Cellular and Molecular Biology

Integrated genomic and molecular insights into astrocyte- and oligodendrocyte-derived amyotrophic lateral sclerosis: focus on miRNAs and extracellular vesicles

2025-07-06T10:21:32+02:00

Motor neurons in the brain and spinal cord begin to die off in Amyotrophic lateral sclerosis (ALS), a disease that can be fatal. Molecular pathways in neurological disease, especially ALS, remain a challenge in the medical sciences. In this disease, a disorder in both astrocytes and oligodendrocytes can cause the disease to progress. This study aimed to investigate the molecular mechanisms and find key elements between these two cells in ALS with a bioinformatics perspective. In this study, using integrated and continuous bioinformatics analytics by various tools and databases, we investigated genes, protein products, and miRNAs between astrocytes and oligodendrocytes. The obtained data were involved in the Cellular senescence, actin cytoskeleton, and cell cycle signaling pathways. Then, after careful evaluation of the information, TP53, MDM2, KRAS, PTPRC, and GSK proteins were candidates, which are regulated by hsa-miR-564, hsa-miR-496-5p, hsa-miR-324-5p, hsa-miR-296-5p, and hsa-miR-4258-3p miRNAs. Finally, the four genes had a more robust and better relationship in this study between astrocyte and oligodendrocyte-derived ALS.

Molecular and cellular effects of chronic low-dose X-ray exposure on thyroid function and blood cell parameters in radiology staff

2025-07-06T10:21:50+02:00

This study investigated the impact of long-term, low-dose occupational exposure to X-rays on thyroid function and hematological parameters in radiology staff. A cross-sectional analysis was conducted on 136 radiology and radiation-related employees. Thyroid status, assessed via ultrasonography and serum T3, T4, and TSH measurements, was compared with complete blood counts (CBC). While no significant gender-based differences in thyroid hormones or structure were observed, TSH levels correlated significantly with occupational group (P=0.016), and T3 levels correlated with working hours (P=0.03). Radiologists exhibited higher RDW-CV compared to radiographers and other staff (P=0.009). Significant gender differences were noted in lymphocyte counts, hemoglobin, red blood cells, and hematocrit. In males, WBC and MCHC fluctuated significantly with increased working hours. These findings suggest that chronic, low-dose X-ray exposure may influence thyroid hormone regulation and hematopoiesis in radiology professionals. Further research is warranted to elucidate the underlying cellular and molecular mechanisms and to refine radiation safety protocols.

Association of inflammatory gene variants with problematic alcohol use in a Colombian population

2025-07-06T10:22:00+02:00

Alcohol dependence is a multifactorial disease that constitutes a significant public health concern and a significant risk for individual, family, and social health. Its genetic component exhibits significant ethnic variation and is closely associated with the personal evolution of the disease. However, although multiple loci have been identified, no functional variants have been identified. In this work, we selected some genes from the inflammatory response pathway and searched for SNV (single nucleotide variants) in their promoter region that could be associated with the disease. We compared cases of problematic alcohol consumption (n=66) with controls (n=73) in a population sample taken at the National University of Colombia, Bogotá headquarters. Peripheral blood DNA extraction was performed. We used PCR and Sanger sequencing to find 28 SNVs and one STR in 10 inflammatory response genes that are connected to alcoholism. Then, using various bioinformatic tools, the analysis of haplotypes, linkage disequilibrium, epistasis and genetic networks was carried out. Allele and genotypic frequencies for this Colombian population were reported for the first time. Additionally, we found haplotypes that could be protective and risk factors for the disease, and gene interactions that have cumulative effects related to the drinker phenotype. The investigation of haplotypes, gene interaction, and gene networks is a highly effective methodology for identifying potential associations in small samples. Additionally, SNCA, IL-6R1, TNFR1, and MIF genes were profiled for further studies.

Anti-Müllerian hormone as a diagnostic and prognostic marker in polycystic ovary syndrome: a clinical study

2025-07-06T10:22:09+02:00

A study was conducted on anti-Müllerian hormone (AMH) and polycystic ovary syndrome (PCOS) in women. Samples were collected from 96 women with PCOS and 91 control women, aged 20 to 45 years for both groups. Levels of AMH, estrogen, progesterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), LH/FSH ratio, endometrial growth rate (WHR), and body mass index (BMI) were measured. The results showed significantly lower AMH levels in women with PCOS compared to healthy women at the probability level (P=0.05). The results also demonstrated a significant positive association between the prevalence of PCOS and AMH levels for estrogen, LH, and LH/FSH. It had an inverse relationship with AMH, progesterone, FSH, endometrial growth rate (WHR), and body mass index (BMI). Furthermore, the risk of AMH deficiency in women with PCOS increases with age, due to decreased fertility and egg production from the ovaries, especially after the age of 30, as well as weight gain. This suggests that age-related declines in AMH concentrations and weight gain are indicative of increased risk factors for PCOS. Finally, this study demonstrated a relationship between PCOS risk factors and AMH concentrations, suggesting that low AMH concentrations increase the risk of PCOS, especially with age. This suggests the potential for incorporating AMH into early detection tests and the development of more effective treatments for this condition.

Functional analysis of intron 3 in the regulation of gene expression of the human lipoprotein lipase gene

2025-07-06T10:22:19+02:00

Lipoprotein lipase (LPL) is a key enzyme that hydrolyzes triglycerides (TGs) into free fatty acids. Several genetic variants of LPL are directly or indirectly associated with variations in lipid levels, causing different lipid metabolic disorders. Previous studies on the LPL gene have shown that exons and introns are essential for gene expression and regulation. However, mechanisms through which introns regulate gene expression and function remain unclear. In this study, we successfully designed a protocol to assess the function of LPL intron 3 in LPL regulation. This was accomplished by constructing luciferase reporter vectors, containing full and partial intron 3 fragments from a healthy human DNA sample. These recombinant constructs facilitated the analysis of transcriptional activity using dual-luciferase reporter assays in cell lines. The results showed that the luciferase activity of the chimeric firefly luciferase reporter construct containing the full-length LPL intron 3 was higher than that of other constructs. In this study, a successful protocol was developed to assess the function of LPL intron 3 in regulation of the LPL gene. This protocol provides a novel method for functional analysis of introns and intronic variants that can be applied to other genes.

CTHRC1 overexpression in gastric cancer patients contributes to a poor prognosis and is related to immune cell infiltration

2025-07-06T10:22:27+02:00

Gastric cancer is a common solid tumor of the digestive system, This research aimed to investigate the relationships among CTHRC1 expression, prognostic values and tumor-infiltrating immune cells (TIICs) in GC patients.The expression of CTHRC1 in gastric cancer patients was analyzed using the GEPIA database and the TCGA database. The relationship between CTHRC1 expression and survival of gastric cancer patients was then explored using the Kaplan-Meier Plotter database and TCGA database.Subsequently, we explored whether there was an associative effect between the expression of CTHRC1 and TIICs in gastric cancer tissues. Then we constructed a prognostic model using immunomodulatory genes related to CTHRC1 and verified the specificity and accuracy of the model.Compared with normal tissues, the expression of CTHRC1 was significantly upregulated in gastric cancer tissues. And the high expression of CTHRC1 was associated with poor prognosis of gastric cancer patients.It is suggested that CTHRC1 is a reliable prognostic indicator for predicting the prognosis of gastric cancer patients.Then we successively used univariate and multivariate COX methods to obtain three high-risk immunomodulatory genes (TGFB2, CXCR4 and NT5E) and one low-risk immunomodulatory gene (TNFRSF18) associated with CTHRC1. These four immunomodulatory genes were used to construct a prognostic model for gastric cancer.Gastric cancer patients with high CTHRC1 expression have a poor prognosis and are associated with immune cell infiltration. Therefore, CTHRC1 can be considered as a potentially reliable prognostic indicator for gastric cancer patients.

Levels of Trimethylamine N-Oxide and Lipopolysaccharides in Vascular and ıdiopathic erectile dysfunction

2025-07-06T10:22:46+02:00

The gut microbiota influences endothelial dysfunction through metabolites like lipopolysaccharides (LPS) and trimethylamine-N-oxide (TMAO), affecting cardiovascular health and contributing to atherosclerosis and hypertension development. We evaluated TMAO and LPS levels in patients with vascular and idiopathic erectile dysfunction (ED). In this study of 151 participants (50 vascular ED, 50 idiopathic ED, 51 healthy controls), patients were categorized using comprehensive clinical assessment including International Index of Erectile Function (IIEF), laboratory tests, and imaging methods. While age (mean 55.15±7.17 years) and TMAO levels showed no significant differences between groups (p>0.05), LPS levels were significantly elevated in the vascular ED group (497.36±87.83) compared to idiopathic ED (430.62±69.72) and control groups (436.98±105.37) (p<0.05). These findings suggest that gut microbiota metabolites, particularly LPS, play a significant role in ED pathophysiology through endothelial dysfunction. Regulating gut microbiota may serve as both a protective factor against ED development and a potential treatment option for existing cases. Further comprehensive studies are warranted to explore these therapeutic possibilities.

Inhibitory potential of probiotic strains against pathogens associated with osteomyelitis of the jaws: a quantitative MIC and MBC analysis

2025-07-06T10:22:56+02:00

Osteomyelitis of the jaw is a significant health burden, often associated with biofilm formation, antimicrobial resistance, and complex bacterial interactions. This study aimed to evaluate the antibacterial efficacy of two probiotic strains, Lactobacillus reuteri DSM 17938 and ATCC PTA 6475, against three key pathogens: Fusobacterium nucleatum, Actinomyces israelii, and Staphylococcus aureus. The focus was on their potential as adjunctive treatments for osteomyelitis of the jaws. This in-vitro study employed tailored standard protocols to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the probiotic strains against the tested pathogens. Probiotic suspensions were prepared in serial dilutions and bacterial strains were inoculated into brain heart infusion media for exposure. The growth inhibition and eradication were confirmed through subculture techniques and microscopic examination of Gram-stained samples. Both probiotic strains exhibited significant antibacterial activity against Fusobacterium nucleatum and Actinomyces israelii at low concentrations, achieving MIC of 0.625 µl and MBC of 1.25 µl for both pathogens. In contrast, Staphylococcus aureus demonstrated greater resistance, with MIC and MBC recorded at 100 µl and 200 µl, respectively. The differential response underscores the variable susceptibility of bacterial species to probiotics and highlights the potential for selective therapeutic application. Within the scope of this study, two Lactobacillus reuteri strains demonstrated promising antibacterial effects against Fusobacterium nucleatum and Actinomyces israelii, suggesting their potential utility in managing osteomyelitis of the jaws. However, higher concentrations or synergistic approaches may be needed to combat Staphylococcus aureus. Further studies including preclinical and clinical trials are essential to translate these findings into effective therapeutic strategies.

Histological changes in dental pulp tissue with age: a comparative study

2025-07-06T10:23:07+02:00

The dental pulp undergoes several histological changes with age. These include a reduction in the number of cells and blood vessels, resulting in a decreased capacity for regeneration and repair. Furthermore, there is an increase in collagen fiber density and the formation of secondary dentin, which reduces the volume of the pulp chamber. This study performed a comparative histological analysis to characterize these age-related changes in dental pulp tissues. A cross-sectional comparative analysis was conducted to examine the histological changes in dental pulp tissue from two distinct age groups. Extracted teeth were thoroughly cleaned and immediately fixed in 10% formalin for preservation. Each sample was then decalcified using 10% formic acid for a duration sufficient to allow sectioning without damaging the tissue. After decalcification, the teeth were embedded in paraffin wax, and serial sections, with a thickness of 4-6 μm, were prepared using a microtome. The sections were then stained with Hematoxylin and Eosin (H&E) to visualize general tissue structure. The histological analysis demonstrated notable differences in dental pulp tissue between the two age groups. Group A (young) samples showed a high cellularity, with numerous fibroblasts and odontoblasts and a clearly defined odontoblastic layer. However, Group B (elderly) samples exhibited a marked reduction in cellularity, with fewer odontoblasts present and evidence of increased fibroblast degeneration. The histological changes observed in our study underscore the impact of aging on dental pulp tissue.

Interplay of upper respiratory tract microbiota, ımmune response, and molecular dynamics in SARS-CoV-2 infection

2025-07-06T10:23:36+02:00

Understanding the interplay between upper respiratory tract microbiota, immune responses, and molecular changes is critical for improving the diagnosis and management of SARS-CoV-2 infections. In this study, we investigated the association between respiratory tract microbiota composition, immune markers, and molecular diagnostic parameters in 123 RT-PCR-confirmed COVID-19 patients. Co-infection rates with Gram-positive and Gram-negative bacteria were high, particularly in the nasopharynx (35.4% and 64.4%, respectively), highlighting the risk of secondary bacterial infections. Diagnostic evaluation showed that RT-PCR cycle threshold (Ct) values and serological markers (IgG, IgM) had high sensitivity and specificity for distinguishing infection status. Lower Ct values correlated with higher viral loads and acute infection, while antibody levels reflected immune response dynamics. Significant correlations were observed between bacterial presence and immune parameters such as ACE-2, FASL, and RBD. These findings underscore the importance of integrated diagnostic approaches that consider microbiota, molecular, and immunological markers for effective management of COVID-19 and its complications.

Urea adulteration alters raw milk composition and induces renal tissue damage: a molecular and histopathological study

2025-07-06T10:23:43+02:00

This study investigates the impact of urea adulteration in raw milk on milk quality, safety, and renal tissue integrity, addressing a significant public health concern, particularly in low-to-middle-income countries where over 76.6% of milk samples have compromised quality and 77.89% are adulterated. Raw milk samples were analyzed for quality parameters, adulterants, and the presence of synthetic by-products using FTIR. In vivo studies in mice were conducted to assess the histopathological effects of urea-adulterated milk on renal and liver tissues. Results revealed that urea adulteration leads to significant alterations in milk composition, including changes in fat content from 5.73% to 0.6% in boiled milk, solids-not-fat from 9.11% to 12.84%, protein from 3.61% to 5.28%, and lactose content from 3.81% to 5.57%, alongside the formation of synthetic compounds such as lactose monohydrate and polyvinyl stearate. Histological examination of mice kidneys exposed to urea-adulterated milk demonstrated morphological, inflammatory, degenerative, congestive, and necrotic changes. This study highlights the cellular-level damage induced by urea adulteration, emphasizing its potential to induce neoplastic changes in renal tissues and underscoring the need for stringent monitoring of milk quality to safeguard public health.

A super-enhancer-related gene signature predicts prognosis and immune microenvironment features in glioma

2025-07-06T10:23:16+02:00

Glioma is the most frequent malignant tumor in the brain. Super-enhancer (SE) is a class of transcriptional activator, which drives gene expression. SE-related genes (SERGs) affect occurrence and development of several tumors. We explored the predictive role of SERGs in the prognosis and immune features of glioma. A total of 1557 glioma patients were collected from four data sets, including The Cancer Genomic Atlas (TCGA, n = 691), the Chinese Glioma Genomic Atlas (CGGA) array (n = 286), the CGGA sequencing (n = 316), and GSE16011 (n = 264) from Gene Expression Omnibus (GEO) database. SERGs were selected from SEdb (http://www.licpathway.net/sedb), a comprehensive human SE database. Survival analysis and visualization were performed using the R packages survival (v3.3-1) and survminer (v0.4.9). Immune subtype classification was conducted with the ImmuneSubtypeClassifier (v0.1.0) R package. A nomogram was generated using the rms (v6.7-1) package. A risk score model based on 13 super-enhancer-related genes (SERGs) was constructed, demonstrating that patients in the low-risk group had significantly better prognosis. The SERGs signature significantly correlated with age, molecular and immune subtypes, IDH mutation, MTMG promoter methylation, 1p19q co-deletion, and expression of immune checkpoint genes in glioma patients. The SERGs signature could predict the prognosis and immune features of glioma, and SERGs might serve as novel immunotherapy options for glioma.

Phalaenopsis orchid flower extract attenuates high glucose-induced senescence via Nrf2/HO-1 activation and promotes wound healing in human dermal fibroblasts

2025-07-06T10:23:52+02:00

Skin aging in diabetic patients is closely associated with delayed wound healing and oxidative stress-mediated fibroblast dysfunction. This study investigated the protective and regenerative effects of a water extract of Phalaenopsis orchid flower (WEPF), an ornamental plant endemic to Taiwan, on high glucose (HG)-induced cellular senescence in human dermal fibroblasts (CCD-966SK), with a focus on the Nrf2/HO-1 antioxidant pathway. Cytotoxicity, cellular senescence, and ROS production were respectively assessed using MTT assay, senescence-associated β-galactosidase (SA-β-gal) staining, and DCFDA-cellular reactive oxygen species assay. Western blotting and ELISA were used to analyze the cellular senescence-related proteins. Fibroblasts treated with WEPF under HG conditions exhibited reduced senescence-associated β-galactosidase (SA-β-gal) activity, lower ROS levels, and attenuated cell cycle arrest. Protein expression profiling revealed suppression of the p53/p21^Waf1, and p16^INK4a/Rb pathways and decreased matrix metalloproteinase-1 (MMP-1) expression. Mechanistically, WEPF exerted its effects by activating the Nrf2/HO-1 axis and restoring the expression of senescence marker protein-30 (SMP30), thereby promoting fibroblast repair and reducing pro-inflammatory signaling. These findings support the potential of WEPF as a botanical therapeutic agent for diabetic wound healing and age-related skin deterioration.

Comparative study of DNA and RNA extraction methods for high-quality nucleic acid isolation from Cullenia exarillata A. Robyns

2025-07-06T10:22:37+02:00

Isolation of high-quality DNA and RNA from plants with high polysaccharide and secondary metabolite content is typically difficult, particularly in the case of trees. Metabolites commonly undergo co-precipitation with RNA and DNA, resulting in degradation of their quality. Cullenia exarillata leaf samples were subjected to various DNA and RNA extraction techniques, and the resulting data were compared and analysed. The isolation of high-quality DNA and RNA is crucial for the advancement of molecular and biotechnological techniques that aim to preserve the endemic status of C. exarillata and this research is to establish an efficient procedure for extracting DNA and RNA from C. exarillata. This method will make molecular and genomic research easier in forestry and conservation fields. In this research, we evaluated various methods for extracting high quality DNA and total RNA from C. exarillata tree, incorporating minor modifications in standard procedure. To acquire DNA and RNA of superior quality, a comparison was made between conventional DNA and RNA extraction methods and a variety of commercial kits thatrevealed the conventional technique yielded DNA samples of superior purity and concentration. It was discovered that combining modified commercial and conventional procedures yielded RNA with exceptionally high concentration and purity. The Agilent 2100 Bioanalyzer and NanoDrop spectrophotometer ensure the impeccable purity of the nucleic acids generated via these procedures. Additionally, the application of agarose gel electrophoresis unveiled unique bands. Further investigation was conducted to validate the purity and amplification of the DNA and RNA that were collected. This study clarifies a method for extracting sufficient and high-quality amounts of DNA and RNA from C. exarillata; future research on this plant will greatly benefit from knowing this information.

CAR-T cell therapy for rheumatoid arthritis: current status and molecular insights

2025-07-06T10:23:26+02:00

Chimeric antigen receptor (CAR)-T cell therapy, a breakthrough in hematological cancer treatment, is now being explored for autoimmune diseases like rheumatoid arthritis (RA). RA, characterized by chronic joint inflammation and autoantibody production, presents a compelling target for CAR-T cell therapy due to its potential for precise targeting of aberrant immune cells and restoration of immune tolerance. This review analyzes current strategies in CAR-T cell therapy for RA, focusing on molecular mechanisms and clinical implications. We discuss approaches such as CD19-targeted B cell depletion, simultaneous targeting of B cells and memory plasma cells, and the use of chimeric autoantibody receptors (CAARs) to target specific autoantigens. Furthermore, we explore the latest advancements in CAR-T cell engineering, including novel costimulatory domains, dual-targeting strategies, and the development of regulatory CAR-T cells (CAR-Tregs). This review provides insights into the efficacy and safety of CAR-T cell therapy for RA, highlighting its potential to revolutionize clinical applications and future directions in the field.