Search Results (431)

This study described TTN gene variants in Ecuadorian patients with hereditary cardiac diseases, integrating genetic ancestry to improve variant interpretation in an underrepresented population. Sixty patients with confirmed hereditary cardiac conditions were analyzed using the TruSight Cardio NGS panel (Illumina, San Diego, CA, USA), which targets 174 cardiac-associated genes. Bioinformatic analyses and classification were performed in accordance with ACMG/AMP guidelines, and ancestry inference was conducted using 46 Ancestry Informative Markers (AIM-InDels). From 4008 detected TTN variants, 29 variants of interest remained after filtering: 27 classified as variants of uncertain significance (VUS) and two as likely pathogenic. All variants were heterozygous and distributed across exons 3–358, primarily in the A-band region, commonly associated with cardiomyopathies and arrhythmic phenotypes. Two truncating variants (exons 267 and 272) met PVS1 criteria, while several missense variants (p.Ser91Gly, p.Pro12140Ser, p.Arg34653Cys) showed possible modulatory effects on hypertrophic or arrhythmic outcomes. Genetic ancestry revealed a predominant Native American background, followed by European and African components. These findings expand the understanding of TTN-related cardiac disease in Latin America, suggesting that TTN functions as a genetic modifier influencing disease expression. Incorporating ancestry information enhances genomic interpretation and supports precision medicine in diverse populations. Full article

CFTR modulators have significantly affected the prognosis for cystic fibrosis, improving the clinical course in most patients with the F508del variant and several other CFTR gene variants. The presence of complex alleles, including more than one variant in the cis position, can change the properties of the protein and the efficacy of modulators. Objective: We aimed to describe the efficacy of CFTR modulators in the treatment of two siblings with the c.[1521_1523delCTT;1399C>T];[54-5940_273+10250del21kb] (L467F;F508del/CFTRdele2,3) genotype in clinical practice and in vitro. This article presents the clinical presentation and results of CFTR channel function assessment and personalized selection of CFTR modulators in monochorionic diamniotic twins with cystic fibrosis and the L467F;F508del/CFTRdele2,3 genotype. This is the first demonstration of the efficacy of a new CFTR modulator in patients with a complex allele and a class I variant in the genotype. The obtained results may be useful for choosing treatment strategies for patients with a complex allele and a class I variant in the genotype. Full article

This study aims to investigate the genotype characteristics of newborns with biallelic GJB2 mutations and their correlation with hearing phenotypes, providing a basis for clinical genetic counseling and hearing management. A retrospective study was conducted on 652 newborns with biallelic GJB2 mutations detected at the Newborn Diseases Screening Center of Shenzhen Maternal and Child Health Care Hospital from January 2022 to December 2024. The differences in mutation types, hearing screening, and diagnostic results were analyzed and compared between the homozygous and compound heterozygous mutation groups to assess their correlation with hearing phenotypes. Genotype analysis identified 543 cases of homozygous mutations, mainly the c.109G>A/c.109G>A genotype (98.90%). Compound heterozygous mutations were identified in 109 cases, with the majority being c.109G>A/c.235delC (76.15%). Following two-stage hearing screening, 227 (34.82%) of the 652 cases were referred, with bilateral failure accounting for the majority (81.94%) of these cases. The referral rates showed no significant difference between the homozygous (35.54%) and compound heterozygous (31.19%) groups (p > 0.05). The overall hearing loss detection rate was 6.90% (45/652); among these, eight infants who had initially passed the newborn hearing screening were later found to have hearing loss between 2.5 and 6 months of age. Among the 45 confirmed deaf children, hearing loss was mainly mild to moderate (87.50%), and profound deafness was only seen in the homozygous mutation group (10.29%, 7/68 ears). Most newborns with biallelic GJB2 mutations passed the two-stage hearing screening, and associated hearing loss was typically mild to moderate. Long-term auditory monitoring remains essential for all genetically confirmed infants to monitor late-onset progression. Full article

Protein–protein interactions (PPIs) take place in many cellular processes, including the activation of cellular cascades, such as the MAPK/ERK (Mitogen-Activated Protein Kinase/Extracellular-Regulated Kinase) pathway. Deregulation of these pathways leads to the development of diseases, such as cancer. DEL-22379 is an ERK2 dimerization inhibitor, which presents anti-tumoral effects, without affecting ERK2 phosphorylation. Our aim was to identify new therapeutic molecules targeting ERK2 dimerization, based on DEL-22379 structure. In this study, we implemented a combination of computational and experimental workflow, which includes in silico techniques, such as scaffold hopping and virtual screening to generate a dataset of candidate compounds, a native PAGE (PolyAcrylamide Gel Electrophoresis) electrophoresis to experimentally screen the potential inhibitors, and a detailed molecular docking and chemical profile prediction to understand the potential mechanism of action of the selected compounds. From an initial dataset of 536 compounds, we obtained two hit molecules that exhibited inhibitory effects on ERK2 dimerization: Drug73 and Drug120. A computational analysis of the mechanism of action, unveiled that Drug73 and Drug120 presented an improved docking score, and better drug-like properties when compared to DEL-22379. This study shows that computational studies, in combination with experimental evaluation, can be useful and efficient to find new therapeutic compounds. Full article

Annona muricata (soursop) produces secondary metabolites with antimicrobial and insecticidal properties. Arbuscular mycorrhizal fungi (AMF) are known to enhance the production of secondary metabolites in medicinal plants. In this study, we aimed to evaluate the insecticidal activity of ethanolic leaf extracts from AMF-colonized soursop trees against the fall armyworm (Spodoptera frugiperda) and the triatomine bug Triatoma pallidipennis, a vector of Chagas disease. Ethanolic leaf extracts were obtained from trees inoculated with two AMF consortia (Cerro del Metate and Agua Dulce), with the species Rhizophagus intraradices and Funneliformis mosseae, and from non-mycorrhizal plants (SM). Extracts were tested in bioassays specific to each insect, including chemical and negative controls, and survival was analyzed using Kaplan–Meier curves. Extracts from plants colonized by F. mosseae exhibited insecticidal activity against S. frugiperda, causing 72% larval mortality, comparable to that of the commercial insecticide. In contrast, extracts from plants inoculated with the Agua Dulce consortium caused 65% mortality in T. pallidipennis adults. These extracts showed significantly higher annonacin content (µg·g⁻¹ DW). Overall, the results demonstrate that AMF colonization can enhance the synthesis of metabolites such as annonacins and contribute to increased insecticidal activity in A. muricata. Our findings suggest AMF-assisted cultivation has the potential to enhance botanical insecticides. Full article

Background/Objectives: Chronic lymphocytic leukemia (CLL) is a hematologic malignancy characterized by uncontrolled accumulation of B lymphocytes. A key feature of CLL is the presence of genetic aberrations, particularly alterations of chromosome 17, such as deletion of 17q and/or mutations in the TP53 gene. Since these abnormalities are highly relevant for therapeutic decision-making, assessment of TP53 mutational status is strongly recommended in routine diagnostics. This study aimed to evaluate the reliability of TP53 sequencing results depending on the type of DNA sample analyzed. Methods: DNA was isolated from two different sample types of the same patient: mononuclear cells (CLL1) and purified CD19+ cells (CLL2). The entire coding region of TP53 (exons 2–11), including splice sites (+/− 10 bp), was analyzed using capture-based next-generation sequencing (NGS). Reads were aligned to the GRCh37/hg19 reference genome, and variants were interpreted using DRAGEN Enrichment (Illumina) and Franklin (QIAGEN). Results: In sample CLL1, the NM_000546.6:c.626_627del mutation (Tier I) was identified with a variant allele frequency (VAF) of 57.06%. The same mutation was confirmed in CLL2, but with a higher VAF of 94.78%. Importantly, an additional Tier I mutation (NM_000546.6:c.825_826del) was detected exclusively in CLL2 at a VAF of 1.59%. Both findings met the required sequencing depth as well as coverage per sample, confirming their validity. Conclusions: The study demonstrates that inadequate starting material for DNA isolation may mask low-frequency TP53 mutations, resulting in false-negative results. Accurate detection requires ensuring sufficient CD19+ cell content, which is critical for reliable diagnostics and supports personalized treatment approaches in CLL. Full article

Background/Objectives: Cystic fibrosis transmembrane conductance regulator modulator (mCFTR) therapy has been proven efficacious in controlled clinical trials for individuals with cystic fibrosis. This post-approval retrospective study aimed to determine the comprehensive effects of elexacaftor/tezacaftor/ivacaftor (ETI) on nutritional status, the respiratory system and quality of life over 12 months of clinical use in paediatric patients treatment-naïve to mCFTR. Methods: A retrospective analysis of records of CF adolescents on ETI therapy was conducted. The selected parameters of anthropometric measurements, body composition assessed by BIA, spirometry, and multiple breath nitrogen washout (MBNW) to measure lung clearance index (LCI), were evaluated before therapy and at 3 and 12 months after treatment initiation. Additionally, children completed the Cystic Fibrosis Questionnaire-Revised (CFQ-R). Results: Over 18 months, data from 58 patients (mean age 14.34 ± 1.70, 50% female; 43% homozygous F508del) on ETI were collected. Body weight increased significantly over 12 months, with a mean gain of 3.33 kg at 3 months (p < 0.001) and 7.10 kg at 12 months (p < 0.001), alongside improvements in BMI z-score, fat-free mass, and fat mass. Significant changes (p < 0.001) were also observed after 3 and 12 months in ppFEV₁ (8.91 ± 8.23; 9.67 ± 8.77) and ppFVC (4.46 ± 5.24; 4.61 ± 5.70), with a decrease in LCI (−1.62 ± 2.15; −1.68 ± 1.89). The CFQ-R Respiratory score increased by 11.75 points and correlated with most of the pulmonary and nutritional parameters. Conclusions: In real-world settings, clinical improvement during ETI therapy reflects a comprehensive impact on nutritional status, body composition, pulmonary function, and quality of life for adolescents with CF. Full article

Background/Objectives: Spinocerebellar ataxia (SCA), or hereditary ataxia, is a progressive neurodegenerative disorder primarily affecting motor control and voluntary muscle coordination due to cerebellar or spinocerebellar dysfunction. While numerous genetic variants have been linked to SCA in both humans and dogs, some cases remain genetically unexplained. This study aimed to describe the clinical and pathological phenotype, and to identify the genetic basis, of an atypical form of SCA observed in a mixed-breed dog presenting with additional clinical signs beyond classic SCA. Methods: Clinical and postmortem examinations were performed to document neurological and systemic pathology. Whole-genome sequencing (WGS) was conducted on the affected dog, and variant filtering was carried out using a control cohort of over 700 unaffected dog genomes to identify candidate variants. Results: In addition to classical SCA features, the affected dog exhibited retinal and optic nerve degeneration and severe, non-regenerative anemia. WGS did not reveal any known SCA-associated variants. Variant filtering identified a novel homozygous 4-base-pair frameshift deletion in CLPX (caseinolytic mitochondrial matrix peptidase chaperone subunit X) [XM_038580726.1:c.1723_1726del; chr30:g.29943285_29943288del]. This variant is predicted to cause a frameshift and premature stop codon within 17 amino acids, truncating approximately 6.64% of the protein. Conclusions: This is the first report associating a CLPX variant with SCA in any species. Given the gene’s high evolutionary conservation and known role in mitochondrial protein homeostasis, this finding may have implications for understanding CLPX-related neurodegeneration and anemia in both veterinary and human medicine. Full article

Background/Objectives: Short-read-sequencing (SRS) is currently the standard for genetic testing in inherited human diseases. Intrinsic limitations include PCR dependency, restricted read length, and challenges in identifying structural variants (SVs), copy number variations (CNVs), and intronic small variants (SNVs/indels). Long-read-sequencing (LRS) enables the sequencing of long DNA molecules, detection of deep intronic variants, rapid testing of few samples, and improved resolution of SVs, CNVs, and SNVs/indels. We therefore aimed to validate Oxford Nanopore Technologies (ONT) LRS for potential clinical application. Methods: We evaluated the ONT’s ability to detect pathogenic/likely pathogenic (P/LP) variants previously identified by SRS and confirmed via Sanger sequencing, Multiplex-Ligation-dependent-Probe-Amplification (MLPA), or quantitative-PCR (qPCR). In total, 509 samples were analyzed, including 393 with P/LP variants and 116 negative controls. We used CE-IVD panels HEVA pro, CARDIO pro, BRaCA panel, and ClinEX pro (4Bases-CH). Sequencing was performed on MinION, GridION, and PromethION-2 platforms. Data were analyzed using the 4eVAR pipeline. Results: ONT successfully identified all P/LP variants across the panels (sensitivity 100%); identified a previously missed CNV in ENG gene; precisely defined the breakpoints of a del(13q) (unsuspected and diagnosed as BRCA2 del ex2–14); improved the coverage profiles in difficult-to-map regions (e.g., ex1 TGFBR1, PSM2CL); expanded the coverage of out-of-target deep intronic regions; and allowed for the set-up of fast-track tests (<24 h) for urgent clinical needs. Conclusions: Our findings demonstrate that ONT LRS provides diagnostic performance comparable to SRS, with significant advantages in resolving complex and previously undetectable variants. Ongoing developments are further increasing read length, expanding detectable targets, and potential clinical applications. Full article

Background: The aim of this study is to identify people with cystic fibrosis (pwCF) at risk for future pulmonary exacerbations (PEx) based on established and unestablished markers of chronic inflammation. There is currently no universal definition of PEx in cystic fibrosis (CF), but it is commonly characterized by clinical deterioration and a drop in FEV1 ≥10% with or without elevations in systemic inflammatory markers. PEx negatively affect clinical outcomes in pwCF; therefore, predicting and preventing PEx is a crucial goal in the treatment of pwCF. Methods: We retrospectively examined pwCF ≥18 years who had ≥2 pulmonary function tests per year for a 3-year period. The first year was marked as the baseline. The follow-up period (FU) was defined as the following two-year period after baseline. PEx were defined as a need for intravenous antibiotic treatment due to clinical deterioration. Various scoring systems and ratios (neutrophil/lymphocyte (NLR), lymphocyte/monocyte (LMR), CRP, CRP/albumin, Glasgow Prognostic Score (GPS), high-sensitivity modified Glasgow Prognostic Score (hs-GPS)) were compared in pwCF with and without PEx during the FU. Logistic regression models were used to determine the best marker for predicting PEx, considering factors such as age, sex, PEx at baseline, BMI, homozygote F508del mutation, diabetes mellitus, chronic bacterial infection, and CFTR (cystic fibrosis transmembrane conductance regulator)-modulator therapy. The results are reported as odds ratios (ORs) with p-values. Results: Out of 283 pwCF, 131 were included in the study. In total, 43.5% were female, and the mean age was 34.0 years. A total of 75 pwCF (57.3%) had PEx during FU. In the multivariate analysis, the following markers at baseline were significantly associated with having a PEx during FU: CRP(log) (OR = 7.29, p = 0.01), CRP/albumin (OR = 1.08, p = 0.006), decreased LMR (OR = 0.51, p = 0.02), increased NLR (OR = 1.52, p = 0.02), and GPS of 1 vs. 0 (OR = 2.75, p = 0.04). The results indicate that the CRP/albumin ratio was the best model for predicting PEx in pwCF during the FU, outperforming other models. Conclusions: While several inflammation-based scoring systems can predict PEx in pwCF, the easily calculated CRP/albumin proved to reliably identify pwCF with an increased risk for PEx, making it a promising tool in clinical practice. Full article

Elexacaftor/tezacaftor/ivacaftor (ETI) is a cystic fibrosis (CF) transmembrane conductance regulator modulator, which has shown efficacy in people with CF (pwCF) carrying the F508del (F) variant, both in homozygosity and heterozygosity with a minimal function (MF) variant. Limited data exist on the effects of ETI in pwCF with advanced lung disease. Our aim was to investigate ETI safety and effectiveness in this patient group in a real-life setting over 2 years. A multicenter observational cohort study was designed to gather real-world information on the effect of ETI treatment on CF patients (aged >12 years, genotype: F/MF mutation) with advanced lung disease as defined by a FEV1 < 40% predicted. Retrospective demographic and clinical data were recorded for the two years preceding and the two years following ETI initiation. The following outcomes were investigated: treatment-associated adverse events (AEs), drug interruptions (temporary or permanent), variations in percent predicted FEV1 (ppFEV1), sweat chloride concentration (SwCl), antibiotic use, body mass index (BMI), and quality of life. A total of 124 (51.6% males) pwCF were treated with ETI over 2 years. The median (IQR) age and ppFEV₁ were 34 (26, 43) years and 34 (29, 41) percentage points, respectively. ETI was discontinued in two pwCF due to lung transplantation, and temporarily interrupted in two because of skin rash, and in three following elevated levels of aminotransferase. Most AEs were mild and short-lasting. In 12.1% pwCF, we registered an increase greater than twice the upper limit of the normal range in alanine aminotransferase, and in 16% we registered an increase in conjugated bilirubin with no increase in aminotransferase. Both increases were recurrent in about half of the subjects. The mean differences (95% CI) for ppFEV₁ and SwCl, assessed as mean values in the pre-ETI and ETI treatment periods, were +11.8 (11.1 to 12.6) and −43.7 (−47.6 to −39.9) mmol/L. A modest increase in ppFEV₁ persisted during the second year of treatment. Number of oral and IV antibiotic cycles/year, as well as hospitalizations/year, decreased significantly from 3.6 to 1.2, from 2.4 to 0.6, and from 2.1 to 0.5 during ETI treatment. A total of 8 of 16 (50%) pwCF were taken off the waiting list for lung transplantation, and significant reductions in the percentages of pwCF using long-term oxygen therapy and non-invasive ventilation were observed. A poor concordance between ppFEV1 and SwCl was found. In only 3/82 (3.7%), subjects with chronic airway infection by Pseudomonas aeruginosa cultures were always negative during ETI treatment. In CF patients with advanced lung disease on ETI treatment, we observed an improvement in a number of clinically significant outcomes over a 2-year study period. However, several additional observations, such as liver dysfunction, variable degrees of lung function improvement, and limited impact on chronic airway infection, underscore the fact that the benefit–risk profile of ETI treatment in cystic fibrosis patients with advanced lung disease has not been fully elucidated and warrants prolonged-term monitoring. Full article

Timely reporting of microbiological results is critical for clinical decision-making, particularly in pediatric hospitals where delays can significantly impact outcomes. Despite advances in laboratory automation, workflow inefficiencies and resistance to change remain barriers to improvement in Latin America. This study aimed to evaluate the effect of implementing a Kaizen-based change management strategy on reducing turnaround time (TAT) in the microbiology laboratory of Hospital Roberto del Río, Santiago, Chile. We conducted a prospective, pre–post intervention study focusing on blood culture processing. The baseline period (July 2022) included 961 cultures processed with the BacT/ALERT^® 3D system. A Kaizen/LEAN intervention was designed, comprising workflow redesign, staff training, and installation of the BACT/ALERT^® Virtuo^® (bioMerieux, Marcy l’Etoile, France) continuous-loading blood culture system. The intervention engaged all technical and professional staff in a five-day Kaizen immersion, followed by eight months of monitoring. Outcomes were assessed by comparing TAT for positive blood cultures before and after implementation (June 2023, 496 samples). Statistical analysis was performed using the Mann–Whitney U test, with p < 0.05 considered significant. The intervention achieved a median reduction in TAT from 68.22 h (IQR 56.14–88.59) pre-intervention to 51.52 h (IQR 41.17–66.57) post-intervention, corresponding to a 24.48% improvement (p < 0.001), surpassing the 20% target. Time to preliminary Gram reporting also decreased, and workflow standardization enhanced staff productivity and culture validation frequency. Implementation of Kaizen principles in a pediatric microbiology laboratory significantly reduced blood culture TAT and improved workflow efficiency. Beyond technological upgrades, active staff engagement and structured change management were key to success. These findings support the applicability of Kaizen-based interventions to optimize laboratory performance in resource-constrained public healthcare systems. Full article

Accessory breasts denote the formation of extra breast tissue along the milk line, and are known to be more prevalent among Black and Asian populations, affecting both genders. This first-ever study aimed to determine the genetic aetiology of accessory breasts in a multiplex family, where all female siblings present with bilateral accessory breasts. The study also ascertained secondary findings (SFs) responsible for comorbidities. Clinical data and saliva samples were obtained from all family members. Ultrasound and histopathology confirmed the diagnosis. Whole-exome sequencing was conducted on DNA samples obtained from the saliva, with variant calling conducted utilising the Sentieon workflow. Variant classification was based on American College of Medical Genetics and Genomics guidelines. After segregation analysis, 12 candidate genes emerged. Among these, PRSS50 and FANCC emerged as top candidates, being implicated in breast diseases. However, two variants in FANCC (c.360del; p.His120GlnfsTer24 and c.355_358del; p.Ser119IlefsTer24) were selected as the most probable causal variants because of the role of this gene in hereditary breast and ovarian cancer syndromes. The remaining ten genes were reported as potentially accounting for comorbidities segregating with accessory breasts. Reported SFs involve TTR and RYR1. In conclusion, pathogenic variants in FANCC cause familial accessory breasts. These novel observations impact pathophysiology, genetic counselling, and personalised medicine. Full article

The palatial architecture of the Nasrid Alhambra in Granada was organized around courtyards that have been restored or transformed over the centuries. This research analyzes and graphically recreates a wooden gallery that was built in the Patio del Cuarto Dorado (Courtyard of the Golden Room) by the Catholic Monarchs, which disappeared around 1872. The methodology is based on the compilation of documentary sources and graphic analysis as the basis for new manual and digital drawings. Although no archival documentation detailing its construction or demolition has been identified, a large set of historical images (plans, views, and photographs) has been gathered, analyzed, and arranged chronologically. From these, freehand sketches were drawn to understand its construction elements, using other preserved galleries as a reference. Using this graphic documentation and measurements of the current courtyard, scale drawings were made. All of this allowed for the creation of a reconstructed digital model, the digital fabrication of a small-scale model, and the development of new representational graphics using advanced media. In this way, the aim is to understand and introduce the gallery that occupied this courtyard for centuries, offering a new view of the complex transformations of an architectural complex included on the UNESCO World Heritage List. Full article

Marine protected areas (MPAs) near the coast are a global concern due to potential impact of anthropogenic activities highly relevant when it comes to trace elements pollution in sediment. This study aims to assess the levels of trace elements in sediment, their potential mobility and the ecological risk in Tremiti Islands, a sensitive and vulnerable MPA. Sediment was analyzed for granulometry, mineralogy, pseudo-metal concentrations and available fractions using BCR method. Statistical analysis and different pollution and ecological risk indices were applied to interpret the results, determine the contamination levels and assess the element availability and their potential impact using Sediment Quality Guidelines. Spatial variability in grain size and mineralogy was found across the sampling sites. The finer quartz-rich sediments exhibiting higher trace element concentrations. Site-specific enrichments were evident for As and Zn at Cala Spido and for Pb at Cala Matano. Cu and Mn showed notable potential bioavailability with residual fractions below 30% at all sites; low Cd concentrations were found, but it was highly available. Cala Spido and Grotta del Sale showed higher contamination-degree, while Pagliai and Cala Matano stood out for their higher ecological risk and availability indexes. These findings demonstrated that even within a Marine Protected Area, site-specific anthropogenic pressures can significantly influence sediment quality and ecological risk. Full article