Wawaimuli Arozal

Acute diarrhea is associated with alterations in the gut microbiota, yet data from Indonesian pediatric populations remain limited. This study characterized the fecal microbiota of children and toddlers with acute diarrhea using 16S rRNA gene amplicon sequencing. A cross-sectional study was conducted on 24 pediatric patients with acute diarrhea (12 toddlers and 12 children). Fecal microbiota composition, alpha diversity, beta diversity, and functional profiles were analyzed using QIIME2 and PICRUSt2. Streptococcus thermophilus and Faecalibacterium duncaniae exhibited higher abundance in toddlers. In contrast, Fusobacterium mortiferum was relatively more abundant in children. Alpha diversity indices showed no significant differences between children and toddlers. Beta diversity analyses also demonstrated substantial overlap between groups, with no significant difference in microbial community composition. Functional prediction identified the most discriminatory pathways as those related to energy and carbohydrate metabolism, amino acid metabolism, cellular biosynthesis, and environmental adaptation. Poor nutritional status (underweight) was associated with a decrease in protective commensal bacteria (Faecalibacterium duncaniae) (p < 0.05) and an increase in dysbiosis-associated bacteria (Parabacteroides distasonis) (p < 0.05). Despite modest species-level differences with age, alpha diversity, beta diversity, and overall bacterial community composition did not differ significantly between toddlers and children, indicating that acute diarrhea itself was the dominant factor shaping gut bacterial community structure in this cohort. Further studies with larger cohorts, external validation, and shotgun metagenomic sequencing are needed to validate these findings and explore candidate microbiota-based markers for pediatric acute diarrhea.

Background/Objectives: Nutritional intake plays an important role in modulating lead absorption and toxicity. In addition to micronutrient status, emerging evidence suggests that body fat distribution may influence heavy metal toxicokinetics, yet this aspect remains poorly explored in occupational settings. This study aimed to investigate the associations of dietary intake of zinc, calcium, vitamin D, and protein, as well as anthropometric indicators, with blood lead levels (BLLs) among lead-exposed male workers. Methods: A cross-sectional study was conducted involving 144 male workers from five areas with varying degrees of environmental lead contamination in Java, Indonesia. Nutrient intake was assessed using a semi-quantitative food frequency questionnaire (SQ-FFQ). Anthropometric measurements included body mass index (BMI) and waist-to-height ratio (WHtR). BLLs were measured using inductively coupled plasma mass spectrometry (ICP-MS). Bivariate and multivariate analyses were performed to identify independent predictors of elevated BLLs. Results: The median BLL was 6.8 µg/dL (Q1–Q3: 4.75–13.08), and 32% of participants had BLLs above 10 µg/dL. BLLs differed significantly across exposure areas (p < 0.001). In bivariate analysis, WHtR, protein intake, zinc intake, and vitamin D intake were significantly associated with BLLs. Workers with higher WHtR had a greater proportion of elevated BLLs (p = 0.023), whereas BMI was not associated. In multivariate analysis, low zinc intake (p = 0.031) and low vitamin D intake (p = 0.021) remained significant predictors of high blood lead levels. Conclusions: Environmental exposure remains the main determinant of BLLs, while low intake of zinc and vitamin D increases the risk of high blood lead levels. Central adiposity, reflected by WHtR, may represent a potential anthropometric marker of lead burden, suggesting a potential role of body fat distribution in lead toxicokinetics that warrants further investigation.

Mitochondrial dysfunction is a critical driver of cardiac remodeling under conditions of chronic stress, such as hypertension and heart failure. Andrographolide, a bioactive diterpenoid from Andrographis paniculata, has demonstrated antioxidative and anti-inflammatory effects; however, its role in mitochondrial quality control within the heart remains unclear. In this study, a network pharmacology approach was applied to explore the molecular targets of andrographolide related to mitochondrial dysfunction in cardiac remodeling. A total of 1763 mitochondrial-associated cardiac remodeling genes were retrieved from CardGenes and intersected with 539 andrographolide-related targets identified using PharmMapper, SwissTargetPrediction, and the comparative toxicogenomics database. Fifty-four overlapping genes were subjected to protein–protein interaction analysis using STRING and Cytoscape. The top-ranked hub gene was identified as TP53. Functional enrichment indicated key involvement in the PI3K-Akt, mitogen-activated protein kinases (MAPK), Forkhead box protein (FOXO), and AGE-RAGE pathways, linking andrographolide to the modulation of oxidative stress, metabolism, and cell survival. Gene ontology (GO) terms supported roles in kinase activity and membrane-associated signaling. Molecular docking showed strong binding affinities between andrographolide and TP53 regulators, particularly ataxia telangiectasia mutated (ATM) (−8.63 kcal/mol, Kd 6.34 × 10−7 M) and checkpoint 2 (CHK2) (−8.47 kcal/mol, Kd 4.68 × 10−7 M). Machine learning predictions indicated favorable LELP and moderate bioactivity for ataxia telangiectasia Rad3-related (ATR), CHK2, and Sirtuin 1 (SIRT1). These findings suggest that andrographolide exerts cardioprotective effects by modulating mitochondrial stress signaling and p53 regulatory networks. Further experimental validation is warranted to confirm its therapeutic potential in cardiac remodeling-related diseases.

Hypertension-related fatigue involves both elevated blood pressure and reduced exercise tolerance, yet its molecular basis remains unclear. This study investigated the repositioned effects of spironolactone on hemodynamic and metabolic parameters in a uninephrectomy–deoxycorticosterone acetate-salt rat model of salt-sensitive hypertension. Male Sprague–Dawley rats were randomized to receive spironolactone (100 mg/kg/day) or vehicle (carboxymethylcellulose) for five weeks. Blood pressure was measured noninvasively, exercise capacity was assessed using a weighted swimming test, plasma sodium was quantified by enzyme-linked immunosorbent assay, and cardiac HIF-1α expression was analyzed by quantitative polymerase chain reaction. Complementary in silico analyses included functional enrichment of spironolactone–HIF-1α targets, molecular docking with factor inhibiting hypoxia-inducible factor-1 (FIH1; PDB ID: 8II0), free energy calculations, and quantum mechanical assessment. Spironolactone prevented increases in systolic, diastolic, and mean arterial pressure, normalized plasma sodium levels, and suppressed cardiac HIF-1α expression. A non-significant trend toward prolonged time-to-fatigue was observed in spironolactone-treated rats. When interpreted alongside metabolic and hypoxia-related molecular findings, this trend suggests a potential antifatigue effect, although definitive conclusions cannot be drawn from the current dataset. Computational analyses identified HIF-1α and glycolytic pathways as central interaction hubs, with spironolactone demonstrating favorable binding to catalytic residues of FIH1, an asparaginyl hydroxylase, consistent with a mechanism that may enhance HIF-1α hydroxylation and attenuate hypoxia-driven glycolytic reprogramming in the hypertensive myocardium. These findings suggest that spironolactone exerts integrated hemodynamic and metabolic benefits and may hold therapeutic potential for hypertension-related fatigue, warranting confirmation in future studies or clinical translation in salt-sensitive populations.

Insulin Resistance (IR) is a complication that frequently occurs in obesity. The inflammation-mediated senescence in White Adipose Tissue (WAT) is important in obesity-induced IR. Centella asiatica (CA) is a potential medicinal plant with anti-aging and anti-obesity properties. Here, we explored the effect of CA on obesity-mediated IR in mice fed with a High Fat-High Fructose (HFHF) diet and treated simultaneously with CA at 150 mg/kgBW (CA150) or 300 mg/kgBW (CA300). The total body mass and visceral WAT weight in both CA groups decreased in comparison with HFHF group alone. We demonstrated that HFHF-diet mice treated with CA300 improved insulin sensitivity and enhanced Irs-1 activation in WAT. CA300, but not CA150, prevented the senescence phenotype in WAT, represented by decreased Senescence-associated beta-galactosidase (SA-β-Gal) activity and diminished Cdkn2a and Cdkn1a expression levels at mRNA level. Mechanistically, CA300 prevented the enhancement of Il6 and Il1b mRNA expression levels and macrophage activity in the immunostaining analysis of WAT. In vitro, RAW264.7 cells stimulated with high glucose and low dose of Lipopolysaccharides (LPS) also confirmed that CA 200 μg/ml alleviated the expression levels of M1 markers such as Ccl2, Il6, Il1b, and Tnf at mRNA level. Our data indicated that CA has therapeutic potential for obesity-mediated IR by suppressing proinflammatory M1 macrophages and preventing inflammation-induced senescence in WAT.

BACKGROUND: Sleep deprivation (SD) affects 20–30% of children and is known to impair cognitive functions, particularly memory. Despite its impact, there is currently no standardized treatment. Evidence from both adult animal and human suggests that N-acetylcysteine (NAC) possesses neuroprotective properties. This study was conducted to evaluate the effects of NAC on memory deficits induced by SD in juvenile rats.METHODS: Juvenile Sprague-Dawley rats were subjected to SD using the modified multiple platform method. NAC was administered intraperitoneally at doses of 100 mg/kgBW or 500 mg/kgBW. Y-maze and novel object recognition (NOR) tests were used for neurobehavioral assessment. Biochemical analyses were conducted to measure cortisol, brain-derived neurotrophic factor (BDNF), and acetylcholine (ACh) levels, using enzyme-linked immunosorbent assay (ELISA). Acetylcholine esterase (AChE) activity was measured by colorimetric method. Western blot analysis was performed to examine cAMP response element-binding protein (CREB) and phosphorylated CREB (p-CREB). Additionally, cytokine mRNA expressions were evaluated using polymerase chain reaction (PCR).RESULTS: Spontaneous alteration and discrimination ratio were decreased in SD group without treatment compared to the normal group (p<0.05). Similarly, BDNF was also decreased compared with normal group (p<0.05). Cortisol level and mRNA expression of TNF-α were increased significantly compared with normal group (p<0.05) along with a slight increase of ACh activity. Interestingly, NAC treatment mainly at the dose of 500 mg/kgBW prevented those pathological features significantly.CONCLUSION: NAC might prevent the SD-induced memory deficits by suppressing the inflammatory markers, activity of AChE, cortisol, and enhancing the level of BDNF.KEYWORDS: SD, NAC, memory, AChE, cortisol, inflammatory markers, BDNF

Cardiac remodeling is a phenotype of heart failure characterized by a molecular, cellular, and interstitial change in the heart, which manifests in the change of size and function of the heart after specific insults with multiple mechanisms. Centella asiatica (CA) and its main bioactive triterpenoid, asiatic acid (AA), pose antioxidant and anti-inflammatory effects. Still, no adequate clinical trials support the potency of CA and AA as anti-cardiac remodeling. Hence, this systematic review aims to provide an in-depth analysis of CA extract and AA in animal studies' prevention or therapy of cardiac remodeling. The search strategies were conducted based on preferred reporting Items for systematic reviews and meta-analysis (PRISMA) protocol through Pubmed, EMBASE, Scopus, and Web of Science using keywords as follows: “Centella asiatica” OR “Asiatic Acid” AND “Cardiac Remodeling” OR “Cardiac Hypertrophy” OR “Cardiac Fibrosis” along with their synonym. The data collected included hemodynamic parameters based on echocardiography, biomolecular tests such as quantitative polymerase chain reaction (qPCR), Western blotting, or biochemistry procedures. The paper quality was assessed using Systematic Review Centre for Laboratory Animal Experimentation (SYRCLE) risk of bias (RoB). The previous selected study has shown that CA and AA might prevent and cure cardiac remodeling by inhibiting various pathways and protein expressions through AMPKα, NOX2/4, PI3K/Akt/mTOR, p70S6K, YAP/TAZ, and IL-1β, IL-6, and IL-18 cytokines. CA and AA, thus, exhibit cardioprotective effects in the animal model, which need to be confirmed in the clinical trials on humans. Keywords: cardiac remodeling, cardiac hypertrophy, cardiac fibrosis, Centella asiatica, asiatic acid

Heart failure is still a global health problem that demands new pharmacological treatments. The Apelin Receptor (APR), a class A (rhodopsin-like) G-Protein Coupled Receptor (GPCR), is one of the cell membrane receptors that potentially become a specific target of heart failure therapy when activated. However, no clinically approved drugs target APR. Phytochemical compounds from Indonesian herbs have become readily available for discovering novel apelin agonists. This study investigates bioactive phytochemicals from ten Indonesian medicinal herbs using computer-aided drug design (CADD) to predict ligand-receptor interactions via molecular docking and bioactivity prediction through machine learning. The selected herbs include Andrographis paniculata, Centella asiatica, Zingiber officinale, Curcuma longa, Curcuma domestica, Morinda citrifolia, Guazuma ulmifolia, Orthosiphon stamineus, Moringa oleifera, and Garcinia mangostana. Their pharmacokinetic and physicochemical profiles were also assessed using web-based predictions. Active metabolites were sourced from the Knapsack Database. Molecular docking using Molegro Virtual Docker assessed binding energy, with more negative MolDockScores indicating stronger interactions. Gambogic acid (-155.1 kJ/mol) from Garcinia mangostana and Procyanidin B2 (-154.5 kJ/mol) from Guazuma ulmifolia exhibited stronger binding than the APR agonist, Azelaprag (-149.9 kJ/mol). This study showed that Gambogic acid, Procyanidin B1, and Procyanidin B2 may predict excellent half maximal effectivity (EC50) against apelin receptors, despite their unideal lipophilicity-ligand efficiency (LELP). Gambogic acid demonstrated favorable pharmacokinetic properties: good bioavailability, minimal blood-brain barrier penetration, no cytochrome P450 (CYP) enzyme interactions, and low toxicity. The study concluded that all five selected compounds exhibited strong interactions and bioactivity as APR agonists, supporting the need for further validation through in-vitro studies.

Objective: This study aims to investigate the possibility of additive cytotoxic effects of cisplatin and Andrographis paniculate (Burm. f.) Nees(AP) via apoptotic, cell cycle and angiogenesis pathways. Methods: CC50 cisplatin, AP and Andrographolide (AG) were determined by the cell viability of SKOV3 after its exposure to these substances. SKOV3 cells were then divided into 6 experimental groups: one negativecontrol group, one with CC50 cisplatin alone, and three where CC50 was combined with CC50 AP, ½CC50 AP, and 1.5CC50 AP, respectively. The additive cytotoxic effect of cisplatin with AP or AG was evaluated through the modulation of several pathways via qRT-PCR of their markers: apoptotic pathways indicated by Bax, BCL2, Caspase 3 and Caspase 9 expression; cell cycle indicated by Cyclin-D expression; angiogenesis pathways by VEGF expression. Results: Cisplatin reduces cell viability to 54%, 37% when combined with AG, and 30%, 23% and 20% with ½CC50 AP, CC50 AP and 1.5CC50 AP, respectively. AG and AP extract decreases SKOV3 cell viability in a dose-dependent manner. Cisplatin combined with AP showed a statistically significant increase in BAX, Caspase 3, Caspase 9 expression and a decrease in BCL2, which indicated synergy in apoptotic pathways. The best result was seen in cisplatin combined with ½CC50 AP. A decrease in Cyclin D and VEGF was seen in all groups, the best seen in ½CC50 AP and CC50 AP, respectively, showing optimal cell cycle arrest and anti-angiogenesis properties when cisplatin is combined with AP extract. Conclusion: Combining cisplatin with AP extract enhanced cell cycle arrest, apoptosis, and anti-angiogenesis properties.

The rapid aging of the global population is a major worldwide issue because of the close relationship between age and the development of several diseases. Aging or senescence is among the most widely studied topics at the moment. However, no pharmaceuticals have been developed that claim to possess anti-senescence properties. Andrographis paniculata, is a medicinal plant found widely throughout tropical and subtropical Asia. This review aims to identify the potential anti- senescence effect of A. paniculata extract and its bioactive compounds. By following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, five databases were used and in vivo and in vitro studies were included in this review. A. paniculata extracts and their bioactive compounds exert anti-senescence properties through their anti-inflammatory and antioxidant properties. This herb and its compounds enhanced memory, cognitive function and behaviour in Alzheimer's disease. The extract also promoted cell cycle progression and proliferation in the skin. In addition, andrographolide exhibited anti-senescence effects in endothelial cells through the activation of PI3K/Akt/Nrf and PI3K/Akt/AP-1 pathways. A. paniculata along with its bioactive compounds including andrographolide and 14-deoxyandrographolide, may have the potential to be used as anti-senescence through anti-inflammatory and antioxidant properties. However, the specific markers to evaluate the senescence are necessary to be conducted. Any clinical trials should be done to establish these findings. Since in clinical settings this potential herbal may be used for long-life time, the safety profile and toxicity of A. paniculata should be considered. Keywords: herbal plants, Andrographis paniculata, andrographolide, bioactive compounds, senescence