Eurofins (France)

Zoonotic Salmonella is one of the most important foodborne zoonoses worldwide, with Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST) being the most important serovars, which may be transmitted via poultry meat and eggs. Vaccination of breeding- and laying-hen flocks using live vaccines administered via drinking water has long been a well-recognized tool and contributed significantly to decreasing cases of salmonellosis in humans and its prevalence in the chicken sector. However, if a vaccine strain is detected in feces from recently vaccinated birds, it is imperative to have methods for reliably distinguishing between field and vaccine strains. In this study, we validated a disc diffusion method based on the antimicrobial resistance patterns of the vaccine strains contained in a live bivalent vaccine conferring protection against SE and ST. Discs from eight suppliers were sourced and tested for their suitability for reliably distinguishing between field and vaccine strains, a requirement fulfilled by discs from four suppliers. This method serves as an alternative approach for laboratories equipped to perform disc diffusion assays, and it is also a more cost-effective solution than real-time PCR assays.

MECOM encodes a developmental and haematopoietic transcription factor associated with a rare early-onset syndrome including bone marrow failure, skeletal and other congenital anomalies. Heterozygous de novo variants are the primary cause. We previously identified MECOM as a candidate gene for paediatric pulmonary arterial hypertension (PAH) using trio exome sequencing. To test the role of MECOM in paediatric PAH and further define the clinical phenotype of MECOM-associated syndrome, we queried GeneMatcher and screened rare disease databases for individuals with predicted deleterious MECOM variants. We analysed the clinical spectrum of patients, performed protein modelling of genetic variants and assessed cardiopulmonary expression. We identified 15 individuals with MECOM variants, including 11 unrelated probands and 8 de novo variants. 11 individuals had severe or mild thrombocytopenia, 9 had skeletal issues, 8 had cardiac anomalies, 6 had PAH and 10 had additional conditions. Three were diagnosed in utero and died in the neonatal period. All missense variants map to the zinc finger 6 or zinc finger 8/9 region, a known hotspot for MECOM-associated syndrome. Protein modelling predicted that both regions are DNA-binding, and that the variants may interfere with binding to a VEGFR2/KDR enhancer. Data from LungMAP showed that MECOM is primarily expressed in pulmonary arterial endothelial cells. Rare MECOM variants are associated with early-onset syndromic PAH. PAH monitoring should be considered for all individuals with rare MECOM variants. We speculate that the pathogenetic mechanism for PAH and cardiac defects may be impaired VEGFR2/KDR signalling.

Being topmost cancer in India, oral cancer management warrants discovery of novel biomarkers, treatment strategies, and targets to help with early diagnosis, treatment, and recovery. To have a continuous supply of cells, the study was aimed at generation and characterization of established cell line from buccal mucosa (BM) tumors from patients of Indian origin which can be developed as a pre-clinical tool for biomedical application. Surgically resected tumor tissue from histo-pathologically confirmed oral cancer were processed for explant culture. TBM-02 cell line was passaged and characterized for morphology and function. Further, the cell line was silenced for inflammasome pathway gene NLRP3 to evaluate its linkage with oral cancer tumorigenesis. TBM-02, successfully established from BM, was maintained up to 100 passages, exhibited epithelioid morphology, high EpCam expression and triploid ploidy with chromosomal aberrations. Novelty and human origin of TBM-02 was authenticated by Short Tandem Repeats profiling and comparison with DSMZ database. TBM-02 revealed tumorigenic potential in vitro and in vivo which was abrogated on silencing NLRP3. Increased expression of NLRP3, hallmark of chronic inflammation in TBM-02, was validated at protein and gene level and in xenograft. TBM-02 demonstrated migratory potential and was found to be a sensitive tool to study drug response. RNA sequencing demonstrated upregulation of oral cancer-associated genes and pathways. Thus, in current study, we have reported development of novel cell line from early-stage buccal mucosa cancer patient which has a strong potential to be developed and to be used as pre-clinical model for improving oral cancer management and therapeutics.Graphical Supplementary InformationThe online version contains supplementary material available at 10.1007/s13577-025-01332-6.

ABSTRACTDuctal plate malformations (DPM) encompass a spectrum of congenital liver disorders characterized by abnormal bile duct development, often associated with conditions such as Caroli disease. Variants in the PKHD1 gene cause a wide spectrum of DPM, but genotype–phenotype correlations remain challenging. We report a couple with two consecutive terminated pregnancies following prenatal detection of hepatic anomalies suggestive of DPM. Genetic analyses revealed compound heterozygous variants in PKHD1 in both fetuses. One variant (c.931A>G) was classified as likely pathogenic, while the second (c.533T>A), initially reported as a variant of uncertain significance, was reclassified as likely pathogenic after recurrence of the phenotype. This case highlights the importance of integrating prenatal imaging, postmortem examination, and whole‐gene sequencing to refine variant classification and improve genetic counseling. Furthermore, it expands the clinical spectrum of PKHD1‐related disorders.

<p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="first" dir="auto" id="d8896998e85">The BioMAP® phenotypic profiling platform provides an unbiased, target-agnostic and data-driven approach to understanding the effects of new therapies on human tissue biology. This is accomplished through use of human primary cells from different tissues in complex (often co-) culture conditions stimulated to recapitulate specific tissue and disease states. These models or systems include endothelial and epithelial cells, fibroblasts, PBMCs, and other cell types under experimental conditions that accurately represent inflammation, fibrosis, immune responses, respiratory, dermal, and cardiovascular biology, and have been qualified using clinical agents where their effects are already known. When novel test agents are evaluated against twelve such systems, the measurement of 148 translational protein biomarkers that are relevant to the tissue biology being modelled generates a phenotypic “fingerprint” profile. Enhanced data analytics and bioinformatics-based approaches allow statistical correlation of a new profile with those generated from the testing of over 4500 substances, allowing mechanism of action identification and clustering, benchmarking with standard of care drugs, and predictive efficacy and potential safety/toxicity determinations. <p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" dir="auto" id="d8896998e87">Nintedanib was originally developed to treat Idiopathic Pulmonary Fibrosis (IPF) but has also been approved to treat various proliferative diseases, including cancer, and is under review as a treatment option for cardiac fibrosis. We will show how anti-fibrotic aspects of nintedanib’s BioMAP profile are similar to those of pirfenidone, a drug that is also approved for treating IPF. However, nintedanib’s additional activities support its efficacy as an anti-cancer drug, which is further underlined by demonstrated similarities to other approved anti-cancer drugs. <p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" dir="auto" id="d8896998e89">Another prominent example is aspirin: originally developed for pain and inflammation, BioMAP data also support its use as an anti-coagulant and show potential for treating cancer, which represent other possible applications for this well-known drug. <p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" dir="auto" id="d8896998e91">Taken together, these examples highlight the suitability of BioMAP for drug repurposing.

Wee1-like protein kinase (Wee1) is a serine/threonine kinase essential for cell cycle regulation and the DNA damage response (DDR), acting as a tumor suppressor in normal cells. Its overexpression in cancers such as melanoma, hepatocellular carcinoma, and glioblastoma underscores its potential as a therapeutic target. Wee1 inhibition has emerged as a strategy to sensitize tumor cells to genotoxic therapies, though no inhibitors have yet received broad clinical approval. In the first round of screening, we employed ESSENCE-Dock virtual screening to identify potential Wee1 inhibitors from the Eurofins-Villapharma library. Of the 12 compounds experimentally validated, six exhibited IC50 values below 100 µM, with compound 1 demonstrating promising activity (IC50 = 240 nM). Based on this initial hit, we synthesized and tested additional compounds, culminating in compound 20, which showed an almost order of magnitude improvement in potency (IC50 = 33 nM). Structure–activity relationship (SAR) analysis highlighted key activity determinants, including the effects of methyl substitution on the phenylpiperazine and double-chlorine halogenation on the terminal phenyl group. Compound 20 compared favorably to known Wee1 inhibitors such as MK-1775, while maintaining a distinct chemical scaffold. To further validate these findings, molecular dynamics simulations were employed to elucidate the binding interactions and conformational dynamics of the high-affinity compounds. These simulations revealed stabilizing interactions that aligned with the observed SAR trends, solidifying compound 20 as a strong lead for further development. In addition, in vitro ADME profiling demonstrated favorable physicochemical properties and drug-likeness, including high solubility and metabolic stability, supporting its potential as a drug candidate. Kinome-wide profiling at 10 µM revealed modest selectivity, highlighting a key consideration for future optimization.

Objective: Ventriculomegaly is the main prenatal imaging feature for diagnosing fetal central nervous system anomalies in humans. Many ventriculomegalies can be related to genetic causes, regardless of their imaging presentations. Among these, MPDZ variants have been reported to cause severe ventriculomegaly inherited in an autosomal recessive manner (OMIM#615219). Several hypotheses have been put forward linking MPDZ variants to ventriculomegaly, but the precise underlying mechanisms, in particular whether its origin is obstructive or non-obstructive, are yet to be elucidated. Methods: To address this question, we retrospectively analyzed pre- and postnatal neuro-imaging and neuropathological data for cases of ventriculomegaly in which MPDZ variants were found through exome or genome sequencing. We performed anti-MPDZ immunostaining on fetal brain samples. Results: We analyzed six cases (four fetuses and two children) of ventriculomegaly of variable severities with MPDZ variants. The precise analysis of brain MRI data, corroborated by fetopathological examinations, demonstrated an obstructive pattern of ventriculomegaly upstream from partial fusion of the thalami, also called diencephalosynapsis, with partial atresia of the third ventricle, which could extend to Sylvius’s aqueduct. Conclusions: The morphological analysis using targeted brain magnetic resonance imaging (MRI) and neuropathological data allowed us to unravel the underlying mechanisms of congenital ventriculomegaly related to MDPZ variants.