Dual Target Anticancer Potential of Exiguobacterium-derived Metabolites from the Maros-Pangkep Karst: An In Silico Study Targeting EGFR and Caspase-3

Background: Cancer is the second leading cause of death worldwide, and its multifactorial nature limits the effectiveness of single-target therapies. Combination treatments may improve efficacy but are often associated with toxicity and drug interactions. Extremophilic bacteria from karst environments remain underexplored as sources of multitarget anticancer compounds. This study aimed to evaluate the dual-target anticancer potential of bioactive metabolites produced by bacteria isolated from the Maros–Pangkep karst region using molecular docking and ADME analysis.Materials and Methods: A bacterial isolate obtained from soil samples from the Maros–Pangkep karst region, Indonesia, was selected based on colony morphology and identified as Exiguobacterium using 16S rRNA gene analysis. Bioactive compounds were characterized by GC-MS, and three major metabolites were docked against EGFR (PDB ID: 1M17) and caspase-3 (PDB ID: 1PAU). ADME analysis was performed to evaluate drug-likeness and toxicity.Results: Docking analysis showed that compound (1) Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl) exhibited the strongest binding affinity toward EGFR (−7.2 kcal/mol), exceeding the native ligand erlotinib (−6.1 kcal/mol). Compound (2) Cyclo(L-prolyl-L-valine) and compound (3) 2,5-piperazinedione,3-methyl-6-(1-methylethyl) derivatives showed binding affinities of −6.4 and −5.9 kcal/mol, respectively. For caspase-3, the compounds displayed binding affinities ranging from −4.5 to −6.0 kcal/mol , exceeding the native ligand Ac-DEVD-CHO (−3.6 kcal/mol) and interacted outside the active site, indicating potential allosteric activation. All compounds complied with Lipinski’s rule of five and showed low toxicity (LD50>800 mg/kg).Conclusion: This study shows that metabolites from Exiguobacterium possess dual-target anticancer potential via EGFR inhibition and caspase-3 modulation, suggesting their use as multitarget drug candidates. Keywords: 16S rRNA, bacteria, biological products, caspase-3, epidermal growth factor receptor, Kart Maros-Pangkep, molecular dockin

Hypoxia-Conditioned MSC Exosomes Upregulate mTORC1 and Suppress MMP-2 in a UV-B–Induced Collagen Loss Rat Model

Background: Ultraviolet-B (UV-B) radiation accelerates photoaging by disrupting extracellular matrix (ECM) homeostasis through dysregulation of mechanistic target of rapamycin complex 1 (mTORC1) and upregulation of matrix metalloproteinase-2 (MMP-2). This study evaluated the effects of exosomes of hypoxia-conditioned mesenchymal stem cells (EH-MSCs) on mTORC1 and MMP-2 expression in a UV-B–induced collagen loss rat model.Materials and Methods: Thirty male Wistar rats were randomized into five groups: healthy control, UV-B + saline, UV-B + hyaluronic acid, UV-B + 200 µL EH-MSCs, and UV-B + 300 µL EH-MSCs. Collagen loss was induced by UV-B irradiation for two weeks (10 sessions, 8 min/session). A single treatment was administered on day 22, and tissue was collected on day 29. Exosomes were isolated from hypoxia-conditioned MSCs and characterized by morphology and surface markers. Gene expression of mTORC1 and MMP-2 was assessed by qRT-PCR and analyzed using one-way ANOVA.Results: UV-B exposure induced collagen loss histologically. EH-MSCs significantly increased mTORC1 expression, highest in the 300 µL group (p < 0.001), and reduced MMP-2 expression, lowest in the 300 µL group.Conclusion: EH-MSCs exert dual regulatory effects by upregulating mTORC1 and suppressing MMP-2 in UV-B–induced collagen loss, suggesting therapeutic potential to mitigate photoaging via anabolic signaling (via mTORC1) and reduced ECM degradation (via MMP-2).Keywords: collagen loss, exosomes, MMP-2, mTORC1, UV-

Fluoroquinolones Inhibit, while β-Lactams Fail Against Biofilm-Producing Bacteria Isolated from Paediatric Syrups

Background: Paediatric syrups are sugar-rich solutions widely preferred in paediatric healthcare due to their palatability, although they are highly prone to microbial contamination. Of particular concern is the presence of biofilm-producing microorganisms; existing studies have focused on contamination while overlooking the enhanced resistance mechanisms conferred by biofilm formation. This study aimed to determine the antibiogram profile of bacterial isolates from commonly prescribed paediatric syrups administered by caregivers to patients at selected healthcare centers.Materials and Methods: A total of 392 syrup sample swabs were collected from hospitals and community sources. Bacterial isolation and identification were performed using standard microbiological methods. Biofilm production was evaluated using the test-tube method, and antibiotic susceptibility was determined via the disk diffusion method.Results: Bacterial counts ranged from 3.0±2.0×10³ to 10.7±3.05×10³ CFU/mL, with community samples showing the highest counts. Bacterial isolates identified included Proteus vulgaris, with the highest frequency of occurrence (18.75%) > Streptococcus agalactiae, Klebsiella pneumoniae, Acinetobacter baumannii (12.59%) > Pseudomonas aeruginosa, Arthrobacter agilis, Enterococcus faecium, Enterobacter cloacae, Escherichia coli (6.25%). All isolates produced biofilms significantly different (p<0.05) from the negative control (broth tube without bacterial cells), except Arthrobacter agilis. Antibiotic susceptibility testing indicated multidrug resistance, particularly against amoxicillin and amoxicillin-clavulanate, while showing comparatively higher susceptibility to fluoroquinolones.Conclusion: P. vulgaris was the most frequent isolate, while K. pneumoniae and A. baumannii produced the strongest biofilms. The highest resistance was observed against amoxicillin and amoxicillin-clavulanic acid, whereas fluoroquinolones remained the most effective. Paediatric syrups can harbor biofilm-producing multidrug-resistant bacteria, underscoring the importance of monitoring and safe handling.Keywords: paediatric syrup, antibiogram profile, biofilm, bacterial isolates, test-tube metho

Hypoxia-Derived Mesenchymal Stem Cell Exosomes Downregulate CXCL12 and Upregulate IL-10 Expression in a Murine Model of Androgenic Alopecia

Background: Androgenic alopecia (AGA) is characterized by hair follicle miniaturization and chronic inflammation mediated by dihydrotestosterone (DHT). Current therapies primarily target hormonal pathways and do not adequately address inflammatory dysregulation. Exosomes derived from hypoxia-conditioned mesenchymal stem cells (EH-MSCs) exhibit enhanced immunomodulatory properties. However, their effects on key inflammatory mediators in AGA remain unclear. This study evaluated the effects of EH-MSCs on CXCL12 and IL-10 expression in a DHT-induced AGA mouse model.Materials and methods: Male C57BL/6 mice were allocated into five groups: healthy control, DHT-induced alopecia without treatment, DHT-induced alopecia treated with topical minoxidil, and EH-MSCs administered at 100 or 200 µL/kgBW. Alopecia was induced by subcutaneous DHT injection for 17 days. Following model validation, treatments were administered on days 25 and 32. CXCL12 and IL-10 expression in dorsal skin tissue was analyzed using RT-PCR on day 39.Results: DHT induction significantly increased CXCL12 expression and reduced IL-10 levels (p<0.05). EH-MSC administration dose-dependently downregulated CXCL12 (1.73±0.57 and 1.54±0.44 fold-change) and upregulated IL-10 expression (3.10±0.75 and 3.29±0.67 fold-change), demonstrating greater immunomodulatory effects compared with minoxidil.Conclusion: EH-MSCs effectively modulated inflammatory biomarkers by suppressing CXCL12 and enhancing IL-10 expression in a DHT-induced AGA model, suggesting their potential as an immunoregenerative therapeutic strategy for androgenic alopecia.Keywords: androgenic alopecia, mesenchymal stem cells, exosomes, hypoxia, CXCL12, IL-1

High Prevalence of dam and fimA , Biofilm Formation, and Antibiotic Resistance in Uropathogenic Escherichia coli

Background: Urinary tract infections (UTIs) are among the most common bacterial infections in women and remain a significant public health problem. Uropathogenic E. coli (UPEC) is the main cause of UTIs and can form biofilms, which lead to recurrent infections and antibiotic resistance. Type 1 fimbriae in UPEC, encoded by the fim operon, facilitate bladder attachment, while the dam an orphan DNA methyltransferase in E. coli, contributes to bacterial colonization and biofilm formation. Data on the association between antibiotic susceptibility, fimA and dam gene prevalence, and biofilm formation in UPEC isolates from UTI patients in Indonesia remain limited. This study aimed to investigate the association of the dam and fimA virulence genes with biofilm formation in UPEC causing UTIs.Materials and Methods: Fifty UPEC isolates were obtained from a clinical microbiology laboratory. Biofilm formation was assessed using the tube method. Antibiotic susceptibility testing was performed using the Kirby–Bauer disk diffusion method with amoxicillin, ciprofloxacin, and gentamicin. The presence of the dam and fimA was determined by PCR.Results: Seventy percent of UPEC isolates were capable of biofilm formation. High resistance rates were observed for amoxicillin (92%), ciprofloxacin (88%), and gentamicin (56%). In UPEC isolates that were positive for the dam, 62% of them had the ability to form biofilms. Meanwhile, in UPEC isolates that were positive for the fimA, 52% of them had the ability to form biofilms.Conclusion: UPEC isolates showed a high prevalence of the dam and fimA genes, which were associated with biofilm formation and increased antibiotic resistance.Keywords: biofilm, antibiotic, dam, fimA, urinary tract infection

Doxycycline Suppresses Hypertension through Renin– Angiotensin System (RAS) Regulation: Insights from Molecular Docking and Renal Gene Expression

Background: Doxycycline, a tetracycline antibiotic known for inhibiting matrix metalloproteinases, has shown potential antihypertensive effects. However, its role in modulating the renin–angiotensin system remains poorly understood. This study aims to specifically evaluate Doxycycline’s effects on key RAS components and blood pressure responses to clarify its underlying mechanism and support its development as a targeted antihypertensive therapeutic candidate.Materials and Methods: This study integrated an in-silico and experimental approach to assess the antihypertensive effects of doxycycline. Bioinformatics analyses were first conducted, including target prediction, gene ontology enrichment, hub-gene identification, PPI network construction, and KEGG pathway analysis, followed by molecular docking and molecular dynamics simulations to predict doxycycline’s interactions with key RAS targets. To validate these computational findings, qRT-PCR was performed to measure the expression of selected genes in kidney tissues from hypertensive rats.Results: Bioinformatics analysis identified six key target genes, including AGT, AGTR1, AGTR2, REN, ACE, and ACE2. Molecular docking showed that doxycycline exhibited stronger binding affinity to AGTR1 (-8.346 kcal/mol) than its native ligand. Molecular dynamics confirmed the stability of the doxycycline–AGTR1 complex at 20 ns. Gene expression analysis of kidney tissues from hypertensive rats revealed a significant reduction in AGTR1 expression in the group treated with doxycycline 15 mg/kg (p<0.05), while no significant change was observed at 30 mg/kg.Conclusion: Low-dose doxycycline may modulate the renin–angiotensin pathway through AGTR1 inhibition, indicating its potential as a candidate for further antihypertensive research and warranting more comprehensive in vivo evaluation.Keywords: Hypertension, doxycycline, molecular docking, gene expression, Renin-Angiotensin Syste

Elevated Maternal Leptin Levels and Higher Birth Weight in Obese Mothers Compared with Normal-weight Mothers

Background: Maternal obesity is associated with disrupted metabolic regulation, including elevated leptin levels that may affect fetal growth. Previous studies have reported mixed findings on maternal and cord blood leptin concentrations, with some showing higher leptin levels in infants born to obese mothers, while others found no significant differences. Associations between maternal obesity and neonatal outcomes have also been inconsistent. This study aims to generate new evidence from an Indonesian cohort by comparing maternal and cord blood leptin levels, as well as neonatal outcomes, between obese and normal-weight mothers.Materials and Methods: Analytical cross-sectional study enrolled 30 obese and 30 normal-weight mothers from two hospitals in Padang, Indonesia. Maternal and cord blood samples were collected at delivery, and leptin levels were measured using ELISA. Ethical approval and informed consent were obtained. Neonatal outcomes included birth weight, length, head circumference, and ponderal index. Data were analyzed using Independent Samples t-test or Mann–Whitney U test, with p<0.05 considered statistically significant. Results: Maternal leptin levels were significantly higher in obese mothers than in normal-weight mothers (70.8±39.0 vs. 44.5±42.5 ng/mL; p=0.002). Cord blood leptin levels did not differ between groups (19.7±14.9 vs. 16.1±17.5 ng/mL; p=0.394). Neonates of obese mothers had higher birth weight (p=0.003), while other anthropometric measures were similar.Conclusion: Maternal obesity was associated with elevated maternal leptin levels and increased neonatal birth weight, while cord blood leptin did not differ significantly between groups. These findings suggest that maternal leptin reflects maternal metabolic status rather than fetal growth.Keywords: maternal leptin, cord blood leptin, obesity, pregnancy, neonatal outcom

Elevated Procalcitonin Levels in Pediatric Severe Bacterial Pneumonia Caused by Klebsiella pneumoniae

Background: Klebsiella pneumoniae is a major pathogen in pediatric pneumonia. Procalcitonin (PCT) distinguishes between Gram positive and Gram negative infections but lacks data on K. pneumoniae infection's relationship with PCT levels in children. Therefore, this study was conducted to investigate the serum PCT levels in children with K. pneumoniae infection.Materials and methods: A cross-sectional study was conducted on 61 pediatric subjects with the age of 2-59 months with severe bacterial pneumonia. Blood and sputum samples were collected and analyzed for PCT and cultured for 24 hours. PCT levels and K. pneumoniae infection were statistical analyzed with chi-square and logistic regression. Receiver operating characteristic (ROC) analysis was used to determine the PCT cut-off specific to K. pneumoniae. Results: K. pneumoniae was identified in 33%. Median PCT levels were significantly higher in the K. pneumoniae group (p<0.05). A PCT cut-off of 0.725 ng/mL yielded 70% sensitivity, 70.7% specificity, negative predictive value (NPV) of 82.9%, and area under the receiver operating characteristic curve (AUROC) of 0.74. Elevated PCT was significantly associated with K. pneumoniae infection with Odds ratio (OR) of 12.08, 95% Confidence Interval (CI): 2.54-57.36; p=0.002).Conclusion: Along with K. pneumoniae infection, serum PCT levels was elevated, supporting its potential as a biomarker for early diagnosis.    Keywords: K. pneumoniae, procalcitonin, bacterial pneumonia, pediatri

UC-MSCs Secretome Induces Proliferation of CD4+ T Cells, CD8+ T Cells, NK Cells, and Increases sPD-1 Levels in Severe COVID-19’s Whole Blood

Background: Clinical features of severe coronavirus disease 2019 (COVID-19) predominantly include respiratory symptoms and exacerbated multi-organ complications, especially in patients with comorbidities. Cellular immunity, including lymphocytes, is a critical factor in combating SARS-CoV-2 infection. However, immune dysregulation occurs in severe COVID-19 patients, characterized by cytokine storm and lymphopenia. The effectiveness of mesenchymal stem cell (MSC) therapies for COVID-19 is being assessed. The secretome released by MSC functions similarly to the cells themselves as an immunomodulator, offering potential advantages in terms of safety and cost-effectiveness. This study was conducted to assess the effect of umbilical cord MSC-derived (UC-MSC) secretome treatment on lymphocyte count and soluble programmed cell death-1 (sPD-1) levels in severe COVID-19 patient's whole blood.Materials and methods: Twelve whole blood samples from healthy individuals and severe COVID-19 patients were analyzed for lymphocyte count and functional activation using flow cytometry, along with sPD-1 level measurement in pre-treatment and post-secretome conditions.Results: The lymphocyte count in severe COVID-19 patients was significantly decreased, particularly for T cells and NK cells, indicating lymphopenia. Following secretome treatment, CD4+ T cell counts significantly increased compared to pre-treatment, although this change was not significant in the negative control group. Additionally, there was a minimal reduction in B cell count and an increase in sPD-1 levels. Elevated sPD-1 may alleviate T cell exhaustion by interfering with PD-1 binding to programmed death-ligand 1 (PD-L1).Conclusion: Administration of UC-MSC secretome to the whole blood of severe COVID-19 patients suggested immune improvement, with significant increases in CD4+ T cell counts, enhanced B cell survival, and elevated sPD-1 levels. Keywords: COVID-19, cellular immunity, lymphocytes, secretome, MS

Mesenchymal Stem Cell-Derived Exosomes Enhance FGF-1 and SDF-1 Expression in Rats with Second Degree Burns

Background: Second-degree burns cause extensive damage to the skin and pose significant health challenges, with current treatments facing limitations such as donor skin shortages and complications. Fibroblast growth factor 1 (FGF-1) and stromal-derived growth factor 1 (SDF-1) are critical for tissue repair. Emerging evidence suggests that mesenchymal stem cell-derived exosomes (E-MSCs) are a promising cell-free therapeutic option for enhancing wound healing through the modulation of FGF-1 and SDF-1. This study investigated the effect of E-MSCs on the expression of FGF-1 and SDF-1 genes in rats with second-degree burns.Materials and methods:  This experimental study used a second-degree burn model in Wistar rats, treated with subcutaneous injections of E-MSCs at doses of 100 µL and 200 µL. Gene expression of FGF-1 and SDF-1 was quantified using qRT-PCR. Histological validation confirmed burn severity, and flow cytometry was used to characterize E-MSCs and exosomes.Results: An increase in FGF-1 and SDF-1 expression was observed in exosome-treated groups compared to the NaCL-treated group. The 200 µL E-MSCs-treated group showed the most significant enhancement in both growth factors, with statistically significant differences (p<0.05). These findings underline the efficacy of E-MSCs in modulating critical genes involved in wound healing.Conclusion: E-MSCs significantly upregulate FGF-1 and SDF-1 expression, promoting tissue repair and regeneration in second-degree burn models. This study highlights the potential of E-MSCs as a non-invasive therapeutic approach.  Keywords: exosomes, FGF-1, mesenchymal stem cells, SDF-