Introduction:

Breast cancer (BC) is highly heterogeneous, with varying molecular characteristics. Autophagy, a critical cellular degradation process, helps cells survive under stress, but its regulation can be influenced by altered microRNA (miRNA) expression. Studying miRNA changes during starvation-induced autophagy in both mammary epithelial cells and BC cells could reveal potential molecular targets.

Methods:

BC cell lines and rat mammary epithelial cells were subjected to starvation using Earle’s Balanced Salt Solution (EBSS) or treated with cell medium (CM). The STRING database identified proteins related to autophagy and starvation, which were cross-referenced with known miRNA interactions. Total RNA from both CM- and EBSS-treated cells was analyzed for miRNA expression changes and validated using the TCGA dataset.

Results:

Starvation induced autophagy, reduced cell proliferation in all cell types, and increased the invasive capacity of BC cells. We identified a miRNA signature related to starvation, comprising 27 miRNAs. Six miRNAs had elevated baseline expression, while another six, including miR-218a-5p, were reduced in BC cells compared to healthy cells. Starvation caused significant miRNA dysregulation, with miR-218a-5p being notably affected. Bioinformatic analysis showed that although miR-218a-5p is less expressed in primary BC tissues compared to normal tissues, its expression increases with advanced N status and is higher in metaplastic BC.

Conclusion:

The response to starvation varies between rat BC cells and normal cells. Baseline miRNA expression related to starvation and autophagy differs between breast cancer and healthy cells. Starvation induces BC-specific miRNA dysregulation, particularly affecting miR-218a-5p, which could have therapeutic implications through autophagy modulation.

This research was funded by the Medical University of Warsaw (69/M/MG/N/23) and the Polish Ministry of Science and Higher Education (SKN/SP/569610/2023).

Nuclear factor-κB (NF-κB) is a pivotal transcription factor family involved in key biological processes such as inflammation, immune response, cell survival, apoptosis, cellular stress reactions, and tumorigenesis. Dysregulation of NF-κB is increasingly recognized as a critical factor in the initiation and progression of various cancers. Polymorphisms in miRNA genes or their target sites (miRSNPs) can significantly impact miRNA activity. While polymorphisms in miRNA genes are rare, studies have shown that SNPs at miRNA target sites can either enhance or reduce the strength of miRNA–target interactions. The objective of this study was to identify miRSNPs in the NF-κB gene and SNPs in miRNA genes targeting their 3'UTR and to evaluate their potential roles in apoptosis and cancer using computational tools. We identified 121 miRNA binding sites corresponding to 101 distinct miRNAs, as well as 16 SNPs located within miRNA binding regions of the NF-κB 3'UTR. Notably, a binding site for miR-6826-5p in the NF-κB 3'UTR harbors an SNP (rs960795970, A/G), and the same miRNA’s genomic sequence contains an SNP (rs6771809, C/T) at the same nucleotide position as rs960795970. Additionally, miR-6826-5p has three other SNPs (rs757908839, A/G; rs746350709, C/T; and rs115693266, A/C), with the first positioned directly at its binding site. This cross-matching between miRSNP (rs960795970) in the NF-κB 3'UTR and an SNP (rs6771809) in the miR-6826-5p genomic sequence in the same binding region suggests potential functional interplay. Moreover, miR-6826-5p was found to target several genes associated with cancer and apoptosis, including HIP1, TRIAP1, GSKIP, NIN, DAP, CAAP1, XIAP, TMBIM1, TMBIM4, TNFRSF10A, RAD21, AKT1, BAG1, and BAG4, despite having no previously established cancer-related interactions. These findings imply that miR-6826-5p may play a critical role in apoptosis through pathways beyond NF-κB, potentially influencing cancer progression via alternative mechanisms.

Ayurvedic medicine offers a holistic approach to managing complex diseases such as Type 2 diabetes mellitus, which is primarily driven by insulin resistance. Bioinformatics has emerged as a powerful tool for understanding the molecular mechanisms underlying the effects of Ayurvedic drugs on insulin resistance pathway targets. This study uses bioinformatics techniques to analyze the pharmacological potential of Ayurvedic herbs traditionally used to treat diabetes, such as Gymnema sylvestre and Withania somnifera. The key bioactive compound in G. sylvestre has been shown to suppress the taste of sugar and reduce glucose absorption in the intestines, promoting better blood sugar control. Also, recent studies highlight W. somnifera role in improving insulin sensitivity and reducing blood sugar levels Using databases like DrugBank, STITCH, and PubChem, active compounds in these herbs were identified, and their interactions with key targets in the insulin signaling pathway, including IRS1, PI3K, and GLUT4, were analyzed. Molecular docking, network pharmacology, and gene expression analyses revealed that these phytochemicals potentially modulate glucose uptake, insulin sensitivity, and inflammation, key processes involved in insulin resistance. Our findings suggest that the synergistic effects of these herbs could offer a complementary approach to managing insulin resistance, warranting further experimental validation. The study paves the way for rational drug design based on traditional knowledge. It contributes to the growing field of systems biology in exploring the efficacy of multi-compound, multi-target therapeutic strategies.

Introduction:

Sepsis is a life-threatening disease caused by an excessive immune response to infection, causing inflammation, tissue damage, and organ failure. Globally, sepsis caused 19.7% of all fatalities in 2017, with India having one of the highest rates, with over 4 million cases annually. MicroRNAs (miRNAs) are small, non-coding RNA molecules that regulate gene expression by binding to messenger RNAs (mRNAs) and blocking their transcription. Identifying miRNAs as predictive biomarkers could significantly reduce the worldwide burden of sepsis by enhancing patient care, management, and treatment. This study aims to discover potential diagnostic biomarkers implicated in the dysregulated immune response and provide insights for mechanistic illness progression research.

Methods:

The Gene Expression Omnibus (GEO) datasets GSE134364, GSE134358, GSE94717, GSE236713 and GSE131761 were used to identify the differentially expressed genes involved in sepsis. Biological targets were identified that coincide with the identified differentially expressed miRNAs. The biological pathways were analyzed using KEGG and GO. Hub genes were identified for further functional and signaling pathway analysis.

Results:

Several differentially expressed miRNA biomarkers were identified between the control and sepsis groups (p<0.05 and fold change >2). MiR-146a regulates the immune response, miR-155 promotes pro-inflammatory cytokine expression, miR-150 regulates immune cell function and inflammation, and miR-21 is involved in apoptosis and inflammation. Moreover, we identified matrix metalloproteinase (MMP8 and MMP9) as being involved in neutrophil degranulation.

Conclusion:

MiRNAs regulating MMP-8 and MMP-9 in sepsis regulate immune responses and inflammation, leading to tissue damage. MiR-21 downregulate MMP-9 expression, while miR-146a indirectly affects MMP expression by targeting upstream signaling molecules. Targeting specific miRNAs that regulate MMPs offers a potential therapeutic strategy for managing sepsis. By modulating the levels of these miRNAs, it may be possible to control the activity of MMPs and reduce the harmful effects of excessive inflammation and tissue damage.

Carbapenem-resistant Acinetobacter baumannii (CRAb) is a priority pathogen, according to the World Health Organization (WHO), that is the most threatening to human health, being responsible for pneumonia, bacteraemia, urinary tract, skin and soft tissue infections, with mortality rates approaching 35%. The aim of this study was to evaluate the occurrence of colistin resistance (mcr1-5) and biofilm genes (surA1, bap, ompA, luxR, epsA) in 26 CRAb isolates collected from hospitalised patients and Acinetobacter baumanii ATCC 19606 as the reference strain. Gene prevalence was determined by multiplex or single PCR (polymerase chain reaction), in a 25 µL mixture (12.5 μL of REDTaq® ReadyMix™ PCR Reaction Mix, 1 μL of each primer, 2 μL bacterial DNA). As a result, 92.3% (24/26) and 96.2% (25/26) of the isolates were mcr1- and mcr3-positive, respectively. None of the isolates possessed the mcr2, mcr4 and mcr5 genes. All CRAb isolates had the surA1, bap, ompA and luxR genes, while 61.5% of isolates (16/26) harboured the epsA gene.

CRAb are highly pathogenic bacteria isolated from infections and are increasingly resistant to most available therapies. Every effort should be made to prevent the spread of these microorganisms, including analysing the presence of new genetic determinants in the CRAb genome, and identifying biofilm-related genes whose expression inhibition could more effectively eradicate these pathogens.

Introduction: Nowadays, infertility represents a multifaceted health issue that significantly impacts numerous couples, giving rise to notable psychological and social complexities. Notably, one in six couples experience infertility, with approximately 50% of cases attributed to male factors. Male infertility, a complex disorder influenced by both environmental factors and genetic predisposition, involves the interplay of various genes contributing to its manifestation. It can be categorized into specific subtypes, including asthenozoospermia. The primary aim of this study was to identify novel variants associated with asthenozoospermia within the Greek population and to elucidate the roles of the genes involved. Materials and Methods: Whole-genome sequencing (WGS) was conducted on both normozoospermic and asthenozoospermic individuals. Following the identification of variants exclusively present in asthenozoospermic men, an extensive range of tools, functional assessments, and predictive algorithms were employed to prioritize these variants. Results: The investigation unveiled numerous polymorphisms, comprising 155 classified as high-impact and 715 classified as moderate-impact. While several of these variants were found within genes previously linked to male infertility, a notable subset was associated with asthenozoospermia for the first time. Furthermore, pathway enrichment analysis and Gene ontology (GO) analyses revealed polymorphisms on genes implicated in teratozoospermia through various mechanisms and pathways. Conclusions: This study reaffirms the involvement of previously studied genes in male infertility, while also shedding light on novel molecular mechanisms. By providing a comprehensive list of variants and candidate genes associated with asthenozoospermia within the Greek population, this research contributes significantly to our understanding of male infertility and paves the road for future studies.

Rheumatoid arthritis (RA) is a chronic autoimmune disease affecting multiple joints and causing adverse effects on organs. Early diagnosis, aggressive treatment, and disease-modifying anti-rheumatic drugs (DMARDs) have improved the management and long-term prognosis of the disease. The global prevalence of RA is increasing, primarily in women, and is associated with disability and mortality. The prognosis depends on the disease stage at diagnosis; early diagnosis can prevent or delay disease progression in 90% of patients, preventing irreversible joint damage and disability. However, non-coding RNA (ncRNA), including microRNA and long ncRNA (lncRNA), which are small, non-coding segments of RNA that regulate gene expression, has been discovered to be deregulated in diseases like RA, potentially contributing to its pathogenesis, progression, and treatment. Pre-clinical RA, characterized by auto-antibodies and biomarkers before clinical RA, is a stage of RA research aimed at the early diagnosis and control of immunological abnormalities. For instance, SNP rs2850711 in lnc00305 links RA susceptibility to the response of miR-10a to methotrexate (MTX) treatment, and ncRNA plays a crucial role in refractory RA by regulating drug sensitivity. Lower miR-20a expression in rheumatoid arthritis synovial fibroblasts (RASFs) activates Janus Kinase (JAK)-mediated inflammation, promoting cell proliferation and apoptosis resistance. This study highlights changes in ncRNAs, disease progression, and novel therapeutic targets and strategies.

The complex nature of aggression and challenges in phenotype description often make it difficult to identify genetic variations associated with aggressive behavior. The use of machine learning algorithms and multivariate analysis makes it possible to construct more accurate diagnostic schemes to characterize the underlying causes of aggression. The aim of this study was to develop a method to differentiate aggression profiles among subjects in a heterogeneous sample and to facilitate the association of the obtained profiles with genetic variations of BDNF rs6265, BDNF rs10835210, HTR2A rs6313, and DRD4 1800955 later.

Participants' aggressiveness was assessed using the Buss–Durkee Hostility Inventory (BDHI). On the basis of the 10 scales of the BDHI, 1013 unique subspaces were formed, in which data were clustered. "Cluster neighborhood" plots were constructed for each participant in the study, allowing us to identify the content of each category of subjects in all of the clusters into which the participant fell. This approach demonstrated two dramatically different phenotypes based on all the possible combinations of characteristics. However, many study participants exhibited mixed phenotypes, which were separated using the DBSCAN spatial clustering algorithm.

Individuals initially classified into a control group, addiction-prone group, or inmate group (convicted of felony) were often adjacent to each other in the newly formed clusters, suggesting the need to reevaluate the sample: 13 individuals from the addiction-prone group and 17 individuals from the inmate group were assigned to the control group, and 24 individuals from the inmate group and 6 individuals from the control group were assigned to the addiction-prone group. A total of 140 individuals belonged to the monotype group, while the remaining 261 individuals belonged to the mixed phenotype group.

Phenotypes defined by indirect traits often create an erroneous picture. It is necessary to identify similar groups of patients based on their psychometric data and then examine their genotypes within these groups.

Chronic kidney disease (CKD) poses a significant global threat, with increased rates of cardiovascular and all-cause mortality. Anemia is common in CKD, and is often associated with the accumulation of uremic toxins in the bloodstream. Previously, we demonstrated that the uremic toxin indoxyl sulfate (IS) has an impact on the regulation of erythropoiesis in both cellular and preclinical CKD models. In this study, on the roles of non-coding RNAs in this toxin/toxic effect, we measured the effect of IS on microRNA expression in the human erythropoietic cell line UT7/EPO, using nanostring technology. We found a significant increase in miR-223 in cells treated with IS. This finding was further validated in human primary CD34+ cells, a more physiological model for human erythropoiesis. Finally, serum levels of miR-223 correlated with representative uremic toxins, including IS, in patients with various stages of CKD, and also with endothelial dysfunction markers, indicating a link with vascular damage. These findings suggest that miR-223 may contribute to the development of anemia in CKD. Further investigation into the involvement of miR-223 in erythropoiesis is needed for a better understanding of the mechanisms underlying anemia in CKD and the potential role of uremic toxins. Ultimately, this may open up new therapeutic possibilities for the management of anemia in CKD.

Background: Congenital heart disease (CHD) is a global health concern, particularly in low- to middle-income countries like India. The renin–angiotensin–aldosterone system (RAAS) plays a crucial role in the development of cardiovascular disorders and hypertension, with the angiotensin-converting enzyme insertion/deletion (ACE I/D) polymorphism being a key genetic factor. Our study aims to elucidate the genetic influence of ACE I/D polymorphism on CHD in a north Indian cohort.

Methods: A total of 667 CHD cases, including 433 individuals with parental data and 104 controls, were enrolled and genotyped by polymerase chain reaction. Case–control association, parental transmission tests, and the association of patients' and parents' clinical parameters with ACE I/D were explored.

Results: The frequencies of the DD, ID, and II genotypes were 0.23, 0.47, and 0.30, respectively. Our findings highlight significant associations, notably the increased CHD risk conferred by the DD genotype in females (p = 0.036; OR = 1.68), its correlation with abnormal hemoglobin (p = 0.049; OR = 1.68), and its impact on primigravida (p = 0.05). Conversely, the II genotype was found to significantly elevate the risk of CHD in the offspring of tobacco-consuming fathers by 2.5-fold (p = 0.029). Notably, cyanotic cases exhibited a heightened prevalence of ACE I/D mutations (p = 0.059), with the Tetralogy of Fallot showing the strongest association (p = 0.024). Additionally, the DD genotype's involvement in conditions such as stenosis (p = 0.026) and pulmonary artery hypertension (p = 0.05) underscores its clinical relevance. The parent-of-origin test showed maternal transmission of the D allele in combined (p = 0.037) and acyanotic cases (p = 0.039) and paternal transmission in ventricular septal defect (p = 0.021).

Conclusion: This is the first study from India and possibly the only study globally that reports a significant association between ACE I/D and CHD, highlighting the importance of genetic factors in CHD susceptibility.