Search Results (471)

Cronobacter spp. are opportunistic foodborne pathogens that can cause neonatal meningitis, necrotizing enterocolitis, and sepsis. This study conducted a systematic contamination survey and whole-genome epidemiological analysis of 562 low-water-activity food samples in Hunan Province of China. The results showed an overall Cronobacter spp. detection rate of 41.99% (236/562), with spices exhibiting the highest contamination rate (60.06%), and with high-level contamination samples (>110 MPN/g) concentrated in this category. The 236 isolates comprised 6 species, 120 sequence types, and 39 clonal complexes, with C. sakazakii being the most frequently isolated species (64.83%) and high-risk clones ST4, ST1, ST148, and ST64 prevailing. Multiple virulence genes (TraJ, fur, rcsAB, rpoS) and antimicrobial resistance genes (qnrS1, blaTEM-1, blaCTX-M-55, blaLAP-2, aac(3)-IId, aadA2, tet(A), floR, mcr-9.1, sul2) were detected. Core genome multilocus sequence typing (cgMLST) identified two clustering patterns: Cluster C, whose genetic clustering was consistent with transmission associated with potential common upstream raw materials across different brands and provinces, and Cluster G, whose clustering suggested potential persistent colonization in the production environment across multiple batches of the same brand. This study elucidates the contamination characteristics of Cronobacter spp. in low-water-activity foods from Hunan Province and provides a basis for WGS-based active surveillance and supply chain traceability. Full article

Background: The detection of JAK2 exon 12 mutations is important for the differential diagnosis of myeloproliferative neoplasms (MPN) and is included in the World Health Organization’s diagnostic criteria for polycythemia vera (PV). We developed and evaluated a pyrosequencing-based technique to detect somatic mutations in JAK2 exon 12 and tested the method in a group of PV patients. Methods: PCR and pyrosequencing primers were designed for region JAK2 exon 12; PCR conditions were optimized for subsequent qualitative and quantitative detection by pyrosequencing. Diagnostic specificity and sensitivity were determined using plasmid controls. Genomic DNA from 145 MPN patients was used to validate the method. Results: The analytical characteristics of the method were as follows: the limit of blank was 1.2–7.4% and the limit of detection was 3.4–9.9% depending on mutation type. DNA samples from Russian patients with clinical evidence of MPNs were analyzed. In 7 of 145 cases (4.8%) JAK2 exon 12 mutations were detected. One patient had the mutation I540-E543insdelTCAGAAATG<AAA. The A-homopolimer insertion in this complex mutation type could not be clearly identified by pyrosequencing, and Sanger confirmation was required. For another patient with N542-E543delAATGAA mutation, we observed the mutant allele burden growing from 15 to 55% between 2014 and 2018 and further reduction during treatment to 11%. Conclusions: The developed pyrosequencing-based method presents a simple solution to qualitative and quantitative JAK2 exon 12 mutations detection. However, some complex mutation types may require manual interpretation and Sanger confirmation. Full article

Background/Objectives: Chronic myeloid leukemia (CML), essential thrombocythemia (ET), primary myelofibrosis (PMF), and polycythemia vera (PV) are myeloproliferative neoplasms (MPNs) that require precise molecular characterization. Although driver mutations such as BCR-ABL1 and JAK2 are diagnostically important, they do not fully explain disease heterogeneity. The NanoString nCounter^® system enables direct multiplex gene expression analysis without RNA amplification and is suitable for degraded bone marrow specimens. This study aimed to analyze cytokine gene expression in bone marrow mononuclear cells of patients with MPNs and controls using NanoString technology, identify differentially expressed genes (DEGs) among MPN subtypes, and investigate their biological significance. Methods: Bone marrow aspirates were collected from 19 patients with MPNs (CML, ET, PMF, and PV) and 6 control patients. Mononuclear cells were isolated, and RNA expression of a 40-gene cytokine panel was analyzed using the NanoString nCounter^® system with strict quality control and normalization. DEGs were identified for each MPN subtype, followed by Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analyses. Results: CML and PV demonstrated 20 and 12 DEGs, respectively. In contrast, ET showed only one DEG (IRAK2), and PMF showed none. Functional analyses revealed enrichment of cytokine signaling, Toll-like receptor (TLR), and JAK-STAT pathways in CML, indicating immune and inflammatory dysregulation. PV DEGs were associated with TLR signaling, IL-17 pathways, and cytokine–cytokine receptor interactions, suggesting active cytokine-mediated inflammation. Conclusions: CML and PV exhibited distinct cytokine-driven transcriptional signatures, whereas ET and PMF exhibited minimal alterations. These findings support the clinical utility of NanoString technology for bone marrow specimens and highlight disease-specific immune pathways as potential diagnostic biomarkers in MPNs. Full article

Myeloid malignancies encompass a heterogeneous group of haematological disorders, primarily including myelodysplastic syndromes (MDSs) and myeloproliferative neoplasms (MPNs). MDS is characterised by defective myeloid cell maturation, while MPNs involve the pathological overproduction of myeloid lineage cells. In the absence of timely diagnosis and effective clinical intervention, both entities carry a substantial risk of progression to acute myeloid leukaemia (AML). Although allogeneic haematopoietic stem cell transplantation remains the only potentially curative therapy, its application is frequently constrained by patient-related factors such as advanced age and comorbid conditions. While currently, hypomethylating agent therapy (azacitidine and decitabine) is mainly used in high-risk MDS patients, and ruxolitinib is primarily used in symptomatic primary myelofibrosis (PMF-MPN), their clinical efficacy remains suboptimal. More recently, focus has turned toward the role of the tumour microenvironment (TME) in disease pathogenesis and whether therapeutically targeting the TME, either alone or in combination with conventional therapy, could present a new treatment option. Emerging evidence underscores the significant influence of TME components, particularly macrophages and T cells, in modulating immune responses and shaping the leukaemic niche to either facilitate or hinder malignant progression. In response, a new generation of immune checkpoint inhibitors are being developed to target the TME, including PD-1/CTLA-4 blockers, macrophage-directed agents including CD47 inhibitors, and T cell-targeting checkpoint inhibitors such as TIM-1 and LAG-3. This review will describe the functional role of key TME constituents in the progression of myeloid malignancies and explore the current landscape and future potential of advanced cellular and molecular immunotherapies in the treatment of these disorders. Full article

Introduction: The concomitant occurrence of myeloproliferative neoplasms (MPNs) and plasma cell dyscrasias is rare and presents significant diagnostic challenges. Accurate distinction between overlapping features is essential, particularly when bone marrow fibrosis (BMF) is present. Case Description: We report a 57-year-old female, with a 10-year history of thrombocytosis managed with antiplatelet therapy, who presented with anemia and severe lumbar pain. Bone marrow biopsy revealed marked fibrosis, and imaging revealed multiple vertebral lesions. Diagnostic workup identified features consistent with myelofibrosis (MF) and coexisting IgG-Kappa multiple myeloma (MM). Although the patient initially fulfilled WHO criteria for MF, the rapid resolution of fibrosis following first-line plasma-cell-directed therapy suggested a secondary, cytokine-mediated process rather than a true concomitant MPN. Conclusions: This case highlights the importance of an integrated diagnostic approach in patients with overlapping features of hematologic malignancies. Differentiating between MM-associated fibrosis and true concurrent MPN and MM is critical, as misclassification may alter both prognosis and therapeutic strategy. In triple-negative cases, the histologic response to plasma-cell-directed therapy can serve as a key discriminating criterion. Awareness of the potential association between MM with fibrosis and extramedullary disease is also essential for clinical management. This case underscores the importance of an integrated diagnostic approach in patients with overlapping hematologic features. Full article

The disinfection process in wastewater treatment is key to the discharge and/or reuse of high-quality effluent. However, disinfection using ultraviolet (UV) light may be inefficient because bacteria possess mechanisms for repairing damaged DNA. This study aimed to assess the photoreactivation and dark repair of total coliform (TC) in wastewater effluent after UV-C disinfection treatment. Four UV-C doses (28.8, 53.1, 57.6, and 106.2 mJ/cm²) and two post-irradiation conditions (light vs. darkness) were applied. Reactivation was monitored after 2, 4, 6 and 24 h (25 °C). Similar TC inactivation efficiencies were observed for the three lowest UV-C doses, whereas the 106.2 mJ/cm² dose achieved the greatest reduction (1.1 Log of TC), decreasing TC concentrations from 3.1 × 10⁵ ± 3.5 × 10⁵ to 1.2 × 10⁵ ± 1.4 × 10⁵ MPN/100 mL. Reactivation assays revealed substantial bacterial recovery after UV treatment, with 24 h survival rates up to 2.3 × 10³ under light and 9.2 × 10² in darkness. Photoreactivation and dark repair assays revealed substantial variability in bacterial recovery after UV treatment depending on UV-C dose, post-irradiation condition and incubation time. In general, bacterial recovery was still detected even at the 106.2 mJ/cm² dose, particularly after 24 h of incubation (178–604%). These findings suggest that effective organic matter removal before UV-C disinfection is critical to improve UV transmittance, reduce shielding effects, and limit subsequent bacterial recovery. Full article

Background/Objectives: Somatic CALR gene insertions/deletions in exon 9, causing frameshift, are a diagnostic sign of myeloproliferative neoplasms (MPNs). Besides the most common somatic mutations of type I (52 bp deletion) and type II (5 bp insertion), there are rare ones whose significance is not always clear. This study evaluates the frequency of rare mutations and demonstrates a germline rather than somatic nature for some of them. Methods: A retrospective analysis of 8417 blood samples subjected to molecular diagnosis of myeloproliferative neoplasm (MPN) was performed. Cases suspected as germline variants were sequenced, and paired samples (when available) of buccal epithelium were analyzed. Results: We have identified 632 CALR gene mutation-positive cases. Most of the cases were typical insertions/deletions (5 bp/52 bp). Non-type I/II frameshift or nonframeshift mutations were observed in 68 cases (11%). The buccal swab samples obtained from 4 patients confirmed the germline nature of these variants. It is worth noting that the MPN diagnosis for three of these patients was made considering the presence of the JAK2 V617F mutation (two cases) or BCR::ABL1 translocation (one case). In one case, the diagnosis of MPN was reclassified to CML. Conclusions: Non-type I/II CALR mutations, according to our data, could be found in 0.8% of MPN-suspected cases, and may not be associated with the diagnosis. The detection of a non-standard CALR mutation with an allelic frequency close to 50% should raise suspicion of the possibility of a germline CALR variant, and such cases should be investigated further. Full article

Surface water degradation has intensified due to anthropogenic pressures, especially in urban areas, where unplanned land use compromises the integrity of aquatic ecosystems. This study investigated the relationship between water quality and land use in a Permanent Preservation Area (PPA) within an urban perimeter in Caçador, Santa Catarina, Brazil. Monthly sampling was conducted throughout 2024 at 11 points distributed along urban and rural sections of the river and its tributaries. Physicochemical and microbiological parameters were evaluated, and the Water Quality Index (WQI) established by the National Sanitation Foundation (NSF) was calculated in order to associate the results with the sampling points, complemented by Principal Component Analysis (PCA) to identify multivariate patterns of spatial variability in water quality across the study area. In parallel, the PPA within the urban perimeter was delimited according to current environmental legislation, and land use was classified using ArcGIS and Google Earth Pro. The results revealed greater water quality degradation in urban stretches of the river, particularly at sampling point SP7, which recorded the lowest dissolved oxygen concentration (3.10 mg L⁻¹), alongside elevated values of biochemical oxygen demand (5.23 mg L⁻¹), total phosphorus (2.94 mg L⁻¹), nitrate (18.75 mg L⁻¹), and thermotolerant coliforms (2759.20 MPN 100 mL⁻¹). The WQI ranged from 40.18 (SP7: bad category) to 73.57 (SP1: good category), reflecting a pronounced spatial gradient of water quality degradation associated with increasing urbanization along the river course. Mapping of the PPAs revealed that only 43.72% of the total area was covered by native vegetation, while the remaining 56.28% was occupied by anthropogenic land uses, including miscellaneous use (30.32%), agriculture (9.09%), buildings (6.09%), roads (4.64%), and railway infrastructure (5.81%). PCA accounted for 89.06% of the total data variance and indicated that greater interaction of sampling points with urbanized areas was consistently associated with reduced water quality, thereby demonstrating the direct influence of anthropogenic activities on the environmental parameters assessed throughout the study area. These findings demonstrate that land use patterns directly affect water quality and reinforce the need for riparian forest restoration, expanded sanitation infrastructure, and more sustainable urban planning. Full article

Drought is a critical constraint for legume production in semi-arid regions, yet breeding for drought tolerance in faba bean through induced mutagenesis remains largely unexplored. To our knowledge, this is the first EMS-derived mutant population in faba bean specifically developed for drought tolerance, comprising 45 M₂/M₃ lines derived from small-seeded cv. Zina and large-seeded cv. Aguadulce Superlonga), evaluated under two irrigation regimes—100% field capacity (well-watered control) and 40% field capacity (severe stress)—over two consecutive growing seasons in a randomized complete block design with three replications. Drought stress caused severe yield losses, reducing mean seed number per plant by 42.2% and mean seed weight per plant by 47.1%. Analysis of variance revealed highly significant effects of genotype, irrigation, and generation/year on both yield components. The non-significant genotype × irrigation interaction indicated similar proportional drought response across genotypes, while the non-significant three-way interaction suggested relatively consistent genotype rankings across generations/growing seasons. Among the ten drought tolerance indices evaluated, seed-number-based mean productivity (MPn) and stress tolerance index (STIn) were the most discriminating, whereas weight-based indices failed to differentiate genotypes due to the inherent seed-size contrast between botanical backgrounds. Dunnett’s comparisons identified genotype 23 (Zina-derived) as the top performer, significantly exceeding its parent for both MPn and STIn; genotypes 22, 24, 12, 3, and 15 similarly outperformed controls. Cluster analysis broadly distinguished three groups: a tolerant cluster dominated by Zina-derived lines, a moderately tolerant cluster (Zina wild-type), and a sensitive cluster of Aguadulce Superlonga-derived lines. These findings suggest that EMS mutagenesis generated potentially heritable and exploitable variation for drought tolerance, with selected lines representing promising candidates for further multi-environment validation. Full article

Myeloproliferative neoplasms (MPNs) are a heterogenous group of myeloid disorders including polycythemia vera (PV), essential thrombocythemia (ET) and myelofibrosis (MF). All these conditions can be associated with significant morbidity including increased risk of thrombotic events as well as reduced survival. Selecting therapy is dependent on patients’ risk assessment and thus prognostication is vital in the management of these conditions. There have been significant developments in prognostic and risk stratification models for MPNs over the last few years, including incorporation of molecular markers. This narrative review article aims to summarize these prognostic models and provide practical advice on how clinicians can utilize these tools to develop personalized treatment strategies for MPN patients. Full article

Bivalve mollusks efficiently bioaccumulate human enteric viruses, posing significant food safety risks. This study assessed the prevalence of Norovirus (NoV GI and NoV GII), Hepatitis A virus (HAV), and Hepatitis E virus (HEV) in 59 samples of live mussels (Mytilus galloprovincialis) collected from the Bulgarian Black Sea coast between July 2022 and July 2023. Viral detection was performed using one-step real-time reverse transcription-polymerase chain reaction (RT-qPCR) following ISO 15216-2 standards, with a mean extraction efficiency of 4.06%. Norovirus GII was the most prevalent pathogen, with detection peaks following intense rainfall events in July 2023. In contrast, all samples tested negative for HAV and HEV. The analysis showed no significant correlation between E. coli contamination levels and the presence of NoV (Mann–Whitney U test, p = 0.565). The viral RNA was detected in several samples that otherwise complied with regulatory bacterial standards for direct consumption (≤230 MPN/100 g). In conclusion, within the limitations of the evaluated sample size and the specific geographically unbalanced sampling design, NoV GII was the predominant genogroup detected. These results suggest that current bacterial indicators may be insufficient to ensure viral safety in these products. In this regard, national control authorities need to undertake timely policies and measures for better and adequate surveillance, control and prevention of viruses in the different parts of the food chain. Full article

Juvenile myelomonocytic leukemia (JMML) is a rare, aggressive myeloproliferative neoplasm of early childhood characterized by constitutive activation of the RAS-MAPK signaling pathway. RASopathies are a heterogeneous group of complex genetic disorders arising from germline mutations that dysregulate RAS-MAPK signaling. Noonan syndrome, CBL syndrome, and neurofibromatosis type 1 (NF1) are the three major RASopathies predisposing to JMML. More than 90% of JMML cases harbor germline or somatic mutations in one of five canonical driver genes—PTPN11, NRAS, KRAS, NF1, or CBL—establishing JMML as the prototypical malignant manifestation of RASopathy biology. The fifth edition of the World Health Organization Classification of Tumours reclassified JMML as a myeloproliferative neoplasm while the International Consensus Classification adopted JMML under pediatric and/or germline mutation-associated disorders, introducing a JMML-like category for cases lacking five canonical mutations but harboring emerging drivers such as SH2B3::LNK alterations and ALK::ROS1 fusions. The distinction between germline and somatic mutations profoundly influences prognosis: e.g., germline PTPN11-associated myeloproliferations and many germline CBL cases undergo spontaneous resolution, whereas somatic PTPN11- and NF1-mutated JMML is more aggressive and requires prompt allogeneic hematopoietic stem cell transplantation. DNA methylation profiling has emerged as the most robust prognostic framework, with consensus defining high-, intermediate-, and low-methylation subgroups that independently predict outcome. Both genotype and DNA methylation subclassification have been integrated into clinical decision-making, incorporating pretransplant azacitidine, watch-and-wait approaches for favorable-risk patients, and emerging targeted therapies including MEK inhibitors. This review synthesizes recent advances in understanding JMML as a bona fide RASopathy; provides a diagnostic algorithm, molecular landscapes, and prognostic models; and highlights opportunities for molecularly targeted therapeutic intervention. Full article

Fe³⁺–polyphenol coordination complexes have attracted growing interest for photothermal applications due to their tunable chemistry and good biocompatibility. However, how pH and the metal-to-ligand ratio collectively affect their photothermal performance remains poorly understood. In this work, we synthesized Fe³⁺–gallic acid (GA) metal–phenolic networks (MPNs) under a wide range of pH conditions and different mixing ratios. The materials were then characterized through electron microscopy, infrared spectroscopy, UV-vis absorption, and photothermal testing. Our results show that a near-neutral pH (around 7) is critical for forming an effective ligand-to-metal charge transfer complex, which appears as a distinct absorption band near 560 nm. Acidic or strongly alkaline environments severely disrupt coordination and weaken light absorption. Among all formulations, the sample prepared at pH 7 with a suitable Fe³⁺/GA ratio gave the best photothermal conversion, reaching a temperature rise of 42.8 °C and a photothermal conversion efficiency of 32.67%. We also found that photothermal heating increases steadily with GA concentration and peaks sharply at neutral pH. These findings demonstrate that optimal photothermal efficiency requires both neutral pH and a well-balanced metal-to-ligand ratio. This work provides a simple and practical set of conditions for developing high-performance Fe³⁺-GA MPNs for applications such as local heating, antibacterial surfaces, and light-triggered drug release. Full article

Background: Myeloproliferative neoplasms (MPNs)—including primary myelofibrosis (PMF), polycythemia vera (PV), and essential thrombocythemia (ET)—are clonal stem-cell disorders driven by JAK–STAT pathway activation within an inflammatory microenvironment. Ruxolitinib (RUX), a JAK1/2 inhibitor, improves splenomegaly and symptom burden in myelofibrosis and serves as second-line therapy in hydroxyurea-resistant/intolerant PV. However, real-world data from Middle Eastern populations remains scarce. This study evaluates the effectiveness, safety, and dosing practices of RUX in a heterogeneous Omani MPN cohort. Methods: Electronic records were reviewed for adults diagnosed with MF, PV, or ET who received RUX at Sultan Qaboos University Hospital or the National Hematology and BMT Center between 2018 and 2025. Only patients with at least six months of follow-up were included. Clinical variables, laboratory results, dosing information, and data on treatment responses and toxicities were extracted. Outcomes focused on spleen size changes, symptom improvement, hematologic responses, adverse events, and reasons for treatment discontinuation. Results: Twenty-eight patients were included (71% PMF; mean age 52.6 years). The median time from diagnosis to RUX initiation was 28 months. Among MF patients, mean spleen size decreased by 35% (p = 0.003); 28% achieved ≥35% reduction. Symptomatic improvement was reported by 60%. LDH levels remained unchanged. PV patients showed spleen control in 75% and symptom benefit in all cases, while ET responses were limited. Cytopenias (21%) and infections (11%) were the most common toxicities; discontinuation occurred in 8/28 of patients, and two patients died while on therapy. Initial dosing was frequently inappropriate, particularly underdosing in MF and overdosing in PV. Conclusions: RUX provided meaningful spleen and symptom improvement in MF and PV; however, toxicity, delayed initiation, and inconsistent dosing practices limited overall outcomes. Enhanced guideline-based dosing, proactive toxicity management, and structured multidisciplinary follow-up are essential to optimize RUX use in regional MPN practice. Full article

Background and Objectives: Peripheral cytopenia occurs in approximately 2% of the population; however, in up to 0.9%, no cause is identified by conventional tests. Next-Generation Sequencing (NGS) detects somatic variants consistent with clonal hematopoiesis (CH). We aimed to determine the prevalence of CH and pre-myelodysplastic syndrome (pre-MDS) using a 51-gene panel with histopathological assessment. Materials and Methods: Bone marrow samples from 96 consecutive patients evaluated for cytopenia were retrospectively analyzed for genetic alterations. Results: In the overall cohort (n = 96), the median follow-up was 8.1 months (range, 1–20). A total of 37 (39%) out of 96 patients were diagnosed with idiopathic cytopenia of undetermined significance (ICUS), 9 (9%) with clonal cytopenia of undetermined significance (CCUS), 9 (9%) with clonal cytopenia and monocytosis of undetermined significance (CCMUS), 34 (36%) with myelodysplastic syndrome (MDS), and 7 (7%) with myelodysplastic/myeloproliferative neoplasm (MDS/MPN). Among 41 patients in whom no cytogenetic abnormalities were detected by fluorescence in situ hybridization (FISH), somatic variants were identified by NGS. In CCUS, 88% of patients had a single variant, most commonly ASXL1 (44%), followed by TET2 (22%). In CCMUS, ASXL1 and DNMT3A (each 25%) were the most frequent variants. The mean variant allele frequency (VAF) was higher in MDS (33.4%) than in CCUS/CCMUS (13.6%). In MDS patients aged 60 years and older, a higher number of variants were found compared to patients aged less than 60 years (p = 0.028). RUNX1 variants (n = 8) were associated with leukopenia (p = 0.012). Patients with SRSF2 variants (n = 4) had significantly poorer progression-free survival (p = 0.001). EZH2 and SETBP1 variants were associated with inferior overall survival (p = 0.04 and p = 0.019, respectively). In MDS patients (n = 34), thrombocytopenia (plt < 100.000) was associated with shorter PFS (p = 0.005). Conclusions: Given that pre-MDS conditions are considered predictors of hematologic malignancies, conventional diagnostic workup may be insufficient to accurately identify these entities, whereas NGS provides significant additional diagnostic value. Full article