Excess of NPM-ALK oncogenic signaling promotes cellular apoptosis and drug dependency.

Most of the anaplastic large-cell lymphoma (ALCL) cases carry the t(2;5; p23;q35) that produces the fusion protein NPM-ALK (nucleophosmin-anaplastic lymphoma kinase). NPM-ALK-deregulated kinase activity drives several pathways that support malignant transformation of lymphoma cells. We found that in ALK-rearranged ALCL cell lines, NPM-ALK was distributed in equal amounts between the cytoplasm and the nucleus. Only the cytoplasmic portion was catalytically active in both cell lines and primary ALCL, whereas the nuclear portion was inactive because of heterodimerization with NPM1. Thus, about 50% of the NPM-ALK is not active and sequestered as NPM-ALK/NPM1 heterodimers in the nucleus. Overexpression or relocalization of NPM-ALK to the cytoplasm by NPM genetic knockout or knockdown caused ERK1/2 (extracellular signal-regulated protein kinases 1 and 2) increased phosphorylation and cell death through the engagement of an ATM/Chk2- and γH2AX (phosphorylated H2A histone family member X)-mediated DNA-damage response. Remarkably, human NPM-ALK-amplified cell lines resistant to ALK tyrosine kinase inhibitors (TKIs) underwent apoptosis upon drug withdrawal as a consequence of ERK1/2 hyperactivation. Altogether, these findings indicate that an excess of NPM-ALK activation and signaling induces apoptosis via oncogenic stress responses. A 'drug holiday' where the ALK TKI treatment is suspended could represent a therapeutic option in cells that become resistant by NPM-ALK amplification.We thank Maria Stella Scalzo for technical support, Dr Emanuela Colombo for kindly providing MEFs that lack NPM1 (MEF NPM−/−p53−/−) and control fibroblasts (MEF p53−/−), Dr Guido Serini for the use of his confocal microscopy unit at the Candiolo Cancer Institute—IRCCS, Torino, Italy. We also thank Ariad Pharmaceutical, Pfizer, Astellas and Novartis that kindly provided all drugs used in this study. This work was supported by the Regione Lombardia (ID14546A) and Fondazione Berlucchi Onlus Grant 2014 (to CGP), and by grants FP7 ERC-2009-StG (Proposal No. 242965—‘Lunely’); Associazione Italiana per la Ricerca sul Cancro (AIRC) Grant IG-12023; Koch Institute/DFCC Bridge Project Fund; Ellison Foundation Boston; Worldwide Cancer Research Association (former AICR) grant 12-0216; the Grant for Oncology Innovation by Merck-Serono and R01 CA196703-01 (to RC).This is the author accepted manuscript. The final version is available from NPG via http://dx.doi.org/10.1038/onc.2015.45

Investigation of mRNA expression levels of DNA damage response genes in Merkel Cell Polyomavirus-positive Merkel Cell Carcinoma: a pilot study

Merkel Cell Polyomavirus (MCPyV) is recognized as the major aetiological agent of Merkel Cell Carcinoma (MCC), an aggressive skin tumor. MCPyV-mediated oncogenesis is strictly dependent on viral integration and the expression of a truncated form of the Large T Antigen (LT). Moreover, like other oncogenic DNA viruses, MCPyV may interfere with the DNA damage response (DDR) machinery, thus promoting genomic instability and tumorigenesis. Therefore, the objective of this study was to characterize MCPyV infection in 7 MCC patients and to elucidate the plausible role of the virus in the DDR pathway. MCPyV DNA was detected in 3/7 MCC patients and, as expected, viral integration and LT truncation were observed in virus-positive MCCs, along with the expression of early genes only. Over-expression of DDR genes such as ATM, ATR and their downstream kinases Chk1 and Chk2 was reported in MCPyV-positive MCCs supporting the potential role of the virus in interfering with DDR. Our findings support the established viral aetiology of MCC, and describe, for the first time, an over-expression of DDR components in MCPyV-positive MCC, laying the basis for future studies aimed at investigating the contribution of this pathway to MCPyV-mediated carcinogenesis and exploring the plausible clinical implications of host DDR factors for the treatment of MCC

Neurobrucellosis Presenting with Motor Damage or Hearing Loss, and Use of Steroids are Associated with a Higher Risk of Sequelae or Relapse: A Systematic Review of Individual Participant Data

Background: Neurobrucellosis presents diverse clinical challenges and risks of long-term complications. Objective: We aimed to assess the relationship between the duration of antibiotic therapy, clinical factors, and the outcome of neurobrucellosis with a case report combined with a systematic review of the literature. Methods: We present a case of a 31 years-old man successfully treated at our Institution. We then searched Ovid MEDLINE, Embase and Scopus for articles that encompassed neurobrucellosis cases, duration of treatment, and outcome. The primary outcome was to assess an association between the duration of treatment and the risk of sequelae or relapses. Univariate, multivariate and sensitivity analysis were carried out to define which variables affect​ed​ the clinical outcome. Quality assessment was performed using a dedicated tool. Results: A total of 123 studies were included, totaling 221 patients. Median duration of treatment was 4 months (IQR 3 - 6), 69% patients recovered without sequelae, 27% had sequelae. Additionally, five patients had a relapse, and 4 patients died. Multivariate analysis found that the duration of treatment, age, and the use of ceftriaxone were not associated with a higher risk of sequelae or relapses. A significant association was found for corticosteroids use (OR 0.39, 95% IC 0.16 - 0.96, p = 0.038), motor impairment (OR 0.29, 95% IC 0.14 - 0.62, p = 0.002), and hearing loss (OR 0.037, 95% IC 0.01 - 0.11, p < 0.001). Conclusions: This study highlights the variability in clinical presentations and treatment approaches for neurobrucellosis. Patients with factors indicating higher sequelae risk require meticulous follow-up

The unseen evidence of reduced Ionicity. The elephant in (the) room temperature ionic liquids

The unambiguous quantification of the proton transfer in Protic Ionic Liquids (PILs) and its differentiation from the concept of ionicity are still unsolved questions. Albeit researchers awfully quickly treat them as synonyms, the two concepts are intrinsically different and imply a dramatic modification in the expected chemical and physical properties of a PIL. Some attempts have been made to shed light on this discrimination, but single-technique-based approaches fail in giving a clear answer. Aiming at definitively figuring out the differentiation between proton transfer and ionicity, we performed a multi-technique analysis (NMR, Raman, IR, thermal and electrochemical analyses, among others). Indeed, thermal and spectroscopic analyses are employed to determine the acid strength's role in ions' complete formation. To overcome the ambiguity between ionicity and formation degree, we introduce a new paradigm where Reduced Ionicity accounts for both the quantities mentioned above. The reduced ionicity directly affects the thermal stability, the phase behavior, and the spectroscopic observations, resulting in particular features in NMR and vibrational spectra. The combination of physical-chemical analyses and Pulsed-Gradient Spin-Echo (PGSE) NMR allows determining the reduced ionicity (and not the ionicity, as reported so far) of the investigated systems. In this context, being the proton transfer not quantitatively accessible directly, the reduced ionicity of a reference series of triethylamine-based PILs is investigated through transport properties as a function of temperature. Our findings point towards a substantial dependence of the reduced ionicity by the acid strength and the anion's coordination power. Furthermore, some interesting insights about the proton transfer are obtained, combining all the findings collected

Trimethoprim-sulfamethoxazole significantly reduces the risk of nocardiosis in solid organ transplant recipients: systematic review and individual patient data meta-analysis

Background: Whether trimethoprim-sulfamethoxazole (TMP-SMX) prophylaxis prevents nocardiosis in solid organ transplant (SOT) recipients is controversial. Objectives: To assess the effect of TMP-SMX in the prevention of nocardiosis after SOT, its dose-response relationship, its effect on preventing disseminated nocardiosis, and the risk of TMP-SMX resistance in case of breakthrough infection. Methods: A systematic review and individual patient data meta-analysis. Data sources: MEDLINE, Embase, Cochrane Central Register of Controlled Trials, Cochrane Database of Systematic Reviews, Web of Science Core Collection, and Scopus up to 19 September 2023. Study eligibility criteria: (a) Risk of nocardiosis between SOT recipients with and without TMP-SMX prophylaxis, or (b) sufficient details to determine the rate of TMP-SMX resistance in breakthrough nocardiosis. Participants: SOT recipients. Intervention: TMP-SMX prophylaxis versus no prophylaxis. Assessment of risk of bias: Risk Of Bias In Non-randomized Studies-of Exposure (ROBINS-E) for comparative studies; dedicated tool for non-comparative studies. Methods of data synthesis: For our primary outcome (i.e. to determine the effect of TMP-SMX on the risk of nocardiosis), a one-step mixed-effects regression model was used to estimate the association between the outcome and the exposure. Univariate and multivariable unconditional regression models were used to adjust for the potential confounding effects. Certainty of evidence was assessed using Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach. Results: Individual data from three case-control studies were obtained (260 SOT recipients with nocardiosis and 519 uninfected controls). TMP-SMX prophylaxis was independently associated with a significantly decreased risk of nocardiosis (adjusted OR = 0.3, 95% CI 0.18-0.52, moderate certainty of evidence). Variables independently associated with an increased risk of nocardiosis were older age, current use of corticosteroids, high calcineurin inhibitor concentration, recent acute rejection, lower lymphocyte count, and heart transplant. Breakthrough infections (66/260, 25%) were generally susceptible to TMP-SMX (pooled proportion 98%, 95% CI 92-100). Conclusions: In SOT recipients, TMP-SMX prophylaxis likely reduces the risk of nocardiosis. Resistance appears uncommon in case of breakthrough infection

SARS-CoV-2 genomic evolution during a severe and long-lasting omicron infection under antiviral therapy

BackgroundProlonged SARS-CoV-2 infection observed in immunocompromised individuals even in the presence of antiviral treatment provides opportunities for viruses to evolve in immune escape and drug-resistant variants.Case presentationA 72-year-old male with IgG4-related disease was admitted to the Emergency Department of a city Hospital in Milan and then transferred to Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico in December 2023, due to respiratory distress due to SARS-CoV-2 infection diagnosed in November 2023. After 117 days since the onset of the infection, and two cycles of sotrovimab/remdesivir combined therapy, the clinical improvement allowed the hospital discharge, notwithstanding the persistent SARS-CoV-2 positivity. Fifteen days later, the patient was re-admitted to the hospital due to worsening clinical conditions. After a third cycle of sotrovimab/remdesivir combined therapy prolonged with nirmatrelvir/ritonavir, nasopharyngeal load dropped and clinical conditions improved, ending with a successful discharge. SARS-CoV-2 whole genome sequences, obtained at six time-points of infection, showed an FL.1.5.1 recombinant form infection and a genetic distance of median (IQR) 0.00052 (0.00041-0.00066) similar to the genetic distance observed among the 43 contemporaneous FL.1.5.1 recombinant forms (p = 0.098). De novo SNPs were observed at all time points, with a peak (n = 70) at day 133 of infection, corresponding to the time of the second hospitalization. Six non-synonymous mutations (three in the RdRp and three in the spike protein, four of them known to be associated with drug resistance) appeared transiently, after the third and fourth course of sotrovimab 500 mg/remdesivir combination. Five de novo SNPs, three of them in the spike protein, were fixed over the long-lasting infection. The spike N856K, associated with reduced fusogenicity and infectivity in Omicron BA.1, was completely replaced by constitutive N at day 136.ConclusionsThis clinical case confirms the intra-host evolution dynamics of SARS-CoV-2 in an immunocompromised, prolonged-infected individual, involving positions associated with drug resistance and fusogenic traits of SARS-CoV-2. These results underscore the importance of the early detection of SARS-CoV-2 infection in immunocompromised individuals, and its rapid containment using highly effective treatment, to limit serious complications and the risk of new and potentially concerning viral variants emergence

Effectiveness of Snail Slime in the Green Synthesis of Silver Nanoparticles

The development of green, low cost and sustainable synthetic routes to produce metal nanoparticles is of outmost importance, as these materials fulfill large scale applications in a number of different areas. Herein, snail slime extracted from Helix Aspersa snails was successfully employed both as bio-reducing agent of silver nitrate and as bio-stabilizer of the obtained nanoparticles. Several trials were carried out by varying temperature, the volume of snail slime and the silver nitrate concentration to find the best biogenic pathway to produce silver nanoparticles. The best results were obtained when the synthesis was performed at room temperature and neutral pH. UV-Visible Spectroscopy, SEM-TEM and FTIR were used for a detailed characterization of the nanoparticles. The obtained nanoparticles are spherical, with mean diameters measured from TEM images ranging from 15 to 30 nm and stable over time. The role of proteins and glycoproteins in the biogenic production of silver nanoparticles was elucidated. Infrared spectra clearly showed the presence of proteins all around the silver core. The macromolecular shell is also responsible of the effectiveness of the synthesized AgNPs to inhibit Gram positive and Gram negative bacterial growth

High rates of anal Merkel Cell Polyomavirus and HPV co-infection among people living with HIV

Knowledge of Human Polyomavirus (HPyV) infection in the anal area and its association with sexually transmitted infections such as Human Papillomavirus (HPV) and Human Immunodeficiency Virus (HIV) remains limited. Therefore, anal specimens from 150 individuals of both sexes were analyzed for screening purposes. HPV DNA was found in 50.7% of cases, with a predominance of high-risk (HR) genotypes. HPyV DNA was found in 39.3% of samples, with Merkel Cell Polyomavirus (MCPyV) being the most common, with a higher viral load than JCPyV and BKPyV. In addition, MCPyV viral load increased in people living with HIV (PLWH) with HPV infection (p < 0.0001)

Wide variability of the definitions used for native vertebral osteomyelitis: walking the path for a unified diagnostic framework with a meta-epidemiological approach

BACKGROUND CONTEXT Native Vertebral Osteomyelitis (NVO) has seen a rise in incidence, yet clinical outcomes remain poor with high relapse rates and significant long-term sequelae. The 2015 IDSA Clinical Practice Guidelines initiated a surge in scholarly activity on NVO, revealing a patchwork of definitions and numerous synonyms used interchangeably for this syndrome. PURPOSE To systematically summarize these definitions, evaluate their content, distribution over time, and thematic clustering. STUDY DESIGN/SETTING Meta-epidemiological study with a systematic review of definitions. PATIENTS SAMPLE An extensive search of multiple databases was conducted, targeting trials and cohort studies dating from 2005 to present, providing a definition for NVO and its synonyms. OUTCOME MEASURES Analysis of the diagnostic criteria that composed the definitions and the breaking up of the definitions in the possible combinations of diagnostic criteria. METHODS We pursued a thematic synthesis of the published definitions with Boolean logic, yielding single or multiple definitions per included study. Using 8 predefined diagnostic criteria, we standardized definitions, focusing on the minimum necessary combinations used. Definition components were visualized using Sankey diagrams. RESULTS The literature search identified 8,460 references, leading to 171 studies reporting on 21,963 patients. Of these, 91.2% were retrospective, 7.6% prospective, and 1.2% RCTs. Most definitions originated from authors, with 29.2% referencing sources. We identified 92 unique combinations of diagnostic criteria across the literature. Thirteen main patterns emerged, with the most common being clinical features with imaging, followed by clinical features combined with imaging and microbiology, and lastly, imaging paired with microbiology. CONCLUSIONS Our findings underscore the need for a collaborative effort to develop standardized diagnostic criteria. We advocate for a future Delphi consensus among experts to establish a unified diagnostic framework for NVO, emphasizing the core components of clinical features and MRI while incorporating microbiological and histopathological insights to improve both patient outcomes and research advancements