Introduction: Different techniques have been used to improve the stability and bioavailability of biomaterials. This study aims to modify the crude stem extract of Spondia mombin used in ethno-medical management of benign prostrate hyperplasia (BPH).

Methods: Spondia mombin stem extract was modified by microencapsulation and freeze drying, and characterised using scanning electron microscopy (SEM), energy dispersive X-ray (EDX), and Fourier transform infrared spectroscopy (FTIR) analysis. The extracts were evaluated for physicochemical properties using micromeritic determination and dissolution studies, and for in vivo pharmacological properties using prostate-specific antigen (PSA), serum testosterone, and prostate index (PI) analysis on male Wister rats.

Results: SEM analysis showed a larger particle size of 50–80 µm for microencapsulated extract compared to >50 µm of crude extract. FT-IR and EDX analysis revealed relatively unchanged spectrum and additional molecules respectively in the modified extract compared to the crude extract. These results are suggestive of chemical compatibility and physical agglomeration in the interaction of the crude extract with microencapsulation excipients. Dissolution studies revealed steady release of extract from the microencapsulated form over 60 minutes, and > 9.76 times increase in peak release compared to release from the crude extract. The micromeritic studies showed improved flow properties for the microencapsulated extract with angle of repose < 27.9°, Hausner’s ratio < 1.06, and compressibility index < 6.06 %. The microencapsulated extract showed peak reduction in PSA at 100 mg / kg, compared to peak reduction of PSA of crude extract at 400 mg / kg. Serum testosterone levels decreased significantly in the microencapsulated extract.

Conclusion: Microencapsulation improved the flow and dissolution properties of Spondia mombin extract, and retained its pharmacological activities. There is a need to conduct human clinical trials to further explore the potentials of this microencapsulated

Coleus aromaticus (Roxb.) Benth. (Lamiaceae) is a species of semi-succulent perennial plant with recognized medicinal and nutritional relevance, widely employed in traditional medicine. While its aromatic fraction and essential oils have been extensively investigated, the non-volatile secondary metabolism of the plant remains poorly characterized. In particular, the free amino acids have never been investigated.

In this study, fresh leaves of C. aromaticus were treated with liquid nitrogen, ground, and subjected to different extraction procedures: hydroalcoholic maceration, maceration in ethyl acetate, and liquid–liquid partitioning yielding an organic phase in dichloromethane and a hydroalcoholic phase. The resulting fractions were analyzed by HPTLC and HPLC to define their metabolic profiles. In addition, the Bligh–Dyer method was applied to extract polar and non-polar metabolites, which were subsequently characterized by NMR spectroscopy.

Chromatographic and spectroscopic analyses revealed the presence of organic acids, polyphenols (notably flavonoids), and proteinogenic amino acids. The detection of both polar and non-polar metabolites provided a detailed chemical fingerprint of C. aromaticus leaves. In particular, the organic acids rosmarinic acid and caffeic acid were found to be the most abundant compounds, with concentrations of 88.89 mg/100 g and 49.66 mg/100 g, respectively. The flavonoids apigenin, luteolin and quercetin were also clearly identified, together with the free proteinogenic amino acids alanine, arginine, asparagine, aspartic acid, glutamic acid, glutamine, and serine. The most abundant amino acid was asparagine, with a concentration of 7.51 mg/100 g of matrix. This integrated approach offered a broader compositional overview of C. aromaticus, expanding its phytochemical profile beyond the well-documented essential oils.

These findings contribute to a deeper understanding of the non-volatile secondary metabolism of C. aromaticus and emphasize its potential as a source of bioactive metabolites with possible nutraceutical and pharmaceutical applications.

Background: Skin aging and pigmentation disorders are influenced by both intrinsic factors (genetics, metabolism, hormones) and extrinsic factors (ultraviolet radiation, pollution, chemicals). These factors contribute to progressive structural and functional alterations of the skin, clinically manifested as wrinkles, loss of elasticity, and hyperpigmentation. Among the key enzymes implicated in these processes are elastase and tyrosinase. In response to the growing demand for safer and more sustainable cosmetic ingredients, plant-derived extracts have attracted considerable interest as sources of natural anti-aging and depigmenting agents.

Methods: The present study investigated in vitro the inhibitory activities of hydroethanolic leaf extracts of Cistus salviifolius (CSLE) and Cistus laurifolius (CLLE) using spectrophotometric enzyme assays. Enzymatic inhibition was assessed using N-succinyl-Ala-Ala-Ala-p-nitroanilide as substrate for elastase and L-DOPA for tyrosinase, with elastatinal and kojic acid employed as reference inhibitors, respectively.

Results: CSLE exhibited notable anti-elastase activity with an IC₅₀ value of 43.0 µg/mL, significantly lower than that of CLLE (IC₅₀ =218.2 µg/mL), while the positive control elastatinal displayed an IC₅₀ of 0.5 µg/mL. Regarding tyrosinase inhibition, CSLE (IC₅₀ = 28.7 µg/mL) and CLLE (IC₅₀ = 30.2 µg/mL) demonstrated comparable activities, both slightly exceeding the efficacy of kojic acid (IC₅₀ = 31.1 µg/mL).

Conclusion: These findings highlight the potential of CSLE and CLLE extracts, particularly CSLE, as valuable natural sources of elastase and tyrosinase inhibitors, thereby supporting their prospective application in the formulation of cosmeceutical products.

Chondroitin sulphate is a sulphated glycosaminoglycan (GAG), abundantly present in connective tissues, particularly articular cartilage. There, it plays a critical role in maintaining structural integrity and elasticity. As a key component of the extracellular matrix, it is involved in various biological processes, including cellular signalling pathways, tissue hydration, and matrix organisation [1,2]. Furthermore, the therapeutic potential of chondroitin sulphate as an agent in joint-related disorders such as osteoarthritis has been studied due to its anti-inflammatory and cartilage-protective properties [3,4]. The therapeutic potential of chondroitin sulphate lies in its ability to reduce cartilage catabolism by inhibiting extracellular matrix degradation [5].

The combination of NMR spectroscopy and density functional theory (DFT) provides valuable insights into the structure, dynamics, and solubility of glycosaminoglycans [6]. In this study, DFT calculations were performed using the MN15/6-311++G(2d,2p)//SMD approach to determine the 3D structures of chondroitin 6-sulphate and chondroitin 4-sulphate tetrasaccharides. Computed proton–proton and proton–carbon coupling constants revealed that the one-bond proton–carbon coupling constants (¹J_C-H) depend strongly on the substitution position (C-6 vs. C-4). However, substitution also affects the ¹J_C-H and ³J_C-H magnitudes in the neighbouring atoms. The ¹J_C-H values were consistent with the experimental NMR values. The ³J_C-H values also revealed differences in the glycosidic linkages in chondroitin tetrasaccharides. These findings demonstrate that the position of sulphate groups on the chondroitin backbone significantly affects electronic distribution, geometry, and ⁿJ_C-H magnitudes, thereby highlighting the role of hydrogen bonding and ionic interactions in the 3D structure of chondroitin tetrasaccharides.

This work was financially supported by Slovak grant agency VEGA 2/0071/22.

• Mizumoto S., Yamada S., Sugahara K. Opin. Struct. Biol. 34 (2015) 35–42.
• Schwartz N., Domowicz M. S. Cell Dev. Biol. 10 (2022) 745372.
• Wildi L. et al. Rheum. Dis. 70 (2011) 982–989.
• Shen Q. et al.: Molecules 28 (2023) 7093–7126.
• Ma Y. et al. Orthoped. Surg. Res. 20 (2025) 11–23.
• Perez S. et al.: JACS Au. 3 (2023) 628–656.

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains the leading cause of infectious mortality worldwide, and the emergence of drug-resistant strains has intensified the search for new therapeutic agents. Marine macroalgae from underexplored ecosystems such as Antarctica represent promising sources of bioactive metabolites. Here, we investigated the antimycobacterial and anti-inflammatory potential of Desmarestia anceps. Crude extracts from eight sites across the Antarctic Peninsula were screened, among which the extract from Penguin Island exhibited the strongest effect against Mtb H37Rv (MIC₅₀ = 6.6 µg/mL) and concomitant reduction of nitric oxide (IC₅₀ = 4.2 ± 1.1 µg/mL) and TNF-α production (IC₅₀ = 21.4 ± 1.1 µg/mL). Bioassay-guided fractionation identified DAF2 as the most promising, displaying potent antimycobacterial effects (MIC₅₀ = 16.9 µg/mL against H37Rv), no cytotoxicity in murine macrophages (CC₅₀ > 100 µg/mL), and suppression of NO levels in LPS-stimulated macrophages, thus suggesting a dual role in infection control and inflammation modulation. High-speed counter-current chromatography (HSCCC) of DAF2 revealed sterols, fatty acids and diterpenoids as major constituents, among which saringosterol—previously shown to have potent antimycobacterial activity—and fucosterol were prominent. Fucosterol, isolated and tested individually, exhibited remarkable activity against Mtb H37Rv (MIC₅₀ = 0.37 ± 0.09 µg/mL) and the hypervirulent strain M299 (MIC₅₀ = 3.05 ± 1.11 µg/mL), with a selectivity index above 10 based on macrophage cytotoxicity (CC₅₀ = 51.45 µg/mL). Altogether, these findings highlight D. anceps as a promising source of natural products with combined antimycobacterial and anti-inflammatory activities, positioning fucosterol as a promising lead for TB drug discovery.

Background:
Alcoholic liver disease (ALD) remains one of the major causes of liver-related morbidity and mortality worldwide. Oxidative stress and inflammation are key mechanisms driving hepatic injury. Sinapic acid (SA), a naturally occurring phenolic compound abundant in cereals, oilseeds, and vegetables, is recognized for its antioxidant and anti-inflammatory properties and has been identified in several traditional medicinal formulations.

Methods:
The present study evaluated the hepatoprotective efficacy of SA in Wistar albino rats exposed to ethanol-induced hepatotoxicity. Liver injury was induced by oral administration of ethanol (40%) for 21 days. SA was administered at doses of 40, 80, and 100 mg/kg p.o., and its protective effects were assessed through biochemical markers (SGOT, SGPT, ALP, total cholesterol, triglycerides, HDL), antioxidant parameters (GSH, MDA, CAT, SOD), proinflammatory cytokines (TNF-α, IL-6), and histopathological examination.

Results:
Ethanol exposure markedly increased ALT (154.2 ± 6.3 IU/L), AST (178.5 ± 8.1 IU/L), and ALP (326.4 ± 10.2 IU/L) compared with normal controls (48.3 ± 3.1, 72.6 ± 4.8, and 162.7 ± 7.5 IU/L; p < 0.001). SA at 80 and 100 mg/kg significantly reduced these levels, with ALT at 82.4 ± 5.2 and 76.8 ± 4.9 IU/L and AST at 102.3 ± 6.1 and 96.7 ± 5.4 IU/L. GSH levels increased from 18.3 ± 1.2 to 32.5 ± 1.8 µmol/mg, while MDA decreased from 7.4 ± 0.5 to 3.2 ± 0.3 nmol/mg (p < 0.001). qRT-PCR revealed downregulation of TNF-α and IL-6 by 68% and 61%, respectively. Histological findings confirmed reduced necrosis and inflammation with near-normal hepatic architecture in treated groups.

Conclusion:
Sinapic acid exhibits strong hepatoprotective effects against alcohol-induced liver injury, likely through attenuation of oxidative stress and inflammatory pathways. These findings support its potential as a natural therapeutic candidate for managing ALD.

Lichens represent a promising source of novel molecules with diverse biological activities. This study aims to evaluate the antioxidant and antibacterial properties of extracts from the lichen Pseudevernia furfuracea to assess its potential for valorization in the pharmaceutical and food industries. Following a preliminary morphological characterization of the thallus, extraction was performed via maceration using three solvents: methanol, ethyl acetate, and dichloromethane. Antioxidant activity was quantified through three distinct methods (DPPH, CAT, and FRAP assays), while antibacterial efficacy was tested against four ATCC-referenced bacterial strains (Enterobacter cloacae ATCC 13047; Klebsiella pneumoniae ATCC 700603; Staphylococcus aureus ATCC 25923 and Staphylococcus aureus ATCC 43300 ) using the agar diffusion technique. The results demonstrated significant antioxidant activity of the ethyl acetate extract (IC50=71,72 µg/ml of DPPH assay and 66,66 mg EAA/g extract of cat assay), whereas the methanolic and dichloromethane extracts exhibited moderate activity. The results of the antibacterial activity assessment revealed that all tested strains exhibited susceptibility to the extracts. Notably, the ethyl acetate extract was found to be the most effective, demonstrating significantly greater efficacy than the other solvents used. The pronounced efficacy of the ethyl acetate extract is attributed to its richness in bioactive secondary metabolites, underscoring its potential for applications in medicinal and cosmetic formulations.

Colibacillosis remains a major threat to the poultry industry of Pakistan, leading to significant economic losses. The disease is caused by avian pathogenic Escherichia coli (APEC), which induces multiple complications leading to prolonged and irrational antibiotic use. This is resulting in the emergence of multidrug-resistant APEC strains, highlighting the urgent need for alternative control strategies. Medicinal plants and their endophytic microbiota represent promising sources of bioactive compounds with antimicrobial potential, as they have co-evolved with their host plants and often mimic or enhance host defense mechanisms. In this study, bacterial endophytes were isolated from Emblica officinalis (Amla), a medicinal plant widely used in traditional medicine, and their extracts were tested for antibacterial activity against pre-confirmed APEC strains using the agar well diffusion method. To provide a comparative analysis, fruit extracts of E. officinalis were also evaluated for antibacterial activity under the same conditions. The endophytic extracts exhibited significantly stronger antibacterial activity (28 mm inhibition zone) than the fruit extracts (16 mm inhibition zone), indicating the superior potency of endophyte-derived metabolites. Partial purification via thin layer chromatography (TLC) confirmed distinct active components responsible for this activity. These findings suggest E. officinalis endophytes as valuable candidates for alternative, sustainable approaches against antibiotic-resistant poultry pathogens.

Introduction: Antibiotic resistance remains a key medical challenge, highlighting the need for alternative antimicrobial agents. Antimicrobial peptides are promising candidates due to their broad activity and high selectivity. Histatins produced by salivary gland cells are notable for their fungicidal, bacteriostatic, wound-healing, and immunomodulatory properties. Recombinant production of histatins may offer a more feasible approach than chemical synthesis. This study aimed to develop a bacterial expression system for histatin 5 (Hst5), including cassette constructs designed to enhance expression.

Methods: Plasmids pH5 and p10xH5 were constructed using the pET30(a) vector. Cloning was performed in E. coli XL1-Blue, followed by plasmid purification and restriction analysis. Peptide expression was performed in E. coli BL21(DE3) with IPTG induction. Peptides were detected by PAGE and purified using SP-Sepharose ion-exchange chromatography. Cassette peptide was hydrolyzed with cyanogen bromide to release Hst5 monomers. Peptide concentration was determined by the Bradford assay. Antimicrobial activity was assessed against E. coli using agar diffusion assay.

Results: Both vectors encoding the monomeric and cassette Hst5 were obtained. Optimal induction conditions were established (200 μM IPTG, 90 min), and Hst5 was eluted at 170–290 mM NaCl. Hydrolysis of the cassette form resulted in Hst5 monomers (280.7 μg/mL). The recombinant peptide displayed pronounced antimicrobial activity comparable to ampicillin.

Conclusions: The results demonstrate the feasibility of producing biologically active Hst5 in a bacterial expression system. This approach offers a cost-effective strategy for generating histatins, supporting the development of novel therapeutics against resistant pathogens. This work was supported by the Ministry of Science and Higher Education of the Russian Federation in 2025 (Agreement No. 075-15-2025-464).

Clitoria ternatea L. (CT) is a perennial herbaceous plant with deep blue flowers native to tropical Asia. Present study aims at characterization of phytochemicals from crude methanolic extract of CT flower and evaluation of antidiabetic and antidepressant effects on 15% fructose induced diabetic mice. GC-MS was employed for identification of chemical constituents from CT flower. Molecular docking and in-silico ADMET analysis were performed to validate the results. Thirty-one phytochemicals were identified. In diabetic mice, metformin reduced glucose from 13.82±0.69 to 11.46±1.03 mmol/L after 1 week and 7.68±0.40 mmol/L after 2 weeks. CT extract at 200 and 400 mg lowered glucose to <10 and <6.8 mmol/L after the 1^st and 2^nd week, respectively. In behavioral test, within 5 minutes, average head-dipping increased from 5.7±1.21 to 11.5±1.64 and for metformin, while it increased to 11.5±1.64 and 13.8±1.83 for 200 mg and 400 mg after 1^st week. All three groups reached ~21 dippings after 2^nd week. In Tail Suspension Test (TST), immobility time 20.9±1.59 decreased to 13.6±1.12 seconds, 14.8±1.45 seconds, and 8.3±0.59 seconds after 1^st week, and to 8.1±1.15 seconds, 8.6±0.79 seconds, and 6.8±1.01 seconds after 2^nd week for metformin, 200 mg and 400 mg, respectively. In Elevated Maze Test (EMT), diabetic mice exhibited 5.8±1.33 exploratory movements within 5 minutes, increasing to 14±1.26, 15±1.26, and 19.3±1.86 after 1 week, and 14.5±0.55, 19.5±2.07, and 23.5±2.26 after 2 weeks for metformin, 200 mg and 400 mg, respectively. Molecular docking studies against five anti-diabetic and five anti-depressant targets further validated the findings, and Stigmasterol and γ-Sitosterol exhibited best results (-7.0 to -10.5 kcal/mol) which also complied with good ADMET profile. Our study connects the outcomes of in-vivo and in-vitro anti-diabetic and anti-depressant activities to the molecular mechanisms involved in treating diabetes and depression by the drug-like constituents present in CT flower.