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© 2023 Published by Elsevier B.V. on behalf of International Federation of Clinical Neurophysiology., Objective: To assess the repeatability and suitability for multicentre studies of MScanFit motor unit number estimation (MUNE), which involves modelling compound muscle action potential (CMAP) scans. Methods: Fifteen groups in 9 countries recorded CMAP scans twice, 1-2 weeks apart in healthy subjects from abductor pollicis brevis (APB), abductor digiti minimi (ADM) and tibialis anterior (TA) muscles. The original MScanFit program (MScanFit-1) was compared with a revised version (MScanFit-2), designed to accommodate different muscles and recording conditions by setting the minimal motor unit size as a function of maximum CMAP. Results: Complete sets of 6 recordings were obtained from 148 subjects. CMAP amplitudes differed significantly between centres for all muscles, and the same was true for MScanFit-1 MUNE. With MScanFit-2, MUNE differed less between centres but remained significantly different for APB. Coefficients of variation between repeats were 18.0% for ADM, 16.8% for APB, and 12.1% for TA. Conclusions: It is recommended for multicentre studies to use MScanFit-2 for analysis. TA provided the least variable MUNE values between subjects and the most repeatable within subjects. Significance: MScanFit was primarily devised to model the discontinuities in CMAP scans in patients and is less suitable for healthy subjects with smooth scans., This study was financially supported by the Lundbeck Foundation, Aage & Johanne Louis-Hansen Foundation, Dagmar Marshall's Foundation, Grosserer L. F. Foghts Foundation and The Jascha Foundation and FONDECYT N° 1201225

Pancreatic cancer is a devastating disease with a five-year survival rate below 10% according to the American Cancer Society. Novel chemotherapeutics and targeted therapies in pancreatic cancer have shown limited success, illustrating the urgent need for new treatments. Lurbinectedin is a chemotherapeutic synthetic tetrahydroisoquinoline alkaloid that inhibits active transcription by binding to guanine rich sequences in the minor groove of DNA. Lurbinectedin has been shown to reduce oncogenic transcription by stalling and degrading RNA Polymerase II while also inducing single- and double-stranded breaks to DNA, causing subsequent apoptosis. Lurbinectedin received accelerated FDA approval in 2020 for metastatic small cell lung cancer on or after platinum-based chemotherapy and is currently undergoing clinical trials in a variety of tumor types. We recently described a synergistic interaction between lurbinectedin and ONC201/TIC10, a compound that induces the TRAIL pathway, in killing SCLC cell lines associated with activation of p-Chk1 and the integrated stress response. We now demonstrate lurbinectedin’s efficient killing of pancreatic tumor cells as a single agent in PANC-1, BxPC-3, and HPAF-II cell lines, with IC-50s corresponding to sub-nanomolar concentrations. We also demonstrate that a combination of lurbinectedin and irinotecan, a topoisomerase I inhibitor with FDA approval for advanced pancreatic cancer, results in synergistic killing of pancreatic tumor cells in vitro. We further demonstrate a combination of lurbinectedin and ONC212, an imipridone, the same class of compounds as ONC201, also results in synergistic killing of pancreatic tumor cells. Cell viability was measured using the CellTiterGlo assay at varying drug concentrations. We hypothesize a combination therapy of lurbinectedin and irinotecan or ONC212 can enhance immune cell killing of pancreatic tumor cells. This is being explored by co-culturing CD8+ T lymphoblast cells with pancreatic tumor cells treated with lurbinectedin, irinotecan, ONC212, and combination, along with molecular mechanisms of cell death and effects on cytokines in the tumor microenvironment. Our results are developing insights regarding molecular mechanisms underlying therapeutic efficacy of a novel combination drug treatment for pancreatic cancer. Citation Format: Tej Tummala, Arielle De La Cruz, Ash Uruchurtu, Nicholas R. Liguori, Abbas E. Abbas, Leiqing Zhang, Christopher G. Azzoli, Lanlan Zhou, Wafik S. El-Deiry. Synergistic combinations of lurbinectedin with irinotecan and ONC212 in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2674.

The American Cancer Society estimates that ~15% of all lung cancers are categorized as small cell lung cancer (SCLC) with an overall five-year survival rate of less than 7%. Due to disease aggressiveness, more other malignancies, the standard of care is based on clinical efficacy rather than helpful biomarkers. Lurbinectedin is a small molecule RNA polymerase II inhibitor that binds the minor groove of DNA to induce double-strand breaks. Lurbinectedin has efficacy towards SCLC cells at sub-nM concentration and received accelerated FDA approval in 2020 for metastatic SCLC that progressed on platinum-based therapy. ONC201/TIC10 is a TRAIL pathway-inducing compound that with demonstrated clinical efficacy in H3K27M-mutated diffuse midline glioma and neuroendocrine tumors, in early phase clinical trials. We hypothesized that combining ONC201 and lurbinectedin may yield synergistic and targeted killing of SCLC cells. SCLC cell lines H1048, H1105, H1882, and H1417 were treated with ONC201 and lurbinectedin and cell viability was determined using a CellTiter-Glo assay using varying drug concentrations. Synergistic growth inhibition of SCLC cells was noted with combination of ONC201 and lurbinectedin. Induction of the integrated stress response mediator ATF4 and CHOP was observed with ONC201 and lurbinectedin along with induction of PARP cleavage indicative of apoptosis in response to cellular stress. Additionally, SCLC lines treated with the combination therapy displayed increased DNA breakage-related proteins such as phosphorylated Chk-1, Wee1 and γ-H2AX. Combination index revealed the most potent synergy occurred at the concentrations of 0.16 μM ONC201 and 0.05 nM lurbinectedin in the H1048 cell line, demonstrating highly efficient and selective killing of these tumor cells in vitro. While these therapies showed potency against the cell lines derived from SCLC patients, it is noteworthy that the combination showed significantly less toxicity to healthy human lung epithelial cells. Future studies could explore the combination of ONC201 and lurbinectedin in SCLC cell lines, SCLC patient-derived organoids, other tumor types, including in vivo studies and clinical translation.

Small cell lung cancer (SCLC) accounts for 15% of lung cancer and is responsive to chemotherapy but most patients relapse with drug-resistant disease and have very poor 5-year survival of less than 7%. One of the most notable advances in the last decade has been the approval of lurbinectedin for platinum resistant SCLC. Lurbinectedin is a small molecule inhibitor, targeting RNA polymerase II, and binding the minor grooves of DNA to induce double-strand breaks. The dismal SCLC survival rate underscores the need for a novel combination to increase patient survival outcomes. ONC201 is a pro-apoptotic TRAIL-inducing compound that activates the integrated stress response and appears to have efficacy in neuroendocrine tumors including paragangliomas. In our preclinical experiments, we explored the combination of ONC201 and lurbinectedin as a potentially effective treatment regimen for SCLC. Lurbinectedin has been shown to have effective cancer cell killing ability in multiple SCLC cell lines (H1048, H1105, H1882, H1417) at sub-nanomolar concentrations, while sparing healthy lung epithelial tissue cells (cell line HSAEC). We hypothesized that combining ONC201 and lurbinectedin will yield synergistic outcomes due to combining drugs that act by different potentially complementary mechanisms. We treated SCLC cell lines with ONC201 and lurbinectedin at increasing concentrations of the drugs. CellTiter Glo was used to perform cell viability assays, and combination index analysis (Combenefit software) revealed the most synergy between the agents occurred at the concentrations of 0.156 uM ONC201 and 0.047 nM lurbinectedin. We are expanding this combination’s efficacy in other SCLC lines and tumor types. Western blotting further examined these synergies and showed that PARP cleavage and expression of ATF4 and TRAIL death receptor DR5 were increased at higher concentrations of these agents. We further noted that combinatorial treatment with these agents induced marked upregulation of Phosphorylated Chk1 and CHOP at the highest treatment concentrations versus lower concentrations and control. These effects indicate effectiveness of the combinatorial treatment in causing DNA damage and instability, inducing double-stranded DNA breaks, as well as initiating the intrinsic apoptosis pathway through the integrated stress response, selectively in the malignant cells. Ongoing directions include testing similar concentrations of both ONC201 and lurbinectedin in additional SCLC cell lines, SCLC patient-derived organoids, and in vivo, as well as exploring immune correlates of the drug treatments. Keywords: SCLC; lurbinectedin; chemotherapy; imipridones; genomics Citation Format: Nicholas R. Liguori, Ashley Fss Uruchurtu, Young Lee, Leiqing Zhang, Abbas Abbas, Varun Prabhu, Lanlan Zhou, Christopher J. Azzoli, Wafik S. El-Deiry. Synergistic activity of Lurbinectidin plus ONC201 in SCLC is associated with ATF4, CHOP and pChk1 induction [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 4064.

Additional file 1: Figure S1. Computerized visual fields: generalized defect in RE (A) and pseudo-altitudinal defect in LE (B) (PPTX 226 kb)

Our objective was to describe incidence and clinical features of ALS from a prospective population-based study in Emilia Romagna Region (ERR). From 2009 onwards, a prospective registry recorded all incident cases of ALS among residents in the ERR (population, 4.4 million inhabitants), involving 17 neurological departments. For each patient, detailed demographic and clinical information was collected by caring physicians. Results showed that from 1 January 2009 to 31 December 2011, 347 patients received a new diagnosis of ALS with a crude incidence rate of 2.63/100,000/year. There was micro-geographic heterogeneity throughout ERR, with higher incidence rates in the low density population (3.27/100,000) (p < 0.01). ALS patients have been more frequently employed in agriculture than the general ERR population (8.64\% vs. 4.6\%, p < 0.01). Clinical features were similar to those described in previous population based studies. In conclusion, we report incidence rates similar to those reported by European registries, reflecting good accuracy of our prospective study. We confirmed previous studies reporting higher incidence rates in rural areas and among agricultural workers. Although genetics has been gaining increasing importance in ALS aetiology, some epidemiological data are still unexplained. Identifying geographical areas or populations with high incidence rates can be a starting point for identifying environmental risk factors. Further studies having this specific aim can shed light on these topics.

Intraneural misfolded α-synuclein (syn) characterized different synucleinopathies such as pure autonomic failure (PAF), idiopathic Parkinson disease (IPD) and dementia with Lewy Bodies (DLB). The aim of this study is to characterize by immunofluorescence the skin intraneural α-synuclein (syn) deposits in PAF, IPD and DLB to ascertain conformation-specific differences which may justify a different clinical phenotype. We identified a total of 21 skin intraneural abnormal syn deposits in PAF (3 patients), 22 in IPD (8 patients) and 40 in DLB (7 patients). Ten healthy subjects were used controls. Skin biopsy was performed on proximal (C7 paravertebral) and distal (thigh and leg) sites. To characterize abnormal syn deposits we used primary antibodies against native α-synuclein (NAC) and against C-terminal α-synuclein epitopes involved in post-translational modifications such as phosphorylation at serine 129 (pS129) and tyrosine 125 (pY125), nitrate α-synuclein at tyr125-133 (nY125-133) and advanced glycation end products (AGEs). Furthermore, the mature amyloid α-synuclein fibrils were characterized by using a non-commercial antibody (Syn-F) and ubiquitin deposits were identified by a specific antibody (UBI). Antibody raised against pS129 disclosed abnormal skin nerves syn deposits in all patients and never in the control group. Abnormal pS129 syn deposits were often (80–90% of all analysed deposits) stained by the antibody recognizing a fibril conformation, seldom (around 40%) by native antibodies and very occasionally by antibodies against nY125-133. Furthermore, abnormal syn deposits were not stained by antibodies against pY125, AGEs and UBI. The immunofluorescence characterization of abnormal syn deposits showed similar findings in IPD, PAF and DLB although in these latter abnormal deposits were quantitatively higher than in IPD. Phosphorylation at serine 129 was the most sensitive and specific epitope to identify skin nerves abnormal syn deposits for the in vivo diagnosis of synucleinopathies; (2) skin syn neuritis showed no relevant immunofluorescence differences in PAF, IPD and DLB suggesting a similar conformation among different clinical phenotypes.

Fludioxonil (FDL) is a 'reduced risk' fungicide recently registered for post-harvest treatments of different horticultural crops in the USA. FDL is a broad spectrum fungicide with a mechanism of action different from other fungicides currently in use for pre- or postharvest treatments of selected fruit crops, including citrus fruits in the US. The effectiveness of postharvest 30-s-dips in aqueous mixtures of (FDL) and 20°C or imazalil (IMZ) containing 300 or 600 mg L -1 (active ingredient, a.i.) at 20°C was compared for controlling green mold caused by Penicillium digitatum. In fruit treated 24h after artificial wounding or wound-inoculating with P. digitatum (10 6 conidia ml -1 ) the effectiveness of FDL was similar or slightly inferior to IMZ, respectively, following conditions of low or high disease pressure. In experiments aimed at assessing the protective activity of FDL with fruit subjected to the fungicidal treatments 24h before inoculation with P. digitatum, the effectiveness of FDL was found to be dependent on the mode of fungal inoculation. Activity of FDL was superior in fruit wounded and inoculated by spray application compared to fruit inoculated by dipping a steel rod into the conidia suspension before puncturing. The lower performance of FDL in fruit inoculated by puncturing was ascribed to the lack of systemic properties of the fungicide. However, during storage it is unlikely that fruit become wounded: re-infections may take place mainly through old wounds caused before or at the time of fungicide treatments. In normal situations FDL can be highly effective as a curative as well as a protective agent against Penicillium decay and can successfully be used in place or in combination/rotation of/with IMZ or thiabendazole to overcome the risk of selection of resistant strains of pathogens.

We present a method to determine the magnitude of the uncorrelated background distribution obtained with the event mixing technique, through the simultaneous observation of the projectile elastic scattering in different detectors, which correspond to random coincidences. The procedure is tested with α – d angular correlation data from the Li 6 + Co 59 reaction at E lab = 29.6 MeV . We also show that the method can be applied using the product of singles events, when singles measurements are available.

The 10B(p,α)7Be reaction is the main responsible for 10B destruction in stellar interior. In such environments this p-capture process occurs at a Gamow energy of ∼10 keV, and takes places mainly through a resonant state ( E x = 8.701 MeV ) of the compound 11C nucleus. Thus, a resonance right in the region of the Gamow peak is expected to significantly influence the behaviour of the astrophysical S(E)-factor. The 10B(p,α)7Be reaction has been investigated by means of the Trojan Horse Method (THM) applied to the 2H(10B,α7Be)n three-body process. The experiment was performed at E b e a m = 27 MeV at the Instituto de Fisica Nuclear de Sao Paulo. Preliminary results concerning the extraction of the bare-nucleus S(E)-factor will be presented.

Nuclear structure effects observed in the sub-Coulomb fusion of heavy-ions demand a careful investigation of the reaction cross section and optical potentials near the Coulomb barrier. The elastic scattering for the 16 O + 46,50 Ti systems was investigated in the bombarding energy range 30 ⩽ E lab ⩽ 70 MeV through an analysis using the Sao Paulo Potential. Experimental results suggest that there is no significant effect due to the differences in structure of the deformed 46 Ti and the neutron magic 50 Ti targets. This result is in agreement with the observation of no significant structure effects in the fusion cross section study for the same systems.

It's well known that iv. immunoglobulins may be useful to overcome habitual abortions, but the mechanisms at the base of a successful outcome and the likelihoods are still unknown.In one hundred and sixty women with habitual abortions and one hundred and sixty healthy mothers, we evaluated blood IgG subclasses; among the patients, sixteen merely showed IgG subclass deficiency, after leaving out any autoimmunity and/or coagulation disorders. All the patients (100%) showed IgG3, twelve (75%) IgG1, eight (50%) IgG4 and six (37,5%) IgG2 deficiency; healthy control people's IgG subclasses fell in normal range in 156 women, but just four women showed IgG2 and IgG4 deficiency with neither immune deficiency's clinical marks nor increased vulnerability to infections. All the patients were treated with whole immunoglobulins iv. infusion (200 mg/kg/monthly) all over the pregnancy.The successful pregnancy rate is very high (90%): 100% out of women showing IgG1 (12/12), 87,5% of IgG3 (14/16), 75% of IgG4 (6/8) and 66% of IgG2 deficiency (4/6) had successful pregnancies. The Odd's Ratio between IgG subclass deficiency and recurrent abortions is 4,33 with confidence interval of 95%; chi square value is 7.68 (p0.025).Low dose immunoglobulin infusion is the only effective way to reach successful pregnancy, despite previous habitual abortions in patients suffering from IgG subclass deficiency without autoimmunity and/or coagulation disorders, likely restoring idiotype-antiidiotype network; showing evidence of IgG subclasses deficiency (mostly IgG1 and IgG3) may help identify patients who can benefit from iv. immunoglobulin treatment.

We present new measurements and a general discussion of the behavior of the fusion, break-up and reaction cross sections of different projectiles on the same target 64 Zn, at near and above barrier energies. The projectiles are the tightly bound 16 O, the stable weakly bound 6 Li, 7 Li and 9 Be and the radioactive very weakly bound 6 He nuclei. We also compare the results with the ones for heavier targets.

We report a case of temporomandibular disorder patient with disc displacement without reduction, myofascial pain, limited opening and a novel, never described, nonsynonimous mutation of catechol-O-methyl-transferase (COMT) gene. COMT is one of the enzymes that metabolizes catecholamines, thereby acting as a key modulator of dopaminergic and adrenergic/noradrenergic neurotransmissions, which play a key role in pain modulation. This novel mutation, p.R58S, changed a codon (58 from arginine to serine) in the COMT protein. The introduction of a serine residue in a highly organised secondary structure, in critical regions of the protein, results in a structural alteration. Therefore, we speculate an influence of the mutation on the high pain sensitivity of the patient.

An external PIGE–PIXE setup was installed on a beam line of the 8 MV tandem Pelletron accelerator of the Open Nuclear Physics Laboratory (LAFN). Proton beam energy was chosen in the 8–12 MeV range, sufficient to get an acceptable gamma ray yield but not so high as to prevent us from measuring X-rays. This also allowed the use of a thick aluminum exit window (0.5 mm) instead of the usual thin and sometimes fragile plastic windows. This external PIXE–PIGE system was used to analyze trace element concentrations in the enamel of human and animal teeth. The main interest was to find compatible human teeth substitutes for dentistry laboratory practice and chemical tests. In spite of their morpho-histological similarity, trace element concentrations in human and animal teeth have not yet been compared. Teeth from humans, cattle and swine collected primary at Sao Paulo region were analyzed. The elements Cu, K, Zn, Fe, Ti, Sr, V, Mn and Zr were detected by high energy external beam PIXE technique. Though preliminary, the results showed that the trace element concentrations observed in the enamel of human and swine are more similar to each other than to cattle teeth.

An elastic recoil detection analysis (ERDA) system using ΔE−E technique was assembled for stoichiometric and depth profile studies of materials formed by light elements. An ionization chamber with a surface barrier detector (SBD) was used in this work. From the energy loss (ΔE) information obtained from the gas and the residual energy (E) obtained by the SBD, it was possible to identify atomic number of the arriving particles at the SBD. An incident beam of 35 Cl of 58 MeV was used for the elastic scattering with the sample components. From the spectra obtained for carbon nitride thin films it was possible to clearly identify the elements carbon and nitrogen. The presence of oxygen in the spectra was probably due to the sample exposure to air. The chemical composition of the films was calculated and compared with that obtained from X-ray photoelectron spectroscopy (XPS) spectra for C 1s and N 1s. A LiNiO2 thin film was also analyzed.

The status and the near future plans for the Brazilian AMS program are described. The 8 MV Tandem accelerator at the University of Sao Paulo (USP) is ready to measure standard AMS samples. A recently installed 1.7 MV Tandem at the University of Rio de Janeiro will have a 14C AMS line. Together with external laboratories, we developed some projects on paleoclimatic and maritime geology. During these studies we have also learned sample preparation procedures.

A new method was developed in order to obtain continuous energy loss curves, combining transmission and backscattering techniques. The method was tested using 16 O projectiles scattered in thick Ta and Nb targets going through a mylar foil. Energy loss curves were obtained for a range from 4 to 25 MeV within 1% precision. Theoretical simulations were performed in order to study the influence of some physical effects in the method.

Angular distributions of the elastic scattering of the ${}^{6,7}\mathrm{Li}$ on ${}^{138}\mathrm{Ba}$ were measured, in the energy range $21l~{E}_{\mathrm{lab}}l~32$ MeV. The analysis of the data was performed via the optical model, using the Woods-Saxon potential, in order to find the energy dependence of the real and imaginary parts of the nuclear potential. The existence of a threshold anomaly for the ${}^{7}\mathrm{Li}$ scattering and its absence for the ${}^{6}\mathrm{Li}$ one was observed. It is interpreted that the breakup of ${}^{6}\mathrm{Li}$, with dissociation energy smaller that its first excited state, is responsible for the vanishing of the threshold anomaly.

We have identified and characterized two transcriptional modules derived from the promoters of liver genes whose expression is specific for proliferating hepatocytes: A) An enhancer module from the HNF1β promoter. HNF1α and β are two trans-acting factors that play a central role in the expression of liver-specific genes. We reported that, during mouse and rat development, HNF1β expression precedes that of HNF1α and is inducible by retinoic acid treatment. Moreover HNF1β (but not HNF1α) is expressed in undifferentiated hepatocytes. A combined strategy of deletions and site-specific mutagenesis allowed us to identify the cis-acting elements of the HNF1β promoter. In our transient expression assay, the -435/+33 region of the promoter is sufficient to reproduce the expression pattern of the endogenous gene. This region can be further subdivided into a tissue-specific enhancer and a proximal promoter element. The transcriptional activity of the enhancer element, as measured by co-transfection assays and band-shift experiments, depends on the binding of two transcriptional activators: the liver-specific HNF-4 factor (-255/-235), and the RARα/RXRα heterodimeric protein, that recognize a RARE element partially overlapping with the HNF-4 binding site. B) The proximal element of the alpha-foetoprotein promoter. The alpha-fetoprotein (AFP) gene is expressed in the prenatal life in the yolk sac and in the foetal liver. At birth, the AFP gene is "shut down" at the transcriptional level, and it can be re-expressed in the adult liver only in pathologic conditions, like hepatic parenchyma regeneration or liver tumours. Therefore, the AFP gene must contain cis acting regulatory elements able to activate transcription only in proliferating hepatocytes. We have isolated and individually analysed the key elements of the AFP transcriptional regulation located in the proximal promoter region, where two partially overlapping AP1 and GRE elements dictate the activation or repression of the gene, respectively. Our results suggest that, while two adjacent HNF1α/β binding sites restrict the AFP promoter activity to liver cell, changes in the GR/AP1 ratio can account for the AFP reactivation in the regenerating hepatocyte. We have constructed a series of artificial promoters by assembling different combinations of these transcriptional modules. The most efficient combinations have been used to construct SV40-derived episomal vectors, that we plan to use for gene therapy of proliferating hepatocytes. These experiments are currently in progress.

Elastic scattering angular distributions for the symmetric ${}^{27}\mathrm{Al}{+}^{27}\mathrm{Al}$ system were measured at bombarding energies from 50 to 70 MeV. The integrated inelastic cross section for the first two excited states was determined by the in-beam $\ensuremath{\gamma}$-ray spectroscopy method. The data were analyzed by the optical model and by coupled channel calculations. The spin-orbit couplings of the ground states of both nuclei are analyzed.