APOLLO 11 Project, Consortium in Advanced Lung Cancer Patients Treated With Innovative Therapies: Integration of Real-World Data and Translational Research

Introduction: Despite several therapeutic efforts, lung cancer remains a highly lethal disease. Novel therapeutic approaches encompass immune-checkpoint inhibitors, targeted therapeutics and antibody-drug conjugates, with different results. Several studies have been aimed at identifying biomarkers able to predict benefit from these therapies and create a prediction model of response, despite this there is a lack of information to help clinicians in the choice of therapy for lung cancer patients with advanced disease. This is primarily due to the complexity of lung cancer biology, where a single or few biomarkers are not sufficient to provide enough predictive capability to explain biologic differences; other reasons include the paucity of data collected by single studies performed in heterogeneous unmatched cohorts and the methodology of analysis. In fact, classical statistical methods are unable to analyze and integrate the magnitude of information from multiple biological and clinical sources (eg, genomics, transcriptomics, and radiomics). Methods and objectives: APOLLO11 is an Italian multicentre, observational study involving patients with a diagnosis of advanced lung cancer (NSCLC and SCLC) treated with innovative therapies. Retrospective and prospective collection of multiomic data, such as tissue- (eg, for genomic, transcriptomic analysis) and blood-based biologic material (eg, ctDNA, PBMC), in addition to clinical and radiological data (eg, for radiomic analysis) will be collected. The overall aim of the project is to build a consortium integrating different datasets and a virtual biobank from participating Italian lung cancer centers. To face with the large amount of data provided, AI and ML techniques will be applied will be applied to manage this large dataset in an effort to build an R-Model, integrating retrospective and prospective population-based data. The ultimate goal is to create a tool able to help physicians and patients to make treatment decisions. Conclusion: APOLLO11 aims to propose a breakthrough approach in lung cancer research, replacing the old, monocentric viewpoint towards a multicomprehensive, multiomic, multicenter model. Multicenter cancer datasets incorporating common virtual biobank and new methodologic approaches including artificial intelligence, machine learning up to deep learning is the road to the future in oncology launched by this project

Acute Delta Hepatitis in Italy spanning three decades (1991–2019): Evidence for the effectiveness of the hepatitis B vaccination campaign

Updated incidence data of acute Delta virus hepatitis (HDV) are lacking worldwide. Our aim was to evaluate incidence of and risk factors for acute HDV in Italy after the introduction of the compulsory vaccination against hepatitis B virus (HBV) in 1991. Data were obtained from the National Surveillance System of acute viral hepatitis (SEIEVA). Independent predictors of HDV were assessed by logistic-regression analysis. The incidence of acute HDV per 1-million population declined from 3.2 cases in 1987 to 0.04 in 2019, parallel to that of acute HBV per 100,000 from 10.0 to 0.39 cases during the same period. The median age of cases increased from 27 years in the decade 1991-1999 to 44 years in the decade 2010-2019 (p < .001). Over the same period, the male/female ratio decreased from 3.8 to 2.1, the proportion of coinfections increased from 55% to 75% (p = .003) and that of HBsAg positive acute hepatitis tested for by IgM anti-HDV linearly decreased from 50.1% to 34.1% (p < .001). People born abroad accounted for 24.6% of cases in 2004-2010 and 32.1% in 2011-2019. In the period 2010-2019, risky sexual behaviour (O.R. 4.2; 95%CI: 1.4-12.8) was the sole independent predictor of acute HDV; conversely intravenous drug use was no longer associated (O.R. 1.25; 95%CI: 0.15-10.22) with this. In conclusion, HBV vaccination was an effective measure to control acute HDV. Intravenous drug use is no longer an efficient mode of HDV spread. Testing for IgM-anti HDV is a grey area requiring alert. Acute HDV in foreigners should be monitored in the years to come

Some new details of the copper-hydrogen peroxide interaction

The addition of neocuproine (NC) or bathocuproine-disulphonate at the end of the autooxidation of Cu-I in phosphate buffer, pH 7.4, regenerates almost entirely the O-2 consumed. Other chelating agents assayed, including o-phenanthroline, cannot replace NC in promoting the O-2 formation. O-2 is also produced by adding NC to a mixture of Cu-II and H2O2 Concomitant with the O-2 evolution, the typical absorbance of the (NC)(2)Cu-I complex appears to account for the complete reduction of Cu-II to Cu-I. It is concluded that the addition of H2O2 with Cu-II produces the equilibrium Cu-II(O2H)(-) (CdO2H)-O-I.. Addition of NC shifts the equilibrium to the right side by binding CuI. The released O-2(.-) then reacts with the remaining Cu-II yielding, in the presence of NC, the net reaction of 4 NC + 2 Cu-II + H2O2 --> 2 (NC)(2)Cu-I + O-2 + 2 H+. O-2 is also released in the absence of added NC provided the H2O2 concentration is increased. In these conditions the Cu-II(O2H)(-) complex undergoes other reactions leading to the copper-catalysed decomposition of H2O2. (C) 1997 Academic Press

EXPRESSION OF PL10 PROTEIN IN MALE GERM CELLS OF PODARCIS SICULA (REPTILIA, LACERTIDAE) DURING THE REPRODUCTIVE-CYCLE PHASES

PL10 is a DEAD-box protein that functions as ATP-dependent RNA helicase. DEAD-box proteins are involved in many processes related to RNA metabolism. In particular, some of them regulate the translation of multiple mRNAs allowing specific molecules to direct the process of differentiation in male and female germ cells. VASA, PL10, and P68 DEAD-box proteins are members of three closely related subfamilies: Vasa, in several animals, is exclusively expressed to the germ cell lineage, while PL10 and P68 expression are documented also in somatic tissues. In mouse, three PL10 related genes are identified: DDX3 and DDX3Y located on chromosomes X and Y and expressed in germ and somatic cells and the autosomal retrogene PL10 specifically expressed in testicular tissues at the pachytene stage of male meiosis. In non-mammal animals, PL10 is the sole member of the subfamily and in Xenopus and in Danio rerio PL10 expression is well documented in male and female germ cells but also in most embryonic and adult tissues. At the moment, there is no available data regarding PL10 in reptiles. We isolated Podarcis sicula PL10 homologue gene (Ps-PL10), developed a specific antibody (anti-Ps-PL10) and analyzed, at confocal microscopy, the expression pattern of PL10 during spermatogenesis in all phases of adult reproductive cycle of P. sicula (1- full gonadal activity in the spring, 2- complete regression in the summer, and 3- slow autumnal recrudescence without spermiation) with the aim to identify when PL10 is expressed during the differentiation process of male germ cells. Moreover, to verify if this protein is also expressed in the somatic tissue of the testis, this analysis was extend to young testes when the walls of the seminiferous tubules were forming. The obtained results show that PL10 expression is present from spermatocytes I to spermatids. During full gonadal activity (spring), PL10 expression increases in spermatids in the final steps of spermiogenesis, the strong immunostaining accumulates in the cytoplasm of residual bodies. No stained is observed in spermatogonia and in spermatozoa as well as in the somatic cells of seminiferous epithelium in all phases of the reproductive cycle analyzed. The specific expression of PL10 in meiotic cells suggests that this protein is involved in the differentiation of germ cells, in particular, during the differentation process from spermatid to spermatozoa, given the massive expression observed in the cytoplasm of residual bodies

Aminoethylcysteine ketimine decarboxylated dimer protects submitochondrial particles from lipid peroxidation at concentration not inhibitory of electron transport.

In contrast with other inhibitors of the NADH dehydrogenase of the respiratory chain, the decarboxylated dimer of aminoethylcysteine ketimine protects bovine heart submitochondrial particles (SMP) from the NADH-Fe(+3)-ADP-induced lipid peroxidation. This effect, measured as inhibition of malondialdehyde formation, is concentration-dependent in the range 0.02-0.2 mM. This range of concentration is not inhibitory on NADH-oxidase activity of SMP. Furthermore the dimer is able to counteract the malondialdehyde formation stimulated by the Complex I inhibitors rotenone and N-methyl-4-phenylpyridinium (MPP+)

Novel findings on the copper catalysed oxidation of cysteine

The oxidation of cysteine (RSH) has been studied by using O-2, ferricytochrome c (Cyt c) and nitro blue tetrazolium (NET) as electron accepters. The addition of 200 mu M Cu-II to a solution of 2 mM cysteine, pH 7.4, produces an absorbance with a peak at 260 nm and a shoulder at 300 nm. Generation of a cuprous bis-cysteine complex (RS-CuI-SR) is responsible for this absorbance. In the absence of O-2 the absorbance is stable for long time while in the presence of air it vanishes slowly only when the cysteine excess is consumed. The neocuproine assay and the EPR analysis show that the metal remains reduced in the course of the oxidation of cysteine returning to the oxidised form at the end of reaction when all RSH has been oxidised to RSSR. Addition of Cu-II enhances the reduction rate of Cyt c and of NET by cysteine also under anaerobiosis indicating the occurrence of a direct reduction of the acceptor by the complex. It is concluded that the cuprous bis-cysteine complex (RS-CuI-SR) is the catalytic species involved in the oxidation of cysteine. The novel finding of the stability of the complex together with the metal remaining in the reduced form during the oxidation suggest sulfur as the electron donor in the place of the metal ion

PL10 DEAD-Box Protein is Expressed during Germ Cell Differentiation in the Reptile Podarcis sicula (Family Lacertidae)

Among genes involved in the regulation of germ cell differentiation, those of DDX4/Vasa and the Ded1/DDX3 subfamilies encode for DEAD-box ATP-dependent RNA helicases, proteins involved in many mechanisms related to RNA processing. For the first time in reptiles, using specific antibodies at confocal microscopy, we analysed the localization pattern of a Ded1/DDX3 subfamily member in testis and ovary of Podarcis sicula (Ps-PL10) during the reproductive cycle. In testis, Ps-PL10 is expressed in the cytoplasm of spermatocytes and it is not detected in spermatogonia. Differently from Ps-VASA, in round spermatids, Ps-PL10 is not segregated in the chromatoid body but it accumulates in the cytoplasm of residual bodies, and mature spermatozoa are unstained. These observations suggest that in males, Ps-PL10 (1) is involved in spermatogenesis and (2) is then eliminated with residual bodies. In the ovary, Ps-PL10 is present with granules in the cytoplasm of early meiotic cells of the germinal bed (GB), while it is not present in oogonia and somatic cells of the GB stroma. In follicular cells of ovarian follicles, Ps-PL10 expression starts after their fusion with the oocyte. Numerous Ps-PL10 spots are visible in pyriform (nurse-like) cells concomitantly with the protein accumulation in the cytoplasm of differentiating oocyte. In pyriform cells, Ps-PL10 spots are present in the cytoplasm and nuclei, as observed for Ps-VASA, and in the nucleoli, suggesting for Ps-PL10 a role in rRNA processing and in the transport of molecules from the nucleus to cytoplasm and from nurse cells to the oocyte

Reversible cyclization of S-(2-oxo-2-carboxyethyl)-L-homocysteine to cystathionine ketimine.

S-(2-oxo-2-carboxyethyl)homocysteine (OCEHC), produced by the enzymatic monodeamination of cystathionine, is known to cyclize producing the seven membered ring of cystathionine ketimine (CK) which has been recognized as a cystathionine metabolite in mammals. Studies have been undertaken in order to find the best conditions of cyclization of synthetic OCEHC to CK and for the preparation of solid CK salt product. It has been found that ring closure takes place at alkaline pH and is highly accelerated in 0.5 M phosphate buffer. The sodium salt of CK has been prepared by controlled additions of NaOH to water-ethanol solution of OCEHC under N2 atmosphere. A solid product is obtained which, dissolved in water, shows the spectral features of CK. Solutions of the sodium salt of CK show the presence of a pH depending reversible equilibrium with the open OCEHC form. Plot of the absorbance at 296 nm in function of pH indicates that at pH 9 the compound is completely cyclized while at pH 6 is totally in the open OCEHC form. At intermediate pHs variable ratios between the two forms occur. According to the results obtained by the spectral analysis, HPLC assays of the sodium salt of CK show different patterns depending on the pH of the elution buffer