L’impact des immunosuppresseurs sur les réservoirs du VIH après transplantation rénale

La thérapie antirétrovirale (TAR) n'éradique pas le VIH de l'organisme. Le VIH persiste grâce à la prolifération de lymphocytes T CD4+ infectés de manière latente. De plus, bien que la plupart des provirus persistent sous une forme latente, un petit nombre de cellules infectées produisent des protéines virales et des virions. Les immunosuppresseurs administrés aux personnes subissant une greffe de rein, pour prévenir le rejet d’organe, inhibent la prolifération des lymphocytes T et l'activation immunitaire. Nous avons émis l'hypothèse que les immunosuppresseurs pourraient réduire à la fois la taille du réservoir viral latent ainsi que les marqueurs associés à la production résiduelle de VIH. Sept participants vivant avec le VIH sous TAR efficace et ayant subi une transplantation rénale suivie de la prise d’un traitement immunosuppresseur ont été recrutés. Des échantillons sanguins longitudinaux ont été prélevés avant et après la greffe (6-7 échantillons/participant, sur 2 ans). Les effets des immunosuppresseurs sur l'activation des lymphocytes T ainsi que les taux d’anticorps plasmatiques dirigés contre l'enveloppe du VIH (un reflet de la production virale résiduelle) ont été évalués par cytométrie en flux et ELISA, respectivement. L'ADN total du VIH et les transcrits d'ARN-gag-LTR associés aux cellules ont été quantifiés par qPCR et la fréquence des cellules p24+ par HIV-Flow. Pour confirmer les résultats obtenus in vivo, nous avons optimisé un protocole de culture cellulaire pour évaluer l'impact des immunosuppresseurs les plus fréquemment utilisés (MMF, tacrolimus, dasatinib) sur les cellules infectées dans un environnement contrôlé. Suite à la transplantation et à l’initiation du traitement immunosuppresseur, nous avons observé une diminution significative des niveaux d'expression des marqueurs d’activation et de prolifération HLA-DR et Ki67 dans les lymphocytes T CD4+, accompagnée d'une augmentation de la fréquence des cellules mémoires centrales et d'une forte diminution de la fréquence des lymphocytes T régulateurs. Les niveaux d'anticorps ciblant l'enveloppe du VIH ont également diminué pendant le traitement immunosuppresseur. Il y avait une diminution modeste et transitoire des niveaux d'ADN et d'ARN du VIH un mois après la transplantation rénale. Les résultats de HIV-Flow n'ont montré aucun impact significatif des immunosuppresseurs sur le réservoir inductible, avec de grandes variations entre les participants. Le modèle de culture cellulaire in vitro a révélé que de faibles doses d'immunosuppresseurs induisent généralement une diminution des marqueurs du VIH, alors que des doses plus élevées conduisent à des variations dépendantes des donneurs. Ainsi, nos résultats montrent que les traitements immunosuppresseurs diminuent la prolifération et l'activation des lymphocytes T CD4+, ce qui est concomitant avec une diminution modeste et transitoire des taux d'ADN et d'ARN du VIH. Nos résultats indiquent que la combinaison des traitements immunosuppresseurs actuels sont toutefois insuffisants pour affecter profondément et durablement le réservoir du VIH et suggèrent que la taille du réservoir est étroitement régulée par des forces homéostatiques difficiles à contrer.Antiretroviral therapy (ART) does not eradicate HIV from the body. HIV persists through proliferation of latently infected CD4+T-cells. Moreover, although most proviruses persist in a latent form, a small number of infected cells produce viral protein and virions. Immunosuppressants given to people undergoing kidney transplants to prevent rejection inhibit T-cell proliferation and immune activation. We hypothesized that immunosuppressants could reduce both the size of the latent viral reservoir as well as markers associated with residual production of HIV. Seven participants living with HIV on suppressive ART who underwent kidney transplantation and initiated immunosuppressive therapy were enrolled. Longitudinal blood samples were collected before and after the transplant (6-7 samples/participant, over 2years). The effects of immunosuppressants on T-cell activation as well as plasma HIV-envelope antibody responses (a surrogate of residual viral production) were assessed by flow cytometry and ELISA, respectively. Total HIV-DNA and cell-associated LTR-gag-RNA transcripts were quantified by qPCR and the frequency of p24+ cells by HIV-Flow in isolated CD4+T-cells. To confirm the results saw in vivo, we optimized a cell culture protocol to assess the impact of frequently given immunosuppressants (tacrolimus, MMF and dasatinib) on infected cells in a controlled environment. Following organ transplant and initiation of immunosuppressive treatment, we observed a significant decrease in the expression levels of HLA-DR and Ki67 in CD4+T-cells, which was accompanied by an increase in the frequency of central memory cells and a sharp decrease in the frequency of regulatory T-cells. Antibody levels targeting HIV envelope also decreased during immunosuppressive therapy. There was a modest and transient decrease in HIV DNA and RNA levels one month following kidney transplantation. HIV-Flow results showed no significant impact of immunosuppressants on the inducible reservoir, with large variations between participants. The in vitro cell culture model showed that low doses of immunosuppressants generally induces a reduction in HIV markers, but higher doses lead to variations between donors. Immunosuppressive therapy decreases the proliferation and activation of CD4+ T-cells which is concomitant with a modest and transient decrease in HIV DNA and RNA levels. Our results indicate that current immunosuppressive drugs are insufficient to profoundly and durably affect the HIV reservoir and suggest that the size of the reservoir is tightly regulated by homeostatic forces that are difficult to counteract

School Infrastructure Spending and Educational Outcomes in Northern Italy

We explore whether investment in public school infrastructure affects students' achievement. We use data on extra funding to public high schools after the 2012 Northern Italy earthquake and apply a quasi-experimental design and an instrumental variable strategy. We find that spending on school infrastructure increases standardized test scores in mathematics and Italian language, and the effect is stronger for lower-achieving students and in mathematics. These results provide evidence in favor of a positive impact of capital spending in improving the learning environment and performances of high school students

No phenotypic differences for polycystic ovary syndrome (PCOS) between women with and without type 1 diabetes mellitus.

CONTEXT: Women with type 1 diabetes mellitus (DM1) have a higher prevalence of polycystic ovary syndrome (PCOS) than the general population. OBJECTIVE: The aim of this study was to clarify, in DM1 women with PCOS (PCOS-DM1), the influence of insulin therapy and glycemic control and evaluate the hormonal and phenotypic differences with age-matched and body mass index (BMI)-matched women with PCOS without diabetes. DESIGN, SETTING, AND PATIENTS: We evaluated 103 DM1 women with and without PCOS treated with intensive insulin therapy; 38 age-matched and BMI-matched women with PCOS without diabetes were compared in a cross-sectional study. OUTCOME MEASUREMENTS: Clinical, anthropometric, and metabolic parameters were evaluated. Hormonal evaluation and ovary ultrasound were performed during the follicular phase of the menstrual cycle. RESULTS: Applying the diagnostic criteria of the Androgen Excess Society, 38 (36.89%) women with DM1 showed PCOS. The 38 PCOS-DM1 women showed no differences in treatment and glycemic control compared with DM1 women without PCOS. The only difference was a higher visceral adiposity index in PCOS-DM1 (1.21±0.70 vs 0.90±0.32; P=.002). PCOS-DM1 showed no phenotypic differences with age-matched and BMI-matched PCOS without diabetes. The hormonal pattern was similar except that higher levels of Δ4androstenedione were found in PCOS-DM1 (12.89±3.49 vs 2.79±1.75 nmol/L; P=.010). CONCLUSIONS: The women with PCOS-DM1 do not exhibit particular phenotypic characteristics compared with nondiabetic women with PCOS. However, this pathological disorder must not be underestimated because it could be an additional cardiovascular risk factor in women with DM1

Pro-apoptotic activity of the phytochemical Indicaxanthin on colorectal carcinoma cells (Caco-2) and epigenetic CpG demethylation of the promoter and reactivation of the expression of p16

Phytochemicals play prominent roles in human diet and nutrition as protective redo-active substances in prevention of several disorders and chronic diseases in humans. Today, their function as potent modulators of the mammalian epigenome-regulated gene expression is rapidly emerging. In the present study antiproliferative effects of Indicaxanthin (Ind) from the fruits of Opuntia ficus-indica (1), and potential influence on DNA methylation has been investigated on Caco-2 cells, a human cell line of colorectal carcinoma. Ind caused a clear dose- and time-dependent decrease of the cell proliferation (IC(50) 50 M) associated to apoptosis as demonstrated by phosphatidylserine externalization and depolarization of mithocondrial membrane. Ind decreased the Go-G1phase whereas increased S and G2-M phases of the cell cycle. The phytochemical did not altered the intracellular ROS levels but decreased the [Ca2+]i. Investigation on DNA methylation using MESAP-PCR (Methylation-Sensitive Arbitrarily-Primed Polymerase Chain Reaction) (2), showed that 100 M Ind induced a slight global demethylation after a 48 h treatment. Analysis of epigenetic changes in the DNA methylation pattern at CpG promoter of p16 (INK4a), using MSRE (Methylation-Sensitive Restriction Endonucleases Multiplex-Polymerase Chain Reaction), showed that Ind caused CpG demethylation. Western blotting analysis carried out with p16 monoclonal antibody, confirmed the reactivation of the protein expression. Present data, suggesting that a long-term exposure to indicaxanthin in diet might potentially affect epigenetic machines of the intestinal cells, preventing or repairing initial derangements/disorders, encourage studies on the mechanism involved

Reverse Electrodialysis with saline waters and concentrated brines: a laboratory investigation towards technology scale-up

The use of concentrated brines and brackish water as feed solutions in reverse electrodialysis represents a valuable alternative to the use of river/sea water, allowing the enhancement of power output through the increase of driving force and reduction of internal stack resistance. Apart from a number of theoretical works, very few experimental investigations have been performed so far to explore this possibility. In the present work, two RED units of different size were tested using artificial saline solutions. The effects of feed concentration, temperature and flowrate on process performance parameters were analysed, adopting two different sets of membranes. These experiments allowed to identify the most favourable conditions for maximising the power output within the presently investigated range, i.e. 0.1 M NaCl as diluate and 5 M NaCl as concentrate at 40°C. Under these conditions a power density equal to 12 W/m2cell_pair was reached, among the highest so far reported in the literature. Increasing the unit size a slight reduction in power density was observed. These results indicate new directions for a successful scale-up and development of the Reverse Eletrodialysis technology

Endoscopic Endonasal Resection of a Schwannoma of the Anterior Cranial Fossa.

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Short- and long-term effects of a cardiac rehabilitation program in patients implanted with a left ventricular assist device

The efficacy of cardiac rehabilitation in heart-failure patients who received a left-ventricular assist device (LVAD) instead of heart transplantation (HTx) is still unclear. This study aims to evaluate whether cardiac rehabilitation is beneficial in LVAD as HTx patients in the short term and whether its effects in LVAD patients persist over time. Twenty-five LVAD patients were evaluated by functional and psychological tests at admission (T0) and discharge (T1) of a 4-week inpatient structured rehabilitation program, and follow-ups 3 (T2), 6 (T3), and 12 months (T4) after discharge. Twenty-five matched HTx patients were also studied from T0 to T1 to compare the improvements in the six-minute walk test (6MWT). The quality-of-life scores substantially improved in LVAD patients and the 6MWT showed the same functional recovery as in HTx patients from T0 to T1. After T1, numerous LVAD patients withdrew from the study. However, the 6MWT outcome increased further from T1 to T3, with a positive trend during the follow-ups. Hemoglobin and the ventilatory performance increased, and the psychological perception of heart-failure symptoms and pain further improved at T2. In conclusion, exercise-based rehabilitation programs provide similar beneficial effects in LVAD and HTx patients, without deterioration in LVAD patients up to 12 months after discharge

A Cloud-Edge Orchestration Platform for the Innovative Industrial Scenarios of the IoTwins Project

The concept of digital twins has growing more and more interest not only in the academic field but also among industrial environments thanks to the fact that the Internet of Things has enabled its cost-effective implementation. Digital twins (or digital models) refer to a virtual representation of a physical product or process that integrate data from various sources such as data APIs, historical data, embedded sensors and open data, giving to the manufacturers an unprecedented view into how their products are performing. The EU-funded IoTwins project plans to build testbeds for digital twins in order to run real-time computation as close to the data origin as possible (e.g., IoT Gateway or Edge nodes), and whilst batch-wise tasks such as Big Data analytics and Machine Learning model training are advised to run on the Cloud, where computing resources are abundant. In this paper, the basic concepts of the IoTwins project, its reference architecture, functionalities and components have been presented and discussed

Proteasome inhibitors in medullary thyroid carcinoma: time to restart with clinical trials?

IntroductionMedullary thyroid cancer (MTC) is a rare thyroid tumour whose management in advanced stages is challenging, despite effective therapeutic options having expanded in recent years. Proteasome inhibitors (PrIn) have shown the ability to improve patient outcomes, including survival and quality of life, in several malignancies, due to their ability to impair cell proliferation and cause apoptosis through the inhibition of the proteasome activity. Consequently, these drugs could represent a useful tool, alone or in combination with other treatments, in MTC patients.Aim of the studyThis review aims to summarize the available in vitro and in vivo data about the role of PrIn in MTC.Materials and methodsWe performed an extensive search for relevant data sources, including full-published articles in international online databases (PubMed, Web of Science, Scopus), preliminary reports in selected international meeting abstract repositories, and short articles published as supplements of international meetings, by using the following terms: medullary thyroid carcinoma, proteasome inhibitors, bortezomib, carfilzomib, ixazomib, delanzomib, marizomib, oprozomib, and MG132. Additionally, we conducted with the same keywords, an in-depth search in registered clinical trials repositories.ResultsOur search revealed in vitro studies in human and murine MTC cell lines, based on the use of PrIns, both alone and in combination with other anticancer drugs, and two pertinent clinical trials.ConclusionWe found a strong discrepancy between the evidence of PrIns effects in preclinical studies, and the scarcity or early interruption of clinical trials. We might speculate that difficulties in enrolling patients, as happens in other rare diseases, may have discouraged trials’ implementation in favor of drugs already approved for MTC. However, given the concrete improvement in the comprehension of the molecular basis of PrIn effects in MTC, new clinical trials with accurate inclusion criteria of enrollment might be warranted, in order to ascertain whether this treatment, alone or in combination with other drugs, could indeed represent an option to enhance the therapeutic response, and to ultimately improve patients’ outcome and survival

Bollettino Sismico Italiano: Analisys of Early Aftershocks of the 2016 MW 6.0 Amatrice, MW 5.9 Visso and MW 6.5 Norcia earthquakes in Central Italy

The Amatrice-Visso-Norcia seismic sequence is the most important of the last 30 years in Italy. The seismic sequence started on 24 August, 2016 and still is ongoing in central Apennines. At the end of February 2017 more than 57,000 events were located, 80,000 events up to the end of September 2017 (Fig. 1). The mainshocks of the sequence occurred on 24 August 2016 (Mw 6.0 and Mw 5.4), 26 October 2016 (Mw 5.4 and Mw 5.9), 30 October 2016 (Mw 6.5), 18 January 2017 (four earthquakes Mw≥ 5.0). In this seismic sequence, all the waveforms recorded by temporary stations deployed by the SISMIKO emergency group (stations T12**; Moretti et al., 2016) where available in real- time at the surveillance room of INGV. Because of the high level of seismicity and the dense seismic network installed in the region, more than 150 events per day were located at the end of February 2017; still 60 events per day were located up to the end of August 2017.The Amatrice-Visso-Norcia is the most important seismic sequence since 2015, the time when the analysis procedures of the BSI group (Bollettino Sismico Italiano) were revised (Nardi et al., 2015). BSI is now available every four months on the web: bulletins contain revised earthquakes (location and magnitude) with ML≥ 1.5, quasi-real time revision of ML≥ 3.5 earthquakes and phase arrivals from waveforms recorded on seismic stations available from the European Integrated Data Archive (EIDA), (Mazza et al., 2012). These last procedures allow the integration of signals from temporary seismic stations (Moretti et al., 2014) installed by the emergency group SISMIKO (Moretti and Sismiko working group, 2016), even when they are not in real time transmission, if they are rapidly archived in EIDA, together with real time signals from the seismic stations of the permanent INGV network. The analysis strategy of the BSI group for the Amatrice -Visso - Norcia seismic sequence (AVN.s.s in the following) was to select the earthquakes located in the box with min/max latitude: 42.2/43.2 - and min/max longitude: 12.4/14.1 to prepare a special volume of BSI on the seismic sequence.PublishedTrieste, Italy1SR. TERREMOTI - Servizi e ricerca per la Societ