Omar S. El-Masry

Background: Experimental animal models are essential for understanding the molecular mechanisms of human leukemia and testing potential therapies. Chemical induction using 7,12‐dimethylbenz[a]anthracene (DMBA) has shown promise in recapitulating features of leukemogenesis in rodents, but its molecular fidelity to human disease remains underexplored. The aim of this study was to evaluate the transcriptomic landscape of a DMBA‐induced leukemia rat model and assess its alignment with known gene expression signatures and oncogenic pathways in human leukemia. Methods: Male Wistar rats were treated with DMBA to induce leukemia. RNA was extracted from whole blood and subjected to RNA sequencing. Differentially expressed genes (DEG) were identified using RNA‐seq by expectation–maximization and NOISeq. Functional enrichment analysis, Gene Ontology mapping, and protein interaction networks were analyzed via STRING. Prognostic relevance of key DEGs was explored using data from the Human Protein Atlas. Results: The DMBA‐induced model showed significant upregulation of leukemia‐associated genes, including FLT3 , NRAS , KRAS , ABL1 , BCR , and NPM1 . The top 10 up‐ and downregulated genes were enriched in pathways related to cell fate, proliferation, apoptosis, and immune regulation. Particularly, genes such as MEIS2 , CXCL9 , and WIF1 were linked to hematopoietic dysregulation and poor prognosis in various cancers. Comparative analysis revealed strong transcriptomic overlap with human leukemia/carcinogenesis gene signatures, supporting the model's translational validity. Conclusion: In conclusion, the DMBA‐induced leukemia rat model exhibits a transcriptomic profile that closely mirrors key oncogenic and prognostic features of human leukemia. This model holds significant promise for preclinical studies targeting leukemogenesis and therapeutic intervention strategies.

Colorectal cancer (CRC) is a major cause of cancer-related morbidity and mortality worldwide, and dysregulation of phosphoinositides signaling pathways plays a critical role in its pathogenesis. The involvement of Set Binding Factor 1 (SBF1; MTMR5), a pseudophosphatase of the myotubularin family in CRC, remains unexplored. This study compiled experimental and bioinformatics approaches to investigate the expression and potential relevance of SBF1 in CRC. Immunohistochemical analysis was performed on CRC tissue specimens and corresponding normal adjacent tissues (NAT), with estimation of SBF1 staining followed by statistical comparison. In parallel, bioinformatics assessed gene ontology and protein-protein interactions, and open CRC datasets were analyzed to assess SBF1 alterations, expression profiles, and survival associations. The results demonstrated a significant overexpression of SBF1 in CRC tissues compared with NAT. Bioinformatics revealed SBF1 as a partner within a functional network enriched in phosphoinositide metabolism. Bioinformatics identified SBF1 genetic alterations in approximately 5% of cases, and survival analysis indicated that wild-type SBF1 could be an unfavorable prognostic marker, as reflected by hazard assessment. Collectively, these findings provided the first experimental evidence that SBF1 is overexpressed in CRC and suggest that it may represent an unrecognized component of phosphoinositide related pathways involved in colorectal tumorigenesis.

Ovarian cancer (OC) remains a major cause of female cancer mortality, with chemoresistance impeding effective treatment. This study investigated the expression of p62/SQSTM1 and Nrf2 in chemoresistant versus chemosensitive OC cell lines and patient tissues. Using quantitative reverse transcription PCR, Western blotting, and immunohistochemistry, we found co-upregulation of p62 and Nrf2 proteins in chemoresistant samples. Proliferation assays revealed that higher p62 expression correlated with reduced cisplatin sensitivity. Sanger sequencing detected synonymous p62 variants in exon 6, with no functional impact. Immunohistochemical scoring showed significantly higher p62 and Nrf2 levels in chemoresistant patients compared to remission and sensitive groups. A strong correlation was observed between p62 expression and cisplatin IC50 values. These findings support the role of the p62/Nrf2 axis in OC chemoresistance and highlight their potential as predictive biomarkers and therapeutic targets. Future studies are warranted to explore targeted interventions that disrupt this pathway to enhance treatment response in OC.

Adaptive, rather than innate, immunity relies mainly on antigen–antibody recognition. This recognition is driven by the binding of specific antibody paratopes to distinct epitopes found on antigens. This interaction is pivotal for immune responses that have been re-purposed for diagnostic and therapeutic purposes. This article focuses on Western blotting, an in vitro technique performed for protein immunodetection. Traditionally, this technique requires separate incubations of both primary and secondary antibodies, for which these antibodies recognize different antigen epitopes (conventional method). We propose a modified protocol combining both antibodies, involving a single incubation step that reduces time and conserves reagents (non-conventional/improved method). This improved protocol will enhance efficiency without compromising detection accuracy. It will support multiplexing, enabling the simultaneous detection of multiple proteins. Despite the positive results found by applying available antibodies, further optimization is required for a more thorough evaluation, to ensure that all antibodies consistently yield successful results in every detection attempt for broader use. Our findings indicate that the tested antibody cocktails remained stable over time, which suggests potential for commercialization of this modified Western blot protocol with a wide scope towards multiplex diagnostic application.

Background: Chronic myeloid leukemia (CML) is a prevalent hematological malignancy known for the presence of the Philadelphia chromosome and activation of the BCR-Abl kinase activity. Although tyrosine kinase inhibitors are widely used as the standard treatment, resistance remains a concern among certain patients. This study aimed to investigate the gene expression profile of a group of CML patients in comparison to a control group in order to identify novel candidate genes associated with the disease. Methods: Whole transcriptome sequencing was performed, and gene expression levels were validated using quantitative real-time PCR. Additionally, single nucleotide and insertion/deletion variants were analyzed in the selected candidate genes among 10 CML patients and 4 healthy control subjects.Results: Analysis revealed a set of differentially expressed genes, whose up- or downregulation was further confirmed by qRT-PCR. Among the upregulated genes in the patient group were ribosomal protein like (RPL) members, specifically RPL9, RPL34, RPL36A, and RPL39, while downregulation was observed in CCDC170, LDB1, and SBF1 compared to the healthy subjects. Furthermore, gene variant studies identified novel genetic changes in these candidate genes, suggesting potential clinical significance in CML. Conclusions: This study highlights RPL9, RPL34, RPL36A, RPL39, CCDC170, LDB1, and SBF1 as potential targets in CML. Additionally, it underscores the importance of investigating these genes and their variants in larger cohort studies to assess their clinical significance in CML patients.

Chronic myeloid leukemia (CML) is a common blood malignancy awaiting decisive cure. Understanding the genomic nature of the disease could help to improve treatment strategies and introduce novel therapeutic targets. The purpose of this study was to sequence the entire transcriptome of a cohort of CML patients to understand differences in gene expression profiles and gene variants compared to healthy controls. RNA was extracted from 10 CML and four healthy control subjects and sequenced by the DNBSEQ platform. Differential gene expression was evaluated and confirmed by the q-RT-PCR technique. Gene variants were also analyzed. The results showed that COX7B and COX7C were upregulated in CML patients than in controls; this was confirmed by quantitative PCR. In addition, novel single nucleotide and insertion/deletion variants were also found in both genes. In conclusion, the results of this study recommend further exploration of COX7B and COX7C and their novel variants in myeloid leukemogenesis.

Diagnostic laboratory tests are frequently overused in healthcare entities, leading to an increased strain on laboratory resources, additional workload,