels, in proximity of the lesion and far from it, in each specimen with a preclinical scanner. We differentiated patients by pathological tumor stage, and we compared the chemical composition of adipose tissue between patient groups, both close the tumor and far from it. RESULTS: Monounsaturated fatty acids (MUFA) close the lesion are increased at higher tumor stages, while MUFA far from the lesion do not follow this trend. This study is really explorative due to the small numbers of subjects and we are cautious on the interpretation of the results. However, the approach of the present work allows a further insight in the tumor characterization. CONCLUSION: Looking at the lipid tissue around a lesion through (1) H-MRS can return important information regarding its genesis and development.

Multifunctional nanoprobes based on upconverting lanthanide doped CaF2: towards biocompatible materials for biomedical imaging.

Water dispersible Gd3+,Yb3+,Er3+ and Gd3+,Yb3+,Tm3+ doped CaF2 nanoparticles (NPs) were prepared by one-pot hydrothermal synthesis using citrate ions as capping agents without the need for any post-synthesis reaction. UC emissions are easily observed in the visible and infrared regions upon NIR diode laser excitation at 980 nm. EPR spectroscopy confirms the substitutional nature of the rare-earth doping, while magnetometric studies reveal that the NPs have a useful magnetization. MRI experiments conducted in vivo show that after 40 min from the injection, the NPs localize in the liver and spleen. Electron microscopy images of liver tissue reveal that the NPs are located in the Kupffer cells, although a small amount is also found in the hepatocytes. An excitation with a 980 nm emission on the excised liver and epithelial tissue induces clearly visible UC emission. The local temperature upon 980 nm irradiation was monitored in situ and it was found to increase slowly with the exposure time, maintaining under 1-2 °C for less than 60 second exposure. The NPs show a low toxicity towards cultured HeLa cells and human primary dendritic cells (DCs), and did not induce pro-inflammatory cytokine secretion by cultured human DCs, indicating that the NPs do not cause relevant adverse reactions in immune cells. Therefore, the present NPs are suitable candidates to be efficiently used in surgery applications, where spatial resolution and lack of harmful effects on human health are important issues.

Proton magnetic resonance spectroscopy: ex vivo study to investigate its prognostic role in colorectal cancer.

BACKGROUND: Proton Magnetic Resonance Spectroscopy (1H MRS) is used for clinical diagnosis in some tumours. The aim of this study is to explore ex vivo the potential of 1H MRS in identifying malignancy through metabolic markers in the perspective of its application in all cases of difficult diagnosis and after neoadjuvant treatment. METHODS: Spectroscopy was performed ex vivo on 29 colorectal specimens. All patients were staged with imaging, underwent radical surgery and then followed-up. Spectral quantification analysis of components expressed in colorectal tumours and in healthy mucosa were evaluated. The MRS-tumour marker (MRS-tm) was calculated for each case. The U-test was used to compare MRS-tm in tumours and in healthy mucosa. In order to select a cut-off for MRS-tm in the tumour and healthy mucosa and to distinguish patients who were disease-free or with recurrence-progression, we performed the ROC curve analysis. RESULTS: In the 24 subjects without neoadjuvant treatment, it was found that MRS-tm is able to discriminate healthy and neoplastic tissue and can discriminate patients with risk of recurrence/progression CONCLUSION: Our data seem to show that 1H MRS may be successfully applied in vivo non-invasively to differentiate tumours from healthy mucosa and could also distinguish patients with different prognoses.

Development of isoxazoline-containing peptidomimetics as dual αvß3 and α5ß1 integrin ligands.

Isoxazoline-containing peptidomimetics, designed to be effective α(v)ß(3) and α(5)ß(1) integrin ligands, were synthesized through an original procedure involving N,O-bis(trimethylsilyl)hydroxyamine conjugate addition to alkylidene acetoacetates, followed by intramolecular hemiketalization. To mimic the RGD recognition sequence, basic and acidic terminal appendages were introduced, and the final products were tested in cell adhesion inhibition assays. All the synthesized compounds proved to be excellent ligands for both integrin receptors, and a strong influence on intracellular signaling and phosphorylation pathways was demonstrated by evaluation of fibronectin-induced phosphorylation of ERK. The molecular basis of the observed inhibitory activity was suggested on the results of docking experiments.

A straightforward route to enantiopure 2-substituted-3,4-dehydro-ß-proline via ring closing metathesis.

The synthesis of unusual cyclic amino acids, that may be envisaged as proline analogs, is an area of great interest for their potential applications as scaffolds for the design of bioactive peptidomimetics or units for the creation of novel foldamers. We have carried out the preparation of cyclic dehydro-ß-amino acids starting from allylic carbonates via a two-step allylic amination/ring closing metathesis (RCM) protocol. The introduction of the allylamino moiety has been carried out either without a catalyst, through an S(N)2' reaction, or in the presence of iridium complexes. The backbone of the allylamino intermediates contains two unsaturations, thus suggesting that RCM could be a valuable tool for the preparation of dihydropyrrole scaffolds. A similar reaction has been already reported in the literature for racemic aromatic-substituted substrates, but no examples of enantiopure derivatives bearing aliphatic chains have been reported. The reaction was optimized by testing different Grubbs' catalysts and carbamate nitrogen protecting groups. Moreover, in view of a future application of these dehydro-ß-amino acids as central core of peptidomimetics, the malonate chain was also used to protect nitrogen prior to RCM.

Investigation of adipose tissues in Zucker rats using in vivo and ex vivo magnetic resonance spectroscopy.

In vivo single-voxel magnetic resonance spectroscopy (MRS) at 4.7T and ex vivo high-resolution proton magnetic resonance spectroscopy (HR-NMR) at 500 MHz were used to study the composition of adipose tissues in Zucker obese and Zucker lean rats. Lipid composition was characterized by unsaturation and polyunsaturation indexes and mean chain lengths. In vitro experiments were conducted in known mixtures of triglycerides and oils in order to validate the method. To avoid inaccuracies due to partial peak overlapping in MRS, peak quantification was performed after fitting of spectral peaks by using the QUEST algorithm. The intensity of different spectral lines was also corrected for T2 relaxation. Albeit with different sensitivity and accuracy, both techniques revealed that white adipose tissue is characterized by lower unsaturation and polyunsaturation indexes in obese rats compared with controls. HR-NMR revealed similar differences in brown adipose tissue. The present findings confirm the hypothesis that obese and lean Zucker rats have different adipose tissue composition.

Synthesis of dehydro-beta-amino esters via highly regioselective amination of allylic carbonates.

The allylic amination of acetates and carbonates affords dehydro-beta-aminoesters, which are useful precursors of biologically active compounds. The uncatalyzed reaction proceeds via a S(N)2' mechanism. On the other hand, under palladium-catalyzed conditions, the reaction shows a strong solvent-dependent regiocontrol, affording exclusively one of the two possible regioisomers with complete transfer of chirality from the substrates to the products.

An efficient diastereoselective route to differentially protected anti-4-amino-1-alken-3-ols.

Zinc-promoted hydroxyallylation of alpha-amidoalkyl arylsulfones 4 using 3-bromo-propenyl methyl carbonate 5 proceeds smoothly in DMF at room temperature to afford high yields of differentially protected anti-1,2-amino alcohols 6.

Pulmonary sarcoidosis: atypical HRTC features and differential diagnostic problems.

PURPOSE: The aim of this paper is to present atypical or unusual appearances of pulmonary sarcoidosis, assessed at HRCT, and to suggest some elements for the differential diagnosis. MATERIALS AND METHODS: The HRCT scans of 31 patients were retrospectively reviewed. The final diagnosis of sarcoidosis was established on the basis of clinical and laboratory data alone in one patient, of clinical and laboratory data supported by bronchoalveolar lavage (BAL) findings in 14 patients, lung biopsy in 15, and liver biopsy in one patient. Fifteen patients presented with the main aspects of atypical manifestations. RESULTS: In the 15 patients who presented with less frequent or atypical manifestations of pulmonary sarcoidosis, the following patterns were identified: 2 cases of large opacities with ill-defined margins, 8 of peripheral "pseudo-plaque" opacities, 2 of punctuate lymph node calcifications, 8 of ground-glass opacities with fine reticulation, and 5 of isolated opacities with dimensions not consistent with the surrounding background, with or without cavitation. CONCLUSIONS: Besides the typical manifestations of pulmonary sarcoidosis, there may be several atypical or nonspecific presentations: it is not by chance that the literature often describes this disease as the "great masquerader". Mediastinal lymphadenopathies and perilymphatic, subpleural or peribronchovascular nodules represent the typical patterns of pulmonary sarcoidosis; interstitial streaks and bilateral para-hilar thickening are also frequently observed. The presence of peripheral "pseudo-plaque" opacities and of ground-glass opacities with fine reticulation is the key to a diagnosis of "atypical" sarcoidosis. Calcifications within hilar and mediastinal lymph nodes, 1 cm or larger in dimensions, with a focal rather than diffuse distribution, should raise a suspicion of sarcoidosis rather than tuberculosis. Upper lobe localisation is rather infrequent in bronchioloalveolar carcinoma. Not all that excavates is tuberculosis: therefore, a differential diagnosis with tuberculosis may prove difficult, and the two diseases may coexist. Pseudoalveolar pulmonary thickening that eventually resolves and disappears does not exclusively entail an inflammatory thickening.

[Differential patterns of contrast enhancement in different focal liver lesions after injection of the microbubble US contrast agent SonoVue].

PURPOSE: To identify differential contrast enhancement patterns in different focal hepatic lesions after injection of the microbubble contrast agent SonoVue using high or low acoustic power imaging. MATERIAL AND METHODS: Forty-seven focal hepatic lesions (1-8 cm) were detected in 45 patients at unenhanced gray-scale ultrasound (US) and evaluated by color Doppler (CD) US with spectral analysis of tumoral vessels. Lesions were subsequently evaluated by US contrast specific modes after IV bolus administration of 2,4-4,8 ml of SonoVue, by intermittent high acoustic power (18 patients) or by continous low acoustic power imaging (27 patients), during arterial, portal and late phase. Subjective evaluation of lesions appearance before and after SonoVue injection was performed. For final diagnosis multiphasic helical CT (21 patients) and/or fine needle US guided biopsy (24 patients) were considered as the reference procedures. RESULTS: Final diagnoses comprised 22 hepatocellular carcinomas (HCCs; 1,5-6 cm), 2 macroregenerative nodules (RNNs; 1-2 cm), 10 metastasis (2-3,5 cm), 10 hemangiomas (2-6 cm) and 3 focal nodular hyperplasias (FNHs; 1-3 cm). On CD evaluation HCCs revealed peripheral basket shaped (12/22) or intranodular (10/22) arterial pattern while, after SonoVue injection HCCs revealed diffuse contrast enhancement during arterial phase with contrast washout during portal and late phase. Metastases did not reveal flow signals on CD or contrast enhancement after SonoVue injection, except for 2 metastases which revealed peripheral and central vessels on CD and a diffuse contrast enhancement during arterial phase, appearing hypoechoic to the adjacent liver during portal and late phase. RNNs revealed dotted contrast-enhancement during portal and late phase with isoechoic appearance to the adjacent liver. Hemangiomas revealed some peripheral venous flows on CD and a peripheral nodular contrast enhancement during arterial phase with a centripetal fill-in during portal and late phase. FNHs revealed low resistance peripheral or central arterial vessels and a diffuse contrast enhancement during arterial phase, preceded or not by central spoke wheel shaped contrast enhancement, and a persistent iso-hyperechogenicity during portal and late phase. CONCLUSIONS: SonoVue injection has showed to identify differential contrast enhancement patterns in different focal hepatic lesions.

US characterization of focal hepatic lesions with intermittent high-acoustic-power mode and contrast material.

RATIONALE AND OBJECTIVES: This study was performed to determine whether ultrasound (US) performed with SonoVue, a contrast agent that contains microbubbles filled with sulfur hexafluoride vapor, depicts differential patterns of contrast enhancement in focal hepatic lesions. MATERIALS AND METHODS: Forty focal hepatic lesions (15 hepatocellular carcinomas [HCCs], 10 metastases, 11 hemangiomas, and four focal nodular hyperplasias) in 39 patients were evaluated by means of US, color Doppler US, and contrast-enhanced US performed by using intermittent high-acoustic-power mode. Contrast-enhanced helical computed tomography (11 patients) and US-guided fine needle aspiration (28 patients) were used as reference procedures. Contrast enhancement patterns were defined by means of both subjective and objective analysis, and baseline and contrast-enhanced US scans were reviewed offline. RESULTS: Thirteen of 15 HCCs, eight of 10 metastases, and all four hemangiomas with an atypical pattern at baseline US were correctly characterized after SonoVue injection. Two of 15 HCCs and two of 10 metastases remained indeterminate, with no characteristic baseline or contrast-enhanced patterns identified. Baseline US was essential in characterizing all hemangiomas with a typical pattern (n = 7), and color Doppler US with spectral analysis of tumoral vessels was essential in characterizing focal nodular hyperplasia. The percentage of diagnostic agreement with reference procedures was significantly increased (P < .001) for contrast-enhanced US compared with baseline US. CONCLUSION: Characteristic patterns of US contrast enhancement with SonoVue help in characterizing and differentiating focal hepatic lesions.