Dinamica virale e risposta immune HIV-1-specifica in neonati sottoposti a terapia antiretrovirale ed immunoricostituzione in bambini infettati da HIV-1 con differente risposta virologica alla terapia

Introduzione. L'infezione perinatale da HIV-1 è acquisita quando il sistema immunitario del bambino è in fase di sviluppo ed è caratterizzata da un'elevata e non controllata replicazione virale. La terapia antiretrovirale altamente efficace (HAART), che genericamente prevede l'utilizzo di inibitori della proteasi e della trascrittasi inversa, riduce efficientemente la carica virale di HIV-1 sotto livelli non rilevabili e aumenta il numero di cellule T CD4+ circolanti nei bambini come negli adulti. Al fine di incrementare le scarse informazioni sul trattamento precoce con HAART e sull'immunoricostituzione in bambini HIV-1-infetti, è stata analizzata la dinamica virale e la risposta immunitaria in bambini trattati precocemente con HAART e l'immunoricostituzione in bambini HIV-1-infetti che hanno mostrato una risposta discordante alla terapia. Metodi. Coorte in HAART precoce: sono stati studiati 6 neonati HIV-1-infetti che hanno iniziato HAART entro i 3 mesi di età. HIV-1 RNA plasmatico, HIV-1 DNA cellula-associato, HIV-1 mRNA unspliced e multiply spliced e anticorpi anti-HIV-1 sono stati analizzati in campioni sequenziali di sangue periferico. La risposta immunitaria cellulare HIV-1-specifica è stata misurata mediante saggio EliSpot. Coorte discordante: le sottopopolazioni cellulari T CD4+ e CD8+ sono state studiate al baseline e dopo circa 2 anni di HAART in 14 bambini HIV-1-infetti che hanno mostrato una soppressione della viremia plasmatica (rispondenti virologici, VR) e in 16 bambini non rispondenti virologici alla terapia (VNR). Risultati. Coorte in HAART precoce. In tutti i bambini è stata osservata una riduzione della viremia plasmatica. In 4 bambini è stato rilevato l’HIV-1 DNA; di questi 2 erano anche positivi per l’HIV-1 mRNA. Solo 2 bambini hanno prodotto propri anticorpi anti-HIV-1 mentre gli altri, dopo la perdita degli anticorpi materni, sono rimasti persistentemente sieronegativi. In nessun paziente è stata osservata una risposta immunitaria cellulare HIV-1-specifica. L’interruzione di terapia è stata effettuata in un paziente HIV-1-sieropositivo ed in uno sieronegativo. L’aumento della viremia plasmatica nel bambino sieronegativo è stato più rapido ed elevato rispetto a quello osservato nel paziente sieropositivo. Coorte discordate. Durante HAART è stato riscontrato un aumento delle cellule T CD4+ sia nei bambini VR sia nei VNR; tale incremento era più elevato nel primo gruppo rispetto a quello rilevato nel secondo. Tutte le sottopopolazioni cellulari T CD4+ (naive, central memory, effector/memory e CD38+) sono aumentate significativamente nei bambini VR mentre nei VNR l’incremento significativo avveniva solo nelle cellule naive. In entrambe i gruppi si è osservato un aumento nelle cellule T CD8+ naive e nella forma epitomale di riarrangiamento del recettore delle cellule T (TREC), un indicatore della funzionalità timica. Le cellule T CD8+CD38+ attivate sono diminuite nei bambini VR mentre sono rimaste elevate nei VNR. I livelli plasmatici di lipopolisaccaride (LPS), un indicatore della translocazione microbica, erano aumentati nei pazienti VNR. Conclusioni. La somministrazione precoce di HAART nel neonato modifica il naturale corso dell’infezione da HIV-1. Tale regime, sebbene controlli la replicazione virale, riduce la viremia plasmatica sotto i livelli soglia necessari ad induce una risposta immunitaria HIV-1-specifica e non previene l’istaurazione di una latenza virale che impedisce l’eradicazione dell’infezione. Un prolungato trattamento con HAART nei bambini HIV-1-infetti permette, inoltre, un aumento delle cellule T naive che è indipendentemente dalla risposta virologica alla terapia. Una viremia persistente, comunque, impedisce l’espansione delle cellule T CD4+ memory suscettibili all’infezione virale e, insieme alla translocazione microbica, contribuisce a mantenere elevati i livelli di immuno-attivazione.Background. Perinatal HIV-1 infection is acquired in the milieu of a developing immune system, leading to high levels of uncontrolled viral replication. Highly active antiretroviral therapy (HAART), which is usually a combination of protease and reverse transcriptase inhibitors, efficiently reduces HIV-1 load to undetectable levels and increases the number of circulating CD4 T cells in children as well as adults. To add to the scarce information available on early HAART treatment and immune reconstitution in HIV-1-infected children, we investigated the viral dynamics and the immunological response in early HAART treated-children and the immune reconstitution in HIV-1-infected children with discordant response to therapy. Methods. Early HAART cohort: 6 HIV-1-infected infants who started HAART within 3 months of age were studied. Plasma HIV-1 RNA, cell-associated HIV-1 DNA, unspliced and multiply spliced HIV-1 mRNAs, HIV-1 antibodies were assessed in sequential peripheral blood samples. HIV-1 cellular immune response was measured by EliSpot assay. Discordant cohort: CD4+ and CD8+ T cell subsets in 14 HIV-1-infected children with suppression of HIV-1 plasma viraemia (virological responders, VR) and in 16 virological non responders (VNR) to therapy were studied at baseline and after approximately 2 years of HAART. Results. Early HAART cohort. All children showed a decline in plasma viraemia to undetectable levels. HIV-1 DNA persisted in 4 children, but only 2 of these had detectable HIV-1 mRNA. All viral parameters remained persistently negative in 2 children. Only 2 children produced HIV-1 antibodies, while the others, after having lost maternal antibodies, remained seronegative. No HIV-1 cellular immune response was observed in any child. Therapy interruption was performed in 2 children: one HIV-1-seropositive and one HIV-1-seronegative with persistently undetectable levels of all viral parameters. Rebound of HIV-1 plasma viraemia in the seronegative child was more rapid and higher than that observed in the seropositive child. Discordant cohort. During therapy, CD4+ T cells increased in both groups, but were higher in the VR than in the VNR group. All CD4+ T cell subsets (naive, central memory, effector/memory and CD38+) increased significantly in VR children, while there was a significant increase only in naive cells in VNR children. Naive CD8+ T cells and T cell receptor rearrangement excision circles (TREC), an indicator of thymic output, increased in both VR and VNR children. Activated CD8+CD38+ T cells decreased in VR but remained high in VNR children. Levels of circulating lipopolysaccharide (LPS), an indicator of microbial translocation, further increased in VNR children. Conclusions. Early ART treatment in infants modifies the natural course of infection by controlling HIV-1 replication and reducing viral load to below the threshold levels required for onset of HIV-1 immune response, but does not prevent the establishment of a reservoir of latently infected cells that precludes virus eradication. Moreover, a long-term HAART treatment induced an increase in naive T cells in all HIV-1-infected children, regardless of their virological response. However, the persistence of viraemia resulted in an impaired expansion of memory CD4+ T cells susceptible to HIV-1 infection, and together with the microbial translocation sustained the persistence of a high level of immune activation

SOCIAL REPORTING PRACTICES IN ITALIAN PUBLIC SECTOR: AN EXPLORATORY STUDY

This paper should be seen in the broad research field that analyses the impact of ICTs on public accountability reports with the objective of understanding whether the institutional websites of public administrations are qualifying and privileged carriers for social reporting and which models they use to draw up their documents. Our analysis refers to the public entities of the Italian regional capital cities that provide the highest number of services for their local communities: municipalities and hospitals. Our research assumption is that, for larger-sized public entities, websites should be the main channel used to meet the accountability expectations of stakeholders, and therefore they are the place where the highest degree of sensitivity to reporting should be observed. The empirical analysis revealed that the approaches to social reporting differ significantly in terms of formal structure, content and communication strength. The analysis seems to suggest that the awareness of the importance of social reporting is still rather scarce, occasional and, in many cases, detached from the criteria set for public accountability processe

Multiscale integration of satellite, airborne and field data for Mediterranean vegetation studies in the natural area of the Castelporziano Estate (Rome)

A new experimental approach to land analysis has recently been developed, based on the integration of information acquired on different scales; it enables the structure and the functionality of the vegetation in natural ecosystems to be analysed. This research aims at assessing the potentiality of the experimental approach by the integration of airborne and satellite remotely sensed data with ground measurements of structural parameters. In July 1999 a joint campaign for the acquisition of airborne (MIVIS, spatial resolution 3 in) and satellite remotely sensed data (Landsat 5TM, spatial resolution 30 in) and measures taken at ground (PAI), was deployed in the Presidential Estate at Castelporziano (Rome, Italy). The spectral signatures of the main vegetational types of the Estate were examined and the PAI were related to NDVI values, calculated by means of satellite and airborne images. The adopted approach enabled PAI maps to be produced. The linear relation between measured PAI and estimated PAI showed a higher coefficient of determination when the MIVIS data were used. The sensor high spectral resolution has moreover allowed to better describe the structural characteristics of the main plant typologies at Castelporziano Estate

KRAS Codons 12 and 13 Mutation Analysis: A Comparative Study between Direct Sequencing and a New Sensitive Real-Time PCR Assay

KRAS somatic mutations are found in 30–40% of colorectal cancer (CRC). Seven mutations in codons 12 and 13 of KRAS (95% of the observed human mutations) preclude the efficacy of anti-EGFR therapy for the treatment of CRC. Assessment of KRAS mutational status has become a standard procedure in the management of patients with CRC. Technically, KRAS mutation testing can be performed with different methods, characterized by distinct sensitivities and specificities. The present study analyzed KRAS in 182 CRC histological samples by using direct sequencing and a new kit based on a Real-Time Sequence-Specific Primers-PCR technology. The kit allowed to recover as positive 17 samples that were negative or unclear by sequencing, with a recovery rate equal to 13.82%. This study proposes a fast, sensitive, and high-throughput system to identify such seven described mutations of KRAS gene in CRC samples

Ecotoxicological Assays with the Calanoid Copepod Acartia tonsa: A Comparison between Mediterranean and Baltic Strains

The use of marine invertebrates in ecotoxicology is important for an integrated approach which takes into consideration physiological responses and chemical levels in environmental matrices. Standard protocols have been developed and organisms belonging to different trophic levels are needed as model organisms to evaluate toxicant bioavailability and assess their impact on marine biota. The calanoid copepod Acartia tonsa is commonly used in ecotoxicology due to its widespread distribution and well-studied biology. However, different strains coming from various geographical areas are available, and possible variations in physiological characteristics raise concerns about the comparability of ecotoxicological results. This study compares the life cycle assessment and sensitivity of Adriatic and Baltic strains of A. tonsa exposed to nickel (Ni2+) in standardized acute and semi-chronic tests. Life cycle assessments revealed differences in egg production, egg-hatching success, and naupliar viability between the strains. The acute toxicity test demonstrated the significantly higher sensitivity of Adriatic strain nauplii to Ni2+ compared to the Baltic strain, whereas the semi-chronic test showed no significant difference in sensitivity between the strains. These findings suggest that while strain-specific differences exist in different geographical populations, responses to toxicants are not significantly different. Particularly, the semi-chronic assessments with both A. tonsa strains emphasized the robustness of this species as a model organism in ecotoxicology

KRAS Codons 12 and 13 Mutation Analysis: A Comparative Study between Direct Sequencing and a New Sensitive Real-Time PCR Assay

KRAS somatic mutations are found in 30–40% of colorectal cancer (CRC). Seven mutations in codons 12 and 13 of KRAS (95% of the observed human mutations) preclude the efficacy of anti-EGFR therapy for the treatment of CRC. Assessment of KRAS mutational status has become a standard procedure in the management of patients with CRC. Technically, KRAS mutation testing can be performed with different methods, characterized by distinct sensitivities and specificities. The present study analyzed KRAS in 182 CRC histological samples by using direct sequencing and a new kit based on a Real-Time Sequence-Specific Primers-PCR technology. The kit allowed to recover as positive 17 samples that were negative or unclear by sequencing, with a recovery rate equal to 13.82%. This study proposes a fast, sensitive, and high-throughput system to identify such seven described mutations of KRAS gene in CRC samples

Microplastics Occurrence in the European Common Frog (Rana temporaria) from Cottian Alps (Northwest Italy)

Microplastics (MPs) pollution is arousing growing attention, yet knowledge about its occurrence in amphibians is scant to date. With this study, we aimed to determine whether plastic (>5000 μm) and MPs (10–5000 μm) could be detected in adult Rana temporaria from a high-mountain ecosystem (the Cottian Alps, northwest Italy). To do this, aquatic compartments and the digestive tract of adult R. temporaria were analyzed. Water, sediment, periphyton, aquatic macroinvertebrates, and tadpoles tested negative for plastic and MPs. Microplastics were detected in all the adult frogs (n = 5); all the identified items (one per specimen) were fibers (size range: 550.91–2355.51 µm). A statistically significant positive correlation between the particle length and frog size was recorded. The predominant fiber color was blue. The chemical composition was polyamide (60%), polyethylene (20%), and polyethylene terephthalate (20%). Since both the biotic and the abiotic freshwater compartments (tadpoles included) revealed the absence of MPs, it can be assumed that adult frogs ingest MPs from the surrounding terrestrial environment.This research was partially funded by Fondazione CRT, the ALPLA II project, grant number 21D03.Peer reviewe

Factor VIII promotes angiogenesis and vessel stability regulating extracellular matrix proteins

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Interrupting the nitrosative stress fuels tumor-specific cytotoxic T lymphocytes in pancreatic cancer

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest tumors owing to its robust desmoplasia, low immunogenicity, and recruitment of cancer-conditioned, immunoregulatory myeloid cells. These features strongly limit the success of immunotherapy as a single agent, thereby suggesting the need for the development of a multitargeted approach. The goal is to foster T lymphocyte infiltration within the tumor landscape and neutralize cancer-triggered immune suppression, to enhance the therapeutic effectiveness of immune-based treatments, such as anticancer adoptive cell therapy (ACT). METHODS: We examined the contribution of immunosuppressive myeloid cells expressing arginase 1 and nitric oxide synthase 2 in building up a reactive nitrogen species (RNS)-dependent chemical barrier and shaping the PDAC immune landscape. We examined the impact of pharmacological RNS interference on overcoming the recruitment and immunosuppressive activity of tumor-expanded myeloid cells, which render pancreatic cancers resistant to immunotherapy. RESULTS: PDAC progression is marked by a stepwise infiltration of myeloid cells, which enforces a highly immunosuppressive microenvironment through the uncontrolled metabolism of L-arginine by arginase 1 and inducible nitric oxide synthase activity, resulting in the production of large amounts of reactive oxygen and nitrogen species. The extensive accumulation of myeloid suppressing cells and nitrated tyrosines (nitrotyrosine, N-Ty) establishes an RNS-dependent chemical barrier that impairs tumor infiltration by T lymphocytes and restricts the efficacy of adoptive immunotherapy. A pharmacological treatment with AT38 ([3-(aminocarbonyl)furoxan-4-yl]methyl salicylate) reprograms the tumor microenvironment from protumoral to antitumoral, which supports T lymphocyte entrance within the tumor core and aids the efficacy of ACT with telomerase-specific cytotoxic T lymphocytes. CONCLUSIONS: Tumor microenvironment reprogramming by ablating aberrant RNS production bypasses the current limits of immunotherapy in PDAC by overcoming immune resistance

Impact of COVID-19 on cardiovascular testing in the United States versus the rest of the world

Objectives: This study sought to quantify and compare the decline in volumes of cardiovascular procedures between the United States and non-US institutions during the early phase of the coronavirus disease-2019 (COVID-19) pandemic. Background: The COVID-19 pandemic has disrupted the care of many non-COVID-19 illnesses. Reductions in diagnostic cardiovascular testing around the world have led to concerns over the implications of reduced testing for cardiovascular disease (CVD) morbidity and mortality. Methods: Data were submitted to the INCAPS-COVID (International Atomic Energy Agency Non-Invasive Cardiology Protocols Study of COVID-19), a multinational registry comprising 909 institutions in 108 countries (including 155 facilities in 40 U.S. states), assessing the impact of the COVID-19 pandemic on volumes of diagnostic cardiovascular procedures. Data were obtained for April 2020 and compared with volumes of baseline procedures from March 2019. We compared laboratory characteristics, practices, and procedure volumes between U.S. and non-U.S. facilities and between U.S. geographic regions and identified factors associated with volume reduction in the United States. Results: Reductions in the volumes of procedures in the United States were similar to those in non-U.S. facilities (68% vs. 63%, respectively; p = 0.237), although U.S. facilities reported greater reductions in invasive coronary angiography (69% vs. 53%, respectively; p < 0.001). Significantly more U.S. facilities reported increased use of telehealth and patient screening measures than non-U.S. facilities, such as temperature checks, symptom screenings, and COVID-19 testing. Reductions in volumes of procedures differed between U.S. regions, with larger declines observed in the Northeast (76%) and Midwest (74%) than in the South (62%) and West (44%). Prevalence of COVID-19, staff redeployments, outpatient centers, and urban centers were associated with greater reductions in volume in U.S. facilities in a multivariable analysis. Conclusions: We observed marked reductions in U.S. cardiovascular testing in the early phase of the pandemic and significant variability between U.S. regions. The association between reductions of volumes and COVID-19 prevalence in the United States highlighted the need for proactive efforts to maintain access to cardiovascular testing in areas most affected by outbreaks of COVID-19 infection