Approach for Predicting Production Scenarios Focused on Cross Impact Analysis

AbstractOne of the most consistent challenges in business is anticipating what the future holds and what impact it may have on current production systems. The scenario technique is a well-established method for developing and forecasting multiple future development paths for companies. However, this method is mostly employed to develop and to support strategic long-term decisions. The core idea of the approach introduced in this paper is to convey the future impact of today's decisions on production systems to employees involved in production planning processes. With the help of immersive visualization, performed in virtual reality (VR) systems, planning participants can perceive how the factory must adapt to fit future demands.In this paper, the focus is on the fourth phase of the scenario technique – so called scenario development – and, in particular, the cross impact analysis. With this methodology, the interrelations, or cross impacts of the different basic elements are determined. The cross impact analysis results serve as a basis for the development of a standardized tool that can be used to create probable production scenarios out of given production systems. This standardized tool will facilitate the usage of the scenario technique for factory planning projects, as it focuses the immense diversity of future uncertainties companies are faced with on the factory level

Assessment of Type I Interferon Signaling in Pediatric Inflammatory Disease

International audiencePURPOSE: Increased type I interferon is considered relevant to the pathology of a number of monogenic and complex disorders spanning pediatric rheumatology, neurology, and dermatology. However, no test exists in routine clinical practice to identify enhanced interferon signaling, thus limiting the ability to diagnose and monitor treatment of these diseases. Here, we set out to investigate the use of an assay measuring the expression of a panel of interferon-stimulated genes (ISGs) in children affected by a range of inflammatory diseases. DESIGN, SETTING, AND PARTICIPANTS: A cohort study was conducted between 2011 and 2016 at the University of Manchester, UK, and the Institut Imagine, Paris, France. RNA PAXgene blood samples and clinical data were collected from controls and symptomatic patients with a genetically confirmed or clinically well-defined inflammatory phenotype. The expression of six ISGs was measured by quantitative polymerase chain reaction, and the median fold change was used to calculate an interferon score (IS) for each subject compared to a previously derived panel of 29 controls (where +2 SD of the control data, an IS of \textgreater2.466, is considered as abnormal). Results were correlated with genetic and clinical data. RESULTS: Nine hundred ninety-two samples were analyzed from 630 individuals comprising symptomatic patients across 24 inflammatory genotypes/phenotypes, unaffected heterozygous carriers, and controls. A consistent upregulation of ISG expression was seen in 13 monogenic conditions (455 samples, 265 patients; median IS 10.73, interquartile range (IQR) 5.90-18.41), juvenile systemic lupus erythematosus (78 samples, 55 patients; median IS 10.60, IQR 3.99-17.27), and juvenile dermatomyositis (101 samples, 59 patients; median IS 9.02, IQR 2.51-21.73) compared to controls (78 samples, 65 subjects; median IS 0.688, IQR 0.427-1.196), heterozygous mutation carriers (89 samples, 76 subjects; median IS 0.862, IQR 0.493-1.942), and individuals with non-molecularly defined autoinflammation (89 samples, 69 patients; median IS 1.07, IQR 0.491-3.74). CONCLUSIONS AND RELEVANCE: An assessment of six ISGs can be used to define a spectrum of inflammatory diseases related to enhanced type I interferon signaling. If future studies demonstrate that the IS is a reactive biomarker, this measure may prove useful both in the diagnosis and the assessment of treatment efficacy

Further insights into Allan-Herndon-Dudley syndrome: characterization of two genetic variants in SLC16A2 gene

Genetics variants in SLC16A2 gene encoding for the monocarboxylate transporter 8 (MCT8) cause a severe X-linked intellectual deficit and neurological impairment known as Allan-Herndon-Dudley syndrome (AHDS). MCT8 promotes cellular uptake and efflux of thyroid hormone and its mutations provoke elevated serum T3 levels in children. Iodothyronine deiodinases (DIO) 1 and 2 are implicated in the conversion of T4 into biologically active T3, while DIO3 converts T4 into the inactive hormone reverse T3 (rT3). Active T3 and retinoid X receptors (RXR) can form heterodimer complexes which bind to hormone response elements (HREs) that leads to activate or repress transcription. Our aim is to investigate the impact of MCT8 mutations on the pathogenetic mechanisms of AHDS. Fibroblasts were obtained from skin biopsies of 2 AHDS and matched controls. To evaluate both MCT8 and thyroid hormone signaling pathway related genes expression, RNA was extracted with TRIzoLTM and assessed by Real-Time PCR. Protein expression was valuated via western blot and immunofluorescence. MTT assay was used to compare cell viability. Live and dead assay was used to discriminate live and dead populations. Lipids were detected via oil red o staining. MTT assay demonstrated a reduced cell viability as consequence of mutations in SLC16A2. We report that SLC16A2 RNA expression in AHDS patients was extremely reduced in comparison with total RNA from healthy controls. Additionally, DIO2, progastricsin, HR and KLF9 RNA expression resulted upregulated, whilst DIO1, DIO2-AS1, DIO3 and TH were downregulated influencing T3 cell entrance. Myelin related genes were significatively reduced. The lipid staining revealed an increasing presence of lipid droplets in AHDS patients. Taken together, our preliminary data emphasize an impairment in AHDS fibroblasts in relation to mutations in MCT8 transporter, increasing our understanding in the pathogenic mechanism of mutation in two patients affected by AHDS

Parental Somatic Mosaicism Uncovers Inheritance of an Apparently De Novo GFAP Mutation

Alexander disease is a leukodystrophy caused by heterozygous mutations of GFAP gene. Recurrence in siblings from healthy parents provides a confirmation to the transmission of variants through germinal mosaicism. With the use of DNA isolated from peripheral blood, next-generation sequencing (NGS) of GFAP locus was performed with deep coverage (>= 500x) in 11 probands and their parents (trios) with probands heterozygous for apparently de novo GFAP mutations. Indeed, one parent had somatic mosaicism, estimated in the range of 8.9%-16%, for the mutant allele transmitted to the affected sibling. Parental germline mosaicism deserves attention, as it is critical in assessing the risk of recurrence in families with Alexander disease

More Than Hypomyelination in Pol-III Disorder

The 4H syndrome (hypomyelination, hypodontia, hypogonadotropic hypogonadism) is a newly recognized leukodystrophy. The classical form is characterized by the association of hypomyelination, abnormal dentition, and hypogonadotropic hypogonadism, but the recent identification of 2 genes responsible for the syndrome demonstrates that these 3 main characteristics can be variably combined among "Pol-III (polymerase III) Yrelated leukodystrophies.'' The pathophysiology of this group of diseases is still to be elucidated, and there are no neuropathologic descriptions of brain tissue. We report the clinical, neuroradiologic, and neuropathologic findings of a patient affected by 4H syndrome with confirmed POLR3A mutations. We found a marked loss of oligodendrocytes, varying in severity in different brain regions, and accompanied by severe loss of myelin, moderately severe loss of axons, and patchy perivascular regions of better preserved white matter. There was relatively mild white matter astrogliosis and microgliosis. A macrophage reaction involving viable normal-appearing oligodendroglia suggests the possibility of an immunologic process in this disorder. Cortical laminar astrogliosis and mineralization of Layers I and II in particular were present. Thus, despite the uniformly hypomyelinating pattern seen on magnetic resonance imaging, neuropathologic examination reveals a complex heterogeneous leukodystrophy with prominent neuroaxonal and glial involvement in this disorder

Functional characterization and transcriptional profiling of fibroblasts from patients with mutations in SLC16A2 gene

Background/Objectives: Genetics variants in SLC16A2 gene encoding for the monocarboxylate transporter 8 (MCT8) cause a severe X-linked intellectual deficit known as Allan-Herndon-Dudley syndrome (AHDS). MCT8 promotes cellular uptake and efflux of thyroid hormones. Active T3 and retinoid X receptors (RXR) can form heterodimer complexes which bind to hormone response elements (HREs) that leads to activate or repress transcription. Our aim is to investigate the impact of MCT8 mutations on the pathogenetic mechanisms of AHDS. Methods: Fibroblasts were obtained from skin biopsies of 2 AHDS mutated patients and matched controls. RNA was extracted with TRIzoLTM. Total RNA sequencing was performed with the CORALL Total RNA-Seq Library Prep Kit using Illumina NextSeq 500 Sequencing. Protein expression was valuated via western blot and immunofluorescence. MTT assay was used to compare cell viability. Live/dead assay discriminated live and dead populations. Lipids were detected via Oil Red O staining. Results: A strong dysregulation in AHDS patients was highlighted by transcriptomic profiling, identifying 396 DEGs when comparing AHDS patients to controls. Moreover, MTT and Live/Dead assays demonstrated a reduced cell viability in AHDS_1 patient with a splicing variant [c.1690G>A (p.Gly564Arg)]. The C-terminal missense variant [c.1690G>A (p.Gly564Arg)] did not affect fibroblasts viability, challenging a personalized in vitro behavior. Target genes expression resulted upregulated in both patients. Furthermore, myelin related genes were significatively reduced. The lipid staining revealed an increasing presence of lipid droplets in AHDS fibroblasts. Conclusion: Our preliminary data emphasize a mutation-specific impairment in patients’ specific primary fibroblasts, used as pre-clinical experimental model of the disease

Role of epigenetics and alterations in RNA metabolism in leukodystrophies

Leukodystrophies are a class of rare heterogeneous disorders which affect the white matter of the brain, ultimately leading to a disruption in brain development and a damaging effect on cognitive, motor and social-communicative development. These disorders present a great clinical heterogeneity, along with a phenotypic overlap and this could be partially due to contributions from environmental stimuli. It is in this context that there is a great need to investigate what other factors may contribute to both disease insurgence and phenotypical heterogeneity, and novel evidence are raising the attention toward the study of epigenetics and transcription mechanisms that can influence the disease phenotype beyond genetics. Modulation in the epigenetics machinery including histone modifications, DNA methylation and non-coding RNAs dysregulation, could be crucial players in the development of these disorders, and moreover an aberrant RNA maturation process has been linked to leukodystrophies. Here, we provide an overview of these mechanisms hoping to supply a closer step toward the analysis of leukodystrophies not only as genetically determined but also with an added level of complexity where epigenetic dysregulation is of key relevance. This article is categorized under: Regulatory RNAs/RNAi/Riboswitches > Regulatory RNA RNA in Disease and Development > RNA in Disease RNA in Disease and Development > RNA in Development

Further insights into Allan-Herndon-Dudley syndrome: a novel SLC16A2 splice site variant

Background Genetics variants in SLC16A2 gene encoding for the monocarboxylate transporter 8 (MCT8) cause a severe X-linked leukoencephalopathy known as Allan-Herndon-Dudley syndrome (AHDS). MCT8 promotes cellular uptake and efflux of thyroid hormone. Iodothyronine deiodinases (DIO) 1 and 2 are implicated in the conversion of T4 into biologically active T3, while DIO3 converts T4 into the inactive hormone reverse T3 (rT3). Active T3 and retinoid X receptors (RXR) can form heterodimer complexes which bind to hormone response elements (HREs) that leads to activate or repress transcription. Aim of the study The aim of this work is to investigate the impact of a novel SLC16A2 splice site variant on the pathogenesis of AHDS. Materials and Methods In silico prediction tools, such as Mutation Taster, were used to assess the pathogenic score of the identified variant, while splicing prediction tools, such as Fruit Fly Splice Predictor, were used to analyze the effect of variant on the splicing mechanism. Fibroblasts were obtained from skin biopsies of both AHDS patient and a matched control. To evaluate MCT8 and thyroid hormone signaling pathway related genes expression, RNA was extracted with TRIzoLTM and assessed by Real-Time PCR. Protein expression was evaluated via western blot and immunofluorescence. MTT assay was used to compare cell viability. Live and dead assay was used to discriminate live and dead populations. Lipid droplets were detected via oil red o staining. Results The identified variant in the SLC16A2 gene causes the breakup of the wild type donor splice site, possibly leading to exon 1 skipping, affecting cell viability. SLC16A2 RNA expression in our AHDS patient was extremely reduced in comparison with total RNA from healthy control. DIO2, progastricsin, HR and KLF9 RNA expression resulted upregulated, whilst DIO1, DIO2-AS1, DIO3 and TH were downregulated influencing T3 cell entrance. Myelin related genes were significatively reduced. The lipid staining revealed an increasing presence of lipid droplets in AHDS patients. Conclusions Our preliminary data emphasize an impairment in AHDS fibroblasts in relation to SLC16A2 splice site variant, increasing our understanding in the pathogenic mechanism of patients affected by AHDS

Telemedicine for Personalized Nutritional Intervention of Rare Diseases: A Narrative Review on Approaches, Impact, and Future Perspectives

Background: Telemedicine represents a growing opportunity to improve ac- cess to personalized care for patients with rare diseases, addressing the challenges of specialized healthcare that is often limited by geographical barriers. The aim of this narrative review is to explore how telemedicine can facilitate tailored nutritional interven- tions for rare diseases, focusing on inherited metabolic diseases, rare neurological disorders, such as leukodystrophies, and neuromuscular disorders, including spinal muscular atrophies. Methods: This narrative review is based on a systematic search of the published literature over the past 20 years, and includes systematic reviews, meta-analysis, retrospective studies, and original articles. References were selected through searches in databases such as PubMed and Scopus, applying predefined inclusion and exclusion criteria. Among the inclusion criteria, studies focusing on pediatric patients aged 0 to 18 years, diagnosed with rare neurological diseases or inherited metabolic disorders, and using telemedicine in addition to in-person visits at their reference center were considered. Among the exclusion criteria, studies involving patients with other pathologies or comorbidities and those involving patients older than 18 years were excluded. Results: A total of 66 documents were analyzed to examine the challenges and specific needs of patients with rare diseases, highlighting the advantages and limitations of telemedicine compared to traditional care. The use of telemedicine has revolutionized the medi- cal approach, facilitating integrated care by multidisciplinary teams. Conclusions: Telemedicine still faces several technical, organizational, and security challenges, as well as disparities in access across different geographical areas. Emerging technologies such as artificial intelligence could positively transform the monitoring and management of patients with rare diseases. Telemedicine has great potential ahead of it in the development of increasingly personalized and effective care, in fact, emerging technologies are important to provide remote care, especially for patients with rare diseases