Maaweya Awadalla

Influenza A viruses remain a major global public health threat due to their high mutation rates, antigenic drift, and periodic antigenic shift, which collectively undermine the effectiveness of seasonal vaccines. Conventional influenza vaccines necessitate frequent reformulation and often result in suboptimal protection. Epitope-based vaccine strategies that focus immune responses toward conserved viral regions represent a promising approach for inducing broader and durable immunity. In this study, reverse vaccinology and immunoinformatics-guided approaches were used to design multi-epitope vaccine constructs derived from conserved regions of Influenza A H3N2 and H1N1 proteins, including hemagglutinin (HA), neuraminidase (NA), nucleoprotein (NP), and matrix proteins (M1 and M2). BALB/c mice were immunized intramuscularly and evaluated for humoral and cellular immune responses. Serum IgG levels were measured by ELISA, while functional antibodies were assessed using hemagglutination inhibition (HAI) assays. Cellular immune responses were evaluated using ELISPOT assays for IFN-γ and TNF-α secretion. Protective efficacy was assessed through homologous viral challenge with H3N2 or H1N1 strains, followed by survival analysis and lung histopathological examination. Immunization with H3N2- and H1N1-derived epitopes elicited robust specific IgG responses and induced functional HAI antibodies. Notably, H3N2-derived epitopes generated higher HAI titers, consistent with enhanced humoral immunogenicity. Vaccinated mice demonstrated strong cellular immune responses, characterized by significantly elevated IFN-γ-producing CD8⁺ T cells compared with mock-immunized controls. Upon viral challenge, all vaccinated mice exhibited 100% survival against H3N2 or H1N1 infection, whereas mock-immunized mice experienced 100% mortality. Histopathological analysis revealed protective immunopathological profiles and preserved alveolar architecture. The protective efficacy of the multi-epitope vaccination was comparable to that of the seasonal quadrivalent influenza vaccine. This study demonstrates that multi-epitope vaccines derived from conserved Influenza A proteins can induce coordinated humoral and cellular immune responses and confer robust protection against H3N2 and H1N1 viral challenge in a mouse model. This suggests support for peptide-based vaccination as an alternative method to conventional strain-matched vaccines, which reduce vaccine effectiveness or universal influenza vaccine development.

Respiratory syncytial virus (RSV) is a major pathogen responsible for lower respiratory tract infections, particularly in infants, young children, older adults, and immunocompromised individuals. Although several RSV vaccines and monoclonal antibody therapies have recently been approved, their protective efficacy is often short-lived and relies mainly on systemic immunity rather than mucosal immunity. As RSV is a mucosal pathogen, there is a compelling need to develop novel vaccine strategies capable of inducing robust mucosal immune responses. Inducible bronchus-associated lymphoid tissue (iBALT) is an ectopic lymphoid structure that plays a critical role in orchestrating the immune response to respiratory pathogens. iBALT is an important target for vaccine development, as it can support both innate humoral and cellular immune responses and contributes to the establishment of immune memory in the respiratory tract. The formation of iBALT is regulated by airway epithelial cells and the lung microenvironment through release of cytokines and chemokines, including interleukin 33 (IL-33) and C-X-C motif chemokine ligand 13 (CXCL13). In this review, we propose a novel mucosal vaccination strategy against RSV based on the combination of the RSV fusion (F) protein with immunomodulatory molecules, IL-33 and CXCL13, which are involved in the induction and regulation of iBALT. These immunostimulatory molecules may act synergistically to enhance the immune response to RSV infections. IL-33 promotes innate, mucosal, and adaptive immune responses and supports T cell activation, whereas CXCL13 facilitates B cell recruitment and organization within lymphoid structures. Although targeting iBALT represents a promising approach to improving RSV vaccine efficacy, further studies are required to determine the optimal regulation of iBALT in the airways.

Urinary tract infections (UTIs) are the most common bacterial infections and are frequently caused by pathogens such as Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis. Increasing antibiotic resistance has intensified the search for alternative antibacterial agents. In this study, extracts and bioactive compounds from Andrographis paniculata were evaluated against GlcN-6-P synthase, a key enzyme involved in bacterial peptidoglycan biosynthesis. Antibacterial activity was assessed using the agar well diffusion method, while molecular docking, molecular dynamics simulation and ADMET analyses were performed to evaluate binding affinity, stability and drug-likeness. The ethyl acetate fraction exhibited the highest antibacterial activity against multidrug-resistant uropathogens (MIC = 6.25 mg/mL; MBC = 12.5 mg/mL). Among the isolated compounds, PubChem IDs 15172, 1110 and 102916 showed the strongest binding affinities of -5.4, -5.2 and -5.1 kcal/mol, respectively. All ligands satisfied Lipinski’s rule of five and demonstrated favourable pharmacokinetic properties without hepatotoxicity. Molecular dynamics simulations identified compound 15172 as the most stable ligand with minimal conformational fluctuations. These findings suggest that the bioactive compounds from A. paniculata possess promising antibacterial and drug-like potential for the development of alternative therapies against UTI-associated pathogens.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) has become a major cause of bloodstream infections and poses serious challenges to clinical management because treatment options are limited. This study aimed to characterize antimicrobial resistance, virulence-associated features, and molecular epidemiology of CRKP bloodstream isolates using integrated phenotypic and genomic approaches. A total of 74 non-duplicate CRKP isolates were collected from bloodstream infections at three tertiary-care hospitals in Riyadh, Saudi Arabia, between 2022 and 2024. All isolates showed classical Klebsiella pneumoniae phenotypic characteristics, including intrinsic resistance to natural and aminopenicillins, and were classified as either multidrug-resistant (MDR) or extensively drug-resistant (XDR). Resistance to imipenem was universal, and resistance to other β-lactams and fluoroquinolones was high. Carbapenemase genes were detected in 96.0% of isolates using the GeneXpert^® Carba-R assay, with bla_OXA-48-like and bla_NDM being most common. Whole-genome sequencing demonstrated predominance of Ambler class D carbapenemases, particularly bla_OXA-232, with additional contributions from bla_NDM-1 and bla_NDM-5. Co-occurrence of carbapenemase genes was observed in a subset of isolates. Virulence analysis showed that 37.8% of isolates exhibited a hypermucoviscous phenotype, and more than half carried at least one virulence-associated determinant linked to capsule regulation or iron acquisition. In contrast, most isolates showed weak or no biofilm-forming capacity. Multilocus sequence typing revealed substantial genetic diversity but clear dominance of high-risk lineages, particularly ST147 and the emerging ST2096, both closely associated with bla_OXA-232 and bla_OXA-48-like genes. Capsular and O-antigen analysis showed a non-random distribution dominated by KL64 and O1/O2. Phylogenetic analysis was consistent with clonal expansion and suggested intra-hospital spread, with the intensive care unit serving as a key reservoir and dissemination to other wards. In conclusion, CRKP bloodstream infections in this setting are largely associated with a limited number of epidemic clones that combine extensive antimicrobial resistance with virulence-associated traits. These findings support the need for ongoing genome-based surveillance, strengthened infection control measures, and antimicrobial stewardship to limit the spread of high-risk K. pneumoniae lineages in healthcare settings.

Background/Objectives: Breast cancer (BC) remains the most frequently diagnosed malignancy among women worldwide and represents a significant global health burden. The genes associated with tumor suppression (p53, BRCA1, and BRCA2), telomere length maintenance (TERT), DNA damage response (FGFR2), and DNA repair (CHD1) are recognized for their intricate function in tumor genesis and progression. However, the prognostic significance of these genes in BC remains an area of research interest. This study aimed to examine the potential association between the presence of the MMTV and the expression patterns of these six genes with patient prognosis and survival outcomes in BC. Methods: This study includes 125 formalin‐fixed, paraffin‐embedded (FFPE) tissue specimens taken from BC patients, in addition to 25 tissue samples of benign breast lesions were incorporated as controls. The mRNA expression levels of six genes, namely p53, BRCA1, BRCA2, TERT, FGFR2, and CHD1, were quantified in FFPE tissue samples using quantitative polymerase chain reaction (qPCR). The correlation between gene expression and prognostic characteristics and the probability of recurrence‐free survival (RFS) and overall survival (OS) were assessed. Results: The results do not indicate an association between MMTV and BC, as the virus was not detected in any of the tissue samples analyzed. We observed a significant differential expression in five of the six studied genes between BC and noncancerous breast tissue, with significant downregulation of BRCA1, BRCA2, CHD1, and TERT, significant upregulation of p53, and unchanged levels of FGFR2. Among BC patients, p53 and BRCA1 expression levels emerged as significant prognostic factors for both RFS (32 vs. 24 months; 34 vs. 26 months) and OS (28.5 vs. 24 months; 31 vs. 28 months), respectively. Kaplan–Meier survival analysis of p53 expression displayed a trend favoring low expression for better survival and showing relatively stable RFS and OS survival curves of p53 until 43 and 54 months of the follow‐up period, respectively. Conclusions: When comparing cancer to noncancer patients, only p53 and BRCA1 expression levels emerged as significant prognostic factors for both RFS and OS in the entire cohort, with p53 displaying a trend favoring low expression for better survival. Although gene expression data provided a prognostic value, future studies should aim at integrating multiomics data and evaluating biomarkers in a broader clinical context to improve the accuracy of prognostic models and guide personalized treatment strategies.

Every two minutes, a woman dies from cervical cancer, which is considered the fourth most common cancer among women worldwide. The dynamic interplay between tumor inflammation, immune crosstalk, oncogenes, and tumor suppressor genes plays a crucial role in tumor development and progression. Using clinical and integrated bioinformatics, the mRNA expression pattern of 168 immune and tumor-related genes in the tumor microenvironment (TME) of HPV-positive cervical squamous cell carcinoma (CSCC) was analyzed. The study identified 94 DEGs, of which 55 genes were remarkably upregulated, including CASP8, ZHX2, BCL2L1, CTNNB1, RB1, BAX, CD274, CCL20, FOXP3, and CCL18. The top three-fold changes were associated with CASP8, ZHX2, and BCL2L1, respectively. In contrast, downregulation was discovered for 39 genes associated with immunity, regulation of cell cycle, and DNA damage response (HRAS, CCND1, ATM, CXCR1, and MIF). Gene-gene interaction and correlation analysis showed positive correlations, including RB1 and CASP8, RB1 and BCL2L1, and CCL20 with CCL18. Notably, six genes exhibited increased expression and showed a strong correlation with enhanced overall survival (OS) and disease-free survival (DFS), indicating their potential utility as prognostic biomarkers. Upregulated genes were positively associated with various immune cells, including B cells, CD8+ and CD4+ T cells, macrophages, neutrophils, and dendritic cells. Functional enrichment analysis revealed involvement in cancer-related processes, inflammatory responses, and cell migration, with key pathways linked to cytokine signaling and chemokine receptor interactions. Through the integration of clinical, experimental, and computational analyses, potential therapeutic targets and prognostic biomarkers were identified that may help improve clinical outcomes. Future studies should focus on the functional assays of identified genes both in vitro and in vivo.

To stop epistaxis, a Saudi medical invention called “Stop Nosebleeds” with a patent was created. It is a silicone-based, adjustable device that is applied externally to the bridge of the nose and speeds up clotting by decreasing the temperature and the compressing effect of the device on the nose This study aims to examine the efficacy of the device in attaining hemostasis in goats with anterior nasal bleeding. An animal experimental nasal-bleeding model was conducted on ten goats in collaboration between King Fahad Medical City and King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia. The left nostril was used as interventional wounds and the right nostril was used as control. Control wounds were treated with manual compression for 20 minutes. Interventional wounds were treated with the Stop Nosebleeds device. The second group of animals was heparinized and 2 wounds were created in each animal, one of which was being treated with the Stop Nosebleeds device, while the other was being treated with manual compression. The mean bleeding duration in the experimental group (both heparinized and non-heparinized goats) was 45.5 seconds (SD = 8.2), compared to 206.0 seconds (SD = 75.7) in the control group (U = 0.00, p < 0.001). The device stopped bleeding in 52.0 seconds and 39.0 seconds in the heparinized and non-heparinized experimental groups. While, the manual compression of epistaxis stopped bleeding in 252.5 seconds (SD = 84.7) and 159.2 seconds (SD = 15.7) in the heparinized and non-heparinized control groups, respectively. The device shows promise for clinical application, however, further research with larger sample sizes and human clinical trials are needed to validate its efficacy and safety. Additionally, the use of a goat model presents limitations in directly translating these results to human clinical practice, and these differences should be careful.