Novel clinical, molecular and bioinformatics insights into the genetic background of autism.

BACKGROUND: Clinical classification of autistic patients based on current WHO criteria provides a valuable but simplified depiction of the true nature of the disorder. Our goal is to determine the biology of the disorder and the ASD-associated genes that lead to differences in the severity and variability of clinical features, which can enhance the ability to predict clinical outcomes. METHOD: Novel Whole Exome Sequencing data from children (n = 33) with ASD were collected along with extended cognitive and linguistic assessments. A machine learning methodology and a literature-based approach took into consideration known effects of genetic variation on the translated proteins, linking them with specific ASD clinical manifestations, namely non-verbal IQ, memory, attention and oral language deficits. RESULTS: Linear regression polygenic risk score results included the classification of severe and mild ASD samples with a 81.81% prediction accuracy. The literature-based approach revealed 14 genes present in all sub-phenotypes (independent of severity) and others which seem to impair individual ones, highlighting genetic profiles specific to mild and severe ASD, which concern non-verbal IQ, memory, attention and oral language skills. CONCLUSIONS: These genes can potentially contribute toward a diagnostic gene-set for determining ASD severity. However, due to the limited number of patients in this study, our classification approach is mostly centered on the prediction and verification of these genes and does not hold a diagnostic nature per se. Substantial further experimentation is required to validate their role as diagnostic markers. The use of these genes as input for functional analysis highlights important biological processes and bridges the gap between genotype and phenotype in ASD.

Functional characterization of cytochrome P450s associated with pyrethroid resistance in the olive fruit fly Bactrocera oleae.

Resistance to pyrethroid insecticides has evolved in Bactrocera oleae populations in Greece, threatening the efficacy of control interventions based on this insecticide class. Here we report the collection of populations from Crete, with resistance levels reaching up to 132-folds, compared to susceptible laboratory strains and show that pyrethroid resistance is substantially suppressed by the PBO synergist, suggesting the involvement of detoxification enzymes. To identify specific candidate genes implicated in resistance, we performed comparative transcriptomic analysis, between the pyrethroid resistant populations from Crete and the susceptible laboratory strains, using both whole bodies and Malpighian tubules. Several genes were found differentially transcribed between resistant and susceptible flies in each comparison, with P450s being among the most highly over-expressed detoxification genes in pyrethroid resistant populations. Four of the over-expressed P450s (Cyp6A61, Cyp6G6, Cyp4P6 and Cyp6G28) were recombinantly expressed in Escherichia coli and in vitro metabolism assays revealed that CYP6A61 is capable of metabolizing alpha-cypermethrin, while CYP6G6, CYP4P6 and CYP6G28 are capable of metabolizing deltamethrin. No metabolism of neonicotinoid insecticides was recorded. We further silenced CYP6G6 in vivo, via RNAi, which led to a small, but significant increase in deltamethrin toxicity. The study provides valuable information towards the development of molecular diagnostics and evidence-based insecticide resistance management strategies.

Dent-2 disease with a Bartter-like phenotype caused by the Asp631Glu mutation in the OCRL gene.

BACKGROUND: Dent disease is an X-linked disorder characterized by low molecular weight proteinuria (LMWP), hypercalciuria, nephrolithiasis and chronic kidney disease (CKD). It is caused by mutations in the chloride voltage-gated channel 5 (CLCN5) gene (Dent disease-1), or in the OCRL gene (Dent disease-2). It is associated with chronic metabolic acidosis; however metabolic alkalosis has rarely been reported. CASE PRESENTATION: We present a family with Dent-2 disease and a Bartter-like phenotype. The main clinical problems observed in the proband included a) primary phosphaturia leading to osteomalacia and stunted growth; b) elevated serum calcitriol levels, leading to hypercalcemia, hypercalciuria, nephrolithiasis and nephrocalcinosis; c) severe salt wasting causing hypotension, hyperaldosteronism, hypokalemia and metabolic alkalosis; d) partial nephrogenic diabetes insipidus attributed to hypercalcemia, hypokalemia and nephrocalcinosis; e) albuminuria, LMWP. Phosphorous repletion resulted in abrupt cessation of hypercalciuria and significant improvement of hypophosphatemia, physical stamina and bone histology. Years later, he presented progressive CKD with nephrotic range proteinuria attributed to focal segmental glomerulosclerosis (FSGS). Targeted genetic analysis for several phosphaturic diseases was unsuccessful. Whole Exome Sequencing (WES) revealed a c.1893C > A variant (Asp631Glu) in the OCRL gene which was co-segregated with the disease in male family members. CONCLUSIONS: We present the clinical characteristics of the Asp631Glu mutation in the OCRL gene, presenting as Dent-2 disease with Bartter-like features. Phosphorous repletion resulted in significant improvement of all clinical features except for progressive CKD. Angiotensin blockade improved proteinuria and stabilized kidney function for several years.

The p.Pro482Ala Variant in the CNNM2 Gene Causes Severe Hypomagnesemia Amenable to Treatment with Spironolactone.

Renal hypomagnesemia syndromes involving CNNM2 protein pathogenic variants are associated with variable degrees of neurocognitive dysfunction and hypomagnesemia. Here, we report a family with a novel CNNM2 p.Pro482Ala variant, presenting with overt hypomagnesemia and mild neurological involvement (autosomal dominant renal hypomagnesemia 6, HOMG6, MIM# 613882). Using a bioinformatics approach, we showed that the p.Pro482Ala amino acid substitution causes a 3D conformational change in CNNM2 structure in the cystathionin beta synthase (CBS) domain and the carboxy-terminal protein segment. A novel finding was that aldosterone inhibition with spironolactone helped to alleviate hypomagnesemia and symptoms in the proband.

SPECT analysis and language profile in Greek speaking patients with subtypes of frontotemporal dementia.

OBJECTIVE: We aimed to examine if single photon emission computed tomography (SPECT) can discriminate between variants of frontotemporal dementia (FTD). As a secondary investigation we identify and establish the linguistic differences between those variants. MATERIALS AND METHODS: Nine patients with semantic variant primary progressive aphasia (svPPA), 8 with non-fluent variant primary progressive aphasia (nfvPPA) and 17 with behavioral variant of frontotemporal dementia (bvFTD) were compared on Addenbrooke's cognitive examination-revised (ACE-R), auditory comprehension, oral expression and verbal fluency. All patients were also compared with healthy controls. Patients were evaluated using technetium-99m-hexamethylproyleneamine oxime (99mTc-HMPAO) brain SPECT as a measure of regional cerebral flow. RESULTS: Significant group differences between all patients and controls were found for ACE-R, auditory comprehension and oral expression. Semantic variant primary progressive aphasia patients performed higher in letter compared to category fluency with significant deficits in auditory comprehension and oral expression. Non-fluent variant primary progressive aphasia patients showed significant deficits in auditory comprehension but not oral expression while performed lightly worse in letter fluency compared to category. Behavioral variant of frontotemporal dementia patients showed deficits in auditory comprehension and oral expression and performed similar in category and letter fluency. Single photon emission computed tomography analysis revealed left frontotemporal hypoperfusion extending to the right frontotemporal region in svPPA patients. Non-fluent variant primary progressive aphasia patients presented left frontotemporal hypoperfusion with participation of the left parietal and right frontotemporal regions. Behavioral variant of frontotemporal dementia patients showed bilateral frontotemporal hypoperfusion compared to parietal and visual cortices. CONCLUSION: Our findings suggest that SPECT may assist in the discrimination of the FTD variants. We also confirmed that bvFTD patients share similar language deficits with svPPA patients.

Characterization of Fungi Associated with Olive Fruit Rot and Olive Oil Degradation in Crete, Southern Greece.

In November 2019, a severe outbreak of fruit rot was observed in olive orchards in Crete, southern Greece. Symptoms appeared primarily on fruits and stalks, resembling those caused by anthracnose. Typical symptoms were fruit rot, shrinkage, and mummification, associated commonly with stalk discoloration and fruit drop. Disease incidence was estimated at up to 100% in some cases, and an unprecedented increase in olive oil acidity reaching up to 8% (percentage of oleic acid) in severely affected olive groves was recorded. Thirty-two olive groves were then surveyed, and samples of fruit, stalk, leaf, and shoot were collected. Visual, stereoscopic, and microscopic observations revealed several fungi belonging to the genera Alternaria, Botryosphaeria, Capnodium, Colletotrichum, Fusarium, and Pseudocercospora. Fungal infection in fruits was commonly associated with concomitant infestation by the olive fruit fly Bactrocera oleae along with increased air temperature and relative humidity conditions that prevailed in October and November 2019. Twenty representative fungal strains isolated from symptomatic fruits and stalks were characterized by morphological, physiological, and molecular analyses. By internal transcribed spacer regions of ribosomal DNA region and translation elongation factor 1-α gene sequencing analysis, these isolates were identified as Alternaria spp., A. infectoria, Botryosphaeria dothidea, Colletotrichum boninense sensu lato, Fusarium lateritium, F. solani species complex and Stemphylium amaranthi. Pathogenicity tests on punctured fruits revealed that all isolates were pathogenic; however, F. solani isolates along with B. dothidea were the most virulent, and wounds were necessary for efficient fungal infection. Moreover, as few as 10 spores of F. solani were sufficient to cause significant infection in punctured fruits. F. solani was also capable of infecting olive fruits in the presence of B. oleae, with no additional wounding, in artificial inoculation experiments. Moreover, it was capable of colonizing and affecting olive blossoms. Further analyses of olive oil extracted from fruits artificially inoculated with F. solani indicated a significant increase in oil acidity, K232, K270, and peroxide value, whereas total phenol content was significantly decreased. To the best of our knowledge, this is the first report of F. solani associated with olive fruit rot and olive oil degradation worldwide.

Semantic fluency difficulties in developmental dyslexia and developmental language disorder (DLD): poor semantic structure of the lexicon or slower retrieval processes?

BACKGROUND: Children with dyslexia and/or developmental language disorder (hereafter children with DDLD) have been reported to retrieve fewer words than their typically developing (TD) peers in semantic fluency tasks. It is not known whether this retrieval difficulty can be attributed to the semantic structure of their lexicon being poor or, alternatively, to words being retrieved more slowly despite semantic structure being intact. AIMS: To test two theoretical models that could potentially account for retrieval difficulties in semantic fluency tasks, namely, the Poor Lexical-Semantic Structure Model and the Slow-Retrieval Model. Both models predict that children with DDLD will retrieve fewer items compared with TD children. However, while the Poor Lexical-Semantic Structure Model predicts a less sophisticated network of semantic connections between words in the lexicon, as evidenced by smaller clusters of related items in children with DDLD, the Slow-Retrieval Model predicts intact inter-item associations in the lexicon, as evidenced by the two groups' clusters being of a similar size. The groups' semantic fluency performance was therefore compared. How semantic fluency performance related to children's language, literacy, and phonological skills was also investigated. METHODS & PROCEDURES: A total of 66 children with DDLD aged 7-12 years and 83 TD children aged 6-12 years, all monolingual Greek speakers, were tested on semantic fluency, using the categories 'animals', 'foods' and 'objects from around the house'. The numbers of correct and incorrect responses, clusters and switches, and the average cluster size were computed. Children were also assessed on non-verbal IQ, language, literacy and phonological tasks. OUTCOMES & RESULTS: In both groups, productivity in semantic fluency tasks correlated strongly with the numbers of clusters and switches, but not with average cluster size. The DDLD group produced significantly fewer correct responses and fewer clusters compared with the TD group, but the two groups showed similar switching and average cluster size. Children's language, literacy and phonological skills significantly predicted the number of correct responses produced, beyond the significant effect of age. CONCLUSIONS & IMPLICATIONS: We conclude that poorer semantic fluency performance in children with DDLD results not from a lexicon with poor semantic structure, but rather from slower retrieval processes from a lexicon with intact semantic structure. The underlying causes of slow lexical retrieval still need further investigation.

Estimating the Necessary Amount of Driving Data for Assessing Driving Behavior.

The aim of this paper was to provide a methodological framework for estimating the amount of driving data that should be collected for each driver in order to acquire a clear picture regarding their driving behavior. We examined whether there is a specific discrete time point for each driver, in the form of total driving duration and/or the number of trips, beyond which the characteristics of driving behavior are stabilized over time. Various mathematical and statistical methods were employed to process the data collected and determine the time point at which behavior converges. Detailed data collected from smartphone sensors are used to test the proposed methodology. The driving metrics used in the analysis are the number of harsh acceleration and braking events, the duration of mobile usage while driving and the percentage of time driving over the speed limits. Convergence was tested in terms of both the magnitude and volatility of each metric for different trips and analysis is performed for several trip durations. Results indicated that there is no specific time point or number of trips after which driving behavior stabilizes for all drivers and/or all metrics examined. The driving behavior stabilization is mostly affected by the duration of the trips examined and the aggressiveness of the driver.

Time reference, morphology and prototypicality: tense production in stroke aphasia and semantic dementia in Greek.

The present study aims at investigating verb inflection in aphasia and semantic dementia. In particular, it addresses the contribution of time reference and morphological complexity. Moreover, it investigates whether the lexical properties of the verb, such as argument structure and lexical aspect interact with the production of tense. Ten individuals with (different types of) stroke aphasia and five individuals with semantic dementia and their respective control groups conducted a sentence completion task. Three tenses were tested: past perfective, past imperfective and present. All tenses had to be produced with three different verb classes, which differed with respect to syntactic and semantic properties: unergative, unaccusative and transitive verbs. The findings imply problems with marking aspect and an interaction between inflection and lexical aspect but no effect of morphological complexity or across the board difficulties with reference to the past in aphasia. Moreover, the results suggest problems with inflection in semantic dementia, an interaction between inflection and lexical aspect and a selective difficulty with imperfective tenses. The study contributes to a better understanding of inflection problems in aphasia and it provides evidence for inflection problems in semantic dementia.

Glutathione-PEGylated liposomal methylprednisolone in comparison to free methylprednisolone: slow release characteristics and prolonged lymphocyte depression in a first-in-human study.

AIMS: Intravenous high-dose free methylprednisolone (MP) hemisuccinate is the primary treatment for an acute relapse in relapsing-remitting multiple sclerosis. However, it is inconvenient and its side effects are undesirable. Both dose and dosing frequency can be reduced by incorporating free MP in glutathione-PEGylated liposomes, creating a slow-release formulation with reduced toxicity and prolonged peripheral efficacy. This first-in-human study was designed to assess the safety, pharmacokinetics and pharmacodynamics of glutathione-PEGylated liposomes containing MP (2B3-201). METHODS: The first part was a double-blind, three-way cross over study in 18 healthy male subjects, receiving ascending doses of 2B3-201, active comparator (free MP) or placebo. Part 2 of the study was an open-label infusion of 2B3-201 (different doses), exploring pretreatment with antihistamines and different infusion schedules in another 18 healthy male subjects, and a cross-over study in six healthy female subjects. MP plasma concentrations, lymphocyte counts, adrenocorticotropic hormone, osteocalcin and fasting glucose were determined. Safety and tolerability profiles were assessed based on adverse events, safety measurements and central nervous system tests. RESULTS: The most frequent recorded AE related to 2B3-201 was an infusion related reaction (89%). 2B3-201 was shown to have a plasma half-life between 24 and 37 h and caused a prolonged decrease in the lymphocyte count, adrenocorticotropic hormone and osteocalcin, and a rise in fasting glucose. CONCLUSION: 2B3-201 is considered safe, with no clinically relevant changes in central nervous system safety parameters and no serious adverse events. In addition, 2B3-201 shows a long plasma half-life and prolonged immunosuppressive effects.

Assessing the immunomodulatory potential of high-molecular-weight extracts from mushrooms; an assay based on THP-1 macrophages.

BACKGROUND: Food is a potential source of immunomodulating compounds that may be used to steer immune responses towards a desired status such as reducing inflammatory disorders. However, to identify and characterize such bioactive compounds, biologically relevant and standardized assays are required. Macrophages play an important role in immunomodulation and are suited for developing cell-based assays. An assay was developed based on macrophages, in a homogeneous differentiation state, using the human monocytic cell line THP-1 previously used to assess immunomodulatory properties of low-molecular-weight allergens, hormones, dietary supplements and therapeutic drugs. RESULTS: Zymosan and mushroom polysaccharide extracts lead to a heterogeneous differentiation state of THP-1 monocytes, and these cells secrete low levels of cytokines upon stimulation. Differentiation into macrophages using a low concentration of phorbol 12-myristate 13-acetate improved responsiveness. Elevated levels of cytokines were secreted by cells in a homogenous differentiation state. In addition, it was determined that the assay performs best when using cells at a concentration of (2.5-5) × 10(5) cells mL(-1). CONCLUSION: An assay was developed suitable to distinguish the immunomodulatory properties of food compounds in a reproducible manner. It was evaluated using eight mushroom species by measuring the secretion of relevant cytokines TNF-α, IL-1ß, IL-6 and IL-10.

Cognitive vs. Linguistic Training in Children with Developmental Language Disorder: Exploring Their Effectiveness on Verbal Short-Term Memory and Verbal Working Memory.

The present study explores comparatively the effectiveness of a cognitive (verbal short-term memory (vSTM), verbal working memory (vWM)) and of a linguistic training (10-week duration each) in the diffusion of gains in cognitive abilities (vSTM and vWM) of in school-aged Greek-speaking children with developmental language disorder (DLD). To this purpose, two computerized training programs i.e., a linguistic and a cognitive one, were developed and applied to three groups (A, B, and C) of children with DLD (N = 49, in total). There were three assessments with two vSTM tasks (non-word repetition and forward digit span) and a vWM task (backward digit span): pre-therapeutically (time 1), where no significant between-group differences were found, post-therapeutically I (time 2), and post-therapeutically II (time 3) and two training phases. In phase Ι, group A received meta-syntactic training, whereas group B vSTM/vWM training and group C received no training. In phase ΙΙ, a reversal of treatment was performed for groups A and B: group A received vSTM/vWM while group B meta-syntactic training. Again, group C received no training. Overall, the results indicated a significant performance improvement for the treatment groups and revealed beneficial far-transfer effects as language therapy can affect vSTM and vWM in addition to direct and near transfer effects. In addition, the intervention type order affected performance as follows: first, better performance on the vSTM task (non-word repetition) was shown when the linguistic treatment was delivered first; second, better performance on the vWM in Time 2 and Time 3 was shown by group B, for which the cognitive treatment was delivered first. Concluding, not only intervention type but also intervention type order can affect performance in DLD.

Morphosyntactic Abilities and Cognitive Performance in Multiple Sclerosis.

While cognitive abilities in people with multiple sclerosis (PwMS) have been studied in detail, little is known about linguistic abilities in PwMS and their relation to cognitive impairment. In this cross-sectional explorative study, we aim to investigate the morphosyntactic abilities of PwMS alongside their cognitive performance. Furthermore, we explore the effect of clinical factors, namely, the disease duration and MS type, on the linguistic and cognitive performance of PwMS. By so doing, we aim to shed light on neurocognitive and clinical correlates of linguistic performance in PwMS. We included 78 patients and 78 age-, sex- and education-matched healthy individuals. All participants were additionally administered the Brief International Cognitive Assessment for Multiple Sclerosis (BICAMS) battery, a verbal short-term memory task (non-word repetition) and questionnaires about mood, fatigue and quality of life. In addition, they underwent examinations with morphology and syntax tasks. PwMS were found to be impaired in morphology (past tense) and selectively impaired in syntax alongside cognitive impairments. Disease duration had the main impact on cognitive abilities. The MS type selectively impacted linguistic abilities, as shown by the remarkably deficient performance of the MS individuals with the progressive disease subtype. Linguistic impairments were predicted by only one measure of the BICAM test, namely, the Symbol Digit Modalities Test (SDMT), a measure of cognitive processing speed. Overall, this study contributes to the better understanding of the linguistic profile of PwMS by reporting selective deficits in their morphological and syntactical abilities. Furthermore, it provides insights into the clinical and cognitive correlates of linguistic performance. By so doing, it suggests clinical implications for the development of intervention programs for PwMS.

An autologous ex vivo model for exploring patient-specific responses to viro-immunotherapy in glioblastoma.

Oncolytic virus (OV) clinical trials have demonstrated remarkable efficacy in subsets of patients with glioblastoma (GBM). However, the lack of tools to predict this response hinders the advancement of a more personalized application of OV therapy. In this study, we characterize an ex vivo co-culture system designed to examine the immune response to OV infection of patient-derived GBM neurospheres in the presence of autologous peripheral blood mononuclear cells (PBMCs). Co-culture conditions were optimized to retain viability and functionality of both tumor cells and PBMCs, effectively recapitulating the well-recognized immunosuppressive effects of GBM. Following OV infection, we observed elevated secretion of pro-inflammatory cytokines and chemokines, including interferon γ, tumor necrosis factor α, CXCL9, and CXCL10, and marked changes in immune cell activation markers. Importantly, OV treatment induced unique patient-specific immune responses. In summary, our co-culture platform presents an avenue for personalized screening of viro-immunotherapies in GBM, offering promise as a potential tool for future patient stratification in OV therapy.

Investigation on the Attitudes and Perspectives of Medical and Nursing Staff About Euthanasia: Data From Four Regional Greek Hospitals.

INTRODUCTION: The good and benefit of the patient are the main drivers of the decisions that health professionals are asked to make. However, the definition of the good and the actions required for benefit are not always simple and self-evident. The intractable ethical dilemma of euthanasia has been the subject of extensive debates over the years, and numerous studies have been carried out in an attempt to record the attitudes and opinions of both health professionals and the general population. METHOD: This research aims to investigate the opinions and perspectives of the medical and nursing staff of the four regional hospitals regarding euthanasia and to detect the factors that advocate for and against it. Two hundred and eighteen medical and nursing staff members from four regional hospitals in Lasithi participated in the research, whose opinions and influencing factors were investigated using a questionnaire consisting of four sections. The first included demographic and general characteristics questions; the second was the Euthanasia Attitude Scale (EAS); the third was the Death Attitude Profile-Revised (DAP-R); and the last was the Daily Spiritual Experience Scale (DSES). The SPSS software version 25.0 (IBM Corp., Armonk, NY) was used to analyse the data. RESULTS: Of the total, 78.0% of the participants were women, with an average sample age of 44.5 years. 65.1% were married, 23.4% were physicians, while 76.6% were nurses. The mean Euthanasia Attitude score (70.89) is moderate, ranging from 30 to 120, with higher scores suggesting more favourable sentiments. Euthanasia was viewed positively by 24.3% of respondents. There was no significant difference in positive attitudes between medical and nursing staff. However, the nursing staff had significantly lower average levels of General Orientation for Euthanasia, for the Role of Healthcare Professionals in Euthanasia, Values & Ethics, or Daily Spiritual Experience, and conversely higher levels of scores on Patients' Rights Issues for Euthanasia or Death Acceptance. CONCLUSIONS: Health professionals were found to have moderate attitudes about euthanasia, with no significant difference between them, as well as moderate degrees of death and everyday spiritual experience. Overall, a more favourable euthanasia attitude was shown to be strongly associated with individuals who were single, divorced, or widowed, with less death acceptance or more neutral acceptance, but not with daily spiritual experience.

The heterogeneous sensitivity of pediatric brain tumors to different oncolytic viruses is predicted by unique gene expression profiles.

Despite decades of research, the prognosis of high-grade pediatric brain tumors (PBTs) remains dismal; however, recent cases of favorable clinical responses were documented in clinical trials using oncolytic viruses (OVs). In the current study, we employed four different species of OVs: adenovirus Delta24-RGD, herpes simplex virus rQNestin34.5v1, reovirus R124, and the non-virulent Newcastle disease virus rNDV-F0-GFP against three entities of PBTs (high-grade gliomas, atypical teratoid/rhabdoid tumors, and ependymomas) to determine their in vitro efficacy. These four OVs were screened on 14 patient-derived PBT cell cultures and the degree of oncolysis was assessed using an ATP-based assay. Subsequently, the observed viral efficacies were correlated to whole transcriptome data and Gene Ontology analysis was performed. Although no significant tumor type-specific OV efficacy was observed, the analysis revealed the intrinsic biological processes that associated with OV efficacy. The predictive power of the identified expression profiles was further validated in vitro by screening additional PBTs. In summary, our results demonstrate OV susceptibility of multiple patient-derived PBT entities and the ability to predict in vitro responses to OVs using unique expression profiles. Such profiles may hold promise for future OV preselection with effective oncolytic potency in a specific tumor, therewith potentially improving OV responses.

Extracellular vesicles as a promising source of lipid biomarkers for breast cancer detection in blood plasma.

Extracellular vesicles (EVs), including exosomes and microvesicles, mediate intercellular communication in cancer, from development to metastasis. EV-based liquid biopsy is a promising strategy for cancer diagnosis as EVs can be found in cancer patients' body fluids. In this study, the lipid composition of breast cancer-derived EVs was studied as well as the potential of blood plasma EVs for the identification of lipid biomarkers for breast cancer detection. Initially, an untargeted lipidomic analysis was carried out for a panel of cancerous and non-cancerous mammary epithelial cells and their secreted EVs. We found that breast cancer-derived EVs are enriched in sphingolipids and glycerophospholipids compared to their parental cells. The initial in vitro study showed that EVs and their parental cells can be correctly classified (100% accuracy) between cancerous and non-cancerous, as well as into their respective breast cancer subtypes, based on their lipid composition. Subsequently, an untargeted lipidomic analysis was carried out for blood plasma EVs from women diagnosed with breast cancer (primary or progressive metastatic breast cancer) as well as healthy women. Correspondingly, when blood plasma EVs were analysed, breast cancer patients and healthy women were correctly classified with an overall accuracy of 93.1%, based on the EVs' lipid composition. Similarly, the analysis of patients with primary breast cancer and healthy women showed an overall accuracy of 95% for their correct classification. Furthermore, primary and metastatic breast cancers were correctly classified with an overall accuracy of 89.5%. This reveals that the blood plasma EVs' lipids may be a promising source of biomarkers for detection of breast cancer. Additionally, this study demonstrates the usefulness of untargeted lipidomics in the study of EV lipid composition and EV-associated biomarker discovery studies. This is a proof-of-concept study and a starting point for further analysis on the identification of EV-based biomarkers for breast cancer.

Accuracy of Artificial Intelligence-Based Technologies for the Diagnosis of Atrial Fibrillation: A Systematic Review and Meta-Analysis.

Atrial fibrillation (AF) is the most common arrhythmia with a high burden of morbidity including impaired quality of life and increased risk of thromboembolism. Early detection and management of AF could prevent thromboembolic events. Artificial intelligence (AI)--based methods in healthcare are developing quickly and can be proved as valuable for the detection of atrial fibrillation. In this metanalysis, we aim to review the diagnostic accuracy of AI-based methods for the diagnosis of atrial fibrillation. A predetermined search strategy was applied on four databases, the PubMed on 31 August 2022, the Google Scholar and Cochrane Library on 3 September 2022, and the Embase on 15 October 2022. The identified studies were screened by two independent investigators. Studies assessing the diagnostic accuracy of AI-based devices for the detection of AF in adults against a gold standard were selected. Qualitative and quantitative synthesis to calculate the pooled sensitivity and specificity was performed, and the QUADAS-2 tool was used for the risk of bias and applicability assessment. We screened 14,770 studies, from which 31 were eligible and included. All were diagnostic accuracy studies with case-control or cohort design. The main technologies used were: (a) photoplethysmography (PPG) with pooled sensitivity 95.1% and specificity 96.2%, and (b) single-lead ECG with pooled sensitivity 92.3% and specificity 96.2%. In the PPG group, 0% to 43.2% of the tracings could not be classified using the AI algorithm as AF or not, and in the single-lead ECG group, this figure fluctuated between 0% and 38%. Our analysis showed that AI-based methods for the diagnosis of atrial fibrillation have high sensitivity and specificity for the detection of AF. Further studies should examine whether utilization of these methods could improve clinical outcomes.

Metabolic profiling stratifies colorectal cancer and reveals adenosylhomocysteinase as a therapeutic target.

The genomic landscape of colorectal cancer (CRC) is shaped by inactivating mutations in tumour suppressors such as APC, and oncogenic mutations such as mutant KRAS. Here we used genetically engineered mouse models, and multimodal mass spectrometry-based metabolomics to study the impact of common genetic drivers of CRC on the metabolic landscape of the intestine. We show that untargeted metabolic profiling can be applied to stratify intestinal tissues according to underlying genetic alterations, and use mass spectrometry imaging to identify tumour, stromal and normal adjacent tissues. By identifying ions that drive variation between normal and transformed tissues, we found dysregulation of the methionine cycle to be a hallmark of APC-deficient CRC. Loss of Apc in the mouse intestine was found to be sufficient to drive expression of one of its enzymes, adenosylhomocysteinase (AHCY), which was also found to be transcriptionally upregulated in human CRC. Targeting of AHCY function impaired growth of APC-deficient organoids in vitro, and prevented the characteristic hyperproliferative/crypt progenitor phenotype driven by acute deletion of Apc in vivo, even in the context of mutant Kras. Finally, pharmacological inhibition of AHCY reduced intestinal tumour burden in ApcMin/+ mice indicating its potential as a metabolic drug target in CRC.

The psychometric properties of the Greek version of the Social Communication Questionnaire.

There is a scarcity of diagnostic assessments and screening tools for autism spectrum disorders (ASD) in Greek. In this study, we examined the psychometric properties of the recently developed Greek version of the Social Communication Questionnaire (SCQ). We used parental responses for 311 children (mean age: 7.54 years old, SD = 1.92), 122 with a diagnosis of ASD (93 boys, 29 girls) and 189 neurotypical children (104 boys, 85 girls), with 167 responses referring to the Lifetime and 144 to the Current form of the SCQ. Both forms presented adequate construct validity based on the four-factor model, while in both forms, autistic children presented higher SCQ total and subscale scores (four factors) than typical children. The forms had excellent internal reliability. An item-response-theory analysis suggested that over 80% of test items fitted adequately a Rasch model, while a preliminary analysis of gender biases suggested that a small number of items (Lifetime: five; Current: six out of 39) were differentially sensitive to autistic symptomatology in boys and girls. A receiver-operating-characteristic analysis showed excellent diagnostic performance based on the SCQ total score (Lifetime: area-under-the-curve/AUC = 0.937, Current: AUC = 0.963), and acceptable to excellent discrimination for the four subscales (AUCs between 0.737 and 0.955). Our preliminary results suggest that the Greek SCQ presents satisfactory psychometric properties and can be used for differentiating children with ASD from typical children in initial assessments within clinical and research settings. LAY SUMMARY: Autism spectrum disorder (ASD or autism) is a lifelong neurodevelopmental condition with a prevalence of ~1.5%-2% and characterized by difficulties in social interaction and communication and repetitive and restricted behaviors. There is increasing concern that research in ASD has focused on a small number of languages and cultural settings and that this bias challenges the identification and diagnosis of the condition in other languages and cultures, which are underrepresented in autism research. One such language is Greek (spoken by ~13.5 million), for which there is a scarcity of standardized instruments for the diagnosis of autism. This study examines the psychometric properties of the recently published Greek version of the Social Communication Questionnaire (SCQ), a widely used screening tool for ASD. We conduct an in-depth psychometric analysis of the Greek SCQ, including both forms in which the instrument is available (Lifetime and Current). This analysis shows that the Greek SCQ can be used for differentiating children with ASD from typical children in initial assessments within clinical and research settings. The findings of this study have implications for clinicians, special educators and researchers working with Greek-speaking individuals with ASD and, more broadly, for cross-cultural autism research.