Pakistan Journal of Biological Sciences

&lt;b&gt;Background and Objective:&lt;/b&gt; The secondary metabolite profile of Indonesian Arabica coffee (&lt;i&gt;Coffea arabica&lt;/i&gt; L.) is highly influenced by environmental factors, especially the altitude of the growing location. Arabica coffee by-products, such as cascara, have strong bioactive potential, including antioxidant activity. However, currently, by-products like cascara, which consist of dried pulp and skin, are often neglected as coffee demand increases. The objective of this study was to investigate and compare the phytochemical composition and antioxidant activity of Arabica coffee cascara (&lt;i&gt;Coffea arabica&lt;/i&gt; L.) from two different altitudes in West Java. &lt;b&gt;Materials and Methods:&lt;/b&gt; Cascara is obtained from two different regions: Low altitude (Leles, 900-1300 masl) and high altitude (Ibun, 1300-1700 masl). The Soxhlet extraction process was carried out using a stepwise method based on the polarity level of the solvents (n-hexane, ethyl acetate and ethanol). The antioxidant activity was determined using DPPH and phytochemical standardization was carried out in the form of Total Phenolic Content (TPC), Total Flavonoid Content (TFC) and caffeine content. Findings were expressed as Mean±SD and analyzed using independent t-test, ANOVA-Tukey (p<0.05) and Pearson correlation. &lt;b&gt;Results:&lt;/b&gt; Cascara extract from Leles shows higher TPC, TFC, caffeine content and antioxidant activity compared to extract from Ibun. Ethanol extract from Leles shows a strong category of antioxidant activity with an IC&lt;sub&gt;50&lt;/sub&gt; value of 35.74 μg/mL. The results of statistical data processing showed the altitude influenced on the content of phenolic compounds, flavonoids, caffeine and antioxidant activity. &lt;b&gt;Conclusion:&lt;/b&gt; The results indicate that the higher total phenols, total flavonoids and caffeine in Arabica cascara extract can be attributed to the lower altitude of the Arabica coffee growing areas in West Java. The antioxidant activity is influenced by the concentrations of these bioactive compounds.

&lt;b&gt;Background and Objective:&lt;/b&gt; Antimicrobial resistance (AMR) drives an urgent need for new antibiotics. A &lt;i&gt;Lysinibacillus&lt;/i&gt; sp. isolate from Indonesian peat soil has shown promising broad-spectrum antibacterial activity &lt;i&gt;in vitro&lt;/i&gt;. This study aims to evaluate the acute toxicity profile of its cell-free supernatant (CFS) as an initial &lt;i&gt;in vivo&lt;/i&gt; safety assessment. &lt;b&gt;Materials and Methods:&lt;/b&gt; A single oral dose acute toxicity test was conducted on female mice (n = 3 per group). Treatment groups received CFS at volumes of 0.2 mL, 0.5 mL and 1.0 mL. Control groups received 0.5% Na carboxy methyl cellulose (CMC) and nutrient broth (NB). All animals were observed for 14 days for mortality, behavior and body weight changes, followed by necropsy on day 15. Statistical analysis was conducted using SPSS version 26, applying GLM repeated measures with Shapiro-Wilk normality, Mauchly's sphericity and Bonferroni &lt;i&gt;post-hoc&lt;/i&gt; tests at a significance level of p<0.05, with results expressed as Mean±SD. &lt;b&gt;Results:&lt;/b&gt; There was no mortality or significant behavioral changes. However, macroscopic necropsy revealed dose-dependent toxicity. At a dose of 0.2 mL, heart swelling and tracheal hemorrhage were found. A dose of 0.5 mL caused lung hemorrhage and a pale trachea. At the highest dose of 1.0 mL, all animals showed kidney swelling, lung hemorrhage and pale digestive tract. The Na CMC control group was normal, while the NB control showed mild abnormalities. &lt;b&gt;Conclusion:&lt;/b&gt; The CFS of &lt;i&gt;Lysinibacillus&lt;/i&gt; sp. did not show LD&lt;sub&gt;50&lt;/sub&gt; toxicity or behavioral changes in mice. Some high doses showed organ-specific toxicity.

&lt;b&gt;Background and Objective:&lt;/b&gt; Extracorporeal shock wave lithotripsy (ESWL) is a widely used non-invasive treatment for renal stones. However, its effectiveness in clearing stones located in the lower pole of the kidney is often limited due to the anatomical challenges that impede the spontaneous passage of fragmented stones. This study evaluate the efficacy and safety of External Physical Vibration Lithecbole (EPVL) when used as an adjunctive therapy following ESWL in patients with lower pole renal stones, with a focus on improving stone clearance rates. &lt;b&gt;Materials and Methods:&lt;/b&gt; This prospective interventional study was conducted at Al Yarmouk Teaching Hospital over two years and included 100 patients with 10-15 mm lower pole renal stones. Patients were randomized into two groups: The ESWL alone and ESWL plus EPVL. All patients received two ESWL sessions (3000 shocks/session). The treatment group additionally received EPVL therapy using a flank-applied vibration device. Follow-up was performed using ultrasound and KUB radiography. Statistical analysis was conducted using appropriate tests with significance set at p<0.05. &lt;b&gt;Results:&lt;/b&gt; Baseline characteristics, including age, gender, weight, stone size and location, were statistically comparable between groups. The mean stone size was 12.4±1.7 mm. A significantly higher stone-free rate was observed in the ESWL+EPVL group compared to the ESWL-only group (66% vs. 48%, p = 0.027). Although the residual stone size showed a numerical difference favoring EPVL, it was not statistically significant (p = 0.11). Complication rates were low, mild and similar in both groups (p = 0.3). Logistic regression analysis revealed that smaller stone size (p<0.001) and lower patient weight (p = 0.030) were significantly associated with successful stone clearance. Subgroup analysis further confirmed that patients with stones <11 mm or a weight <70 kg had the highest stone-free rates. &lt;b&gt;Conclusion:&lt;/b&gt; In this initial evaluation, EPVL demonstrated a safe and effective adjunct to ESWL in enhancing stone clearance for lower-pole renal stones. Its application was associated with improved treatment outcomes without increasing complication rates. Further studies with extended EPVL sessions and longer follow-up are warranted.

&lt;b&gt;Background and Objective:&lt;/b&gt; &lt;i&gt;Penaeus monodon&lt;/i&gt; is a robust shrimp species capable of surviving under varying water conditions and seasons. Despite its size and potential socio-economic benefits, it has not been cultivated in the Karang Mumus River due to concerns about water pollution. This study aimed to assess the water quality characteristics of the Karang Mumus River as a potential culture medium for &lt;i&gt;P. monodon&lt;/i&gt; during both the rainy and summer seasons. &lt;b&gt;Materials and Methods:&lt;/b&gt; An analytical observational study with a cross-sectional design was conducted. Data were collected twice, in July and October 2023. Shrimp body weight was measured weekly, and water quality parameters, including Biochemical Oxygen Demand (BOD), Chemical Oxygen Demand (COD), and Total Suspended Solids (TSS) were analyzed at multiple river locations. The survival rate data were arcsine-transformed before analysis, and all data were analyzed using SPSS, with significance determined at p<0.05 using Chi-square tests for categorical variables. &lt;b&gt;Results:&lt;/b&gt; Significant differences were observed in TSS and COD between seasons, while BOD and COD values remained within standard threshold limits. Weekly measurements showed progressive increases in shrimp body weight, with the third sampling location demonstrating significantly higher growth compared to other sites. &lt;b&gt;Conclusion:&lt;/b&gt; The findings indicate that &lt;i&gt;P. monodon&lt;/i&gt; can be successfully cultured in the Karang Mumus River. Seasonal variations in TSS and COD exist but remain within acceptable water quality standards, confirming the river's potential as a shrimp culture medium.

&lt;b&gt;Background and Objective:&lt;/b&gt; &lt;i&gt;Moringa oleifera&lt;/i&gt;, a plant rich in flavonoids and phenolic compounds, is widely recognized for its antioxidant potential. This study aimed to evaluate the antioxidant activity of underutilized parts of &lt;i&gt;M. oleifera&lt;/i&gt; (twigs and stems) compared to leaves (well-utilized part), expressed as ascorbic acid equivalents. Additionally, the relationships between total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activity were investigated. &lt;b&gt;Materials and Methods:&lt;/b&gt; TPC and TFC were determined using UV-Visible spectrophotometry. Antioxidant activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Cupric Ion Reducing Antioxidant Capacity (CUPRAC) assays. Correlations among TPC, TFC and antioxidant activity were analyzed using Pearson's correlation, principal component analysis (PCA) and hierarchical cluster analysis (HCA). Flavonoid compounds were further characterized using TLC densitometry. Statistical significance among the data was analyzed using One-Way ANOVA in Minitab software at a significance level of p<0.05. &lt;b&gt;Results:&lt;/b&gt; The ethanolic extract of &lt;i&gt;M. oleifera&lt;/i&gt; leaves exhibited the highest antioxidant activity in both DPPH assays, while the ethyl acetate of &lt;i&gt;M. oleifera &lt;/i&gt;twigs extract showed the highest activity using the CUPRAC assay. It is possible to develop twigs &lt;i&gt;M. oleifera &lt;/i&gt;as a potential antioxidant agent resource. &lt;b&gt;Conclusion:&lt;/b&gt; Among the tested plant parts, leaves demonstrated the highest antioxidant activity compared to twigs and stems. However, twigs also showed promising potential activity to be developed as source of antioxidant agent.

&lt;b&gt;Background and Objective:&lt;/b&gt; Changes in rhizosphere microbial populations have been reported in response to drought, temperature fluctuations, CO&lt;sub&gt;2&lt;/sub&gt; levels and other environmental factors. However, the structure of the root-associated microbes in local North Sulawesi rice using a metagenomic approach has not yet been investigated. This study examined the microbial community structure in local North Sulawesi rice (cv. Superwin) under drought (water deficit) conditions compared to well-watered conditions at the vegetative phase. &lt;b&gt;Materials and Methods:&lt;/b&gt; Rice plants were grown in polybags filled with a 5:1:1 mixture of garden soil, compost and rice husks and were allowed to grow until the four-fully-expanded leaf stage. They were then subjected to two treatments for 14 days: well-watered conditions (irrigated to 100% field capacity) and water deficit conditions (0% field capacity). Root samples were collected for next-generation sequencing analysis to assess molecular response of Superwin rice to water deficit. &lt;b&gt;Results:&lt;/b&gt; During drought, several root-associated microbes were more prevalent, including &lt;i&gt;Nitrospirota&lt;/i&gt; at the phylum level, &lt;i&gt;Rubrobacteria&lt;/i&gt; at the class level, &lt;i&gt;Micrococcales&lt;/i&gt; at the order level, Gaiellaceae at the family level, &lt;i&gt;Gaiella&lt;/i&gt; at the genus level and &lt;i&gt;Gaiella occulta&lt;/i&gt; at the species level. &lt;b&gt;Conclusion:&lt;/b&gt; Root-associated microbes, including taxa &lt;i&gt;Nitrospirota&lt;/i&gt;, &lt;i&gt;Rubrobacteria&lt;/i&gt;, Micrococcales, Gaiellaceae, &lt;i&gt;Gaiella&lt;/i&gt; and &lt;i&gt;Gaiella occulta&lt;/i&gt;, have a higher relative abundance in rice plants under water deficit. &lt;i&gt;Gaiella occulta&lt;/i&gt; serves as sensitive indicator of water deficit in North Sulawesi local rice, i.e. Superwin.

<b>Background and Objective:</b> Rice weevils, <i>Sitophilus oryzae</i> (L.), are major pests of stored grains, causing substantial post-harvest losses. This study evaluated the efficacy of selected medicinal plant extracts for the control and repellency of rice weevils as eco-friendly alternatives to synthetic pesticides. <b>Materials and Methods:</b> Twenty-one medicinal plants were initially screened and four plants were selected for extraction using 70% ethanol and distilled water. Contact toxicity was assessed at concentrations of 5, 10 and 15% (w/v) using plastic cup bioassays. Repellency tests were conducted to determine the effectiveness of extracts over time. Data were analyzed using one-way ANOVA followed by Tukey's HSD test (p<0.05) in SPSS. <b>Results:</b> Among ethanolic extracts, tobacco leaves at 15% showed the highest insect mortality (93.3±3.3%), significantly higher than clover flowers (29.9±15.3%). All concentrations of aqueous tobacco extract resulted in 100% mortality, while lemon and long pepper extracts demonstrated moderate toxicity (>50%). In repellency assays, ethanolic clover flowers (10%) and aqueous clover flowers (15%) exhibited the highest average repellent efficiencies of 95.6±2.9 and 92.2±4.8%, respectively, with maximum repellency observed within 30 min. <b>Conclusion:</b> The findings indicate that selected medicinal plant extracts, particularly tobacco and clover, are effective and eco-friendly agents for the control and repellency of rice weevils. These plant-based extracts show promise as sustainable alternatives to synthetic pesticides for stored grain protection.

&lt;b&gt;Background and Objective:&lt;/b&gt; Amaranth microgreens are increasingly recognized as valuable functional foods due to their rapid growth, short cultivation cycle, high nutritional density and rich phytochemical content. This study investigated how cultivar and harvest stage affect growth performance and the accumulation of key phytochemical compounds in amaranth microgreens grown under greenhouse conditions. &lt;b&gt;Materials and Methods:&lt;/b&gt; Two amaranth cultivars, red amaranth and small red-centered amaranth, were grown under greenhouse conditions and harvested at 7, 10 and 14 days after sowing. Growth parameters, including plant height, fresh weight and dry weight, were measured. Chlorophyll content, dietary fiber, vitamin C, total phenolic content and antioxidant activity were also determined. Data was analyzed using analysis of variance and treatment means were compared at p<0.05. &lt;b&gt;Results:&lt;/b&gt; Harvest stage significantly affected most growth and biochemical traits. In both cultivars, plant height, dry weight, chlorophyll content, total phenolic content and antioxidant activity increased with plant age. At 14 days after sowing, red amaranth showed the highest chlorophyll content (0.49 mg/g fresh weight), total phenolic content (4.51 mg GAE/g fresh weight) and antioxidant activity (52.27%). Small red-centered amaranth showed slightly higher dietary fiber (3.18%) and vitamin C content (94.44 mg/g fresh weight) at the same harvest stage. &lt;b&gt;Conclusion:&lt;/b&gt; Both cultivar and harvest stage influenced the nutritional and phytochemical quality of amaranth microgreens. Harvesting at 14 days after sowing provided the most favorable balance between growth and bioactive compound accumulation, with red amaranth showing superior overall phytochemical performance.

&lt;b&gt;Background and Objective:&lt;/b&gt; Pathogenic microorganisms such as bacteria and parasites cause disease and ill health in domestic pets, including cats. However, few studies have examined the composition of the bacterial community in cat faeces containing helminth eggs. To bridge this knowledge gap, study aimed to investigate bacterial microbiota in cat faeces containing nematode eggs. &lt;b&gt;Materials and Methods:&lt;/b&gt; Bacterial analysis was performed on cat faeces targeting the V3-V4 region of 16S rDNA using next-generation sequencing technology. Samples of cat faeces that contained roundworm eggs were pooled as Group 1, whereas faeces without roundworm eggs placed in Group 2. Results were analysed using R (v3.3.1), SPSS (Chicago, IL, USA) and LEfSe (v1.0) for visualization, statistical testing and differential taxa abundance analysis, respectively (p<0.05). &lt;b&gt;Results:&lt;/b&gt; Groups 1 and 2 shared 306 OTUs (Operational Taxonomic Units). In different OTUs, Group 1 contained 324 and Group 2 was 185 OTUs. At the phylum level, Bacteroidota dominated the community composition of Groups 1 and 2. Firmicutes were abundant in both groups. Campylobacterota were more abundant in Group 1 than in Group 2. However, Fusobacteriota was more predominant in Group 2 than in Group 1. At the genus level, &lt;i&gt;Helicobacter&lt;/i&gt; was higher in Group 1 than in Group 2. In contrast, &lt;i&gt;Bacteroides&lt;/i&gt;, &lt;i&gt;Fusobacterium&lt;/i&gt; and &lt;i&gt;Collinsella&lt;/i&gt; were more abundant in Group 2 than in Group 1. &lt;b&gt;Conclusion:&lt;/b&gt; These results provide the first evidence of the bacterial community and bacterial core in cat faeces containing roundworm eggs.

&lt;b&gt;Background and Objective:&lt;/b&gt; The rapid development and deployment of COVID-19 vaccines constituted a critical global public health response; however, vaccine hesitancy posed a substantial challenge, particularly in low- and middle-income countries such as Iraq. Mistrust, misinformation and sociopolitical factors were reported to influence vaccine uptake. This study aimed to assess public perception, acceptance and hesitancy toward COVID-19 vaccines in Iraq during the pandemic and to identify key sociodemographic and informational determinants influencing vaccination behavior. &lt;b&gt;Materials and Methods:&lt;/b&gt; A cross-sectional survey was conducted among 1,747 participants across Iraq. Vaccine acceptance was defined as receipt of at least one vaccine dose, while hesitancy included delayed or refused vaccination due to concerns regarding safety, efficacy, or other factors. Public perception reflected attitudes and beliefs toward vaccination irrespective of vaccination status. Data were analyzed using descriptive statistics and inferential tests to examine associations between sociodemographic variables, exposure to misinformation and vaccine acceptance, with statistical significance set at p<0.05. &lt;b&gt;Results:&lt;/b&gt; Among the participants, 42.7% were vaccinated and 57.3% were unvaccinated. Vaccine acceptance was significantly higher among men, older individuals and participants with higher educational attainment (p<0.001). The most commonly reported reasons for vaccine hesitancy were concerns about long-term side effects (72.4%) and distrust in vaccine manufacturers (41.9%). Exposure to misinformation via social media platforms showed a significant association with increased vaccine hesitancy (p = 0.03). &lt;b&gt;Conclusion:&lt;/b&gt; The findings identify critical sociodemographic and informational factors influencing COVID-19 vaccine acceptance in Iraq. Targeted public health interventions focusing on women, younger populations and individuals with lower educational levels are warranted. Strengthening evidence-based communication strategies to counter misinformation and rebuild public trust may improve vaccine uptake. Future studies should explore longitudinal trends and intervention effectiveness to address persistent vaccine hesitancy.

&lt;b&gt;Background and Objective:&lt;/b&gt; Onion (&lt;i&gt;Allium cepa&lt;/i&gt; L.) is one of the most important commercial vegetable crops worldwide with high economic value. However, fungal diseases seriously threaten onion production by reducing yield and bulb quality. For sustainable disease management, the use of biological control agents has emerged as an environmentally friendly and effective alternative to chemical fungicides. This study aimed to select potential antifungal bacteria that can promote plant growth in onion. &lt;b&gt;Materials and Methods:&lt;/b&gt; Bacterial isolates were isolated from rhizosphere soil samples, screened and characterized for antifungal activities. Strong antagonistic bacterial strains were selected. These bacterial strains were identified and characterized by their biochemical features. In addition, three selected strains were evaluated for their growth-promoting capacity by pot assay. All experiments were conducted in triplicate; data were expressed as mean±SD and analyzed using one-way ANOVA followed by Tukey's test (p<0.05) in GraphPad Prism 9.3.1. &lt;b&gt;Results:&lt;/b&gt; Three bacterial. strains exhibited strong antagonistic activity against &lt;i&gt;Sclerotium rolfsii&lt;/i&gt; and &lt;i&gt;Curvularia lunata&lt;/i&gt; through the production of diffusible compounds and volatile organic compounds (VOCs). In addition, physiological and biochemical characterization revealed that all strains belonged to the genus &lt;i&gt;Bacillus.&lt;/i&gt; Moreover, soil amendment with the culture of &lt;i&gt;Bacillus&lt;/i&gt; sp. T3, T33 and T36 significantly enhanced onion growth. &lt;b&gt;Conclusion:&lt;/b&gt; These findings suggest that the selected &lt;i&gt;Bacillus&lt;/i&gt; sp. strains represent promising candidates for the development of biocontrol agents against fungal pathogens.

&lt;b&gt;Background and Objective:&lt;/b&gt; Diabetes mellitus, characterized by chronic hyperglycemia, disrupts multiple physiological systems. Streptozotocin (STZ)-induced diabetic models replicate these disturbances, including elevated glucose, liver and kidney dysfunction, and altered lipid profiles. Vitamin D, known for its role in bone metabolism, also modulates biochemical and inflammatory pathways, potentially offering protective effects against diabetes-induced metabolic changes. This study aimed to evaluate the effects of vitamin D supplementation on biochemical parameters in STZ-induced diabetic Wistar rats. &lt;b&gt;Materials and Methods:&lt;/b&gt; Male Wistar rats were divided into four groups: control (non-diabetic), non-diabetic+vitamin D, diabetic, and diabetic+vitamin D. Diabetes was induced by intraperitoneal administration of STZ (35 mg/kg). After the confirmation of hyperglycemia (≥250 mg/dL), the supplemented groups received vitamin D (cholecalciferol, 80 IU/day) for 30 days. Glucose, urea, creatinine, amylase, total cholesterol, HDL-cholesterol, triglycerides, AST, ALT, and total bilirubin and fractions were evaluated. Differences were considered statistically significant when p < 0.05. &lt;b&gt;Results:&lt;/b&gt; The diabetic group exhibited a significant increase (p < 0.05) in glucose, urea, creatinine, triglycerides, AST, ALT, and total bilirubin and fractions, as well as a reduction in HDL-cholesterol, when compared to the control group. Vitamin D supplementation promoted a reduction in glucose, urea, creatinine, triglyceride, and AST levels and an increase in HDL-cholesterol in the diabetic+vitamin D group. &lt;b&gt;Conclusion:&lt;/b&gt; Vitamin D showed potential as a modulator of biochemical parameters altered by STZ, indicating a hepatorenal protective effect and improvement of lipid and glycemic profiles.

&lt;b&gt;Background and Objective:&lt;/b&gt; Rhizosphere fungi play a crucial role in nutrient cycling and plant protection, yet most are difficult to cultivate using conventional methods. Consequently, their ecological functions remain largely unknown. Therefore, metagenomic approaches allow for comprehensive and accurate mapping of fungal taxonomic profiles without the need for cultivation and this study investigated the variation of rhizosphere fungi across different rice cultivation systems to elucidate their diverse potentials. &lt;b&gt;Materials and Methods:&lt;/b&gt; A metagenomic approach was employed to identify fungi originating from the rhizosphere of rice cultivated in various field conditions, including irrigated, rainfed and organic rice fields. The diversity of fungi from rhizosphere samples was assessed to comprehend the relationships and metrics within the rice cropping systems utilized by farmers. &lt;b&gt;Results:&lt;/b&gt; The findings indicated that the rhizosphere fungal index from organic rice fields exhibited the highest Shannon and Simpson index values compared to those from irrigated and rainfed rice fields. &lt;b&gt;Conclusion:&lt;/b&gt; Metagenomic analysis revealed that the most dominant fungal diversity at the family level was Trichocomaceae, at the genus level was Talaromyces and at the species level was &lt;i&gt;Talaromyces wortmannii&lt;/i&gt;.

&lt;b&gt;Background and Objective:&lt;/b&gt; Biochar is widely recognized for its porous structure, stability and large surface area, making it a promising carrier for beneficial soil microorganisms. However, limited information exists regarding its efficiency in supporting actinomycete enrichment at varying bacterial densities. This study addressed this gap by evaluating the colonization potential of biochar enriched with actinomycetes. The objective was to determine the optimal bacterial density and dilution level for maximizing colony formation under controlled laboratory conditions. &lt;b&gt;Materials and Methods:&lt;/b&gt; The experiment was conducted from July to October 2024 at the Laboratory of Plant Biotechnology, Centre for Research Activities (CRA), Hasanuddin University, using a randomized group design with three replications. Actinomycetes were inoculated onto biochar at three bacterial densities (10&lt;sup&gt;4&lt;/sup&gt;, 10&lt;sup&gt;8&lt;/sup&gt; and 10&lt;sup&gt;12&lt;/sup&gt; CFU/mL) and three dilution levels (10&lt;sup&gt;5&lt;/sup&gt;, 10&lt;sup&gt;6&lt;/sup&gt; and 10&lt;sup&gt;7&lt;/sup&gt;). Colony observations were recorded weekly for four weeks. Visual colony morphology was documented and environmental conditions (pH, temperature and humidity) were monitored. Data were tabulated and analyzed using ANOVA and significant treatment effects (p<0.05 or p<0.01) were further compared using Tukey's HSD test at α = 0.05. &lt;b&gt;Results:&lt;/b&gt; Actinomycete isolates consistently exhibited round, raised colonies with irregular edges and smooth to wrinkled surfaces. The highest average colony count occurred at pH 7.93, 24.8°C and 66% humidity. The 10&lt;sup&gt;8&lt;/sup&gt; CFU/g enrichment showed the greatest colony formation during the first week, reaching 4.5×10&lt;sup&gt;9&lt;/sup&gt; CFU/g, significantly higher than the 10&lt;sup&gt;4&lt;/sup&gt; and 10&lt;sup&gt;12&lt;/sup&gt; CFU/g treatments. Among dilution levels, 10&lt;sup&gt;7&lt;/sup&gt; produced the highest colony count (4.5×10&lt;sup&gt;9&lt;/sup&gt; CFU/g), showing a significant difference compared with 10&lt;sup&gt;6&lt;/sup&gt; and 10&lt;sup&gt;5&lt;/sup&gt;. &lt;b&gt;Conclusion:&lt;/b&gt; Biochar demonstrated strong potential as a suitable carrier medium for actinomycete enrichment, with the 10&lt;sup&gt;8&lt;/sup&gt; CFU/g density and 10&lt;sup&gt;7&lt;/sup&gt; dilution yielding optimal growth. These findings highlight biochar's promise as an effective microbial inoculant for enhancing soil health and supporting sustainable agriculture. Further research should explore field-scale validation and long-term microbial stability.

Plasma vitamin D, measured as 25-Hydroxyvitamin D [25(OH)D], is influenced by genetics, lifestyle, age, gender, skin pigmentation, sunlight exposure, latitude, sunscreen use, diet and supplementation. Globally, vitamin D deficiency is frequently reported among healthy populations, yet comprehensive analyses summarizing its prevalence are limited. This systematic review and meta-analysis aimed to assess 25(OH)D status among healthy adults across different regions worldwide. We systematically searched PubMed, Web of Science, ScienceDirect and Google Scholar for observational studies published in English between 2013 and 2024. Heterogeneity among studies was evaluated using Cochran's Q test and the I&lt;sup&gt;2&lt;/sup&gt; statistic, with significance set at p<0.10; I&lt;sup&gt;2&lt;/sup&gt; values were interpreted as low (0-40%), moderate (30-60%), substantial (50-90%) and considerable (75-100%). Due to significant heterogeneity, a random-effects model was employed to estimate the prevalence of vitamin D deficiency. A total of 394,548 participants from 56 studies across 30 countries were included. The pooled prevalence of vitamin D deficiency was 55% (95% CI: 49.0-60.0%), with considerable heterogeneity (Q p<0.001; I&lt;sup&gt;2&lt;/sup&gt; = 100%). Subgroup analysis showed the highest deficiency in the Middle East (62%; 95% CI: 54-70%), followed by Asia (54%; 95% CI: 39-69%), Africa (43%; 95% CI: 24-63%), and Europe (43%; 95% CI: 23-63%). Weighted mean vitamin D levels ranged from 8.4±0.5 ng/mL to 37.7±10.8 ng/mL, with an overall mean of 20.14±6.7 ng/mL. Regionally, the Middle East had the lowest recalculated weighted mean level (17.8 ng/mL), followed by Asia (20.4 ng/mL), Europe (26.8 ng/mL), "Others" (28.0 ng/mL) and Africa (29.2 ng/mL, influenced by one high outlier study). These findings reveal a high prevalence of vitamin D deficiency among healthy adults, particularly in the Middle East, but with the lowest mean levels observed in the Middle East and marked regional disparities, highlighting marked regional disparities.

&lt;b&gt;Background and Objective:&lt;/b&gt; Plant-derived compounds that disrupt bacterial quorum sensing (QS) provide alternatives to combat antibiotic resistance. This quorum quenching (QQ) strategy reduces virulence without inhibiting growth, thus minimizing resistance. Kelakai (&lt;i&gt;Stenochlaena palustris&lt;/i&gt; (Burm.) Bedd.), an endemic Kalimantan plant traditionally used by the Dayak people, was evaluated for QQ activity against &lt;i&gt;Pseudomonas aeruginosa&lt;/i&gt;. &lt;b&gt;Materials and Methods:&lt;/b&gt; Ethanolic extract of &lt;i&gt;S. palustris&lt;/i&gt; leaves was prepared by maceration with 96% ethanol. The extract was tested for inhibition of &lt;i&gt;P. aeruginosa &lt;/i&gt;motility, pyocyanin production and LasA activity. Antibiofilm effects were assessed using the microtiter plate method, EPS quantified, chemical profiling performed by Liquid Chromatography-High-Resolution Mass Spectrometry (LC-HRMS) and acute toxicity evaluated in &lt;i&gt;Rasbora lateristriata&lt;/i&gt; embryos. The results from three independent trials were analyzed in SPSS using one-way ANOVA followed by Dunn's test, with significance set at p<0.05. &lt;b&gt;Results:&lt;/b&gt; At 0.25 mg/mL, the extract inhibited swarming and twitching by 58.04±0.46% and 64.98±0.25%, reduced LasA activity to 58.27±0.70% and reduced pyocyanin to 49.96±1.15%. Although antibacterial activity was weak, at a concentration of 0.25 mg/mL the extract caused 52.47±0.58% inhibition of &lt;i&gt;P. aeruginosa&lt;/i&gt; biofilm formation and &lt;i&gt;P. aeruginosa&lt;/i&gt; EPS was reduced to 67.40±0.54% of the untreated control. At a concentration of 1 mg/mL, pre-formed biofilm was eradicated as much as 58.52±0.23%. The LC-HRMS identified (-)-Strychnine, D-(+)-Pipecolinic acid and Ethyl oleate as major constituents. Toxicity testing revealed developmental toxicity in embryos at 0.06 mg/mL. &lt;b&gt;Conclusion:&lt;/b&gt; &lt;i&gt;Stenochlaena palustris&lt;/i&gt; ethanol extract showed strong QQ activity against &lt;i&gt;P. aeruginosa&lt;/i&gt;, underscoring its potential as a natural anti-infective. Further isolation of active compounds with reduced toxicity is required for therapeutic development.

Osseointegration, the biological integration of dental implants with bone, is critical for the long-term success of implant-supported restorations. Despite advances in implant materials and techniques, achieving rapid and reliable bone healing remains a challenge. Melatonin, a biologically active molecule with antioxidant, anti-inflammatory and immunomodulatory properties, has shown potential to enhance bone regeneration and implant stability. This study aims to evaluate the role of melatonin in promoting osseointegration and to explore its potential clinical applications in implant dentistry. A narrative review of laboratory, animal and preliminary clinical studies investigating the effects of melatonin on bone healing and implant stability was conducted. Studies were selected based on the use of melatonin in local application, systemic supplementation or incorporation into implant surfaces. Key outcomes assessed included osteoblast activity, osteoclast inhibition, vascularization, bone density and healing time. Laboratory and animal studies demonstrated that melatonin enhances osteoblast proliferation, reduces osteoclast-mediated bone resorption and promotes vascularization through the regulation of vascular endothelial growth factor. Preliminary clinical studies indicated improved bone density on radiographic assessment and reduced healing time following local melatonin application. However, evidence remains limited and heterogeneity in dosing, administration methods and follow-up periods was observed. Melatonin shows promising potential to enhance osseointegration and accelerate bone healing in dental implant procedures. While initial findings are encouraging, optimal dosing, delivery methods and long-term safety remain unclear. Well-designed, controlled clinical trials are necessary before melatonin can be recommended as a standard adjunct in implant dentistry.

&lt;b&gt;Background and Objective:&lt;/b&gt; Payangka fish (&lt;i&gt;Ophieleotris aporos&lt;/i&gt;), an underutilized endemic species from Lake Tondano, North Sulawesi, Indonesia, represents a potential resource for sustainable food fortification. Despite comprising 35% of the local catch, its biochemical valorization remains unexplored. This study aimed to optimize the enzymatic hydrolysis of Payangka fish muscle protein using papain and bromelain to produce bioactive hydrolysates suitable for functional food applications. &lt;b&gt;Materials and Methods:&lt;/b&gt; Fish protein hydrolysis was performed at 55°C with a solid-to-liquid ratio of 1:4, using varying concentrations of papain and bromelain. The resulting hydrolysates were characterized for degree of hydrolysis (DH), proximate composition, color, amino acid profile and antioxidant activity. Statistical analysis was conducted to assess the effect of enzyme type and concentration on hydrolysis and bioactivity. Differences were considered statistically significant at p<0.05. &lt;b&gt;Results:&lt;/b&gt; Increasing enzyme concentration positively correlated with DH. Papain achieved optimal DH (86.5±0.01% to 93.13±0.01%) within 2 hrs, whereas bromelain reached 71.94±0.01% to 84.01±0.01% after 8 hrs. Papain-derived hydrolysates (2 hrs, 4% enzyme) showed increased protein content (0.52-0.68%) and the highest antioxidant activity (85.76%). Total essential and non-essential amino acids were 74.47 ppm and 86.30 ppm, respectively, with lysine (26.65 ppm), leucine (15.02 ppm) and valine (11.01 ppm) being dominant essential amino acids and glutamic acid (26.34 ppm), aspartic acid (17.53 ppm) and alanine (9.83 ppm) as dominant non-essential amino acids. &lt;b&gt;Conclusion:&lt;/b&gt; Fish protein hydrolysate from Payangka, particularly using 4% papain, demonstrates high bioactivity and favorable amino acid composition, highlighting its potential as a functional ingredient for food fortification. Future research could explore its incorporation into food products and long-term bioactivity.

&lt;b&gt;Background and Objective:&lt;/b&gt; Platelet aggregation plays a critical role in hemostasis and thrombosis and its dysregulation can lead to cardiovascular disorders such as stroke and myocardial infarction. Phytochemicals derived from plants have shown potential in modulating platelet function, but the molecular mechanisms remain unclear. This study aimed to investigate the interactions of selected phytochemicals with key proteins involved in platelet aggregation pathways to explore their potential antiplatelet effects. &lt;b&gt;Materials and Methods:&lt;/b&gt; In this &lt;i&gt;in silico&lt;/i&gt; study, five phytochemicals-caffeic acid, chlorogenic acid, coumaric acid, gallic acid and salicylic acid-were docked onto four target proteins: Prostaglandin-Endoperoxide Synthase 1 (PTGS1), prostacyclin synthase (PGIS), Glycoprotein VI (GPVI) and Protease-Activated Receptor 1 (PAR1). Computational molecular docking techniques were used to evaluate binding modes and affinities, providing insights into potential modulatory effects on platelet function. &lt;b&gt;Results:&lt;/b&gt; Coumaric acid, caffeic acid and chlorogenic acid exhibited significant interactions with all four target proteins, demonstrating favorable binding affinities and stable docking conformations. These interactions suggest their potential to modulate platelet aggregation pathways. Notably, coumaric acid showed the strongest binding to PTGS1 and GPVI, indicating a possible mechanism for its antiplatelet activity. &lt;b&gt;Conclusion:&lt;/b&gt; The study provides molecular-level evidence supporting the antiplatelet potential of selected phytochemicals, particularly coumaric, caffeic and chlorogenic acids. These findings lay the groundwork for future experimental and clinical investigations into their therapeutic applications in cardiovascular disease prevention and treatment.

&lt;b&gt;Background and Objective:&lt;/b&gt; Chronic hyperglycemia causes oxidative stress, immunological dysfunction and decreased angiogenesis, compromising wound healing. Wound healing requires VEGF, which stimulates angiogenesis and tissue regeneration. The aim of this study was to assess the effect of the ethanolic extract of &lt;i&gt;Gynura procumbens&lt;/i&gt; leaves on VEGF expression in diabetic wound tissue using &lt;i&gt;in vivo&lt;/i&gt; and &lt;i&gt;in silico&lt;/i&gt; approaches. &lt;b&gt;Materials and Methods:&lt;/b&gt; This study divided 30 alloxan-induced rats into five groups: Diabetic controls and those given Bioplacenton ointment and topical &lt;i&gt;G. procumbens&lt;/i&gt; extract at 100, 200 and 300 mg/kg bw. Histology included hematoxylin-eosin staining and VEGF immunohistochemistry. The active chemicals of &lt;i&gt;G. procumbens&lt;/i&gt; were examined using LC-MS/MS and VEGF was tested &lt;i&gt;in silico&lt;/i&gt; utilizing molecular docking and dynamics. Data were analyzed using Kruskal-Wallis and Mann-Whitney tests, with results expressed as Mean±SD and significance set at p<0.05. &lt;b&gt;Results:&lt;/b&gt; &lt;i&gt;Gynura procumbens&lt;/i&gt; extract increased epithelial thickness, collagen density and balanced the number of fibroblasts in diabetic wounds, with the best results at a dose of 300 mg/kg body weight. VEGF expression increased significantly in group A3 compared to the untreated diabetic group. &lt;i&gt;In silico&lt;/i&gt; tests showed that Mahuannin F had the highest binding affinity for VEGF (-7.2 kcal/mol) and was able to stabilize the VEGF structure disrupted by alloxan through hydrogen and hydrophobic interactions. Molecular dynamics simulations supported the conformational stabilization of VEGF by Mahuannin F. &lt;b&gt;Conclusion:&lt;/b&gt; Diabetes wounds with topically administered&lt;i&gt; G. procumbens &lt;/i&gt;extract had higher VEGF expression and better skin tissue architecture. Active substances like Mahuannin F, VEGF structural stabilizers, promote this chemical process.