Human Cytomegalovirus Inhibitor AL18 Also Possesses Activity against Influenza A and B Viruses

AL18, an inhibitor of human cytomegalovirus DNA polymerase, was serendipitously found to also block the interaction between the PB1 and PA polymerase subunits of influenza A virus. Furthermore, AL18 effectively inhibited influenza A virus polymerase activity and the overall replication of influenza A and B viruses. A molecular model to explain the binding of AL18 to both cytomegalovirus and influenza targets is proposed. Thus, AL18 represents an interesting lead for the development of new antivirals

Unraveling pedestrian mobility on a road network using ICTs data during great tourist events

Tourist flows in historical cities are continuously growing in a globalized world and adequate governance processes, politics and tools are necessary in order to reduce impacts on the urban livability and to guarantee the preservation of cultural heritage. The ICTs offer the possibility of collecting large amount of data that can point out and quantify some statistical and dynamic properties of human mobility emerging from the individual behavior and referring to a whole road network. In this paper we analyze a new dataset that has been collected by the Italian mobile phone company TIM, which contains the GPS positions of a relevant sample of mobile devices when they actively connected to the cell phone network. Our aim is to propose innovative tools allowing to study properties of pedestrian mobility on the whole road network. Venice is a paradigmatic example for the impact of tourist flows on the resident life quality and on the preservation of cultural heritage. The GPS data provide anonymized georeferenced information on the displacements of the devices. After a filtering procedure, we develop specific algorithms able to reconstruct the daily mobility paths on the whole Venice road network. The statistical analysis of the mobility paths suggests the existence of a travel time budget for the mobility and points out the role of the rest times in the empirical relation between the mobility time and the corresponding path length. We succeed to highlight two connected mobility subnetworks extracted from the whole road network, that are able to explain the majority of the observed mobility. Our approach shows the existence of characteristic mobility paths in Venice for the tourists and for the residents. Moreover the data analysis highlights the different mobility features of the considered case studies and it allows to detect the mobility paths associated to different points of interest. Finally we have disaggregated the Italian and foreigner categories to study their different mobility behaviors

Analysis of the common genetic component of large-vessel vasculitides through a meta- Immunochip strategy

Giant cell arteritis (GCA) and Takayasu's arteritis (TAK) are major forms of large-vessel vasculitis (LVV) that share clinical features. To evaluate their genetic similarities, we analysed Immunochip genotyping data from 1,434 LVV patients and 3,814 unaffected controls. Genetic pleiotropy was also estimated. The HLA region harboured the main disease-specific associations. GCA was mostly associated with class II genes (HLA-DRB1/HLA-DQA1) whereas TAK was mostly associated with class I genes (HLA-B/MICA). Both the statistical significance and effect size of the HLA signals were considerably reduced in the cross-disease meta-analysis in comparison with the analysis of GCA and TAK separately. Consequently, no significant genetic correlation between these two diseases was observed when HLA variants were tested. Outside the HLA region, only one polymorphism located nearby the IL12B gene surpassed the study-wide significance threshold in the meta-analysis of the discovery datasets (rs755374, P?=?7.54E-07; ORGCA?=?1.19, ORTAK?=?1.50). This marker was confirmed as novel GCA risk factor using four additional cohorts (PGCA?=?5.52E-04, ORGCA?=?1.16). Taken together, our results provide evidence of strong genetic differences between GCA and TAK in the HLA. Outside this region, common susceptibility factors were suggested, especially within the IL12B locus

Colorectal Cancer Stage at Diagnosis Before vs During the COVID-19 Pandemic in Italy

IMPORTANCE Delays in screening programs and the reluctance of patients to seek medical attention because of the outbreak of SARS-CoV-2 could be associated with the risk of more advanced colorectal cancers at diagnosis. OBJECTIVE To evaluate whether the SARS-CoV-2 pandemic was associated with more advanced oncologic stage and change in clinical presentation for patients with colorectal cancer. DESIGN, SETTING, AND PARTICIPANTS This retrospective, multicenter cohort study included all 17 938 adult patients who underwent surgery for colorectal cancer from March 1, 2020, to December 31, 2021 (pandemic period), and from January 1, 2018, to February 29, 2020 (prepandemic period), in 81 participating centers in Italy, including tertiary centers and community hospitals. Follow-up was 30 days from surgery. EXPOSURES Any type of surgical procedure for colorectal cancer, including explorative surgery, palliative procedures, and atypical or segmental resections. MAIN OUTCOMES AND MEASURES The primary outcome was advanced stage of colorectal cancer at diagnosis. Secondary outcomes were distant metastasis, T4 stage, aggressive biology (defined as cancer with at least 1 of the following characteristics: signet ring cells, mucinous tumor, budding, lymphovascular invasion, perineural invasion, and lymphangitis), stenotic lesion, emergency surgery, and palliative surgery. The independent association between the pandemic period and the outcomes was assessed using multivariate random-effects logistic regression, with hospital as the cluster variable. RESULTS A total of 17 938 patients (10 007 men [55.8%]; mean [SD] age, 70.6 [12.2] years) underwent surgery for colorectal cancer: 7796 (43.5%) during the pandemic period and 10 142 (56.5%) during the prepandemic period. Logistic regression indicated that the pandemic period was significantly associated with an increased rate of advanced-stage colorectal cancer (odds ratio [OR], 1.07; 95%CI, 1.01-1.13; P = .03), aggressive biology (OR, 1.32; 95%CI, 1.15-1.53; P < .001), and stenotic lesions (OR, 1.15; 95%CI, 1.01-1.31; P = .03). CONCLUSIONS AND RELEVANCE This cohort study suggests a significant association between the SARS-CoV-2 pandemic and the risk of a more advanced oncologic stage at diagnosis among patients undergoing surgery for colorectal cancer and might indicate a potential reduction of survival for these patients

Infected pancreatic necrosis: outcomes and clinical predictors of mortality. A post hoc analysis of the MANCTRA-1 international study

: The identification of high-risk patients in the early stages of infected pancreatic necrosis (IPN) is critical, because it could help the clinicians to adopt more effective management strategies. We conducted a post hoc analysis of the MANCTRA-1 international study to assess the association between clinical risk factors and mortality among adult patients with IPN. Univariable and multivariable logistic regression models were used to identify prognostic factors of mortality. We identified 247 consecutive patients with IPN hospitalised between January 2019 and December 2020. History of uncontrolled arterial hypertension (p = 0.032; 95% CI 1.135-15.882; aOR 4.245), qSOFA (p = 0.005; 95% CI 1.359-5.879; aOR 2.828), renal failure (p = 0.022; 95% CI 1.138-5.442; aOR 2.489), and haemodynamic failure (p = 0.018; 95% CI 1.184-5.978; aOR 2.661), were identified as independent predictors of mortality in IPN patients. Cholangitis (p = 0.003; 95% CI 1.598-9.930; aOR 3.983), abdominal compartment syndrome (p = 0.032; 95% CI 1.090-6.967; aOR 2.735), and gastrointestinal/intra-abdominal bleeding (p = 0.009; 95% CI 1.286-5.712; aOR 2.710) were independently associated with the risk of mortality. Upfront open surgical necrosectomy was strongly associated with the risk of mortality (p < 0.001; 95% CI 1.912-7.442; aOR 3.772), whereas endoscopic drainage of pancreatic necrosis (p = 0.018; 95% CI 0.138-0.834; aOR 0.339) and enteral nutrition (p = 0.003; 95% CI 0.143-0.716; aOR 0.320) were found as protective factors. Organ failure, acute cholangitis, and upfront open surgical necrosectomy were the most significant predictors of mortality. Our study confirmed that, even in a subgroup of particularly ill patients such as those with IPN, upfront open surgery should be avoided as much as possible. Study protocol registered in ClinicalTrials.Gov (I.D. Number NCT04747990)

A Large-Scale Genetic Analysis Reveals a Strong Contribution of the HLA Class II Region to Giant Cell Arteritis Susceptibility

We conducted a large-scale genetic analysis on giant cell arteritis (GCA), a polygenic immune-mediated vasculitis. A case-control cohort, comprising 1,651 case subjects with GCA and 15,306 unrelated control subjects from six different countries of European ancestry, was genotyped by the Immunochip array. We also imputed HLA data with a previously validated imputation method to perform a more comprehensive analysis of this genomic region. The strongest association signals were observed in the HLA region, with rs477515 representing the highest peak (p = 4.05 × 10−40, OR = 1.73). A multivariate model including class II amino acids of HLA-DRβ1 and HLA-DQα1 and one class I amino acid of HLA-B explained most of the HLA association with GCA, consistent with previously reported associations of classical HLA alleles like HLA-DRB1∗04. An omnibus test on polymorphic amino acid positions highlighted DRβ1 13 (p = 4.08 × 10−43) and HLA-DQα1 47 (p = 4.02 × 10−46), 56, and 76 (both p = 1.84 × 10−45) as relevant positions for disease susceptibility. Outside the HLA region, the most significant loci included PTPN22 (rs2476601, p = 1.73 × 10−6, OR = 1.38), LRRC32 (rs10160518, p = 4.39 × 10−6, OR = 1.20), and REL (rs115674477, p = 1.10 × 10−5, OR = 1.63). Our study provides evidence of a strong contribution of HLA class I and II molecules to susceptibility to GCA. In the non-HLA region, we confirmed a key role for the functional PTPN22 rs2476601 variant and proposed other putative risk loci for GCA involved in Th1, Th17, and Treg cell function

SVILUPPO DI NUOVE STRATEGIE ANTIVIRALI DIRETTE CONTRO L' RNA POLIMERASI DEL VIRUS DELL'INFLUENZA

Influenza (flu) is an airborne highly-infectious disease, characterized by high morbidity and significant mortality, especially in at-risk population (young children, elderly people, patients with cronical disease). Annually, 20% of general population is affected by seasonal epidemics sustained by genetic variants of already circulating influenza virus strains. For this reason the vaccine has to be changed every year with a considerable economic impact. Furthermore, the influenza virus is responsible for occasional pandemics affecting million of people worldwide. The migratory water bird are the natural reservoir for the avian flu. The recent avian flu virus that affected humans suggests that a new flu epidemic could be able to overcome the species barrier and spread from human to human. For these reasons there is an urgent need to develop a new drug able to target a conservative flu motif. To date, there are a few commercially available drugs that target some envelope viral proteins: neuraminidase (NA) and the M2 ion channel. Oseltamivir (Tamiflu) and zanamivir are NA inhibitors, which prevent the release of the viral particle from infected cells and are active against flu A and flu B. Amantadine and rimantadine inhibit the protein M2 and viral uncoating. These drugs have important drawbacks including neurological toxicity, they can't be administrated to subjects younger than 12 year-old, finally, they have to be taken at the time of onset of symptoms. Furthermore, they can very frequentely cause the induction of drug-resistant strains. Moreover, although an anti-flu vaccine is available, it has to be changed every year because the virus is prone to antigenic modifications. For all of the above, there is still an urgent need for effective anti-flu compounds with a different mechanism of action from the current anti-flu drugs. A possible novel anti-influenza strategy is rapresented by the RNA polymerase encoded by flu virus; in fact, the influenza virus polymerase is a conserved element among different viral strains, as it is not subjected to genetic variability. The influenza virus RNA polymerase is a heterocomplex which consists of three proteins (PB1, PB2, and PA) that interact each other, in particular PB1 interacts with PB2 and PA, while PA and PB2 do not interact. It has been demonstrated that the PA/PB1 interaction is crucial for flu virus replication: in fact, the inhibition of this interaction cause the inhibition of flu replication. Thus, the inhibition of such interactions may represent a feasible strategy for the development of anti-flu compounds able to block the replication of the virus independently from the viral strain. The main aim of this project is to identify small molecules able to block the interaction between the PA and PB1 subunits of the viral RNA polymerase and consequently to inhibit virus replication. To this aim, an in silico screening has been conducted on approximately 3 millions of virtual compounds taking advantage of the crystallographic structure of a truncated form of PA bound a PB1-derived peptide. In this screening we identified 32 compounds potentially able to inhibit the PA/PB1 interaction. We then tested these compounds using an ELISA to verify their capability to disrupt the physical interaction between PA and PB1 in vitro. From this analysis, we identified 3 compounds able to block the PA/PB1 interaction with an IC50 comparable to that of a peptide corresponding to aa 1-15 of PB1, that rapresent the PA-binding domain. In addition, the cytotoxicity of the active compounds has been evaluated in a panel of cell lines. Analogs of the most promising compounds have been synthesized to increase antiviral activity and decrease the toxicity. The compound activity has been further characterized by the "minireplicon assay", a cellular assay that evaluates the compounds effects on flu A and flu B polymerase activity. We identified some compounds able to inhibit the polymerase activity with an IC50 value comparable to that of the PB11-15 peptide. We also analised the ability of the compounds to inhibit the viral replication by a plaque reduction assay. We found that 3 compounds inhibit flu A/PR/8/34 replication with EC50 values comparable to that of the reference compound ribavirin (RBV). Furthermore, we evaluated the activity of these compounds on the replication of a flu B strain (B/Malaysia/2506/4) and of some flu A strains (A/WSN/67/05, A/Solomon Island/3/06, and Tamiflu-resistant A/PARMA/24/09). Compounds 1 and 34 showed an antiviral activity against these flu strains comparable with that of RBV (EC50 18-22,5 µM for compound 1; EC50 17-25 µM for compound 34; EC50 7-18 µM for RBV). The antiviral activity of the compound 34 has been also characterised at different time post-infection. We found that this molecule has higher antiviral activity until 12 hours post-infection (1-2 µM). Finally, its activity was evaluated also on the intranuclear traslocation of the PA/PB1 complex and on viral protein expression. Compound 34 showed antiviral activity also in these assays. Studies to characterise further its activity and to identify more potent analogues are in progress. Finally, another line of research characterized at the same time, some Stealth siRNAs directed against PB1 mRNA and evaluated their capability to inhibit the flu polymerase activity. Previously data reported an antiviral activity for one of these siRNAs in infected cells; we have confirmed its antiviral activity by minireplicon assays (80% of inhibition at 30 nM). We can conclude that we identified two compounds, compound 1 and 34, and a siRNA that are a good candidate for development of a new anti-flu compound. In the future, we are going to characterize the antiviral activity of the selected compounds and identify new analogues compounds to increase their antiviral activity.L'influenza è un'infezione respiratoria, di origine virale, caratterizzata da elevata morbidità  e mortalità  soprattutto nella popolazione ad alto rischio (bambini, anziani, pazienti con malattie croniche/debilitanti, ecc.). Ogni anno il 20% della popolazione è affetto da un'epidemia causata dalla variabilità antigenica dei diversi ceppi di influenza, questo comporta che il vaccino anti-influenza debba essere riformulato di anno in anno con un considerevole impatto economico. Inoltre il virus dell'influenza è responsabile di pandemie occasionali che colpiscono milioni di persone in tutto il mondo e che si verificano ogni 10-30 anni con incidenza di morte dell'ordine del milione. Gli uccelli selvatici (uccelli acquatici, anatre selvatiche, etc.) sono il serbatoio naturale del virus aviario, e i recenti casi di influenza aviaria che hanno colpito l'uomo indicano che una nuova epidemia influenzale potrebbe essere in grado non solo di saltare le barriere tra le specie ma anche di trasmettersi da uomo a uomo causando milioni di morti. Per questi motivi esiste la necessit  di trovare un nuovo farmaco che abbia come bersaglio un elemento virale conservato. Attualmente esistono in commercio pochi farmaci anti-influenza, che hanno come bersaglio alcune proteine virali di superficie: neuraminidasi (NA) e una proteina che costituisce un canale ionico (M2). Oseltamivir (Tamiflu) e zanamivir sono inibitori di NA e prevengono il rilascio delle particelle virali dalle cellule infettate (per flu A e flu B). Amantadina e rimantadina invece hanno come bersaglio la proteina M2, e agiscono sulla scapsidazione del virus. Questi farmaci presentano però numerosi svantaggi: possono avere effetti collaterali causando tossicità  a livello neurologico, non possono essere somministrati al di sotto dei 12 anni di età , infine per essere efficaci devono essere assunti entro le 48 ore dall'insorgenza dei sintomi; inoltre, per tali farmaci si è registrata la comparsa di molti ceppi virali resistenti. Il vaccino anti-influenza che viene messo a disposizione per la popolazione ha lo svantaggio di dover essere riformulato ogni anno a causa dell'elevata variabilità  antigenica dei diversi ceppi. Di fatto, non esiste una terapia efficace e persiste quindi una necessità  reale di sviluppare nuovi farmaci anti-influenza soprattutto dotati di un meccanismo d'azione diverso da quello dei farmaci attualmente a disposizione. Un possibile bersaglio per lo sviluppo di nuovi farmaci antivirali è l'RNA polimerasi codificata dal virus dell'influenza, in quanto è un elemento conservato fra i diversi ceppi virali e non è soggetto a riassortimento genico. Le tre subunità  proteiche che costituiscono la RNA polimerasi virale (un eterotrimero costituito da PB1, PB2 e PA in rapporto stechiometrico 1:1:1) interagiscono fisicamente tra loro e, in particolare, la subunità  PB1 interagisce sia con PB2 che con PA, ma non c'è interazione tra PB2 e PA. E' stato dimostrato che l'interazione PA/PB1 è essenziale per la replicazione del virus dell'influenza: infatti l'inibizione di questa interazione causa anche inibizione della replicazione virale. L'utilizzo della polimerasi come target farmacologico permetterebbe quindi di inibire la replicazione virale ovviando al problema della variabilità  antigenica e si otterrebbe un farmaco attivo contro moltissimi, se non tutti, i ceppi del virus dell'influenza. Il fatto che il sito di legame PA-PB1 sia molto conservato e che l'interazione sia cruciale per molte funzioni virali fa sì che esso rappresenti un potenziale bersaglio per lo sviluppo di nuovi composti antivirali. Scopo di questo progetto è stato quello di identificare small molecules in grado di dissociare l'interazione fisica tra le subunità  PB1 e PA della RNA polimerasi del virus dell'influenza e quindi la replicazione virale. E' stato condotto uno screening in silico di molecole, a partire da circa 3 milioni di composti virtuali, utilizzando la struttura cristallografica del complesso PA/PB1 e ne sono state identificate 32 potenzialmente capaci di inibire questa interazione. Questi 32 composti sono stati inizialmente saggiati mediante ELISA, per verificare la loro effettiva capacità  di dissociare in vitro l'interazione PA/PB1. Da questa prima analisi sono stati identificati 3 composti in grado di dissociare l'interazione con un valore di IC50 comparabile al peptide corrispondente ai 15 aa di PB1, che rappresenta la regione di interazione con PA e che è stato dimostrato essere sufficiente per inibire l'interazione PA/PB1. Successivamente è stata valutata la citotossicità  delle molecole e sono stati disegnati dei composti analoghi a quelli maggiormente attivi per cercare di aumentarne l'attività  antivirale e/o diminuirne la citotossicità . L'attività  dei composti è stata confermata mediante minireplicon assay, un saggio cellulare che permette di determinare l'effetto delle molecole sull'attività  polimerasica sia del virus dell'influenza A che B. Sono stati identificati alcuni composti in grado di inibire l'attività  polimerasica virale con un valore di IC50 comparabile a quello del peptide di controllo. E' stata valutata, a questo punto, la capacità  dei composti di inibire la replicazione virale mediante saggio di riduzione delle placche. Quello che è emerso è che 3 composti inibiscono la replicazione del virus A/PR/8/34 con valori di EC50 comparabili a quelli della ribavirina (RBV), utilizzata come controllo, e a quelli dei farmaci attualmente utilizzati in terapia. Inoltre, è stata valutata la capacità  di questi composti di inibire anche la replicazione di un virus dell'influenza B (B/Malaysia/2506/4) e altri ceppi di influenza A (A/WSN/67/05, A/Solomon Island/3/06 e A/PARMA/24/09 Tamiflu-resistente). E' stato osservato che i composti 1 e 34 sono attivi contro questi ceppi di influenza in maniera analoga al composto di riferimento RBV (EC50 18-22,5 µM composto 1; EC50 17-25 µM composto 34; EC50 7-18 µM RBV). Per il composto 34 è stata anche valutata l'attività  antivirale nel tempo; quello che è emerso è che una elevata attività  è mantenuta fino a 12 h post-infezione (1-2 µM). L'attività  antivirale dei composti 1 e 34 è stata valutata anche caratterizzando il loro effetto sull'espressione delle proteine virali. Entrambi i composti hanno mostrato attività  antivirale. Inoltre à stato determinata l'attività  del composto 34 sulla traslocazione intranucleare del complesso PB1/PA; il composto 34 ha mostrato attività antivirale anche in questo saggio. Studi per caratterizzarne ulteriormente l'attività e per identificarne dei nuovi analoghi possibilmente più attivi sono in corso. In parallelo sono stati saggiati alcuni Stealth siRNA disegnati contro il trascritto di PB1 per valutare la loro capacità  di inibire l'attività  polimerasica. Dati precedenti hanno dimostrato attività  antivirale per uno di questi siRNA in cellule infettate; la capacità  di inibire anche l'attività  polimerasica è stata confermata mediante minireplicon assay (inibizione dell'80% a concentrazione 30 nM). Possiamo quindi concludere di aver identificato due composti, composto 1 e 34, e un siRNA che possono essere considerati dei buoni candidati per lo sviluppo di un farmaco anti-influenza. Ci proponiamo di caratterizzare ulteriormente l'attività  antivirale dei composti identificati, e, per quanto riguarda i composti selezionati mediante lo screening in silico, di identificare dei composti analoghi in modo da aumentare la loro attività  antivirale

Comment on: Rituximab therapy for takayasu arteritis: A seven patients experience and a review of the literature: Reply

Comment on: Rituximab therapy for Takayasu arteritis: a seven patients experience and a review of the literature: repl

Recent advances in the diagnosis and treatment of polymyalgia rheumatica

Polymyalgia rheumatica is one of the most common rheumatic inflammatory disorders in people older than 50 years characterized by aching and prolonged morning stiffness in the shoulder and pelvic girdle and neck.. Areas covered: In this review, we will focus on recent advances on the diagnosis and management of PMR. Expert commentary: Controversy exist whether PMR represent a single entity disease or is an umbrella term that comprises a clinical presentation common to a range of related conditions (polymyalgic syndrome). To date there are no specific diagnostic tests, and the diagnosis remains clinical, although ultrasonography, positron emission tomography scan and the recent ACR/EULAR classification criteria may help to confirm the clinical diagnosis. A step-wise process for the diagnosis of PMR has been proposed. Low-dose steroids are highly effective in the majority of patients and remain the mainstay of treatment, but relapses occur in about 50% of patients and glucocorticoid related adverse event are common. The steroid sparing effects of the immunosuppressive treatment evaluated to date are unclear

L30. Assessment of vasculitis extent and severity

Review on the assessment of vasculitis extent and severity