Mycobiota composition and changes across pregnancy in patients with gestational diabetes mellitus (GDM)

The gut mycobiota has never been studied either during pregnancy or in patients with gestational diabetes (GDM). This study aimed to analyze the fecal mycobiota of GDM patients during the second (T2) and third (T3) trimester of pregnancy and to compare it with the mycobiota of pregnant normoglycemic women (controls). Forty-one GDM patients and 121 normoglycemic women were studied. GDM mycobiota was composed almost exclusively by the Ascomycota phylum; Basidiomicota accounted for 43% of the relative frequency of the controls. Kluyveromyces (p < 0.001), Metschnikowia (p < 0.001), and Pichia (p < 0.001) showed a significantly higher frequency in GDM patients, while Saccharomyces (p = 0.019), were more prevalent in controls. From T2 to T3, a reduction in fungal alpha diversity was found in GDM patients, with an increase of the relative frequency of Candida, and the reduction of some pro-inflammatory taxa. Many associations between fungi and foods and nutrients were detected. Finally, several fungi and bacteria showed competition or co-occurrence. Patients with GDM showed a predominance of fungal taxa with potential inflammatory effects when compared to normoglycemic pregnant women, with a marked shift in their mycobiota during pregnancy, and complex bacteria-fungi interactions

Tumor stiffening reversion through collagen crosslinking inhibition improves T cell migration and anti-PD-1 treatment

Only a fraction of cancer patients benefits from immune checkpoint inhibitors. This may be partly due to the dense extracellular matrix (ECM) that forms a barrier for T cells. Comparing five preclinical mouse tumor models with heterogeneous tumor microenvironments, we aimed to relate the rate of tumor stiffening with the remodeling of ECM architecture and to determine how these features affect intratumoral T cell migration. An ECM-targeted strategy, based on the inhibition of lysyl oxidase, was used. In vivo stiffness measurements were found to be strongly correlated with tumor growth and ECM crosslinking but negatively correlated with T cell migration. Interfering with collagen stabilization reduces ECM content and tumor stiffness leading to improved T cell migration and increased efficacy of anti-PD-1 blockade. This study highlights the rationale of mechanical characterizations in solid tumors to understand resistance to immunotherapy and of combining treatment strategies targeting the ECM with anti-PD-1 therapy

Transcranial magnetic stimulation of the precuneus enhances memory and neural activity in prodromal Alzheimer's disease

Memory loss is one of the first symptoms of typical Alzheimer's disease (AD), for which there are no effective therapies available. The precuneus (PC) has been recently emphasized as a key area for the memory impairment observed in early AD, likely due to disconnection mechanisms within large-scale networks such as the default mode network (DMN). Using a multimodal approach we investigated in a two-week, randomized, sham-controlled, double-blinded trial the effects of high-frequency repetitive transcranial magnetic stimulation (rTMS) of the PC on cognition, as measured by the Alzheimer Disease Cooperative Study Preclinical Alzheimer Cognitive Composite in 14 patients with early AD (7 females). TMS combined with electroencephalography (TMS-EEG) was used to detect changes in brain connectivity. We found that rTMS of the PC induced a selective improvement in episodic memory, but not in other cognitive domains. Analysis of TMS-EEG signal revealed an increase of neural activity in patients' PC, an enhancement of brain oscillations in the beta band and a modification of functional connections between the PC and medial frontal areas within the DMN. Our findings show that high-frequency rTMS of the PC is a promising, non-invasive treatment for memory dysfunction in patients at early stages of AD. This clinical improvement is accompanied by modulation of brain connectivity, consistently with the pathophysiological model of brain disconnection in AD

Etude du rôle de la nucleoline dans l’adénocarcinome pancréatique ductal

Le microenvironnement de l'adénocarcinome ductale pancréatique (PDAC) est hautement fibrotique et hypoxique, avec une faible infiltration des cellules immunitaires; La combinaison de thérapies ciblées et d'immunothérapie avec la chimiothérapie n'a jusqu'à présent pas permis d'améliorer la survie des patients atteints de PDAC. La nucléoline (NCL) est une protéine nucléolaire surexprimée dans le cancer et notre groupe a montré qu'un niveau élevé de NCL dans PDAC est en corrélation avec une faible survie globale. L'inhibition de NCL par N6L, un pseudopeptide synthétique multivalent breveté par notre laboratoire, diminue la croissance de PDAC et normalise les vaisseaux tumoraux dans des modèles murins de PDAC améliorant l'administration de médicaments et une diminution de l'hypoxie tumorale.Étant donné que la normalisation des vaisseaux tumoraux et la reprogrammation immunostimulante pourraient se réguler mutuellement, nous avons étudié les effets de l'inhibition des NCL sur le microenvironnement immunitaire des modèles murins PDAC. (mPDAC) N6L a réduit spécifiquement la proportion de lymphocytes T régulateurs et de cellules myéloïdes suppressives (MDSC) spécifiquement dans le microenvironnement tumoral mais pas dans la rate et les ganglions lymphatiques des souris porteuses de tumeurs. De plus, N6L a augmenté l'infiltration et l'activation des TIL. Par analyse RNAseq du mPDAC traité par N6L, nous avons trouvé et validé que N6L diminuait l'expansion des CAF et l'expression de l'IL-6 dans les tumeurs, dans le plasma et in vitro. Le traitement du mPDAC par un anticorps bloquant l'IL-6, la proportion de Treg et de MDSC, telle que l'augmentation de l'infiltration de TIL, imitait les effets de N6L. Alors que l'anti-VEGF à faible dose normalisait le vaisseau tumoral du mPDAC, mais n'était pas suffisant pour moduler le micro-environnement immunitaire du mPDAC. De plus, une thérapie combinée avec l'immonucheck-point inhibitor anti-PD1 a été testée. La combinaison n'a pas réussi à montrer de meilleurs résultats que N6L sur le volume de la tumeur sur le modèle mPDAC.En conclusion, ces résultats démontrent que l'inhibition de la NCL bloque l'amplification des cellules immunosuppressives lymphoïdes et myéloïdes et normalise le microenvironnement tumoral PDAC grâce à un nouveau mécanisme d'action dépendant de la régulation du stroma tumoral.Pancreatic ductal adenocarcinoma (PDAC) microenvironment is highly fibrotic and hypoxic, with poor immune cell infiltration; combination of targeted therapies and immunotherapy with chemotherapy has failed to show any improvement in PDAC patient survival so far. Nucleolin (NCL) is a nucleolar protein overexpressed in cancer and our group showed that high NCL level in PDAC correlates with a low overall survival. NCL inhibition by N6L, a multivalent synthetic pseudopeptide patented by our laboratory, impairs PDAC growth and normalizes tumour vessels in mouse models of PDAC improving drug delivery and a decreasing tumour hypoxia.Since evidence tumour vessel normalization and immunostimulatory reprogramming could mutually regulate, we investigated the effects of NCL inhibition on the immune microenvironment of PDAC mouse models. (mPDAC) N6L specifically reduced the proportion of lymphocyte T regulatory cells and myeloid-derived suppressor cells (MDSCs) specifically in the tumour microenvironment but not in the spleen and lymph nodes of tumour bearing mice. Moreover, N6L increased the infiltration and the activation of TILs. By RNAseq analysis of mPDAC treated by N6L we found and validated that N6L decreased CAFs expansion and IL-6 expression in tumours, in plasma and in vitro. Treating mPDAC by an antibody blocking IL-6 the proportion of Tregs and MDSCs such as the increase of TIL infiltration mimicked the effects pf N6L. While low dose anti-VEGF normalized mPDAC tumour vessel but was not sufficient to modulate immune microenvironment of mPDAC. Moreover, a combo therapy with the anti check-point ihibitor anti-PD1 has been tested. The combination failed to show any better results than N6L on tumor volum on mPDAC model.In conclusion, these results demonstrate that NCL inhibition block the amplification of lymphoid and myeloid immunosuppressive cells and normalizes the PDAC tumour microenvironment through a new mechanism of action dependent on the regulation of tumour stroma

The role of diabetes mellitus and BMI in the surgical treatment of ankle fractures

Open reduction and internal fixation is the standard treatment for displaced ankle fractures. However, the presence of comorbidities such as diabetes mellitus and body mass index (BMI) are associated with poor bone quality, and these factors may predict the development of postoperative complications. The study aim was to assess the role of diabetes mellitus and BMI in wound healing in patients younger than 65 years who were surgically treated for malleoli fractures

Theta burst stimulation of the precuneus modulates resting state connectivity in the left temporal pole.

It has been shown that continuous theta burst stimulation (cTBS) over the precuneus acts on specific memory retrieval abilities. In order to study the neural mechanisms beyond these findings, we combined cTBS and resting-state functional magnetic resonance imaging. Our experimental protocol involved stimulation and sham conditions on a group of healthy subjects, and each condition included a baseline and two follow-up acquisitions (5 and 15 min after baseline) after cTBS. We analysed brain functional connectivity by means of graph theoretical measures, with a specific focus on the network modular structure. Our results showed that cTBS of the precuneus selectively affects the left temporal pole, decreasing its functional connectivity in the first follow-up. Moreover, we observed a significant increase in the size of the module of the precuneus in the second follow-up. Such effects were absent in the sham condition. We observed here a modulation of functional connectivity as a result of inhibitory stimulation over the precuneus. Such a modulation first acts indirectly on the temporal area and then extends the connectivity of the precuneus itself by a feed-back mechanism. Our current findings extend our previous behavioural observations and increase our understanding of the mechanisms underlying the stimulation of the precuneus

Transport Infrastructure SHM Using Integrated SAR Data and On-Site Vibrational Acquisitions: “Ponte Della Musica–Armando Trovajoli” Case Study

This work presents the first results obtained by applying in situ and remote-sensing methodologies to monitor the Ponte della Musica-Armando Trovajoli located in Rome, within the activities of the WP6 “Structural Health Monitoring and Satellite Data” 2019-21 Reluis Project. In particular, the use of remote-sensing Differential Synthetic Aperture Radar (SAR) Interferometry (DInSAR) measurements provided a spatial map of the displacement of the investigated infrastructure and the corresponding time-series, with the aim of monitoring deformation phenomena, focusing on the local scale analysis, which produces suitable results for urban monitoring and damage assessment. The DInSAR results have been integrated with the identification of the dynamic characteristics of the bridge, performed through an experimental campaign of ambient vibration measurements carried out in October 2020 and with the local-scale definition of the engineering geological setting of the foundation soil. The subsoil of the bridge is constituted by more than 50 m of recent alluvial deposits resting on Pliocene stiff clay acting as a geological bedrock. A substantially stable behavior of the bridge structural elements has been observed based on the analysis of both satellite and velocimetric data. This case represents a good example about how the integration of in situ sensors with remotely sensed data and the exploitation of a detailed knowledge regarding the on-site conditions represent a key factor for a sustainable structural and infrastructural monitoring and can support the planning both of maintenance and safety management

Validated LC-MS/MS Assay for the Quantitative Determination of Fenretinide in Plasma and Tumor and Its Application in a Pharmacokinetic Study in Mice of a Novel Oral Nanoformulation of Fenretinide

We describe the development and validation of a HPLC-MS/MS method to assess the pharmacokinetics and tumor distribution of fenretinide, a synthetic retinoid chemically related to all-trans-retinoic acid, after administration of a novel oral nanoformulation of fenretinide, called bionanofenretinide (BNF). BNF was developed to overcome the major limitation of fenretinide: its poor aqueous solubility and bioavailability due to its hydrophobic nature. The method proved to be reproducible, precise and highly accurate for the measurement of the drug and the main metabolites. The lower limit of quantification resulted in 1 ng/mL. The curve range of 1–500 ng/mL and 50–2000 ng/mL, for plasma and tumor homogenate, respectively, was appropriate for the analysis, as demonstrated by the accuracy of between 96.8% and 102.4% for plasma and 96.6 to 102.3% for the tumor. The interdays precision and accuracy determined on quality controls at three different levels were in the ranges of 6.9 to 7.5% and 99.3 to 101.0%, and 0.96 to 1.91% and 102.3 to 105.8% for plasma and tumor, respectively. With the application of the novel assay in explorative pharmacokinetic studies, following acute and chronic oral administration of the nanoformulation, fenretinide was detected in plasma and tumor tissue at a concentration higher than the IC50 value necessary for in vitro inhibitory activity (i.e., 1–5 µM) in different cancer cells lines. We were also able to detect the presence in plasma and tumor of active and inactive metabolites of fenretinide

Dynamic reorganization of TMS-evoked activity in subcortical stroke patients

Since early days after stroke, the brain undergoes a complex reorganization to allow compensatory mechanisms that promote functional recovery. However, these mechanisms are still poorly understood and there is urgent need to identify neurophysiological markers of functional recovery after stroke. Here we aimed to track longitudinally the time-course of cortical reorganization by measuring for the first time EEG cortical activity evoked by TMS pulses in patients with subcortical stroke. Thirteen patients in the sub-acute phase of ischemic subcortical stroke with motor symptoms completed the longitudinal study, being evaluated within 20 days and after 40, 60 and 180 days after stroke onset. For each time-point, EEG cortical activity evoked by single TMS pulses was assessed over the motor and parietal cortex of the affected and unaffected hemisphere. We evaluated global TMS-evoked activity and TMS-evoked oscillations in different frequency bands. These measurements were paralleled with clinical and behavioral assessment. We found that motor cortical activity measured by TMS-EEG varied across time in the affected hemisphere. An increase of TMS-evoked activity was evident at 40 days after stroke onset. Moreover, stroke patients showed a significant increase in TMS-evoked alpha oscillations, as highlighted performing analysis in the time-frequency domain. Notably, these changes indicated that crucial mechanisms of cortical reorganization occur in this short-time window. These changes coincided with the clinical improvement. TMS-evoked alpha oscillatory activity recorded at baseline was associated to better functional recovery at 40 and 60 days' follow-up evaluations, suggesting that the power of the alpha rhythm can be considered a good predictor of motor recovery. This study demonstrates that cortical activity increases dynamically in the early phases of recovery after stroke in the affected hemisphere. These findings point to TMS-evoked alpha oscillatory activity as a potential neurophysiological markers of stroke recovery and could be helpful to determine the temporal window in which neuromodulation should be potentially able to drive neuroplasticity in an effective functional direction