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Succinic Semialdehyde Dehydrogenase Deficiency (SSADHD) is an ultra-rare autosomal recessive neurometabolic disorder caused by ALDH5A1 mutations presenting with autism and epilepsy. Here, we report the generation and characterization of human induced pluripotent stem cells (hiPSCs) derived from fibroblasts of three unrelated SSADHD patients – one female and two males with the CRISPR-corrected isogenic controls. These individuals are clinically diagnosed and are being followed in a longitudinal clinical study.

Succinic semialdehyde dehydrogenase deficiency (SSADHD) is a neurometabolic disorder caused by ALDH5A1 mutations presenting with autism and epilepsy. SSADHD leads to impaired GABA metabolism and results in accumulation of GABA and γ-hydroxybutyrate (GHB), which alter neurotransmission and are thought to lead to neurobehavioral symptoms. However, why increased inhibitory neurotransmitters lead to seizures remains unclear. We used induced pluripotent stem cells from SSADHD patients (one female and two male) and differentiated them into GABAergic and glutamatergic neurons. SSADHD iGABA neurons show altered GABA metabolism and concomitant changes in expression of genes associated with inhibitory neurotransmission. In contrast, glutamatergic neurons display increased spontaneous activity and upregulation of mitochondrial genes. CRISPR correction of the pathogenic variants or SSADHD mRNA expression rescue various metabolic and functional abnormalities in human neurons. Our findings uncover a previously unknown role for SSADHD in excitatory human neurons and provide unique insights into the cellular and molecular basis of SSADHD and potential therapeutic interventions.

Advances in the clinical management of pediatric B-cell acute lymphoblastic leukemia (B-ALL) have dramatically improved outcomes for this disease. However, relapsed and high-risk disease still contribute to significant numbers of treatment failures. Development of new, broad range therapies is urgently needed for these cases. We previously reported the susceptibility of ETV6-RUNX1+ pediatric B-ALL to inhibition of signal transducer and activator of transcription 3 (STAT3) activity. In the present study, we demonstrate that pharmacological or genetic inhibition of STAT3 results in p53 induction and that CRISPR-mediated TP53 knockout substantially reverses susceptibility to STAT3 inhibition. Furthermore, we demonstrate that sensitivity to STAT3 inhibition in patient-derived xenograft (PDX) B-ALL samples is not restricted to any particular disease subtype, but rather depends on TP53 status, the only resistant samples being TP53 mutant. Induction of p53 following STAT3 inhibition is not directly dependent on MDM2 but correlates with degradation of MDM4. As such, STAT3 inhibition exhibits synergistic in vitro and in vivo anti-leukemia activity when combined with MDM2 inhibition. Taken together with the relatively low frequency of TP53 mutations in this disease, these data support the future development of combined STAT3/ MDM2 inhibition in the therapy of refractory and relapsed pediatric B-ALL.

IntroductionClassical Hodgkin Lymphoma (HL) is a lymphoproliferative disease typically diagnosed in the young. The excellent results obtained with current treatment lead to long survival with age-related complications affecting patients’ survival and quality of life. One issue affecting HL patients is infertility. This problem can be easily overcome in males with seminal liquid cryopreservation, however, in females it is more complex either in terms of the quality of the cryopreserved material or the patients’ age at diagnosis. Moreover, not all chemo- or radio-therapies have the same negative impact on fertility.The main objectives of this study was to collect epidemiological information on HL patients involved in fertility preservation counseling and to analyze the impact of ABVD (adriamycin, bleomycin, vinblastine, and dacarbazine), the standard treatment for HL, on ovarian function, hormonal levels and ovarian and uterine tissue morphologies. Patterns of fertility preservation were also reported.MethodsData were obtained from 270 female patients at HL onset who were interested in fertility counseling prior to therapy initiation. Each patient was assessed at HL diagnosis for levels of Anti-Mullerian Hormone (AMH), Follicle Stimulating Hormone (FSH), and 17β-oestradiol (17β-oe), with additional assessments at 6 and 12 months after chemotherapy. Patients were evaluated with ultrasound scans to study the number of ovarian follicles and the degree of uterine thickness at the same timepoints.ResultsThe average patient AMH level showed a statistically significant reduction at 6 months after chemotherapy (p=0.05) and by the 12 month time point returned to near pre-chemotherapy values. FSH and 17β-oe levels did not significantly vary throughout the study period. ABVD chemotherapy was associated with a significant reduction of both ovarian follicles and endometrial thickness at the 6 month time point followed by a recovery at the 12 time point in both ovaries. Different results were observed when patients changed treatment to a more intensive one.DiscussionBased on the results from the hormonal measurements and the follicle echography, it appears that the toxic effect of ABVD on fertility is transient, whereas, in contrast, more intensive therapies may potentially be more harmful and long-lasting.

BACKGROUND:Platinum-based drugs such as Cisplatin are commonly employed for cancer treatment. Despite an initial therapeutic response, Cisplatin treatment often results in the development of chemoresistance. To identify novel approaches to overcome Cisplatin resistance, we tested Cisplatin in combination with K+ channel modulators on colorectal cancer (CRC) cells. METHODS:The functional expression of Ca2+-activated (KCa3.1, also known as KCNN4) and voltage-dependent (Kv11.1, also known as KCNH2 or hERG1) K+ channels was determined in two CRC cell lines (HCT-116 and HCT-8) by molecular and electrophysiological techniques. Cisplatin and several K+ channel modulators were tested in vitro for their action on K+ currents, cell vitality, apoptosis, cell cycle, proliferation, intracellular signalling and Platinum uptake. These effects were also analysed in a mouse model mimicking Cisplatin resistance. RESULTS:Cisplatin-resistant CRC cells expressed higher levels of KCa3.1 and Kv11.1 channels, compared with Cisplatin-sensitive CRC cells. In resistant cells, KCa3.1 activators (SKA-31) and Kv11.1 inhibitors (E4031) had a synergistic action with Cisplatin in triggering apoptosis and inhibiting proliferation. The effect was maximal when KCa3.1 activation and Kv11.1 inhibition were combined. In fact, similar results were produced by Riluzole, which is able to both activate KCa3.1 and inhibit Kv11.1. Cisplatin uptake into resistant cells depended on KCa3.1 channel activity, as it was potentiated by KCa3.1 activators. Kv11.1 blockade led to increased KCa3.1 expression and thereby stimulated Cisplatin uptake. Finally, the combined administration of a KCa3.1 activator and a Kv11.1 inhibitor also overcame Cisplatin resistance in vivo. CONCLUSIONS:As Riluzole, an activator of KCa3.1 and inhibitor of Kv11.1 channels, is in clinical use, our results suggest that this compound may be useful in the clinic to improve Cisplatin efficacy and overcome Cisplatin resistance in CRC.

Abstract Background Improving the poor prognosis of infant leukaemias remains an unmet clinical need. This disease is a prototypical fusion oncoprotein‐driven paediatric cancer, with MLL (KMT2A)‐fusions present in most cases. Direct targeting of these driving oncoproteins represents a unique therapeutic opportunity. This rationale led us to initiate a drug screening with the aim of discovering drugs that can block MLL‐fusion oncoproteins. Methods A screen for inhibition of MLL‐fusion proteins was developed that overcomes the traditional limitations of targeting transcription factors. This luciferase reporter‐based screen, together with a secondary western blot screen, was used to prioritize compounds. We characterized the lead compound, disulfiram (DSF), based on its efficient ablation of MLL‐fusion proteins. The consequences of drug‐induced MLL‐fusion inhibition were confirmed by cell proliferation, colony formation, apoptosis assays, RT‐qPCR, in vivo assays, RNA‐seq and ChIP‐qPCR and ChIP‐seq analysis. All statistical tests were two‐sided. Results Drug‐induced inhibition of MLL‐fusion proteins by DSF resulted in a specific block of colony formation in MLL‐rearranged cells in vitro, induced differentiation and impeded leukaemia progression in vivo. Mechanistically, DSF abrogates MLL‐fusion protein binding to DNA, resulting in epigenetic changes and down‐regulation of leukaemic programmes setup by the MLL‐fusion protein. Conclusion DSF can directly inhibit MLL‐fusion proteins and demonstrate antitumour activity both in vitro and in vivo, providing, to our knowledge, the first evidence for a therapy that directly targets the initiating oncogenic MLL‐fusion protein.

Laparoscopic liver resections have gained widespread popularity among hepatobiliary surgeons and is nowadays performed for both standard and more complex hepatectomies. Given the increased technical challenges, preoperative planning and intraoperative guidance is pivotal in laparoscopic surgery to safely carry out complex and oncologically safe hepatectomies. Modern tools can help both preoperatively and intraoperatively and allow surgeons to perform more precise hepatectomies. Preoperative 3D reconstructions and printing as well as augmented reality can increase the knowledge of the specific anatomy of the case and therefore plan the surgery accordingly and tailor the procedure on the patient. Furthermore, the indocyanine green retention dye is an increasingly used tool that can nowadays improve the precision during laparoscopic hepatectomies, especially when considering anatomical resection. The use of preoperative modern imaging and intraoperative indocyanine green dye are key to successfully perform complex hepatectomies such as laparoscopic parenchymal sparing liver resections. In this narrative review, we discuss the aspects of preoperative and intraoperative tools that are nowadays increasingly used in experienced hepatobiliary centers.

We introduce a novel all-optical assay for functional studies of biological neural networks in vitro. We created a novel optogenetic construct named OptoCaMP which is a combination of a channelrhodopsin variant (CheRiff) and a red genetically encoded calcium indicator (GECI) (jRCaMP1b). It enables simultaneous optical stimulation and recording from large population of neurons with single-cell readout. Additionally, we have developed a spatio-temporal all-optical assay to simultaneously stimulate a sub-section of a neural network and record evoked calcium activity, in both stimulated and non-stimulated neurons, thus allowing the investigation of the spread of excitation through an interconnected network. Finally, we demonstrate the sensitivity of this assay to the change of neural network connectivity.

Fedeli, Bortolotto, Sannazaro, Ghedini, Slavazzi, Cecchi, Gasparoli

È piuttosto evidente che le strutture archeologiche sono soggette a significative condizioni di rischio per ragioni connesse alla loro vulnerabilità e alla pericolosità ambientale, essendo in genere costituite da strutture ruderizzate, spesso in avanzato stato di degrado e direttamente esposte agli agenti atmosferici. Per tenere adeguatamente sotto controllo i fattori di rischio, al fine di evitare l’ineluttabile perdita dei manufatti archeologici, o quanto meno per allungare il loro ciclo di vita nelle migliori condizioni di conservazione, è necessario un costante monitoraggio e interventi di prevenzione, protezione e “cura” assidui e proseguiti nel tempo. Negli ultimi anni, sulla base di una consistente fase sperimentale sviluppata su casi di eccellenza, sono stati messi a punto strumenti operativi efficaci che hanno consentono di definire procedure dirette alla attivazione di processi di attività ispettiva (controlli) e di manutenzione preventiva e programmata. Le esperienze maturate, inoltre, hanno messo in evidenza come la conservazione dei beni culturali, anche dei beni archeologici, richiede attenzioni e interventi per i quali, spesso, non necessariamente si deve fare ricorso a raffinate competenze di carattere archeologico, ma più semplicemente al buonsenso e alle conoscenze tecniche derivanti dalle regole dell’arte e dalle buone pratiche del costruire, già ampiamente note e sperimentate. Senza sottovalutare, dunque, l’importanza delle competenze specialistiche, come condizione essenziale per consentire il riconoscimento di valore, e da qui derivare le opportune cautele nell’operare, si deve però considerare che le condizioni di degrado, il più delle volte causate da carenti o errate manutenzioni, non sono in genere dovute a limiti nelle conoscenze scientifiche o tecniche, ma a difetti e omissioni di tipo operativo, previsionale, organizzativo e pianificatorio.

The workers’ village of Crespi d’Adda, registered in the UNESCO World Heritage List in 1995, consists of a large industrial complex of approximately 90,000 square metres, abandoned in 2004 and made up of around a hundred residential buildings which were once inhabited by local workers. The management of the transformation of a “living” urban system of unquestionable historical and documental value, like Crespi d’Adda, is a challenging issue, highly uncertain and controversial but, at the same time, absolutely urgent and necessary. The complexity of the site and the delicate balance between its constitutive elements means we have to consider the protection of the values and the tension towards change not as parts of irreconcilable conflict, but as different needs which must be integrated within a coherent management tool. One of the main points of this strategy, outlined in the Management Plan, is the experimental and normative research on the redevelopment of residential buildings.

The research aims to identify "physical" and "perceptual" wear factors of the historic center of Venice (with particular attention to the effects of anthropogenic pressure related to the phenomenon of tourism) and to assess the damages they could produce to identify criteria and tools of mitigation and control. Research is part of the Action Plan "Protection and conservation of the heritage", established by the Plan of Management for the UNESCO Site of Venice and its Lagoon. Through a systemic reading and analysis of the forces of change in place, this study identifies the " macro-emergencies", i.e. the main factors that adversely affect the site’s safeguarding. The research project "Evaluation of wear processes and critical factors of the City of Venice and its lagoon, and its impact on the site’s protection" is part of the actions set out by the Management Plan of the UNESCO site of Venice and its Lagoon. The main objective of the project is the identification of physical and perceptual factors of wear, which threaten the conservation of the historical and artistic heritage of the historic center of Venice, with a particular focus on the effects of anthropogenic pressure linked to tourism, and the evaluation of their level of danger. A further objective is the recognition of measurable parameters (indicators) for monitoring and, subsequently, mitigation strategies for the most significant phenomena.

The formation of new compounds The study of model systems allows us to reconstruct the complex mosaic of reactions which make up the frying process. In particular such systems, if made up of single oils or their components, verify the transformation of the triglyceride matrix by thermo-oxidizing treatment. Should this then have added specific substrates (protein, starch) they reconstruct the oil-substrate interaction and the influence of the same substrate in the transformation. The contribution of the information is equally distributed between the volatile products and the complex products; in fact both contribute to indicate the reactions in play, supporting the previous considerations for investigation at an analytical level. The present work discusses the newly formed products (volatile or nonvolatile), verifying in particular the formation of reaction products with the substrates.

This research develops various reflections on the role and repercussions of planned preventive maintenance activities in historic heritage sites as an opportunity for their valorisation. The first step is to define the terminology, underscoring reciprocal relationships and the need to organise structured activities and applications that outline contents, procedures and processes for interventions, with innovative strategies that promote the preservation and valorisation of cultural heritage sites. In this sense, this paper proposes the experimental results of work projects conducted on significant cases.

Succinic Semialdehyde Dehydrogenase Deficiency (SSADHD) is an ultra-rare autosomal recessive neurometabolic disorder caused by ALDH5A1 mutations presenting with autism and epilepsy. Here, we report the generation and characterization of human induced pluripotent stem cells (hiPSCs) derived from fibroblasts of three unrelated SSADHD patients – one female and two males with the CRISPR-corrected isogenic controls. These individuals are clinically diagnosed and are being followed in a longitudinal clinical study. [ABSTRACT FROM AUTHOR]

This paper describes the elaboration of the management plan for the UNESCO World Heritage site of Crespi d’Adda, a late-nineteenth-century company town that was inscribed in the World Heritage List in 1995 due to the integrity of its urban structure and the functional continuity of its productive and residential system. At the time of inscription, a management plan was not mandatory, and inscription did not produce a real development strategy able to counter the risks arising from de-industrialisation. The management plan preparation, launched in 2007, was assumed to represent an opportunity to ensure both protection and sustainable evolution. The process of elaboration made it necessary to promote coordination and dialogue among different institutions. This permitted enhancement of local governance. Moreover, the management plan acted as a framework within which to set guidelines for intervention on the external and internal features of the buildings, aimed at preserving the values of the site through the implementation of good practices. [ABSTRACT FROM PUBLISHER]