A novel thermostable protein-tag: optimization of the Sulfolobus solfataricus DNA- alkyl-transferase by protein engineering

In the last decade, a powerful biotechnological tool for the in vivo and in vitro specific labeling of proteins (SNAP-tag™ technology) was proposed as a valid alternative to classical protein-tags (green fluorescent proteins, GFPs). This was made possible by the discovery of the irreversible reaction of the human alkylguanine-DNA-alkyl-transferase (hAGT) in the presence of benzyl-guanine derivatives. However, the mild reaction conditions and the general instability of the mesophilic SNAP-tag™ make this new approach not fully applicable to (hyper-)thermophilic and, in general, extremophilic organisms. Here, we introduce an engineered variant of the thermostable alkylguanine-DNA-alkyl-transferase from the Archaea Sulfolobus solfataricus (SsOGT-H5), which displays a catalytic efficiency comparable to the SNAP-tag™ protein, but showing high intrinsic stability typical of proteins from this organism. The successful heterologous expression obtained in a thermophilic model organism makes SsOGT-H5 a valid candidate as protein-tag for organisms living in extreme environments

Imported Loa Loa Filariasis: Three Cases and a Review of Cases Reported in Non-Endemic Countries in the Past 25 Years

Summary Objectives The aim of this study was to highlight the increasing chance of Western physicians encountering patients (both immigrants and expatriates/travelers) seeking help for loiasis. Methods We describe three cases of imported loiasis observed at two hospitals in Italy and France, and present a review of all previously published cases in the medical literature in the last 25 years (1986–2011). The search was performed using PubMed and Scopus databases using the terms " Loa loa " AND "loiasis". Results We reviewed 101 cases of imported loiasis of which 61 (60.4%) were reported from Europe and 31 (30.7%) from the USA. Seventy-five percent of infestations were acquired in three countries: Cameroon, Nigeria, and Gabon. Overall, peripheral blood microfilariae were detected in 61.4% of patients, eosinophilia in 82.1%, eye worm migration in 53.5%, and Calabar swellings in 41.6%. However, Calabar swellings and eosinophilia were more common among expatriates/travelers, whereas African immigrants were more likely to have microfilaremia. Eye worm migration was observed in a similar proportion in the two groups. Only 35 patients (including the three described here) underwent clinical follow-up for a median period of 10.5 months (range 1–84 months); clinical relapse occurred in three of these patients and persistence or reappearance of blood microfilaria in another two. Conclusions Due to increasing travel and the migration of people from the endemic countries of West Africa to Europe and the USA, we speculate on the possible emergence of loiasis. Western physicians should be aware of the typical (eye worm migration and Calabar swellings) as well as unusual clinical presentations

SARS-CoV-2 infection in health workers: analysis from Verona SIEROEPID Study during the pre-vaccination era

Background: To report the baseline phase of the SIEROEPID study on SARS-CoV-2 infection seroprevalence among health workers at the University Hospital of Verona, Italy, between spring and fall 2020; to compare performances of several laboratory tests for SARS-CoV-2 antibody detection. Methods: 5299 voluntary health workers were enrolled from 28 April 2020 to 28 July 2020 to assess immunological response to SARS-CoV-2 infection throughout IgM, IgG and IgA serum levels titration by four laboratory tests. Association of antibody titre with several demographic variables, swab tests and performance tests (sensitivity, specificity, and agreement) were statistically analyzed. Results: The overall seroprevalence was 6%, considering either IgG and IgM, and 4.8% considering IgG. Working in COVID-19 Units was not associated with a statistically significant increase in the number of infected workers. Cohen's kappa of agreement between MaglumiTM and VivaDiagTM was quite good when considering IgG only (Cohen's kappa = 78.1%, 95% CI 74.0-82.0%), but was lower considering IgM (Cohen's kappa = 13.3%, 95% CI 7.8-18.7%). Conclusion: The large sample size with high participation (84.7%), the biobank and the longitudinal design were significant achievements, offering a baseline dataset as the benchmark for risk assessment, health surveillance and management of SARS-CoV-2 infection for the hospital workforce, especially considering the ongoing vaccination campaign. Study results support the national regulator guidelines on using swabs for SARS-CoV-2 screening with health workers and using the serological tests to contribute to the epidemiological assessment of the spread of the virus

Natalizumab affects T-cell phenotype in multiple sclerosis: implications for JCV reactivation

The anti-CD49d monoclonal antibody natalizumab is currently an effective therapy against the relapsing-remitting form of multiple sclerosis (RRMS). Natalizumab therapeutic efficacy is limited by the reactivation of the John Cunningham polyomavirus (JCV) and development of progressive multifocal leukoencephalopathy (PML). To correlate natalizumab-induced phenotypic modifications of peripheral blood T-lymphocytes with JCV reactivation, JCV-specific antibodies (serum), JCV-DNA (blood and urine), CD49d expression and relative abundance of peripheral blood T-lymphocyte subsets were longitudinally assessed in 26 natalizumab-treated RRMS patients. Statistical analyses were performed using GraphPad Prism and R. Natalizumab treatment reduced CD49d expression on memory and effector subsets of peripheral blood T-lymphocytes. Moreover, accumulation of peripheral blood CD8+ memory and effector cells was observed after 12 and 24 months of treatment. CD4+ and CD8+ T-lymphocyte immune-activation was increased after 24 months of treatment. Higher percentages of CD8+ effectors were observed in subjects with detectable JCV-DNA. Natalizumab reduces CD49d expression on CD8+ T-lymphocyte memory and effector subsets, limiting their migration to the central nervous system and determining their accumulation in peripheral blood. Impairment of central nervous system immune surveillance and reactivation of latent JCV, can explain the increased risk of PML development in natalizumab-treated RRMS subjects

Sistema Integrato Multicentrico di Indicatori. Rapporto 2005. Provincia di Pescara

The SIMI (Integrated System of Indicators multicenter) Project contributes to the development of an integrated management of informative data streams related to drug addicted persons. This report analyzes the phenomenon of addiction on the territory of Pescara province through a description of the network services that provide care and rehabilitation of those addicted. Besides the characteristics of users of local services for addictions, has been developed the analysis of the subjects reported to the prefectures for use of illegal drugs and any action taken. Standard methods of estimation were also applied to quantify the proportion of users of substances that do not relate to services and to identify certain characteristics.Il Progetto SIMI (Sistema Integrato Multicentrico di Indicatori) intende contribuire allo sviluppo di una gestione integrata e sinergica dei flussi informativi relativi ai consumatori di sostanze stupefacenti afferenti alle diverse amministrazioni dello Stato. In linea con quanto proposto dall\u27Osservatorio europeo di Lisbona, per la descrizione e analisi del fenomeno connesso all\u27uso/abuso di sostanze, risulta di fondamentale importanza la possibilit? di ottenere informazioni esaustive e comparabili sulle persone che usano e/o abusano di sostanze psicotrope. Il presente rapporto analizza il fenomeno delle dipendenze sul territorio della provincia di Pescara attraverso la descrizione della rete dei servizi preposti alla cura e riabilitazione dei soggetti tossicodipendenti. Accanto alle caratteristiche degli utenti dei servizi territoriali per le dipendenze, ? stata sviluppata l\u27analisi dei soggetti segnalati alle Prefetture per uso di sostanze illegali e degli eventuali provvedimenti adottati. Sono state inoltre applicate metodologie standard di stima per quantificare la quota parte di utilizzatori di sostanze che non afferiscono ai servizi e per identificarne alcune caratteristiche

Upper limb function in Duchenne muscular dystrophy: 24 month longitudinal data

The aim of the study was to establish 24 month changes in upper limb function using a revised version of the performance of upper limb test (PUL 2.0) in a large cohort of ambulant and non-ambulant boys with Duchenne muscular dystrophy and to identify possible trajectories of progression. Of the 187 patients studied, 87 were ambulant (age range: 7\u201315.8 years), and 90 non-ambulant (age range: 9.08\u201324.78). The total scores changed significantly over time (p<0.001). Non-ambulant patients had lower total scores at baseline (mean 19.7) when compared to the ambulant ones (mean 38.4). They also had also a bigger decrease in total scores over 24 months compared to the ambulant boys (4.36 vs 2.07 points). Multivariate model analysis showed that the Performance of Upper Limb changes reflected the entry level and ambulation status, that were independently associated to the slope of Performance of Upper Limb changes. This information will be of help both in clinical practice and at the time of designing clinical trials

Earthquake and People: The Maltese Experience of the 1908 Messina Earthquake

On December 28, 1908 at 5:20 a.m. local time, a devastating earthquake (Mw = 7.2) struck Southern Italy along the Messina Strait (Fig. 1). This event caused severe ground shaking throughout the region and triggered a local tsunami. As result the cities of Messina along Sicily’s coast and Reggio di Calabria were completely destroyed (Baratta, 1910) causing more than 120,000 fatalities and with many left without shelter. The effects of the earthquake were felt within a 300-kilometer radius. Rescuers searched through the rubble for weeks, and people were still being pulled out alive days later, but thousands remained buried there. The 1908 earthquake had a significant impact on buildings and people and local communities which were displaced. The Maltese experience of the Messina 1908 earthquake relied on communication which reached Malta after the event. The assessment of the Maltese experience of the Messina Earthquake has so far been carried out with reference to published newspaper reports and other brief accounts including Herbert Ganado’s Rajt Malta Tinbidel. Alfons Maria Galea a Maltese author and filanthropist published a book in Maltese on the earthquake and its devastating effects in the popular educational series il-Kotba tal-Mogħdija taż-Żmien just a few weeks after the event. The book is a vivid account of the destruction caused by the earthquake, the suffering of the survivors and the reaction of the population in reviving the city. The document presents first-hand accounts of the events in sufficient detail to give a clear picture of the severity of the event, extents of the damage and impact on the population. It is mostly based on accounts received by Galea from persons in institutions including religious orders in Sicily who he knew. Newspaper reports in Malta and other countries together with Galea’s book present clear first-hand accounts of this event and provide information on the building deficiencies and damage, limitations of communication infrastructure during that period, limits to timely emergency response to support the population and emergency action at the beginning of the 20th century.peer-reviewe

How the First Year of the COVID-19 Pandemic Impacted Patients’ Hospital Admission and Care in the Vascular Surgery Divisions of the Southern Regions of the Italian Peninsula

Background: To investigate the effects of the COVID-19 lockdowns on the vasculopathic population. Methods: The Divisions of Vascular Surgery of the southern Italian peninsula joined this multicenter retrospective study. Each received a 13-point questionnaire investigating the hospitalization rate of vascular patients in the first 11 months of the COVID-19 pandemic and in the preceding 11 months. Results: 27 out of 29 Centers were enrolled. April-December 2020 (7092 patients) vs. 2019 (9161 patients): post-EVAR surveillance, hospitalization for Rutherford category 3 peripheral arterial disease, and asymptomatic carotid stenosis revascularization significantly decreased (1484 (16.2%) vs. 1014 (14.3%), p = 0.0009; 1401 (15.29%) vs. 959 (13.52%), p = 0.0006; and 1558 (17.01%) vs. 934 (13.17%), p < 0.0001, respectively), while admissions for revascularization or major amputations for chronic limb-threatening ischemia and urgent revascularization for symptomatic carotid stenosis significantly increased (1204 (16.98%) vs. 1245 (13.59%), p < 0.0001; 355 (5.01%) vs. 358 (3.91%), p = 0.0007; and 153 (2.16%) vs. 140 (1.53%), p = 0.0009, respectively). Conclusions: The suspension of elective procedures during the COVID-19 pandemic caused a significant reduction in post-EVAR surveillance, and in the hospitalization of asymptomatic carotid stenosis revascularization and Rutherford 3 peripheral arterial disease. Consequentially, we observed a significant increase in admissions for urgent revascularization for symptomatic carotid stenosis, as well as for revascularization or major amputations for chronic limb-threatening ischemia