<p>The most widely accepted technique for the confirmation of semen presence in forensic biology is the microscopic identification of sperm cells. The process of manual microscopic examination, however, can often be time consuming, particularly where sperm numbers are low or where sperm cells are absent, particularly when in the presence of numerous non-sperm cells. The findings of this study demonstrate that automated microscopy utilising Deep Neural Network (DNN) allows for an improvement of the efficiency of examination times associated with microscopy. Additionally, the use of instrument and slide data as diagnostic tools permits rapid detection of invalid results. Metafer with Sperm Detection, using DNN v3.1.1, successfully identified sperm cells present in both high and low semen concentrations in mixed-cell substrates, often with a higher accuracy than manual microscopic examinations where very low sperm numbers were present (1\u8211?5 sperm cells on an entire slide). Whilst there were instances of false positive and false negative sperm cell classifications observed, the overall sensitivity, specificity and accuracy of automated microscopy using Metafer with Sperm Detection (DNN v3.1.1) was 0.9999, 0.9996 and 0.9999, respectively. The conversion of Metafer sperm counts to an existing sperm grading system allows compatibility with manual microscopy outcomes and allows decisions regarding sample triage and submission for specific DNA extraction protocols in routine casework to be made with confidence.</p>

Digital object identifier (DOI): 10.1016/j.scijus.2025.101370

<p>Platelets play a critical role in coagulation and are implicated in pathological clotting. This study analyzes platelet activation and morphology using the cellular fraction remaining after whole blood centrifugation. We briefly examine whether the addition of platelet inhibitors, such as Indomethacin and Prostaglandin E1 (PGE-1), affects platelet activation status during sample preparation. Additionally, we introduce an automated analysis method using the MetaCyte Metafer classifier to enhance objectivity and precision in evaluating platelet fragility and activation. Our disease model includes platelets from participants with Long COVID compared to healthy controls. The automated analysis provided consistent, unbiased data, and detailed metrics such as area, count, and roundness, indicating a significant improvement in platelet analysis. We found that platelet inhibitors did not influence platelet activation during sample processing, suggesting that platelets in Long COVID patients are substantially activated in circulation. Automated analysis using the MetaCyte Metafer classifier offers a reliable approach to identifying subtle morphological changes in platelets, including activation size ranges, which can provide valuable insights into the progression of platelet pathology in Long COVID and other thrombo-inflammatory conditions. Future work should focus on refining algorithms and incorporating artificial intelligence (AI) to enhance automated data interpretation and clinical relevance.</p>

Digital object identifier (DOI): 10.1007/s11239-025-03144-9

<p>We conducted a study to validate the automated scoring of Fluorescent in Situ Hybridization (FISH) in a routine cytogenetics laboratory using selected CD138-positive cells in samples from patients with multiple myeloma. A workstation was optimized based on the manufacturer's configurations. Six commercial probes (CDKN2C/CKS1B, RB1/DLEU1/LAMP1, TP53/CEN17, FGFR3::IGH, CCND1::IGH, and IGH::MAF) were examined to detect gains, losses, and rearrangements of genes across a total of 180 slides. We used reference values proposed by the European Myeloma Network (10% for rearrangements and 20% for gains and losses) to compare the accuracy of manual, automated, and semi-automated (automated coupled with manual revision) scoring. The time spent by the biologist to perform semi-automated and manual scoring was compared. Automated scoring was not effective and lacked validation. In contrast, semi-automated scoring proved to be efficient and highly accurate for all probes and offered time-saving benefits for deletion/gain probes. These findings suggest that semi-automated iFISH scoring for multiple myeloma is feasible and could become a routine practice in cytogenetics laboratories.</p>

Digital object identifier (DOI): 10.1002/jemt.24844

<p>Small RNAs interacting with PIWI (piRNAs) play a crucial role in regulating transposable elements and translation during spermatogenesis and are essential in male germ cell development. Disruptions in the piRNA pathway typically lead to severe spermatogenic defects and thus male infertility. The HENMT1 gene is a key player in piRNAs primary biogenesis and dysfunction of HENMT1 protein in meiotic and haploid germ cells resulted in the loss of piRNA methylation, piRNA instability, and TE de-repression. Henmt1-knockout mice exhibit a severe oligo-astheno-teratozoospermia (OAT) phenotype, whereas patients with HENMT1 variants display more severe azoospermia phenotypes, ranging from meiotic arrest to hypospermatogenesis. Through whole-exome sequencing (WES) of infertile patient cohorts, we identified two new patients with variants in the HENMT1 gene presenting spermatozoa in their ejcaulate, providing us the opportunity to study spermatozoa from these patients. Investigate the spermatozoa of two patients harboring an HENMT1 variant to determine whether or not these scarce spermatozoa could be used with assisted reproductive technologies. HENMT1 variants identified by WES were validated through Sanger sequencing. Comprehensive semen analysis was conducted, and sperm cells were subjected to transmission electron microscopy for structural examination, in situ hybridization for aneuploidy assessment, and aniline blue staining for DNA compaction status. Subsequently, we assessed their suitability for in vitro fertilization using intracytoplasmic sperm injection (IVF-ICSI). Our investigations revealed a severe OAT phenotype similar to knockout mice, revealing altered sperm concentration, mobility, morphology, aneuploidy and nuclear compaction defects. Multiple IVF-ICSI attempts were also performed, but no live births were achieved. We confirm the crucial role of HENMT1 in spermatogenesis and highlight a phenotypic continuum associated with HENMT1 variants. Unfortunately, the clinical outcome of these genetic predispositions remains unfavorable, regardless of the patient's phenotype. The presence of spermatozoa is insufficient to anticipate ICSI pregnancy success in HENMT1 patients.</p>

Digital object identifier (DOI): 10.1111/andr.13730

<p><strong>Background: </strong>Acute myeloid leukemia (AML) is a complicated disease with uncontrolled hematopoietic precursor proliferation induced by various genetic alterations. Runt-related transcription factor-1 (<em>RUNX1</em>) is commonly disrupted by chromosomal translocations in hematological malignancies.</p> <p><strong>Aim: </strong>To characterize <em>RUNX1</em> gene rearrangements and copy number variations in newly diagnosed adult AML patients, with an emphasis on the impact of clinical and laboratory features on the outcome.</p> <p><strong>Methods: </strong>Fluorescence in situ hybridization was used to test <em>RUNX1</em> gene alterations in 77 newly diagnosed adult AML cases. <em>NPM1</em>, <em>FLT3/ITD</em>, <em>FLT3/TKD</em>, and <em>KIT</em> mutations were tested by PCR. Prognostic clinical and laboratory findings were studied in relation to <em>RUNX1</em> alterations.</p> <p><strong>Results: </strong><em>RUNX1</em> abnormalities were detected by fluorescence in situ hybridization in 41.6% of patients: 20.8% had translocations, 22.1% had amplification, and 5.2% had deletion. Translocations prevailed in AML-M2 (<em>P</em> = 0.019) with a positive expression of myeloperoxidase (<em>P</em> = 0.031), whereas deletions dominated in M4 and M5 subtypes (<em>P</em> = 0.008) with a positive association with CD64 expression (<em>P</em> = 0.05). The modal chromosomal number was higher in cases having amplifications (<em>P</em> = 0.007) and lower in those with deletions (<em>P</em> = 0.008). <em>RUNX1</em> abnormalities were associated with complex karyotypes (<em>P</em> &lt; 0.001) and were mutually exclusive of <em>NPM1</em> mutations. After 44 months of follow-up, <em>RUNX1</em> abnormalities affected neither patients' response to treatment nor overall survival.</p> <p><strong>Conclusion: </strong><em>RUNX1</em> abnormalities were mutually exclusive of <em>NPM1</em> mutations. <em>RUNX1</em> abnormalities affected neither patients' response to treatment nor overall survival.</p>

The Emergence of Artificial Intelligence-Guided Karyotyping: A Review and Reflection.

<p>Artificial intelligence (AI) has entered the medical subspecialty of cytogenetics with the recent introduction of AI-guided karyotyping into the clinical laboratory. Karyotyping is an essential component of the cytogenetic analysis process; however, it is both labor-intensive and time-consuming. The introduction of AI algorithms into karyotyping software streamlines this process to provide accurate and abundant auto-karyotyped images for laboratory professionals to review and, also, alters the paradigm for chromosome analysis. Herein, we provide an overview of the AI-guided karyotyping products currently available for clinical use, discuss their utilization in the cytogenetics laboratory, and highlight changes AI-guided karyotyping has brought for early users. Finally, we reflect on our own laboratory observations and experience to discuss issues and practices that may need to adapt to best utilize this promising new technology.</p>

Digital object identifier (DOI): 10.3390/genes16060685

<p>The microtubule-associated protein Tau is a driver of neuronal dysfunction in Alzheimer's disease and other tauopathies. In this process, Tau initially undergoes subtle changes to its abundance, subcellular localization and a vast array of post-translational modifications including phosphorylation that progressively result in the protein's somatodendritic accumulation and dysregulation of multiple Tau-dependent cellular processes. Given the various loss- and gain-of-functions of Tau in disease and the brain-wide changes in the proteome that characterize tauopathies, we asked whether targeting Tau would restore the alterations in proteostasis observed in disease. Therefore, by phage display, we generated a novel pan-Tau antibody, RNJ1, that preferentially binds human Tau and neutralizes proteopathic seeding activity in multiple cell lines and benchmarked it against a clinically tested pan-Tau antibody, HJ8.5 (murine version of tilavonemab). We then evaluated both antibodies, alone and in combination, in the K3 tauopathy mouse model, showing reduced Tau pathology and improvements in neuronal function following 14 weekly treatments, without obtaining synergy for the combination. These effects were more pronounced in female mice. To investigate the molecular mechanisms contributing to improvements in neuronal function, we employed quantitative proteomics, phosphoproteomics and kinase prediction analysis to first establish alterations in K3 mice relative to wild-type controls at the proteome level. In female K3 mice, we found 342 differentially abundant proteins, which are predominantly involved in metabolic and microtubule-associated processes, strengthening previously reported findings of defects in several functional domains in multiple tauopathy models. We next asked whether antibody-mediated Tau target engagement indirectly affects levels of deregulated proteins in the K3 model. Importantly, both immunotherapies, in particular RNJ1, induced abundance shifts towards a restoration to wild-type levels (proteostasis). A total of 257 of 342 (∼75%) proteins altered in K3 were closer in abundance to wild-type levels after RNJ1 treatment, and 73% after HJ8.5 treatment. However, the magnitude of these changes was less pronounced than that observed with RNJ1. Furthermore, analysis of the phosphoproteome showed an even stronger restoration effect with RNJ1, with ∼82% of altered phosphopeptides in K3 showing a shift to wild-type levels, and 75% with HJ8.5. Gene set over-representation analysis further confirmed that proteins undergoing restoration are involved in biological pathways affected in K3 mice. Together, our study suggests that a Tau immunotherapy-induced restoration of proteostasis links target engagement and treatment efficacy.</p>

Digital object identifier (DOI): 10.1093/brain/awae254

<p>Tuberculosis (TB) remains a global health challenge, with timely and accurate diagnosis being critical for effective disease management and control. Recent advancements in the field of TB diagnostics have focused on the identification and utilization of blood-based biomarkers, offering a non-invasive, rapid, and scalable approach to disease detection. This review provides a comprehensive overview of the latest progress in blood-based biomarkers for TB, highlighting their potential to revolutionize diagnostic strategies. Furthermore, we explore emerging technologies such as NGS, PET-CT, Xpert and line probe assays, which have enhanced the sensitivity, specificity, and accessibility of biomarker-based diagnostics. The integration of artificial intelligence (AI) and machine learning (ML) in biomarker analysis is also examined, showcasing its potential to improve diagnostic accuracy and predictive capabilities. This review underscores the need for multidisciplinary collaboration and continued innovation to translate these promising technologies into practical, point-of-care solutions. By addressing these challenges, blood-based biomarkers and emerging technologies hold the potential to significantly improve TB diagnosis, ultimately contributing to global efforts to eradicate this devastating disease.</p>

Digital object identifier (DOI): 10.3389/fcimb.2025.1567592

<p>Intestinal parasites pose a significant global public health burden. Approximately 3.5 billion people are affected by intestinal parasitic infections globally, with more than 200,000 deaths reported annually. While diagnostic methods such as PCR and serology have their advantages, they may not be as sensitive or specific as microscopy, which remains the gold standard for identifying parasitic micro-organisms in biological samples. The foundation of this statement remains true, as long as the proficiency of the laboratory technician performing the analysis is maintained to a high standard. In this regard, external quality assurance (EQA) programs are essential for medical laboratories as they help ensure the accuracy and reliability of laboratory testing results and assess the proficiency of the operator. Providing EQA material for parasitology proficiency testing programs (PTPs) has historically presented many challenges. Sourcing samples of sufficient quality and quantity to satisfy the requirements of a parasitology PTP is often a major difficulty. Once a suitable sample has been sourced, additional challenges arise in maintaining sample stability and homogeneity to ensure all participants receive a sample representative of the desired final interpretation. The Royal College of Pathology of Australasia Quality Assurance Programs (RCPAQAP) has begun a pilot study to assess the feasibility of using virtual microscopy to resolve some common problems encountered while procuring and planning parasitology PTPs. Virtual microscopy allows digitisation of parasitology microscope slides for viewing and analysis on a computer or other digital device. RCPAQAP uses the MetaSystems VSlide slide scanner paired with the Zeiss Axio Imager Z2 for virtual slide scanning. This apparatus can capture over 200,000 images per slide across 17 individual focal planes, stitched together into a final ultra-high resolution virtual slide image using Metasystems Metafer software. The final product is a virtual slide that effectively emulates visual microscopy of a physical sample while ensuring each participant is assessed equally. This technology is perfectly suited for this type of sample. It provides solutions to issues related to standardisation across all PTP participants and the transport and storage of EQA materials.</p>

Digital object identifier (DOI): 10.1016/j.ijid.2024.107762

<p>Potable water contain various chemicals, compounds and disinfection by-products. The presence of these substances can result in mutagenic risk to the public, highlighting the need for surveillance. This study evaluated the mutagenic potential of source water and drinking water from two South African drinking water treatment plants. The study also investigated the high-throughput scoring of the <em>Salmonella typhimurium</em> Ames mutagenicity assay with frameshift and base-pair mutations. Two different scoring approaches were used including visual manual scoring and using the automatic image scanning platform. No mutagenic risk was detected for both TA98 and TA100 bacteria regardless of metabolic activation. Grab sampling may have missed any transient mutagenic events. Despite the limitations, automatic scanning of the microtiter plates ensured consistent, reliable and accurate results that can be reviewed. The outcomes of the study show effective mitigation of mutagenic risk by the treatment plants and deliver public reassurance of drinking water. The advantageous combination of automated scoring technologies, as demonstrated in this study, provides a scalable and standardized monitoring programme for mutagenic risk. Regulatory frameworks would benefit from a mutagenic risk monitoring programme, given the myriad health risks involved in exposure to environmental mutagens.</p>

Digital object identifier (DOI): 10.21203/rs.3.rs-7883227/v1

<p>Dicentric chromosome analysis (DCA) has limitations in its use for the evaluation of the radiation dose upon the development of medical radiation equipment due to its time/labour-consuming procedure and the requirement of highly trained experts. Therefore, we aimed to construct a dose–response curve using a semi-automatic cytokinesis-block micronucleus (CBMN) analysis method that can be easily analysed and utilised by anyone. CHO-K1 cells were exposed to gamma rays at various doses (0–4 Gy). For the CBMN assay, the bi-nucleated cells were selected and captured, and micronuclei (MN) scoring was automatically performed using the Metafer4 system. The MN scores were manually confirmed and corrected by analysts. Using the frequency distributions of MN according to the radiation dose, the dose–response calibration curve was generated using Dose Estimate v5.2 software. The equation of dose–response calibration curve is Y = 0.0299 (±0.0057) + 0.1502 (±0.0151) × D + 0.0111 (±0.0048) × D2. The goodness-of-fit parameters were also calculated (chi-squared [χ 2] = 39.45, degrees of freedom = 5, p = 0.0000). The semi-automated CBMN assay consist of two steps: the automated MN capture/scoring step and the manual confirmation/correction step. Using an established dose–response calibration curve and the procedure of the semi-automated CBMN assay, the dose-estimation of gamma-irradiated (0.5 or 2 Gy) CHO-K1 cells were performed by two analysts individually, and it was inter-compared to verify the accuracy, the results showed that the estimated doses were a good fit the applied doses of radiation. The CBMN assay using CHO-K1 cells can be easily used as a biodosimetry tool for dose assessment of medical radiation equipment due to the advantage of being simple, easy, and quick to measure the dose.</p>

Digital object identifier (DOI): 10.14293/genint.15.1.003

<p>The dicentric chromosome assay is a well-established biodosimetric method used to assess absorbed ionizing radiation doses by detecting dicentric chromosomal aberrations. Here, we present a detailed, reproducible protocol for applying the dicentric chromosome assay for in vitro evaluation of radioprotective agents, including novel piperazine derivatives compared with amifostine and its active metabolite WR-1065. The protocol covers all key steps—blood sample preparation, in vitro irradiation, lymphocyte culture, metaphase preparation, and scoring of dicentric chromosomes. It highlights critical stages that affect data quality and reproducibility. Integrating manual scoring with automated analysis using the Metafer system ensures accurate and efficient assessment. Thus, this protocol bridges the fields of biological dosimetry and preclinical screening of radioprotective agents, providing a reliable framework for emergency radiation dose estimation and the development of new radiation medical countermeasures.</p>

Digital object identifier (DOI): 10.1093/biomethods/bpaf058

<p>This article shows the development of a dose–effect calibration curve for X-ray exposures ranging from 0 to 4 Gy using the cytokinesis-block micronucleus assay and automated analysis—the first effort of its kind reported in Latin America. This work establishes a regional benchmark for high-throughput methodologies in cytogenetic biodosimetry, highlighting their potential to improve operational efficiency and reduce response times in radiological emergencies. Methods: Blood samples from six healthy donors were irradiated with X-rays at seven dose levels (0–4 Gy) using a calibrated 6 MV linear accelerator. Two blind samples (1.5 and 3 Gy) were included for validation. The CBMN assay was performed following IAEA protocols, DAPI-stained slides were analyzed using a AxioImager.Z2 automated microscope integrated with MetaSystems Metafer4 and the MNScoreX classifier software. A negative binomial regression model (NB1) was used for model fitting, accounting for overdispersion in micronucleus (MN) frequency. Results: Automated scoring of binucleated lymphocytes showed a dose-dependent increase in MN frequency. The fitted model followed a linear–quadratic relationship: Y = 0.0545 + 0.0448·D + 0.0145·D², with all coefficients statistically significant (p &lt; 0.001). Dose estimates for blinded samples (1.5 and 3 Gy) matched the true doses within 95% confidence intervals, with all z-scores &lt; |3|. Conclusions: The resulting linear–quadratic dose–response curve enabled accurate estimation of blinded sample doses, with all z-scores falling within acceptable fitness-for-purpose thresholds. These results underscore the value of combining automated microscopy with robust statistical modeling to achieve reliable dose assessment, particularly in high-throughput settings and radiological emergency scenarios.</p>

Digital object identifier (DOI): 10.15392/2319-0612.2025.2908

<p>This study investigated the long-term stability of cytogenetic and morphological markers, including dicentric chromosomes (DC), unbalanced translocation (UT), balanced translocation (BT), and Pseudo Pelger-Huët Anomaly (PPHA), in a radiation worker exposed to an acute dose of 60Co-γ radiation. Initial dose assessment, one week after exposure via Thermoluminescent dosimeters (TLDs) and DC, yielded a physical dose of 438.8 mGy and a biological dose of 398 mGy respectively. A follow-up biodosimetry evaluation, conducted 24 years postexposure, yielded a dose estimate of 449 mGy on the basis of BT, closely matching the initial TLD measurement (+ 2.3% relative error). In contrast, the DC, UT, and micronuclei (MN) frequencies fell within the background range, confirming their instability over time. We also assessed the presence of PPHA in blood smears from the same volunteer. PPHA, a morphological marker of neutrophils originating exclusively in vivo from bone marrow progenitor cells, demonstrated a more than twofold increase in frequency compared to background levels, suggesting an association with radiation exposure. This is the first report of concurrent BT and PPHA persistence in the same individual, demonstrating the suitability of these biomarkers for retrospective detection of past radiation exposure. BT offered reliable dose reconstruction decades after exposure. We could not translate the PPHA yield into an absorbed dose, as no in-house dose‒response curve was established.No abstract available.</p>

Digital object identifier (DOI): 10.1038/s41598-025-14858-4

<h3>Objective</h3> <p>To study the feasibility of automated analysis by FISH technique in the determination of the 1p and/or 19q polysomy in oligodendrogliomas (OGs) and to explore its prognostic value.</p> <h3>Methods</h3> <p>We analyzed a retrospective monocentric series of 145 consecutive OGs with IDH mutation and 1p/19q codeletion. For all cases, automated FISH analyses were performed to determine 1p and/or 19q polysomy status and results were compared to manual analysis to verify the concordance of the two methods. Polysomic status was then compared to clinical and histological data, the CDKN2A deletion status when available, event free survival (EFS) and overall survival (OS).</p> <h3>Results</h3> <p>Our study comprised 79 grade 2 OGs (O2) and 66 grade 3 OGs (O3). Polysomy of 1p and/or 19q was observed in 58 cases (40% of whole cohort) with a significant enrichment in the high grade cohort (59% versus 24%; p &lt; 0,0001) and recurrent cases (55%). A majority of polysomic cases were copolysomic for 1p and 19q (75% of the polysomic cohort) rather than 1p or 19q single polysomy (21% and 4% respectively). Polysomy was correlated to high grade histological criteria of high mitotic and Mib1 proliferative indices (p = 0,002 and p = 0,0005 respectively) and to vascular proliferation (p = 0,0003). Univariate and multivariate analysis showed a significant correlation betwen polysomy and a shorter EFS and OS (p = 0,02 and p = 0,016 respectively). Concordance between manual and automated analysis was almost perfect for both 1p and 19q analysis (96 and 98% respectively, κ = 0,92 and 0,95 respectively). Automated analysis revealed that the large majority of polysomic signatures are represented by a small number of R/G signals (mainly 7 signatures) allowing a very easy implementation to pre-existent FISH platforms analysis software.</p> <h3>Conclusion</h3> <p>1p and/ or 19q polysomy status represent a prognostic factor in OGs and can be easily determined by automated analysis. Our study supports the clinical interest to determine the polysomic status in all primitive or recurrent OGs and underline the benefits of automated analysis which offers a better archive storage and facilitates multicentric comparison.</p>

Digital object identifier (DOI): 10.1371/journal.pone.0322809

<p>Several fusion oncogenes showing a higher incidence in pediatric acute myeloid leukemia (AML) are associated with heterogeneous megakaryoblastic and other myeloid features. Here we addressed how developmental mechanisms influence human leukemogenesis by ETO2::GLIS2, associated with dismal prognosis. We created novel ETO2::GLIS2 models of leukemogenesis through lentiviral transduction and CRISPR-Cas9 gene editing of human fetal and post-natal hematopoietic stem/progenitor cells (HSPCs), performed in-depth characterization of ETO2::GLIS2 transformed cells through multiple omics and compared them to patient samples. This led to a preclinical assay using patient-derived-xenograft models to test a combination of two clinically-relevant molecules. We showed that ETO2::GLIS2 expression in primary human fetal CD34 hematopoietic cells led to more efficient in vivo leukemia development than expression in post-natal cells. Moreover, cord blood-derived leukemogenesis has a major dependency on the presence of human cytokines, including IL3 and SCF. Single cell transcriptomes revealed that this cytokine environment controlled two ETO2::GLIS2-transformed states that were also observed in primary patient cells. Importantly, this cytokine sensitivity may be therapeutically-exploited as combined MEK and BCL2 inhibition showed higher efficiency than individual molecules to reduce leukemia progression in vivo. Our study uncovers an interplay between the cytokine milieu and transcriptional programs that extends a developmental window of permissiveness to transformation by the ETO2::GLIS2 AML fusion oncogene, controls the intratumoral cellular heterogeneity, and offers a ground-breaking therapeutical opportunity by a targeted combination strategy.</p>

Digital object identifier (DOI): 10.1186/s12943-024-02110-y

<p>Upon infecting its vertebrate host, the malaria parasite initially invades the liver where it undergoes massive replication, whilst remaining clinically silent. The coordination of host responses across the complex liver tissue during malaria infection remains unexplored. Here, we perform spatial transcriptomics in combination with single-nuclei RNA sequencing over multiple time points to delineate host-pathogen interactions across Plasmodium berghei-infected liver tissues. Our data reveals significant changes in spatial gene expression in the malaria-infected tissues. These include changes related to lipid metabolism in the proximity to sites of Plasmodium infection, distinct inflammation programs between lobular zones, and regions with enrichment of different inflammatory cells, which we term 'inflammatory hotspots'. We also observe significant upregulation of genes involved in inflammation in the control liver tissues of mice injected with mosquito salivary gland components. However, this response is considerably delayed compared to that observed in P. berghei-infected mice. Our study establishes a benchmark for investigating transcriptome changes during host-parasite interactions in tissues, it provides informative insights regarding in vivo study design linked to infection and offers a useful tool for the discovery and validation of de novo intervention strategies aimed at malaria liver stage infection.</p>

Digital object identifier (DOI): 10.1038/s41467-024-51418-2

<p>Human exposure to radiofrequency electromagnetic fields (RF-EMF) is restricted to prevent thermal effects in the tissue. However, at very low intensity exposure "non-thermal" biological effects, like oxidative stress, DNA or chromosomal aberrations, etc. collectively termed genomic-instability can occur after few hours. Little is known about chronic (years long) exposure with non-thermal RF-EMF. We identified two neighboring housing estates in a rural region with residents exposed to either relatively low (control-group) or relatively high (exposed-group) RF-EMF emitted from nearby mobile phone base stations (MPBS). 24 healthy adults that lived in their homes at least for 5 years volunteered. The homes were surveyed for common types of EMF, blood samples were tested for oxidative status, transient DNA alterations, permanent chromosomal damage, and specific cancer related genetic markers, like MLL gene rearrangements. We documented possible confounders, like age, sex, nutrition, life-exposure to ionizing radiation (X-rays), occupational exposures, etc. The groups matched well, age, sex, lifestyle and occupational risk factors were similar. The years long exposure had no measurable effect on MLL gene rearrangements and c-Abl-gene transcription modification. Associated with higher exposure, we found higher levels of lipid oxidation and oxidative DNA-lesions, though not statistically significant. DNA double strand breaks, micronuclei, ring chromosomes, and acentric chromosomes were not significantly different between the groups. Chromosomal aberrations like dicentric chromosomes (p=0.007), chromatid gaps (p=0.019), chromosomal fragments (p&lt;0.001) and the total of chromosomal aberrations (p&lt;0.001) were significantly higher in the exposed group. No potential confounder interfered with these findings. Increased rates of chromosomal aberrations as linked to excess exposure with ionizing radiation may also occur with non-ionizing radiation exposure. Biological endpoints can be informative for designing exposure limitation strategies. Further research is warranted to investigate the dose-effect-relationship between both, exposure intensity and exposure time, to account for endpoint accumulations after years of exposure. As established for ionizing radiation, chromosomal aberrations could contribute to the definition of protection thresholds, as their rate reflects exposure intensity and exposure time.</p>

Digital object identifier (DOI): 10.1016/j.ecoenv.2024.116486

<p>Astronauts travelling in space will be exposed to mixed beams of particle radiation and photons. Exposure limits that correspond to defined cancer risk are calculated by multiplying absorbed doses by a radiation-type specific quality factor that reflects the biological effectiveness of the particle without considering possible interaction with photons. We have shown previously that alpha radiation and X-rays may interact resulting in synergistic DNA damage responses in human peripheral blood lymphocytes but the level of intra-individual variability was high. In order to assess the variability and validate the synergism, blood from two male donors was drawn at 9 time points during 3 seasons of the year and exposed to 0-2 Gy of X-rays, alpha particles or 1:1 mixture of both (half the dose each). DNA damage response was quantified by chromosomal aberrations and by mRNA levels of 3 radiation-responsive genes FDXR, CDKN1A and MDM2 measured 24 h post exposure. The quality of response in terms of differential expression of alternative transcripts was assessed by using two primer pairs per gene. A consistently higher than expected effect of mixed beams was found in both donors for chromosomal aberrations and gene expression with some seasonal variability for the latter. No synergy was detected for alternative transcription.</p>

Digital object identifier (DOI): 10.1038/s41598-024-62313-7

<p>Manual microscopy of Gram stains from positive blood cultures (PBCs) is crucial for diagnosing bloodstream infections but remains labor intensive, time consuming, and subjective. This study aimed to evaluate a scan and analysis system that combines fully automated digital microscopy with deep convolutional neural networks (CNNs) to assist the interpretation of Gram stains from PBCs for routine laboratory use. The CNN was trained to classify images of Gram stains based on staining and morphology into seven different classes: background/false-positive, Gram-positive cocci in clusters (GPCCL), Gram-positive cocci in pairs (GPCP), Gram-positive cocci in chains (GPCC), rod-shaped bacilli (RSB), yeasts, and polymicrobial specimens. A total of 1,555 Gram-stained slides of PBCs were scanned, pre-classified, and reviewed by medical professionals. The results of assisted Gram stain interpretation were compared to those of manual microscopy and cultural species identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The comparison of assisted Gram stain interpretation and manual microscopy yielded positive/negative percent agreement values of 95.8%/98.0% (GPCCL), 87.6%/99.3% (GPCP/GPCC), 97.4%/97.8% (RSB), 83.3%/99.3% (yeasts), and 87.0%/98.5% (negative/false positive). The assisted Gram stain interpretation, when compared to MALDI-TOF MS species identification, also yielded similar results. During the analytical performance study, assisted interpretation showed excellent reproducibility and repeatability. Any microorganism in PBCs should be detectable at the determined limit of detection of 10 CFU/mL. Although the CNN-based interpretation of Gram stains from PBCs is not yet ready for clinical implementation, it has potential for future integration and advancement.</p>

Digital object identifier (DOI): 10.1128/jcm.00876-23