Background: Accurate and rapid identification of Mycobacterium species is essential for timely diagnosis and appropriate treatment of tuberculosis and nontuberculous mycobacterial (NTM) infections. Conventional methods are time-consuming and can have limited sensitivity, while Matrix-assisted Laser Desorption Ionization–Time of Flight (MALDI-TOF) mass spectrometry offers the potential for fast and precise species-level identification. Methods: This is a cross-sectional study conducted in the laboratory of a tertiary care hospital. The study included all the mycobacterial culture-positive samples identified by MALDI-TOF over 6 months. The effectiveness of MALDI-TOF Mass Spectrometry (MALDI TOF MS) was compared with the MPT64 antigen detection test and multiplex polymerase chain reaction. Results: Out of 836 samples received for acid-fast bacilli culture, 72 samples tested positive for the culture, with 68 (94.4%) accurately identified as various species of Mycobacterium and Nocardia using MALDI-TOF. Four samples could not be identified, as they failed to grow on Löwenstein-Jensen media on subculture. Among the identified samples, 41 were Mycobacterium tuberculosis complex (MTBC). Other findings included 24 NTMs: 10 Mycobacterium abscessus, 7 Mycobacterium fortuitum, 2 Mycobacterium intracellulare, 3 Mycobacterium gordonae, 1 each Mycobacterium oxidans, Mycobacterium margaritense, and 2 Nocardia cyriacigeorgica and 1 Nocardia farcinica. MALDI TOF MS showed high specificity at the species level, as it reliably differentiated MTBC and NTM and Nocardia Species with no evidence of misidentification. Conclusion: MALDI-TOF offers a rapid, easy, and cost-effective method for identifying mycobacterial species, typically requiring specialized laboratories. This technique involves minimal handling and reduces working hours, thereby lowering the risk of laboratory-acquired infections. Our study substantiates that MALDI-TOF is a reliable and fast option for NTM identification. However, standardizing the procedure with liquid media inoculated with clinical specimens is essential to further reduce diagnosis time.
Background: Antibacterial resistance is distributed worldwide, such as in the United States, killing about 23,000 patient/year. It occurs when bacteria evolve to evade the effect of antibiotics through neutralization of antibiotics, modifying their outer structure or receptor, or through mutation of their genetic materials. Previous studies demonstrate that cinnamon oils can change bacterial membrane permeability, which can lead to the destruction of bacteria. Other studies showed that Salvia officinalis contains phenolic essential oils (EOs), which act as inhibitors of bacterial growth. In this study, we investigate the effects of combined Ceylon cinnamon and S. officinalis to evaluate their effectiveness against specific types of bacteria. Methods: This is a cross-sectional laboratory-based study; in this study, 60 isolated bacteria were investigated. The EOs of Ceylon cinnamon and S officinalis were extracted by the Soxhlet method, and the total flavonoids were determined using liquid chromatography–electrospray ionization–mass spectrometry analysis. On the other hand, the minimal inhibition concentration (MIC) and minimal bactericidal concentration (MBC) of the extracts were determined using a 96-well plate technique according to the method described by the Clinical and Laboratory Standards Institute. Two-fold serial dilutions of the extracts were prepared and added to sterile 96-well plates with the concentration of 0.0625 (mg/ml), 0.125 (mg/ml), 0.25 (mg/ml), 0.5 (mg/ml), 1 (mg/ml), and 2 (mg/ml), and dimethyl sulfoxide was used as a control. The altered strains were then inoculated at a preliminary concentration of 106 CFU/mL. Then, the OD600 was checked after 16 h of incubation. The lowest concentration at which the extract inhibited the growth of the strain was selected as the MIC. The MBC was identified by streaking of the broth from clear wells onto Mueller–Hinton agar plates and incubating for 24 h. The lowest concentration of an antimicrobial agent at which all the cells were killed was defined as the MBC. Data were structured in Microsoft Excel and transfered to a Statistical Package for Social Sciences (SPSS) (SPSS version 26) for analysis. The test is considered significant if P < 0.05. Results: Escherichia coli represented the most frequent bacteria isolated with 60%; the MIC and MBC were determined as 2.08 and 16.67 for E. coli, 4.13 and 33.33 for Staphylococcus aureus, and 8.33 and 16.77 for Streptococcus agalactiae, whereas 4.0 and 20.0 for Staphylococcus epidermidis. The extract showed high sensitivity to Staphylococcus epidermidis with 91.7%, then S. agalactiae with 80.0%, followed by E. coli, which represented 74.3%, while it gave low sensitivity to S. aureus, 50%. The extract demonstrates more antibacterial activity 78% compared to ciprofloxacin P = 0.002, and less activity than amikacin antibiotics P = 0.03. Conclusions: Depending on our findings, it can be concluded that cinnamon and S. officinalis (sage) extract have a likely antibacterial activity through inhibition of bacterial growth and effect on membrane permeability, especially against S. epidermidis, S. agalactiae, and E. coli, respectively.
Background: Histidine decarboxylase (HDC) and SLIT and NTRK-like family member 1 (SLITRK1) are genes implicated in Tourette syndrome (TS) through roles in histaminergic neurotransmission, synaptic signaling, and neurodevelopment. This study aimed to characterize the structural and functional properties of their encoded proteins and assess their potential pharmacological relevance using an integrative in silico approach. Methods: Human HDC and SLITRK1 sequences and variant data were obtained from public databases and analyzed using computational tools for physicochemical profiling, secondary-structure prediction, protein interaction analysis, structural modeling, binding-site prediction, molecular docking, and adsorption, distribution, metabolism, excretion, and toxicity (ADMET) assessment. Three-dimensional protein models were generated using AlphaFoldDB-assisted template modeling with refinement and validation. A panel of clinically relevant neuroactive compounds was screened for predicted interactions with both proteins. Results: HDC and SLITRK1 displayed distinct physicochemical and structural features consistent with their biological functions. Variant analysis identified multiple missense and clinically reported variants in both genes, supporting their functional significance. Network analyses linked HDC mainly to histamine synthesis and metabolic pathways, whereas SLITRK1 was more strongly associated with synaptic organization and neuronal signaling. Structural validation supported the suitability of the predicted models for exploratory analyses. Docking studies identified several compounds with favorable predicted binding affinities for both targets, while ADMET profiling revealed differences in pharmacokinetic and toxicity properties among the screened compounds. Conclusion: HDC and SLITRK1 are promising candidate proteins in TS. This integrative computational framework may aid prioritization of variants and compounds for future investigation, although experimental validation remains necessary.
COG6-congenital disorder of glycosylation type IIL (CDG2L) is a rare multisystemic autosomal recessive disorder caused by variants in the COG6 gene, a core component of the conserved oligomeric Golgi (COG) complex. To date, approximately 50 affected individuals with only 20 variants have been reported worldwide. We investigated a pregnant mother with a history of CDG2L in three affected neonates (two females and one male), who died within the 1st month of life due to severe multisystem manifestations of the disease. The first affected girl presented with microcephaly and congenital heart defects. The second affected girl manifested microcephaly, clubfoot, and congenital heart defects. The third affected boy presented with microcephaly, clubfoot, imperforated anus, hydronephrosis, congenital heart defects, and seizures. Whole-exome sequencing of the most recent affected male neonate (12 days old) identified a novel homozygous missense variant (c.1074G>C) in the COG6 gene. During the subsequent pregnancy, Sanger sequencing was done at 12 weeks of gestation using chorionic villus sampling (CVS), which detected the same homozygous variant in the fetus. Functional analysis using reverse transcription-polymerase chain reaction from the CVS showed that this variant causes abnormal splicing, resulting in skipping of exon 11. The parents received appropriate genetic counseling for future reproductive planning. Following prenatal diagnosis of the affected fetus, the pregnancy was terminated. Preimplantation genetic testing was later performed, which resulted in the birth of a healthy male child. This report expands the variant spectrum of COG6 and underscores the importance of functional RNA analysis for accurate variant interpretation. To our knowledge, this is the first study to report a deleterious missense variant in the COG6 gene causing aberrant splicing and lethal CDG2L.
Background: The group specific antigen (Gag) polyprotein of human immunodeficiency virus-1 is an essential protein for the formation of virus particles and has emerged as a target for potential therapeutic interventions. Identification of viable linear B-Cell and cytotoxic T-Cell epitopes within intrinsically disordered regions within the Gag polyprotein can be achieved through immunoinformatics and related computational tools. The identified epitopes can be incorporated into a synthetic peptide vaccine construct designed in silico. Methods: Linear B-Cell and cytotoxic T-Cell epitopes were identified through B-Cell and cytotoxic T-Cell epitope prediction tools found in the Immune Epitope Database. These were assessed for antigenicity, allergenicity, toxicity, cross-reactivity, and sequence conservation in silico. Disorder analysis was conducted and cross-referenced with predicted epitopes. These were combined into a vaccine construct along with adjuvants and linkers and tested for potential immunogenicity. Results: A total of 9 B-Cell and 2 T-Cell epitopes were identified as viable candidates, with both T-Cell epitopes and 7 B-Cell epitopes found in disordered regions. Different portions of a vaccine construct containing these epitopes, RS09, pan HLA DR-binding epitope, and the appropriate linkers binds spontaneously with Toll-like receptor 4 and induced increases and maintenance in memory B-Cell and memory helper T-Cell populations, with corresponding increases of cytotoxic T-Cell populations as well as antibody and cytokine concentrations. Conclusions: A synthetic peptide vaccine construct from B-Cell and T-Cell epitopes found in disordered regions that induces immune responses in simulations was designed in silico. As such, this could warrant further investigations of the vaccine construct in vitro and in vivo.
Background: While the Bacillus Calmette–Guérin vaccine provides early-life protection, its waning efficacy in adults remains a critical barrier to global tuberculosis (TB) control. Despite several candidates targeting region of difference (RD) loci proteins entering clinical trials, a successful replacement has yet to emerge. Consequently, the design of multi-valent, multi-epitope peptide vaccine can elicit the robust humoral and cell-mediated immune responses required for sustained, long-term protection against Mycobacterium tuberculosis. Methods: To develop the vaccine, 38 RD-loci proteins from the H37Rv strain were screened for CD4 + and CD8 + epitopes based on their binding affinity to common Human Leukocyte Antigen (HLA) allele. The construct was validated in silico for stability, safety, and nonallergenicity. The vaccine’s efficacy was further evaluated through molecular docking followed by molecular dynamics (MD) simulations to confirm robust binding affinity and stability with the toll-like receptor (TLR) 2 and TLR4 receptor. Results: A total of 7 B-cell, 9 cytotoxic T-lymphocyte (CTL), and 11 helper T lymphocyte epitopes were predicted and strategically integrated with the adjuvant human beta defensin-3 and neutrophil peptide-1 at N-and C-termini using flexible linkers to enhance immunogenicity. Comprehensive in silico profiling validated the chimeric construct as nontoxic, nonallergenic, and highly antigenic. Furthermore, molecular docking analysis demonstrated a robust interaction with the TLR2 receptor, characterized by a significant binding affinity, suggesting high potential for initiating a potent immune response. Conclusion: Collectively, these in silico findings demonstrate that the designed multi-epitope vaccine possesses the necessary structural and immunogenic properties to elicit robust humoral and cellular immune responses. The construct’s high antigenicity, combined with its stable interaction with key immune receptors, positions it as a highly promising candidate for TB prevention.
Background: Ferroptosis is an iron-dependent form of regulated cell death driven by lipid peroxidation and oxidative stress and emerging as a promising target for cancer therapy. Taxanes such as docetaxel (DTX) are widely used chemotherapeutics but require solvents such as polysorbate-80 and ethanol, which can cause hypersensitivity reactions. Nanosomal DTX lipid suspension (NDLS), a lipid-based formulation, improves drug stability, solubility, and permeability. Methods: Human breast cancer cells (Michigan Cancer Foundation [MCF]-7) were treated with increasing concentrations of NDLS (1–50 μM) for 48 h. Cell viability was determined by MTT assay to derive the half-maximal inhibitory concentration (IC50). Ferroptosis-associated parameters, including intracellular ferrous iron (Fe2+), reduced glutathione (GSH), and reactive oxygen species (ROS) levels, were quantified using standard biochemical assays. Cell death profiles were analyzed by flow cytometry using Annexin V-Fluorescein Isothiocyanate (FITC)/7-aminoactinomycin D (7-AAD) dual staining. All assays were conducted in biological triplicate (n = 3), and data are presented as mean ± standard deviation. Statistical comparisons were performed using one-way analysis of variance followed by Dunnett’s multiple comparison test (P < 0.05). Results: NDLS treatment showed a concentration-dependent reduction in MCF-7 cell viability after 48 h with an IC50 of 26 µM. NDLS exposure significantly increased intracellular Fe2⁺ levels and ROS generation. Intracellular GSH levels showed a marked dose-dependent depletion compared with those of untreated controls. Flow cytometric analysis using Annexin V-FITC/7-AAD staining demonstrated a substantial reduction in viable cells with a concomitant increase in Annexin V-/7-AAD-positive populations. Conclusions: These findings suggest that NDLS induces oxidative and biochemical alterations associated with ferroptosis in MCF-7 breast cancer cells.
Background: Idiopathic short stature (ISS) is characterized by a height below the mean for age, sex, and population, without identifiable systemic, endocrine, nutritional, or chromosomal abnormalities. Dysregulation of the growth hormone/insulin-like growth factor-1 (IGF-1) axis, particularly IGF-1 bioavailability, is a suspected contributor. Stanniocalcin-2 (STC2) inhibits pregnancy-associated plasma protein-A2, a protease that releases IGF-1 from its binding proteins. To investigate the association between circulating STC2 level, free IGF-1 bioavailability, and linear growth in children with ISS. Methods: This case–control study included 88 participants (45 children with ISS, 43 controls) aged 4–18 years. Serum STC2, total IGF-1, free IGF-1, and insulin-like growth factor-binding proteins (IGFBP)-3 levels were measured by the ELISA. Results: Children with ISS exhibited significantly lower free IGF-1 (213 vs. 291 pg/mL, P = 0.001) and IGFBP-3 (5.17 vs. 6.18 ng/mL, P = 0.003) levels than controls, while total IGF-1 remained comparable (P = 0.415). The IGF-1/IGFBP-3 ratio was reduced (27.4% vs. 32%, P = 0.039), as was the free/total IGF-1 ratio (0.14% vs. 0.18%, P = 0.021). STC2 was elevated in ISS patients aged ≥11 years (50.2 vs. 33.7 pg/mL, P = 0.034). Correlation analysis revealed negative associations between STC2 and both height (rs = −0.351, P = 0.004) and free IGF-1 (rs = −0.463, P < 0.05). Mediation analysis confirmed that free IGF-1 mediates STC2 effect on height (indirect effect β = −0.073). Multivariate logistic regression identified free IGF-1 as an independent predictor of ISS (Odds ratio = 0.99, P = 0.0016). Conclusion: Impaired growth in ISS was more closely associated with reduced IGF-1 bioavailability than with deficient production. Elevated STC2 levels, particularly in older children, contribute to a decrease in free IGF-1 levels.
Background: Gastritis is among the most common gastrointestinal disorders worldwide and is associated with serious complications such as gastritis disease and gastric cancer. This study investigated the role of interleukin-8 (IL-8) and microRNA-155 (miRNA-155) expression in patients with Helicobacter pylori-associated gastritis and examined their relationship with disease progression. Methods: A case–control study was conducted on 150 participants, including 100 patients diagnosed with gastritis and 50 healthy controls. Samples were collected from patients presenting with gastritis symptoms at Al-Marjan Gastroenterology Hospital between January and April 2026. H. pylori infection was confirmed by polymerase chain reaction (PCR) targeting the glmM gene. Serum IL-8 concentrations were measured using enzyme-linked immunosorbent assay, whereas miRNA-155 expression was assessed by reverse transcription PCR (RT-PCR) combined with bioinformatics analysis. Results: The study involved 150 participants, including 100 gastritis patients and 50 healthy controls. Age and gender showed no significant differences between groups, whereas urban residence was significantly associated with gastritis. Patients with gastritis demonstrated significantly higher IL-8 levels compared with controls (17.78 vs. 9.05 pg/mL, P < 0.001), reflecting enhanced inflammatory activity. Histopathological findings revealed greater involvement of the corpus, cardia, and other gastric regions among patients, whereas controls showed predominance in the antrum (P = 0.04). Urban residents exhibited a higher prevalence of gastritis, suggesting possible environmental and lifestyle contributions. In addition, miRNA-155 expression was markedly reduced in patients (0.47 ± 0.22) compared with controls (1.07 ± 0.43) (P < 0.0001). Strong positive correlations were found between H. pylori bacterial load, IL-8 levels, and miRNA-155 expression, suggesting enhanced inflammatory and immune responses. H. pylori copy number was identified as the main predictor of gastritis severity. Both IL-8 and miRNA-155 demonstrated excellent diagnostic performance, with IL-8 showing slightly superior accuracy. Power analysis confirmed an adequate sample size and strong statistical reliability. Conclusions: H. pylori-associated gastritis is linked to increased inflammatory responses, altered gastric tissue distribution, and significant downregulation of miRNA-155. These findings indicate that miRNA-155 may serve as a promising biomarker and contribute to the pathogenesis of gastritis.
Background: Seborrheic dermatitis (SD) is a chronic inflammatory dermatosis affecting approximately 5% of the global population; however, diagnosis relies entirely on clinical assessment without objective laboratory confirmation. We evaluated shotgun metagenomics as a laboratory tool for characterizing skin microbiome dysbiosis in SD, with emphasis on diagnostic performance, antimicrobial resistance (AMR) profiling, and clinical utility for treatment guidance. Methods: This cross-sectional study analyzed 115 facial skin samples from 78 Vietnamese participants (38 SD patients and 40 healthy controls) using shotgun metagenomics. We performed taxonomic profiling, functional pathway analysis, AMR gene detection, and virulence factor identification. Random Forest classification evaluated diagnostic accuracy, while MaAsLin2 identified severity-associated biomarkers. Results: We discovered a novel three-phase functional gradient (Control → Non-SD peak → SD decline) suggesting a “biological alert state” in clinically unaffected skin. Malassezia restricta dominated SD lesions (90.92% vs. 81.87%; P < 0.001) with 53.5% reduction in fungal richness. Bacterial profiling revealed Staphylococcus epidermidis expansion and aerobic-to-anaerobic community transition. AMR profiling demonstrated selective enrichment of fusB (fusidic acid resistance) and blaR1 (beta-lactam sensing), providing treatment guidance. The virulence factor iron-regulated surface determinant E showed significant severity association (q = 6.83 × 10−3). Diagnostic accuracy reached 75.64% (bacterial) and 66.67% (fungal profiles). Conclusion: This first Vietnamese shotgun metagenomic study reveals multilayered dysbiosis encompassing taxonomic, functional, AMR, and virulence dimensions in facial SD. The novel functional gradient offers early detection potential, while population-specific AMR profiles support personalized antimicrobial therapy selection. These findings substantially advance understanding of SD pathogenesis beyond previous amplicon-based studies.
Background: Circulating microRNAs (miRNAs) have emerged as promising molecular indicators of exercise-induced physiological adaptation. However, individual miRNAs have shown variable discriminatory performance across exercise contexts. miR-21 is of particular interest because of its proposed involvement in vascular, inflammatory, metabolic, and adaptive responses to exercise. Methods: This exploratory observational proof-of-concept study included 20 observations: 10 postexercise (EX) and 10 preexercise/nonexercise (NON). The dataset included age, sex, body mass index, dietary habits, smoking status, alcohol consumption, medical history, exercise type, duration, intensity, frequency, and miR-21 folding values. Univariate analysis of miR-21 folding was performed using Welch’s independent-samples t-test and the Mann–Whitney U-test. Three supervised machine-learning classifiers were applied to the integrated multivariable profile: L2-regularized Logistic Regression, Support Vector Machine with Radial Basis Function (SVM-RBF), and Gaussian Naive Bayes. Model performance was evaluated using repeated stratified 5-fold cross-validation. Results: miR-21 folding values alone did not clearly separate EX and NON observations, with substantial overlap between the two groups and nonsignificant univariate results. In contrast, the integrated multivariable profile achieved high internal classification performance across the selected models. Mean accuracy was 99.0% for Logistic Regression, 98.5% for SVM-RBF, and 99.5% for Gaussian Naive Bayes. The mean area under the curve (AUC) values ranged from 0.995 to 1.000, indicating strong internal discriminatory performance within the exploratory dataset. Conclusions: The findings suggest that miR-21 should not be interpreted as a standalone classifier of exercise status. Instead, miR-21 may contribute to a broader multivariable exercise-response signature when combined with demographic, lifestyle, clinical, and exercise-related variables.
Background: Passion fruit peel (PFP) is an underutilized agricultural by-product rich in bioactive polysaccharides. This study aimed to optimize the ultrasound-assisted extraction (UAE) of polysaccharides from PFP (PFPPs) and evaluate their physicochemical, functional, and biological properties. Methods: A Box–Behnken design combined with response surface methodology (RSM) was employed to optimize extraction parameters (ultrasonic power, temperature, and time). The physicochemical characterization included monosaccharide composition analysis by high-performance anion-exchange chromatography with pulsed amperometric detection, as well as water-holding capacity (WHC) and oil-holding capacity (OHC). Antioxidant activities were assessed using 2,2-diphenyl-1-picrylhydrazyl, superoxide anion, hydroxyl radical scavenging activity, and reducing power activity. Antibacterial activity was evaluated against Salmonella enterica, Escherichia coli, Micrococcus luteus, and Bacillus cereus. Results: The optimal extraction parameters were established as 82°C, 151 min, and 175 W, resulting in a maximum yield of 9.11 ± 0.013%. PFPPs were identified as an acidic heteropolysaccharide primarily composed of glucose (33.24%) and galacturonic acid (32.63%), exhibiting substantial WHC (2.84 ± 1.27 g/g) and OHC (3.35 ± 1.08 g/g). PFPPs demonstrated robust in vitro antioxidant activities across all assays, alongside significant antibacterial efficacy against the tested pathogens. Conclusions: Moderate ultrasonic degradation enhances the accessibility of active functional groups in PFPPs, promoting their potential as high-value functional ingredients for the food and pharmaceutical industries.
Background: Before conducting challenge tests, cholera toxin B subunit (CTB) was incorporated into the highly conserved region (HCR) through a genetic fusion strategy. CTB will facilitate interaction between the HCR spike glycoprotein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and ganglioside GM1 (GM1), facilitating antigen uptake and promoting mucosal immune responses. An immunoinformatic approach was subsequently employed to characterize the antigen. Methods: Antigen structure prediction was conducted using Iterative Threading ASSEmbly Refinement, Robetta, and 3Dpro, furthermore validated with ERRAT and PROCHECK. Antigenic potential was predicted using VaxiJen and ANTIGENpro. T-cell epitope and B-cell epitope prediction was conducted using Immune Epitope Database, antigenic.pl, Support vector machine–based tripeptide epitope predictor (SVMTriP), ABCpred. Allergenicity was assessed using AllerCatPro and AlgPred. Molecular docking was performed using AutoDock Vina. Results: The optimal structure was generated by Robetta, showing an overall quality factor of 92.434. The antigenic potential score is 0.4054, classified as a “probable antigen.” The top five residue scores for major histocompatibility complex Class I (MHC-I) epitopes ranged from 0.825045 to 0.938498, while major histocompatibility complex Class II (MHC-II) epitope scores ranged from 0.8355 to 0.8601. B-cell epitopes recommended by SVMTriP scored 1.000 and 0.975. AllerCatPro predicted the construct to be “nonallergenic.” Molecular docking of HCR spike glycoprotein of SARS-CoV-2–CTB with ganglioside GM1 yielded the best ΔG value of − 6.8 kcal/mol. Conclusions: Immunoinformatics of the HCR spike glycoprotein of SARS-CoV-2 fused with CTB suggests its potential as a safe, nonallergenic vaccine candidate with the ability to interact with the GM1 ligand and stimulate cellular adaptive immune responses. These findings highlight its potential as a SARS-CoV-2 vaccine candidate, warranting further validation through experimental studies.
Background: Graves’ disease (GD) is an autoimmune thyroid disorder in which the roles of interleukin-40 (IL-40) and IL-41 remain poorly understood. We characterized the serological, transcriptional, and genetic profiles of these cytokines in an Iraqi GD cohort. Methods: In this exploratory case–control study, 50 GD patients and 50 healthy controls were recruited. Serum IL-40 and IL-41 were quantified by enzyme-linked immunosorbent assay (ELISA), peripheral-blood mRNA expression by quantitative real-time polymerase chain reaction, and three IL40 single-nucleotide polymorphisms (SNPs; rs2310998, rs2004339, rs2310999) plus one IL41 SNP (rs2038043973) by Sanger sequencing. P values were Bonferroni-corrected, and multivariable logistic regression was adjusted for age, body mass index, and treatment duration. Results: Serum IL-40 was significantly lower (P < 0.0001) and IL-41 significantly higher (P = 0.010) in GD than in HC. IL40 mRNA was upregulated 2.5-fold (P = 0.005) and IL41 mRNA downregulated 0.65-fold (P < 0.0001). After Bonferroni correction and adjustment for confounders, the rs2310998-GA genotype, rs2310999-TG/GG genotypes and G allele, and the AAG, GAG, and AGT haplotypes were significantly associated with GD susceptibility. The rs2004339 individual-genotype effects and the GAT haplotype were nominally significant but did not survive correction. Conclusion: IL-40 and IL-41 show opposing serological and transcriptional associations with GD, and several IL40 polymorphisms are associated with disease susceptibility in this Iraqi cohort. Functional validation in larger, multiethnic cohorts is required before clinical translation.
Background: Human hair follicle dermal papilla cells (hHF-DPCs) play a critical role in hair follicle regeneration. Under conventional two-dimensional (2D) culture conditions, DPCs progressively lose the expression of key biological markers, suggesting that three-dimensional (3D) culture may better preserve their characteristic properties. Methods: In this study, hHF-DPCs were seeded into 0.2 mL polypropylene microtubes at three cell densities – 5 × 104, 105, and 2 × 105 cells/mL – to evaluate forced aggregation as a 3D culture approach. Aggregate formation and diameter were monitored over time. Viability was assessed by acridine orange/propidium iodide (AO/PI) staining and Trypan blue exclusion, while gene expression of alkaline phosphatase (ALP), versican, alpha-smooth muscle actin (α-SMA), and CD133 was quantified by reverse transcription–quantitative polymerase chain reaction (RT-qPCR). Results: Results demonstrated that hHF-DPC aggregates formed within 24 h across all densities. At 5 × 104 cells/mL, spheroids formed but disaggregated by day 3, whereas higher densities (105 and 2 × 105 cells/mL) produced compact, architecturally stable spheroids maintained for 7–10 days. Necrotic cores were detected from day 3 at the two higher densities, accompanied by a reduction in viable cell number. Compared with 2D culture, 3D spheroids exhibited significant upregulation of ALP, versican, and CD133, while α-SMA expression was downregulated at both high densities (P < 0.05). Conclusions: Microtube-based forced aggregation successfully generated hHF-DPC spheroids at densities of 105 and 2 × 105 cells/mL within 24 h, with stable morphology maintained for up to 10 days and enhanced expression of DPC-specific markers compared to conventional 2D culture.
Background: Zinc oxide nanoparticles (ZnONps) are safe, affordable, and versatile, with strong ultraviolet (UV) absorption and visible light transparency, making them excellent sunscreen agents. Plant extract-mediated synthesis of ZnONps provides an eco-friendly substitute for conventional chemical approaches by avoiding toxic reagents and provides bioactive properties. The study aims to develop non-comedogenic cream formulation using green synthesized ZnONps using the extract of Senna alata and Zingiber officinale-Allium sativum together which possess antibacterial, UV absorption, antioxidant, and anticancer properties. Methods: The characterization was performed using UV-visible spectroscopy, Field Emission Scanning Electron Microscopy, X-ray diffraction, and Fourier Transform Infrared (FTIR) spectroscopy. Antibacterial activity evaluated against Escherichia coli and Staphylococcus aureus. The antioxidant activity was determined by the 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) assay. The anticancer properties were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay using HeLa cells. The Sun Protection Factor (SPF) of the cream formulated by the incorporation of ZnONps was determined. Results: The crystalline sizes of ZnONps synthesized using S. alata leaf and a mixture of Z. officinale-A. sativum extract were 16.98 nm and 92.30 nm, respectively. The FTIR and DPPH assay results showed the presence of phytochemical residue. The antibacterial activities of synthesized ZnONps from the combination of A. sativum and Z. officinale extract have displayed better inhibition (S. aureus: 14.67 ± 0.35 mm) against gram-positive bacteria and gram-negative bacteria (E. coli: 22.63 ± 0.62 mm) than those made with S. alata extract (E. coli 14 ± 0.32 mm and S. aureus 19.4 ± 0.17 mm). The formulated cream showed skin-friendly pH with SPF more than 15. Conclusion: The biosynthesized ZnONPs exhibited antimicrobial, anticancer, and antioxidant potential and can be exploited in bioactive cosmetic formulations.
Background: The emergence of pesticide resistance in phytopathogenic fungi significantly impacts global crop yields. In this study, three series of 39 9-substituted acridine derivatives were synthesized (13 each of 9-methyl, 9-formyl, and 9-formaldehyde oxime derivatives), and their in vitro and in vivo antifungal activities against plant pathogenic fungi were evaluated. Methods: Three series of 39 9-substituted acridine derivatives were synthesized through Ullmann coupling, intramolecular cyclization, oxidation, and oximation reactions. Primary screening via antifungal rate determination against four phytopathogenic fungi identified promising candidate. Selected compound subsequently underwent antifungal evaluation employing mycelial growth rate assays, fluorescent staining, and in vivo experiments, while exploring its potential mechanism of action. Results: Three series of 39 9-substituted acridine derivatives were synthesized using 2-aminoacetophenone and various substituted iodobenzenes as starting materials. At 50 μg/mL, four compounds (5j, 4g, 5i, 4h) exhibited significant inhibitory activity. Notably, compound 4h demonstrated potent inhibition against Ampelomyces humuli. The half-maximal effect concentration value for 4h was 1.227 μg/mL, comparable to the positive control chlorothalonil (1.615 μg/mL) and superior to Famoxadone (14.11 μg/mL) and Carbendazim (20.03 μg/mL). Using the mycelial growth rate method, inhibition rates were 65.47% at 50 μg/mL. Mechanistic studies revealed that 4h (100 μg/mL) induced significant membrane damage (90.95%) in A. humuli and concomitantly suppressed its sporulation. In addition, in vivo studies confirmed that compound 4h exerts both curative and protective effects against A. humuli infection in plants. Conclusions: These results collectively establish the acridine scaffold as a promising candidate for developing novel antifungal agents.
Background: Prostate cancer (PCa) remains a major clinical challenge due to the limited ability of current diagnostic tools to distinguish indolent from aggressive tumors. MicroRNAs (miRNAs) have emerged as promising biomarkers given their stability, tissue specificity, and regulatory roles in carcinogenesis. This study analyzed miRNA expression and clinical data from The Cancer Genome Atlas–Prostate Adenocarcinoma (TCGA-PRAD) to identify differentially expressed miRNAs and evaluate their association with tumor aggressiveness. Methods: miRNA expression profiles and clinical variables from TCGA-PRAD were examined. A total of 550 samples (498 tumors, 52 adjacent normal) and 1,881 miRNAs were analyzed using the differential expression sequencing 2 workflow. Differential expression was assessed using adjusted significance thresholds. Highly deregulated miRNAs were identified based on log2 fold change ≥2 and false discovery rate (FDR) <0.01. Associations with the Gleason score were evaluated using Spearman correlation with multiple testing correction. Results: We identified 137 significantly deregulated miRNAs (FDR < 0.05), with no global trend toward over-or underexpression. Applying stringent filters yielded 24 miRNAs with high-magnitude changes. Correlation analysis revealed 25 miRNAs significantly associated with the Gleason score. The miR-449a, miR-184, and miR-891a showed both strong deregulation and significant correlation with aggressiveness. miR-449a exhibited the greatest tumor overexpression, whereas miR-184 and miR-891a displayed expression patterns consistent with pathways previously linked to proliferation, invasion, and tumor progression. Conclusion: miR-449a, miR-184, and miR-891a emerge as strong candidates for PCa progression biomarkers. Their combined deregulation and association with the Gleason score support their potential as prognostic indicators and therapeutic targets. Validation in independent cohorts and functional studies will be essential to confirm their clinical applicability.
Background: Solid-state fermentation (SSF) has emerged as an effective bioprocess for converting agro-industrial residues into high-value bioactive compounds. In this study, the production of total phenolic compounds and antioxidant-related enzymes by Basidiomycete and Ascomycete fungi was evaluated using banana peel, an abundant food-waste substrate. Four Basidiomycete and four Ascomycete species were assessed and compared on banana peel and potato dextrose agar media. Methods: Fungal cultures were subjected to SSF followed by quantification of enzymatic and antioxidant parameters, including laccase activity (ABTS assay [2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)], catalase activity [CAT] [titanium dioxide, TiO₂ reagent method], total phenolic content (Folin–Ciocalteu assay), reducing power assay, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity. Statistical comparisons were performed using the one-way analysis of variance. Results: Basidiomycota species demonstrated markedly higher laccase activity than Ascomycota species across both media. Clitopilus prunulus BV18 exhibited the highest laccase activity (900.63 U/mL). Conversely, Ascomycota species produced greater CAT, with Xylaria polymorpha BV5 achieving the highest level (6.76 µmol H₂O₂/mL). Phenolic compound biosynthesis varied significantly among species; Basidiomycetes exhibited overall superiority, particularly on banana peel medium, where Pleurotus pulmonarius BV13 and C. prunulus BV18 produced the highest total phenolic contents (388.67 and 287.33 µg/mL, respectively). DPPH radical scavenging activity was also highest in P. pulmonarius BV13, C. prunulus BV18, and X. polymorpha BV5 on banana peel substrate. Conclusions: Basidiomycota fungi exhibit strong inherent potential for the biotransformation of food waste into phenolic-rich antioxidant metabolites under SSF conditions. Their markedly higher phenolic production and antioxidant activity, compared with Ascomycetes, highlight their suitability for sustainable bioprocessing applications aimed at valorizing agro-industrial waste streams.
Background: Foodborne pathogens, particularly Escherichia coli and Salmonella spp., present significant health risks, exacerbated by the growing resistance to antibiotics. This study explores the potential of clove (Syzygium aromaticum) essential oil and oleoresin as natural antimicrobial agents against these pathogens. Methods: Essential oil and oleoresin were analyzed using gas chromatography-mass spectrometry, revealing eugenol, eugenyl acetate, and caryophyllene as the dominant compounds. Results: Antibacterial efficacy was assessed using minimum inhibitory concentration and minimum bactericidal concentration assays, showing clove essential oil to be more potent than oleoresin. The inhibitory effects were further confirmed through time-kill analysis, where clove essential oils exhibited potent bactericidal effects, whereas oleoresins demonstrated bacteriostatic activity. Morphological changes in E. coli and Salmonella spp., including elongated cell forms, suggested a disruption in cell division. Clove essential oil also effectively inhibited biofilm formation by up to 80%, compared to 60% by oleoresins. When applied to artificially contaminated food matrices (chicken and prawns), clove essential oil demonstrated up to a 2-log reduction in Salmonella spp. and a 1-log reduction in E. coli colony counts. Moreover, the essential oil influenced virulence gene expression in both pathogens, indicating its ability to suppress bacterial pathogenicity. Conclusion: The findings suggest that clove essential oil, with its potent antimicrobial properties, could be an effective natural preservative, offering a safer alternative to synthetic agents in food safety applications.
