Acute myeloid leukemia (AML) is a heterogeneous hematologic malignancy characterized by uncontrolled proliferation of leukemic cells. Emerging evidence suggests that dysregulation of cell-cycle regulators may interact with immune checkpoint pathways; however, this relationship remains insufficiently explored in AML. To evaluate the expression of CDC27 and PD-L1 in adult AML patients and assess their clinicopathological and prognostic significance. This case - control study included 80 participants: 40 newly diagnosed adult AML patients and 40 age- and sex-matched healthy controls. Gene expression levels of CDC27 and PD-L1 in bone marrow mononuclear cells were quantified using real-time polymerase chain reaction (RT-PCR). Survival analysis was performed and Patients were stratified according to median expression levels. Univariate and multivariate Cox proportional hazards regression analyses were conducted to identify independent prognostic factors. CDC27 and PD-L1 expression levels were significantly higher in AML patients compared with controls (p = 0.001 and p = 0.005, respectively). Both markers were associated with adverse clinical features and reduced overall survival. Multivariate analysis identified CDC27 expression as an independent predictor of mortality. Conclusion: CDC27 and PD-L1 overexpression is associated with aggressive disease characteristics and poor survival in AML patients.
Acute myeloid leukemia (AML) remains a difficult hematologic malignancy that requires more selective therapeutic strategies. Here, we present the in silico design and comprehensive characterization of a recombinant immunotoxin composed of an anti-CD33 single-chain variable fragment (scFv) fused to the LukS-PV pore-forming toxin. Structural modeling and refinement confirmed correct immunoglobulin-domain folding, preserved LukS-PV architecture, and high stereochemical quality. Physicochemical and mRNA analyses indicated favorable stability, solubility, and strong translational compatibility with Escherichia coli. Protein - protein docking predicted specific and high-affinity binding of the scFv to CD33, while molecular-dynamics simulations supported the overall structural stability and functional flexibility of the fusion construct. Codon optimization yielded a high CAI and balanced GC content, suggesting efficient bacterial expression. Immunogenicity analyses predicted non-allergenic and nontoxic properties. Together, these computational findings identify the LukS-PV - anti-CD33 fusion as a promising candidate for targeted AML therapy and establish a framework for subsequent experimental validation.
The multifactorial primary immune thrombocytopenia (ITP) is an acquired autoimmune disease highlighted by dysregulation of immune responses that lead to both increased platelet destruction and impaired platelet production with reduced blood platelet count. The pathophysiology of autoimmune illnesses such as ITP has been linked to regulatory non-coding RNAs, such as circular RNAs (circRNAs) and microRNAs (miRNAs). Our goal was to assess circTUBD1 and miR-146a gene expression in Egyptian ITP patients to correlate them with clinicopathological characteristics. In this study, two groups of forty Egyptian participants were established: twenty individuals diagnosed with chronic ITP and twenty healthy individuals matched in terms of sex and age, recruited from the Internal Medicine department at Menoufia University Hospitals in Egypt. Blood samples were taken from each individual to analyze the plasma expression levels of circTUBD1 and miR-146a using real-time PCR analysis. There was an upregulation of circTUBD1 and a downregulation of MiR-146a levels that showed a significant difference between the control group and the ITP patient group (p < 0.001 for both biomarkers). The expression level of circTUBD1 and MiR-146a exhibits a significant inverse relationship (r = -0.495and p = 0.026). MiR-146a and circTUBD1 can be promising noninvasive biomarkers of chronic ITP and disease activity.
Breast cancer (BC) is the most frequent cancer among women globally. CD14 high expression levels are associated with more aggressive behavior. The study aims to assess CD14 expressions in locally recurrent breast cancer and corresponding primary tumors whenever available, both intratumorally and peritumorally, and to analyze its association with clinicopathological features. This retrospective cross-sectional study included 55 cases of women with locally recurrent breast cancer. They were immunohistochemically stained using anti-CD14 antibody. Statistical associations between CD14 expression and available clinicopathological data were done. Immunoexpression of CD14 was positive peritumorally in (50.9%) of the recurrent breast cancer cases. In the 18 available corresponding primary breast cancer, CD14 was positive only in 2 cases (11.1%) peritumorally. There was significant association between CD14-positive peritumoral immunohistochemical expression with molecular subtype (luminal B), Ki67 (≥14) and the early recurrence time (≤5 years), P values were (0.002, <0.001 & 0.042) respectively. Peritumoral CD14 positivity was significantly higher in recurrent breast cancer cases (50.9%) than in available primary ones (11.1%). CD14 positivity was higher in recurrent than in primary breast cancer, and its statistically significant association with early recurrence and high Ki67 suggests a potential role in breast cancer progression.
Hashimoto's thyroiditis (HT) is an autoimmune thyroid disorder that includes thyroid parenchymal lymphocytic infiltration. Progranulin (PRGN) is an anti-inflammatory glycoprotein encoded by granulin precursor gene (GRN) and involved in the pathogenesis of several diseases. Neutrophil and platelet lymphocyte ratios (NLR, PLR) are established inflammatory markers. The study aimed to evaluate PRGN, GRN rs3785817 variant, NLR and PLR in hypothyroid HT. Complete blood count, serum TSH, free T4, anti-TPO, PRGN, and GRN rs3785817 genotyping were performed for 45 hypothyroid HT patients and 45 healthy controls. Results showed slightly significantly decreased serum PRGN levels in hypothyroid HT patients compared to controls (p = 0.048). NLR displayed significant higher levels in patients in comparison to controls (p = 0.036). The genotype distribution of GRN rs3785817 showed no significant variation between patients and controls with the predominance of the AA genotype in both groups. However, among patients, there was a significantly increased serum PRGN level in patients with AG+GG genotypes compared to those with AA genotype (p = 0.014). In conclusion, GRN rs3785817 is neither a risk nor a protective factor associated with hypothyroid HT. Nevertheless, PRGN variability between the rs3785817 genotypes may suggest their potential roles in hypothyroid HT.
BACKGROUND:MALAT1 (long non-coding RNA) is associated with growth and proliferation of breast cancer (BC). Functional single-nucleotide polymorphisms in MALAT1 may regulate an individual's susceptibility to cancer. Little is known regarding MALAT1 polymorphisms and the risk of BC in Egyptian women. OBJECTIVES:This study aims to determine the relationship between MALAT1 polymorphisms (rs619586 and rs3200401) and the risk of BC among Egyptian women, as well as clinicopathology and survival rates. METHODS:A case-control study was conducted involving 50 women with confirmed BC and 50 age-matched controls. ELISA was used to measure serum CA 15-3 and carcinoembryonic antigen (CEA). A TaqMan discrimination assay was performed to study MALAT1 SNPs (rs619586 and rs3200401). RESULTS:The G allele at rs619586 and the AG genotype were significantly more prevalent in controls and were associated with a reduced risk of breast cancer. On the other hand, there was no significant link between rs3200401 genotypes and alleles and the risk of BC. The AA genotype of the rs619586 was associated with higher tumor grade. CONCLUSION:These findings suggest an association between rs619586 and the risk of BC in Egyptian women; however, larger, multicenter studies evaluating functional effects should be conducted.
The therapeutic efficacy of antibody-drug conjugates (ADCs) is governed not only by target binding but also by the efficiency of receptor-mediated internalization, intracellular trafficking, and lysosomal degradation. However, most conventional assays provide static or endpoint measurements, limiting their ability to resolve these dynamic, multistep processes. Here, we report a quantitative, dual-probe live-cell imaging platform that simultaneously tracks antibody internalization and lysosomal degradation with spatiotemporal resolution. By combining an internalization tracer with a cathepsin-activated fluorogenic sensor, this assay enables direct, real-time measurement of lysosomal routing following cellular uptake. Using trastuzumab and clinically approved HER2-targeting ADCs as model systems, we demonstrate that internalization and lysosomal degradation are temporally uncoupled and occur with distinct kinetics. Moreover, ADCs sharing the same antibody backbone exhibited markedly different processing profiles, highlighting the influence of linker chemistry and payload properties on their intracellular fate. These results underscore the importance of evaluating ADCs as integrated molecular systems rather than as modular components. This platform provides a scalable and mechanistically informative tool for ADC development, enabling candidate ranking, linker screening, and rational payload - linker optimization based on functional intracellular behavior rather than uptake alone. As such, it offers a framework for data-driven design of next-generation ADCs with improved therapeutic index.
BACKGROUND:Colorectal cancer is identified as the third-most prevalent cancer globally and the second leading cause of cancer mortality. METHODS:This study investigated the immunohistochemical expression of Cyclooxygenase-2 and Hypoxia-Induced Factor 1 alpha & correlated their expression with clinicopathological parameters in 100 retrospectively selected cases of colorectal adenocarcinoma from Port Said Governorate, Egypt. RESULTS:COX-2 was positively expressed in 67% of cases, and HIF-1α was positive in 57%. COX-2 positivity was statistically significantly associated only with lymph node metastasis (p = 0.019). HIF-1α positivity showed significant associations with multiple indicators of aggressive disease, including high tumor grade (p < 0.001), advanced tumor stage (T3-T4, p = 0.047), lymph node metastasis (p < 0.001), and advanced TNM stage (p = 0.006). Crucially, a statistically significant positive correlation was detected between the expression of COX-2 and HIF-1α (p = 0.039). CONCLUSION:COX-2 and HIF-1α are often expressed in colorectal carcinoma and are significantly associated with aggressive clinicopathological features, notably lymph node metastasis and advanced tumor stage. The significant co-expression of these two markers supports a biological constructive collaboration, suggesting that their simultaneous presence drives tumors toward a more invasive and metastatic phenotype, predicting a poorer prognosis. Detection of COX-2 and HIF-1α is recommended as an effective index for assessing prognosis and identifying high-risk patients.
Hepatitis B virus (HBV) infection remains a major concern in Nigeria. Nationally, increasing infection rates continue to pose a significant challenge, which may be due to inadequate diagnosis. Known markers, such as Hepatitis B Surface Antigen (HBsAg) and Hepatitis B e antigen (HBeAg), are good at indicating exposure rates, but generally poor at reflecting active viral replication. The hepatitis B core-related antigen (HBcrAg) may provide a clearer picture of the viral activity and treatment response. We conducted a cross-sectional study involving 450 participants aged 16-65 years from hospitals in Osun and Plateau States. HBsAg screening was performed using rapid diagnostic tests, confirmed by enzyme-linked immunosorbent assay (ELISA), and further evaluated for HBeAg and HBcrAg levels. Correlation analysis was used to test the relationships between these markers. Of the 450 participants screened, 188 (41.7%) were confirmed positive for HBV. HBcrAg was observed in adults aged 17-45 years, suggesting replication activity in symptom-free individuals. Atypical serological profiles, such as HBsAg and HBsAb co-positivity, were also identified. Incorporation of HBcrAg in HBV screening, especially in endemic countries, may be useful in assessing HBV replication stage, reactivation, and staging of chronic infection, especially in resource-limited settings.
BACKGROUND:Rheumatoid arthritis (RA) has a complex etiology that involves environmental and genetic variables. The identification of new genetic connections contributes to accelerating the development of customized medications to treat or delay the progression of diseases. OBJECTIVE:We aimed to assess the relationship between rheumatoid arthritis susceptibility and genetic variations of the Farnesyl Diphosphate Synthase (FDPS) and methyl CpG binding protein 2 (MECP2) genes. PATIENTS AND METHODS:100 patients with rheumatoid arthritis and 100 healthy control subjects participated in this case-control research. Genetic analyses for single-nucleotide polymorphisms (SNPs) in the FDPS and MeCP2 genes were performed on the patients. RESULTS:In terms of FDPS genetic polymorphisms, the sick group had greater frequencies of the TG and GG genotypes, whereas the control group had a higher frequency of the TT genotype. The CC genotype was more common in the control group, but the TC and TT genotypes were more common in the sick group concerning MECP2 genetic polymorphisms. CONCLUSION:For rheumatoid arthritis, the FDPS (rs2297480) TT genotype seemed to be protective. An increased risk of rheumatoid arthritis was linked to the MECP2 (rs2734647) TT genotype, but the CC genotype seemed to be protective against the condition.
The incidence of multiple sclerosis (MS) has increased in recent years. Its pathogenesis involves the interaction between various elements, with interleukin 27 (IL-27) playing a key role in autoimmunity. The presence of the IL-27 receptor on astrocytes emphasizes its involvement in the disease's progression. PURPOSE:The study aims to investigate possible associations between IL27 rs181206, serum level of IL27, and the development of MS. METHODS:The study comprised 70 MS patients and 70 seemingly healthy controls. They were genotyped for IL27 rs181206 using the Taqman allelic discrimination approach, and their serum IL27 levels were estimated using ELISA. RESULTS:The frequency of TT genotype, T allele, and IL27 serum level were significantly higher among MS patients compared to controls. There was no significant difference between IL27 serum levels among different genotypes in both MS patients and controls; however, individuals with TT genotype showed higher levels of IL27 than those with CC genotype. CONCLUSION:TT genotype and T allele can increase the risk of developing MS. On the other hand, carrying the C allele may be associated with a lower risk of MS development. Understanding IL27 genetics and epistatic interactions can help clarify IL27's role in MS pathogenesis and utilize it as a therapeutic target.
Tuberculosis (TB) is caused by an infection with Mycobacterium tuberculosis. The Bacille Calmette-Guérin (BCG) vaccine is used to treat TB. However, its efficacy varies in different countries. A new TB vaccine is urgently required to inhibit the spread of TB infection. DNA vaccines are promising for providing an immune response against the disease. The resuscitation-promoting factors B and D (rpfB/rpfD) genes are promising vaccine candidates. In this study, the vaccine candidates pcDNA3.1-rpfB and pcDNA3.1-rpfD were evaluated for their ability to inhibit M. tuberculosis infection in BALB/c mice. Epitope analysis indicates that the recombinant protein sequences of RpfB and RpfD used in this study possess epitopes recognized by T and B lymphocytes. Although the presence of M. tuberculosis cells in lung tissue was not detected, histopathological analysis revealed the absence of lymphoid aggregates in mice vaccinated with pcDNA3.1-rpfB or pcDNA3.1-rpfD, in contrast to those administered with phosphate-buffered saline or pcDNA3.1. In addition, analysis of the humoral immune response showed the highest IgG2a antibody titer in mice immunized with both vaccine candidates. These results support our previous findings, which indicate that pcDNA3.1-rpfB and pcDNA3.1-rpfD have considerable potential as TB vaccine candidates.
Enzyme-linked immunosorbent assay (ELISA) is a rapid, sensitive, and economical tool for detecting hormones in diverse biological matrices, making it valuable in forensic endocrinology. This review summarizes ELISA's principles, applications, advantages, and limitations in medico-legal practice. ELISA enables quantification of hormones such as cortisol, testosterone, estrogen, melatonin, thyroid hormones, progesterone, and human chorionic gonadotropin (hCG). It supports postmortem toxicology, sexual assault investigations, doping control, and forensic psychiatry. Cortisol may help reconstruct perimortem stress, while hCG detection assists in pregnancy confirmation in assault or maternal death cases. In sports, ELISA screens for anabolic steroids, erythropoietin, and growth hormone, with LC-MS/MS required for confirmation. Its compatibility with blood, saliva, urine, and hair enhances versatility. Key challenges include antibody cross-reactivity, matrix interference, degradation, variability among commercial kits, and limited multiplexing. False positives, hook effects, and inconsistent validation affect admissibility, necessitating strict quality assurance, ISO/IEC 17025 compliance, and confirmatory testing. ELISA is unsuitable for paternity determination; DNA profiling remains the legal standard. Emerging advances - digital ELISA, nanotechnology, AI, and biosensors - promise greater sensitivity and automation but face regulatory, cost, and training barriers. ELISA should be regarded as a complementary, high-throughput screening method integrated into validated, multimodal forensic workflows.
Filgrastim, a therapeutic protein for the treatment of neutropenia, exerts its biological activity by interacting with the granulocyte colony-stimulating factor receptor. However, the affinity of filgrastim to its receptor is usually overlooked during characterization and comparability tests, since its biological activity is assessed only by in vitro proliferation assays. In this work, we propose the use of a cell-based colorimetric method to determine the relative affinity of filgrastim to its receptor expressed in murine myeloid leukemia cells. After performing a validation exercise, the method proved to be accurate, specific, and linear within an affinity interval of 75 to 130%; precise with a confidence interval of 93.6 to 114.2% and a coefficient of variation of 12.6%. The validation exercise proved that the proposed cell-based method is a viable alternative to evaluate the biological activity of filgrastim via the affinity for its receptor and it is suitable to be included as part of its biological characterization exercises as well as in comparability exercises for products coming from distinct manufacturing processes.
Psoriasis is characterized by increased levels of pro-inflammatory cytokines, including TNF-α, IL-1β, IL-17A, and NF-κB, as well as keratinocyte hyperproliferation and epidermal thickening. Its pathophysiology is significantly influenced by oxidative stress and abnormal activation of redox-sensitive signaling pathways, such as NF-κB and MAPK. The present study examined the potential of Syringaldehyde (SYD) in the imiquimod (IMQ) induced psoriasis model. Psoriasis Area Severity Index (PASI) scores, back skin thickness, and spleen hypertrophy decreased at dose levels of SYD 25, 50, and 100 mg/kg. Furthermore, a significant reduction in PASI scores following SYD administration indicated a marked improvement in the disease severity and lesion morphology. High-affinity binding of SYD to NF-κB (PDB ID: 4KIK; -34.76 kcal/mol) and MAPK (PDB ID: 1A9U; -32.87 kcal/mol) was found by molecular docking, indicating interference with nuclear translocation and phosphorylation processes. The treatment groups 50 mg/kg and 100 mg/kg indicated restoration of normal histological features. The biochemical evaluation showed decrease in NF-κB, IL-1β, TNF-α, and IL-17 on treatment with SYD. Thus, SYD appears to be a potential therapeutic option for psoriasis, but additional research is needed to confirm its efficacy and safety.
The diagnosis of Barrett's esophagus (BE) is challenging in the absence of goblet cells (GC). Immunohistochemistry (IHC) may be useful to highlight intestinal differentiation in samples lacking GC. In this study, we aimed to describe the IHC expression of CDX2 in columnar cells of BE and to assess its diagnostic and prognostic utility. This retrospective study, conducted from 2010-2015,included cases suspicious of BE examined in our pathology department. Only cases positive for GC (Alcian Blue positive) were diagnosed as BE. An automated IHC analysis was performed using CDX2. Nuclear CDX2 expression was evaluated in IM zones(goblet cells) and adjacent tissue (columnar cells). OF 39 cases, the diagnosis of BE was confirmed in 34 cases. The mean age of patients was 53 years with a male-to-female ratio of 2.4.Endoscopically, 20 patients had short-segment BE (59%), 8 had ultra-short-segment BE (23%) and 6 had long-segment BE (18%).Histologically, an associated adenocarcinoma was found in two cases. In non-neoplastic BE, nuclear CDX2 expression was observed in both GC (88.2%) and adjacent columnar cells(26.5%). A statistically significant association was found between CDX2 expression and GC (p < 0.005).Both adenocarcinoma cases were CDX2-positive in BE areas but CDX2-negative in tumor foci. CDX2 has a high sensitivity and specificity for IM and its expression is associated with GC. However, its low expression in adjacent columnar cells limits its benefit in BE specimens lacking GC.
The identification of biomarkers for pulmonary exacerbations in cystic fibrosis (CF) is inevitable. We aimed to evaluate the serum levels of substance p (SP), neuropeptide Y (NPY), and Pituitary Adenylate Cyclase Activating Polypeptide (PACAP), at time of pulmonary exacerbation and after treatment with antibiotics. Twenty cystic fibrosis patients with mean age of 8.83 years (±4.37) were enrolled. Serum samples were taken at the time of admission and two weeks post-antibiotic therapy. Serum levels of target markers were determined using ELISA. Serum SP, NPY, and PACAP levels were significantly higher at exacerbation (229.22 ± 73.86 pg/ml, 1869.89 ± 787.14 pg/ml, and 32,261.51 ± 22283.78 fg/ml, respectively) than post-antibiotic therapy (206.29 ± 77.83 pg/ml, 1412.95 ± 647.09 pg/ml, and 17,359.39 ± 10105.39 fg/ml, respectively; p < 0.05). Positive correlations were observed between serum levels of SP and PACAP (r = 0.515, p = 0.020) and NPY (r = 0.779, p < 0.001), and between NPY and PACAP (r = 0.513, p = 0.021). A negative correlation was found between NPY and BMI z-score (r = -0.503, p = 0.024). As a conclusion, serum SP, NPY and PACAP levels are potential biomarkers for CF pulmonary exacerbations and response to antibiotic therapy.
Oral squamous cell carcinoma (OSCC) accounts for over 90% of all oral malignancies and has a 50-60% five-year survival rate, despite advances in treatment. Ki67, a nuclear protein involved in cellular proliferation, has been studied as a prognostic marker in various malignancies, including OSCC. However, its link with different histological grades of OSCC is uncertain. The study aimed to evaluate Ki-67 expression in well-differentiated (WDOSCC), moderately differentiated (MDOSCC), and poorly differentiated (PDOSCC) tumors to ascertain its correlation with tumor differentiation and aggressiveness. Forty OSCC cases were classified using Broder's histological grading system. Ki67 immunohistochemical staining was performed, and three groups were categorized based on Ki67 expression: low (1-25%), moderate (26-50%), and high ( > 50%) proliferation. Statistical analysis assessed the significance of Ki67 expression across OSCC grades. Among 40 cases, 35% were WDOSCC, 45% MDOSCC, and 20% PDOSCC. Low Ki67 proliferation was seen in 64% of WDOSCC, moderate in 61% of MDOSCC, and high in 75% of PDOSCC. There was a significant (p < 0.001) association between OSCC grading and Ki67 expression. Ki67 expression correlates with OSCC histological grades, increasing with tumor proliferation. These findings support Ki67 as a prognostic marker, warranting further large-scale validation studies.
This study explores the relationship between oxidative - antioxidative balance and stromal cell-derived factor-4 (SDF-4) levels in individuals with Generalized Anxiety Disorder (GAD), aiming to clarify their roles in the disorder's pathophysiology. By examining these biomarkers, the research investigates whether oxidative stress mechanisms contribute to GAD. A cross-sectional study was conducted with 43 GAD patients and 40 age- and sex-matched healthy controls. Participants were assessed using the Beck Anxiety Inventory (BAI) and the Structured Clinical Interview for DSM-5. Serum levels of SDF-4, total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) were measured. GAD patients showed significantly higher levels of SDF-4, TAS, TOS, and OSI compared to controls (p < 0.001). No significant differences were found in other hematological parameters. In the GAD group, SDF-4 levels were not significantly correlated with anxiety severity or oxidative markers. ROC analysis indicated good diagnostic performance of SDF-4 for GAD (AUC = 0.870). The findings suggest that oxidative - antioxidative imbalance and elevated SDF-4 levels may play a role in GAD. SDF-4 might represent a compensatory response to oxidative stress. Elevated TAS levels may indicate enhanced antioxidant defense. Further studies are needed to clarify the mechanistic and therapeutic implications.
