Benzofuroxan derivatives N-Br and N-I induce intrinsic apoptosis in melanoma cells by regulating AKT/BIM signaling and display anti metastatic activity in vivo

Abstract\ud \ud Background\ud Malignant melanoma is an aggressive type of skin cancer, and despite recent advances in treatment, the survival rate of the metastatic form remains low. Nifuroxazide analogues are drugs based on the substitution of the nitrofuran group by benzofuroxan, in view of the pharmacophore similarity of the nitro group, improving bioavailability, with higher intrinsic activity and less toxicity. Benzofuroxan activity involves the intracellular production of free-radical species. In the present work, we evaluated the antitumor effects of different benzofuroxan derivatives in a murine melanoma model.\ud \ud \ud Methods\ud B16F10-Nex2 melanoma cells were used to investigate the antitumor effects of Benzofuroxan derivatives in vitro and in a syngeneic melanoma model in C57Bl/6 mice. Cytotoxicity, morphological changes and reactive oxygen species (ROS) were assessed by a diphenyltetrasolium reagent, optical and fluorescence microscopy, respectively. Annexin-V binding and mitochondrial integrity were analyzed by flow cytometry. Western blotting and colorimetry identified cell signaling proteins.\ud \ud \ud Results\ud Benzofuroxan N-Br and N-I derivatives were active against murine and human tumor cell lines, exerting significant protection against metastatic melanoma in a syngeneic model. N-Br and N-I induce apoptosis in melanoma cells, evidenced by specific morphological changes, DNA condensation and degradation, and phosphatidylserine translocation in the plasma membrane. The intrinsic mitochondrial pathway in B16F10-Nex2 cells is suggested owing to reduced outer membrane potential in mitochondria, followed by caspase −9, −3 activation and cleavage of PARP. The cytotoxicity of N-Br and N-I in B16F10-Nex2 cells is mediated by the generation of ROS, inhibited by pre-incubation of the cells with N-acetylcysteine (NAC). The induction of ROS by N-Br and N-I resulted in the inhibition of AKT activation, an important molecule related to tumor cell survival, followed by upregulation of BIM.\ud \ud \ud Conclusion\ud We conclude that N-Br and N-I are promising agents aiming at cancer treatment. They may be useful in melanoma therapy as inducers of intrinsic apoptosis and by exerting significant antitumor activity against metastatic melanoma, as presently shown in syngeneic mice.The present work was supported by Fundação de Amparo a Pesquisa do\ud Estado de São Paulo (FAPESP) and the Conselho Nacional de Desenvolvimento\ud Científico e Tecnológico (CNPq)

Inquadramento clinico-strumentale dei pazienti con disabilità conseguente a COVID-19

Introduzione La malattia da SARS-CoV2 (COVID-19) è nota principalmente per l’interessamento dell’apparato respiratorio che porta, nei casi più gravi, allo sviluppo di una polmonite bilaterale richiedente ospedalizzazione e, nei pazienti critici, supporto ventilatorio meccanico per insufficienza respiratoria acuta. Sono tuttavia sempre più numerose le evidenze che la Coronavirus Disease-19 non sia un’affezione solo respiratoria, ma una malattia multisistemica. Tra le complicanze extrapolmonari più frequenti vi sono i disturbi neurologici centrali e periferici, tra cui ictus, encefalite e sindrome di Guillain-Barré. Un altro apparato coinvolto è quello muscolo-scheletrico, come dimostrato dal riscontro nei pazienti con COVID-19 di elevati livelli ematici di creatinfosfochinasi e dalla manifestazione di mialgie e artralgie. Il medico fisiatra, chiamato a valutare le necessità riabilitative dei pazienti affetti da COVID-19 che hanno superato la fase acuta di malattia, deve identificare possibili esiti disabilitanti a carico di questi sistemi, in modo da impostare un adeguato percorso di recupero funzionale. Obiettivo di questo studio è elaborare una procedura clinico-diagnostica che permetta di individuare eventuali problematiche neuromuscolari associate agli esiti respiratori dell’infezione da SARS-CoV2, attraverso l’utilizzo di esami elettrofisiologici, spirometria, test di tolleranza all’esercizio fisico, sistemi di analisi del movimento e pletismografia optoelettronica. Materiali e Metodi Sono stati reclutati dieci pazienti con diagnosi di COVID-19, clinicamente guariti (real-time reverse transcriptase-polymerase chain reaction su tampone nasofaringeo per SARS-CoV2 negativa), afferenti all'Unità Operativa di Riabilitazione Specialistica dell’Ospedale “Luigi Sacco” di Milano. I criteri di inclusione sono stati: pregressa COVID-19 con necessità di incubazione orotracheale ed età inferiore ai 65 anni. Sono stati esclusi i soggetti non in grado di percorrere una distanza di almeno 5 metri in autonomia senza ausili al momento del reclutamento e i pazienti affetti da deficit cognitivi che compromettono la collaborazione. Il campione arruolato è stato sottoposto a valutazione elettrofisiologica (ENG ed EMG), spirometria semplice, valutazione della forza della muscolatura respiratoria tramite MIP e MEP e 6-Minutes Walking Test presso la struttura ospedaliera, mentre ha eseguito baropodometria e Gait Analysis presso il Laboratorio di analisi del movimento di ORThesys (Milano) ed effettuato pletismografia optoelettronica (OEP) presso il Dipartimento di Bioingegneria del Politecnico di Milano. Risultati Dalla valutazione elettrofisiologica è emersa una sofferenza acuta del sistema nervoso periferico, tra cui: mononeuropatie a carico degli arti inferiori e del nervo frenico e polineuropatie, in otto pazienti su dieci esaminati. L’esame spirometrico ha evidenziato un quadro compatibile con sindrome restrittiva lieve in due soggetti. La MIP e la MEP sono risultate inferiori ai limiti di norma in quattro pazienti. Il test del cammino non ha rilevato episodi di desaturazione in nessuno dei pazienti e il livello di dispnea percepito al termine della prova è stato tra 1 e 2 alla scala di Borg CR10. Alla Gait Analysis si sono evidenziate anomalie a carico degli arti inferiori sia cinematiche, sia dinamiche. In particolare, l’escursione articolare di anca, ginocchio e caviglia risulta complessivamente diminuita in otto pazienti e l’intero campione, ad eccezione di un soggetto, presenta una ridotta erogazione di potenza di caviglia. La valutazione tramite OEP non ha evidenziato anomalie di rilievo nel pattern respiratorio a riposo, mentre si è riscontrata una riduzione della Capacità Vitale in un paziente. Il livello di dispnea percepito al termine di una serie di esercizi con incentivatore di flusso è risultato compreso tra 0 e 4 alla scala di Borg CR10. Conclusioni Questo studio clinico “quasi-sperimentale” mette in risalto l’importanza di effettuare un attento esame clinico-strumentale del paziente affetto da COVID-19, poiché la prevalenza di esiti neuromuscolari è elevata. A questo scopo risultano utili la valutazione elettrofisiologica e i sistemi di analisi del movimento; al fine di elaborare un progetto riabilitativo mirato, incentrato sul recupero stenico globale e il ricondizionamento all’esercizio fisico attraverso l’attività aerobica. I risultati di spirometria, MIP e MEP, 6-Minutes Walking Test e OEP suggeriscono, invece, in questa tipologia di paziente, limitate necessità riabilitative in ambito respiratorio. Pertanto, l’inquadramento clinico e strumentale dei pazienti con sindrome post-COVID, deve interessarsi sia delle problematiche respiratorie (che sembrano manifestarsi in misura minore rispetto alle previsioni), sia delle problematiche neuromotorie, che possono condizionare il benessere del paziente e il pieno recupero dell’autonomia nella deambulazione e nello svolgimento delle attività della vita quotidiana. Bibliografia • Ghannam M, Alshaer Q, Al-Chalabi M, Zakarna L, Robertson J, Manousakis G. Neurological involvement of coronavirus disease 2019: a systematic review. J Neurol. Published online 2020. • Mao L, Jin H, Wang M, et al. Neurologic Manifestations of Hospitalized Patients with Coronavirus Disease 2019 in Wuhan, China. JAMA Neurol. 2020;77(6):683-690. • Disser NP, De Micheli AJ, Schonk MM, et al. Musculoskeletal Consequences of COVID-19. J Bone Jt Surg Am Vol. 2020;102(14):1197-1204

A matter of sex—persistent predictive value of MECKI score prognostic power in men and women with heart failure and reduced ejection fraction: a multicenter study

Background: A sex-based evaluation of prognosis in heart failure (HF) is lacking. Methods and results: We analyzed the Metabolic Exercise test data combined with Cardiac and Kidney Indexes (MECKI) score registry, which includes HF with reduced ejection fraction (HFrEF) patients. A cross-validation procedure was performed to estimate weights separately for men and women of all MECKI score parameters: left ventricular ejection fraction (LVEF), hemoglobin, kidney function assessed by Modification of Diet in Renal Disease, blood sodium level, ventilation vs. carbon dioxide production slope, and peak oxygen consumption (peakVO2). The primary outcomes were the composite of all-cause mortality, urgent heart transplant, and implant of a left ventricle assist device. The difference in predictive ability between the native and sex recalibrated MECKI (S-MECKI) was calculated using a receiver operating characteristic (ROC) curve at 2 years and a calibration plot. We retrospectively analyzed 7,900 HFrEF patients included in the MECKI score registry (mean age 61 ± 13 years, 6,456 men/1,444 women, mean LVEF 33% ± 10%, mean peakVO2 56.2% ± 17.6% of predicted) with a median follow-up of 4.05 years (range 1.72–7.47). Our results revealed an unadjusted risk of events that was doubled in men compared to women (9.7 vs. 4.1) and a significant difference in weight between the sexes of most of the parameters included in the MECKI score. S-MECKI showed improved risk classification and accuracy (area under the ROC curve: 0.7893 vs. 0.7799, p = 0.02) due to prognostication improvement in the high-risk settings in both sexes (MECKI score >10 in men and >5 in women). Conclusions: S-MECKI, i.e., the recalibrated MECKI according to sex-specific differences, constitutes a further step in the prognostic assessment of patients with severe HFrEF

Blockade of MIF-CD74 signalling on macrophages and dendritic cells restores the anti-tumour immune response against metastatic melanoma.

Mounting an effective immune response against cancer requires the activation of innate and adaptive immune cells. Metastatic melanoma is the most aggressive form of skin cancer. While immunotherapies have shown a remarkable success in melanoma treatment, patients develop resistance by mechanisms that include the establishment of an immune suppressive tumor microenvironment. Thus, understanding how metastatic melanoma cells suppress the immune system is vital to develop effective immunotherapies against this disease. In this study, we find that macrophages (MOs) and dendritic cells (DCs) are suppressed in metastatic melanoma and that the Ig-CDR-based peptide C36L1 is able to restore MOs and DCs’ antitumorigenic and immunogenic functions and to inhibit metastatic growth in lungs. Specifically, C36L1 treatment is able to repolarize M2-like immunosuppressive MOs into M1-like antitumorigenic MOs, and increase the number of immunogenic DCs, and activated cytotoxic T cells, while reducing the number of regulatory T cells and monocytic myeloid-derived suppressor cells in metastatic lungs. Mechanistically, we find that C36L1 directly binds to the MIF receptor CD74 which is expressed on MOs and DCs, disturbing CD74 structural dynamics and inhibiting MIF signaling on these cells. Interfering with MIF–CD74 signaling on MOs and DCs leads to a decrease in the expression of immunosuppressive factors from MOs and an increase in the capacity of DCs to activate cytotoxic T cells. Our findings suggest that interfering with MIF–CD74 immunosuppressive signaling in MOs and DCs, using peptide-based immunotherapy can restore the antitumor immune response in metastatic melanoma. Our study provides the rationale for further development of peptide-based therapies to restore the antitumor immune response in metastatic melanoma

With or without ν\nu? Hunting for the seed of the matter-antimatter asymmetry

The matter-antimatter asymmetry underlines the incompleteness of the current understanding of particle physics. Neutrinoless double-beta ($0\nu \beta\beta$) decay may help explain this asymmetry, while unveiling the Majorana nature of the neutrino. The CUORE experiment searches for $0\nu \beta\beta$ decay of $^{130}$Te using a tonne-scale cryogenic calorimeter operated at milli-kelvin temperatures. We report no evidence for $0\nu \beta\beta$ decay and place a lower limit on the half-life of T$_{1/2}$ $>$ 3.8 $\times$ 10$^{25}$ years (90% C.I.) with over 2 tonne$\cdot$year TeO$_2$ exposure. The tools and techniques developed for this result and the 5 year stable operation of nearly 1000 detectors demonstrate the infrastructure for a next-generation experiment capable of searching for $0\nu \beta\beta$ decay across multiple isotopes

Twelve-crystal prototype of Li2_2MoO4_4 scintillating bolometers for CUPID and CROSS experiments

An array of twelve 0.28 kg lithium molybdate (LMO) low-temperature bolometers equipped with 16 bolometric Ge light detectors, aiming at optimization of detector structure for CROSS and CUPID double-beta decay experiments, was constructed and tested in a low-background pulse-tube-based cryostat at the Canfranc underground laboratory in Spain. Performance of the scintillating bolometers was studied depending on the size of phonon NTD-Ge sensors glued to both LMO and Ge absorbers, shape of the Ge light detectors (circular vs. square, from two suppliers), in different light collection conditions (with and without reflector, with aluminum coated LMO crystal surface). The scintillating bolometer array was operated over 8 months in the low-background conditions that allowed to probe a very low, $\mu$Bq/kg, level of the LMO crystals radioactive contamination by $^{228}$Th and $^{226}$Ra.Comment: Prepared for submission to JINST; 23 pages, 9 figures, and 4 table

A first test of CUPID prototypal light detectors with NTD-Ge sensors in a pulse-tube cryostat

CUPID is a next-generation bolometric experiment aiming at searching for neutrinoless double-beta decay with ~250 kg of isotopic mass of $^{100}$Mo. It will operate at $\sim$10 mK in a cryostat currently hosting a similar-scale bolometric array for the CUORE experiment at the Gran Sasso National Laboratory (Italy). CUPID will be based on large-volume scintillating bolometers consisting of $^{100}$Mo-enriched Li$_2$MoO$_4$ crystals, facing thin Ge-wafer-based bolometric light detectors. In the CUPID design, the detector structure is novel and needs to be validated. In particular, the CUORE cryostat presents a high level of mechanical vibrations due to the use of pulse tubes and the effect of vibrations on the detector performance must be investigated. In this paper we report the first test of the CUPID-design bolometric light detectors with NTD-Ge sensors in a dilution refrigerator equipped with a pulse tube in an above-ground lab. Light detectors are characterized in terms of sensitivity, energy resolution, pulse time constants, and noise power spectrum. Despite the challenging noisy environment due to pulse-tube-induced vibrations, we demonstrate that all the four tested light detectors comply with the CUPID goal in terms of intrinsic energy resolution of 100 eV RMS baseline noise. Indeed, we have measured 70--90 eV RMS for the four devices, which show an excellent reproducibility. We have also obtained outstanding energy resolutions at the 356 keV line from a $^{133}$Ba source with one light detector achieving 0.71(5) keV FWHM, which is -- to our knowledge -- the best ever obtained when compared to $\gamma$ detectors of any technology in this energy range.Comment: Prepared for submission to JINST; 16 pages, 7 figures, and 1 tabl

Perspectives on global leadership and the Covid-19 crisis

As the world struggled to come to grips with the Covid-19 pandemic, over twenty scholars, practitioners, and global leaders wrote brief essays for this curated chapter on the role of global leadership in this extreme example of a global crisis. Their thoughts span helpful theoretical breakthroughs to essential, pragmatic adaptations by companies

Half-Life and Precision Shape Measurement of the 2νββ Decay of Te130

We present a new measurement of the 2νββ half-life of ^{130}Te (T_{1/2}^{2ν}) using the first complete model of the CUORE data, based on 1038 kg yr of collected exposure. Thanks to optimized data selection, we achieve a factor of two improvement in precision, obtaining T_{1/2}^{2ν}=(9.32 _{-0.04}^{+0.05}stat _{-0.07}^{+0.07}syst)×10^{20}  yr. The signal-to-background ratio is increased by 70% compared to our previous results, enabling the first application of the improved 2νββ formalism to ^{130}Te. Within this framework, we determine a credibility interval for the effective axial coupling in the nuclear medium as a function of nuclear matrix elements. We also extract values for the higher-order nuclear matrix element ratios: second-to-first and third-to-first. The second-to-first ratio agrees with nuclear model predictions, while the third-to-first ratio deviates from theoretical expectations. These findings provide essential tests of nuclear models and key inputs for future 0νββ searches

CUPID, the Cuore upgrade with particle identification

CUPID, the CUORE Upgrade with Particle Identification, is a next-generation experiment to search for neutrinoless double beta decay (0νββ) and other rare events using enriched Li2100MoO4 scintillating bolometers. It will be hosted by the CUORE cryostat located at the Laboratori Nazionali del Gran Sasso in Italy. The main physics goal of CUPID is to search for 0νββ of 100Mo with a discovery sensitivity covering the full neutrino mass regime in the inverted ordering scenario, as well as the portion of the normal ordering regime with lightest neutrino mass larger than 10 meV. With a conservative background index of 10-4 cts/(keV·kg·yr), 240 kg isotope mass, 5 keV FWHM energy resolution at 3 MeV and 10 live-years of data taking, CUPID will have a 90% C.L. half-life exclusion sensitivity of 1.8·1027 yr, corresponding to an effective Majorana neutrino mass (mββ) sensitivity of 9–15 meV, and a 3σ discovery sensitivity of 1·1027 yr, corresponding to an mββ range of 12–21 meV