The peel of Citrus hystrix DC. (kaffir lime), an underexploited agro-industrial by-product, may have bioactives. This work tested the ethanolic extract of C. hystrix peel's (EeChP) phytochemicals by GC-MS profiling, and the antidiabetic effect was evaluated using the alpha-amylase inhibitory test, and the anti-inflammatory activity was done using the human red blood cell (HRBC) membrane model. The antibacterial effect was determined using the agar well diffusion method against Staphylococcus aureus and Escherichia coli. Major constituents subjected to pharmacokinetic and ADMET filtering, molecular docking, Gene Ontology (GO), and KEGG pathway analysis for mechanistic plausibility. The GC-MS detected 44 phytoconstituents, including limonene, γ-sitosterol, and β-amyrin. EeChP inhibited α-amylase (IC50 = 80.97 µg/mL) and the stabilized HRBC membrane (IC50 = 64 µg/mL). The extract demonstrated substantial antibacterial action against S. aureus (IZD = 6 mm at 200 µg/well) but failed against E. coli. The β-amyrin and scoparone were found to be non-toxic. Scoparone and carbonic anhydrase 12 were connected with β-amyrin and protein tyrosine phosphatase non-receptor type 1. EeChP has anti-diabetic, multi-target, modulatory, and anti-inflammatory effects. The results recommend C. hystrix peel's bioresource potential, but enzyme kinetics, antimicrobial MIC, and in vivo research are necessary. The research has contributed to accomplishing SDG 3 and SDG 12.

Amoebiasis, caused by Entamoeba histolytica, remains a global health concern due to its asymptomatic nature and potential for severe complications such as liver abscesses. To investigate molecular recognition mechanisms, this study focused on the binding affinity and detection limit of aptamers targeting E. histolytica actin (EhActin), a cytoskeletal protein selected as a target molecule. A genomic tRNA-based sequence library was computationally screened to identify potential high-affinity aptamer candidates, followed by molecular docking and molecular dynamics simulations to evaluate interaction profiles. Recombinant EhActin (rEhActin) was expressed in Escherichia coli LEMO21 using a pET28a(+) vector system and used in ELONA to quantify the interaction between a biotinylated DNA aptamer and rEhActin. The limit of detection was also assessed using ELONA with E. histolytica protein extract. Among the shortlisted aptamers, APT29 demonstrated favorable in silico and in vitro binding characteristics. Molecular dynamics analysis supported overall structural stability of the APT29-EhActin complex, although hydrogen bond interactions fluctuated over time. ELONA showed a dissociation constant (Kd) of 44.72 nM for APT29, and this aptamer achieved a limit of detection of 0.4 µg/well of E. histolytica protein extract. These findings suggest APT29 as a promising molecular probe for further biophysical and structural studies of aptamer-target interactions involving E. histolytica, with potential relevance to future diagnostic development for amoebiasis.

Increasing incidences of acne and skin infections, antibiotic resistance, and side effects associated with synthetic dermatology have necessitated the need for safe, sustainable treatment approaches. This work describes the environmentally friendly preparation of Zinc Oxide Nanoparticles (ZnO NPs) through green chemistry by utilizing the leaf extract of Acalypha indica L. plant and further applying the nanoparticles in the preparation of nanogel, herbal soap, and face serum. The biosynthesized zinc oxide nanoparticles were analysed using UV-Visible spectroscopy, Fourier Transform Infra-red (FTIR), X-Ray diffraction (XRD), and Scanning electron microscopy (SEM). From UV-Visible spectra, an excitonic absorption band peak was observed at 384 nm, which confirms the presence of nanoparticles. FTIR spectra show a vibrational peak of Zn-O bonds at 476 cm⁻¹ and 512 cm⁻¹ as well as surface-bonded phenolic, aliphatic, and carbonyl functional groups due to flavonoids, terpenoids, and tannins. XRD confirmed a highly crystalline hexagonal wurtzite structure (JCPDS No. 36-1451) with an average crystallite size of 18.5 ± 3.2 nm. SEM revealed polydisperse, moderately agglomerated nanoparticles with irregular, porous morphology. ZnO NPs displayed strong bactericidal action against nine clinical strains of bacteria, with the highest zone of inhibition observed as 25 mm against Staphylococcus aureus and Salmonella typhi at 75 µg/well concentration. These particles were also found to possess excellent radical scavenging ability, IC₅₀ being recorded as 62.4 ± 2.1 µg/mL for DPPH free radical. The pH of all the three preparations was compatible with the skin, viz., 5-6. Also, these formulations were physically stable and had good spreadability, i.e., nanogel preparation - 28 mm; soap preparation - 47 mm; serum preparation - 21 mm at 150 g. The herbal soap proved ideal for daily hygiene, while the serum enabled rapid spot treatment. This study establishes that Acalypha indica-mediated green synthesis produces stable, crystalline, multifunctional ZnO nanoparticles with substantial antimicrobial and antioxidant properties. The nanogel formulation represents a scalable, cost-effective, natural-origin alternative for acne and skin infection management, addressing antibiotic resistance and synthetic chemical toxicity.

Avapritinib (AVA) is a breakthrough targeted therapy as the first potent inhibitor approved for gastrointestinal stromal tumors (GIST) with PDGFRA D842V mutations. It lacks convenient analytical methods for routine quality control. This study addresses this gap by developing and validating two novel, green, high-throughput microwell spectrophotometric methods (MW-SPMs) for AVA quantification in pharmaceutical tablets-representing the first employment of charge-transfer complexation for AVA analysis. The methods exploit rapid charge-transfer complex (CTC) formation between AVA (electron donor) and two π-acceptors: 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) and chloranilic acid (CLA). The reactions, conducted in 96-well plates using only 200 µL total volume, yielded stable colored complexes with absorption maxima at 460 nm and 520 nm for the reactions with DDQ and CLA, respectively. Following ICH-compliant optimization and validation, both methods demonstrate excellent linearity (3.13-100 µg/well for DDQ; 6.25-100 µg/well for CLA), precision (RSD ≤ 1.8%), and accuracy (recoveries: 98.0-101.7%). Job's method confirmed 1:1 stoichiometry, while density functional theory (DFT) uniquely revealed the molecular basis for the stronger binding affinity of DDQ over CLA through dominant π-π stacking and hydrogen-bonding interactions, demonstrating how computational analysis can guide reagent selection in analytical method development. A comprehensive multi-metric sustainability assessment-utilizing ten tools including AES, AGREE, BAGI, RGB, and RAPI-confirmed the methods' excellent greenness (e.g., AES score = 90), superior practicality, and robust analytical performance, achieving a high White Index of 94.2%. With a substantially higher throughput of ~ 500 samples/hour and minimal solvent consumption (200µL/well), the proposed MW-SPMs offers a rapid, sustainable, and cost-effective alternative to conventional chromatographic and spectrofluorimetric techniques for routine pharmaceutical analysis, aligning with multiple United Nations Sustainable Development Goals.

The search for high-efficiency antimicrobial nanomaterials has intensified due to the escalating threat of antibiotic resistance. In this study, titanium dioxide nanoparticles (TiO2 NPs) were synthesized using aqueous leaf extract of Pterolobium hexapetalum. Physicochemical characterization confirmed the formation of a pure nanocrystalline anatase phase (XRD) with a primary crystallite size of 10-15 nm and an optical band gap of 3.18 eV. While UV-Visible spectroscopy showed characteristic absorption bands at 209.5 and 270.5 nm, DLS and Zeta potential (+ 9 mV) measurements indicated that the nanoparticles exist as aggregated colloidal clusters with a hydrodynamic diameter of 399.2 nm. FTIR and SEM verified the presence of plant-derived functional groups and a clustered, rough surface morphology. Biological evaluation via agar well diffusion, MIC, and MBC assays demonstrated that the TiO2 NPs significantly outperformed the parent leaf extract. At a dose of 100 µg/well, the TiO2 NPs produced inhibition zones of 22.67 mm for Staphylococcus aureus and 27.67 mm for Escherichia coli, surpassing the absolute zones produced by the ciprofloxacin (5 µg/well) control. The MIC values (31.25-62.5 µg/mL) and an MBC/MIC ratio of 2 indicate a potent, primarily bactericidal mode of action, achieving a two- to four-fold increase in potency over the crude extract. These findings establish P. hexapetalum-mediated TiO2 NPs as a promising platform for antimicrobial applications in wound care and decontamination technologies.

Palmyra palm sprout skin was extracted using ethanol to test its antibacterial activity against isolated urinary tract infection (UTI) pathogenic bacteria. The extract demonstrated excellent inhibitory activity at the concentration of 500 µl/well. It was observed that all the isolated UTI-causing bacteria were highly resistant to selected commercially available antibiotics. The ethanol extract of the palmyra palm sprout skin showed excellent antioxidant, anti-inflammatory, antidiabetic, and antiproliferative activities. Before boiling the Panam kilangu, the sprout skin is usually peeled from it and thrown away as it is considered of no use. However, our study demonstrates a waste-to-wealth based alternative use for the palmyra palm sprout skin. Based on studies, these by-products could be used as a source of valuable bioactive compounds. It showed excellent inhibitory activity at the concentration of 500 µg/well against UTI bacteria. It also showed to possess various secondary metabolites, such as flavonoids, saponins, etc. Gas chromatography-mass spectrometer analysis revealed the presence of various bioactive metabolites.

Engineered stone (ES) fabrication generates respirable dust containing crystalline silica (CS), linked to accelerated silicosis outbreaks. Mechanisms underlying this toxicity, particularly the role of particle aging, remain unclear. In the occupational setting, workers are exposed to engineered stone dust (ESD) upon generation by cutting and grinding ES; however, ESD-initiated toxicity is frequently studied in labs using aged particles. This study aimed to compare radical generation and in vitro cytotoxicity of fresh versus aged ESD. Three different respirable ES types (ES A: 60% CS; B: 20%; C: 0%), granite (30%), and Min-u-Sil 5 (MS5, 99.5%) were generated using an automated cutting system and analyzed either freshly stored under N2 at -80°C or after aging in air at room temperature for 2 weeks. RAW 264.7 macrophages were exposed to particles (10 µg/well, 100 µg/ml, 31.25 µg/cm[2], 24 hr), and viability, apoptosis, necrosis, and intracellular reactive oxygen species (ROS) were measured. Fresh ESD/granite exhibited significantly higher electron paramagnetic resonance (EPR) radical signals than aged counterparts and MS5. Fresh ES/granite reduced macrophage viability, while aged materials/MS5 did not. Apoptosis increased with all particles where fresh/aged difference occurred only in ES B. Necrosis rose markedly with fresh ES A. Intracellular ROS was elevated by some materials, but N-acetylcysteine (NAC) antioxidant failed to prevent cytotoxicity induced by fresh particles. In conclusion, freshly generated ESD displayed greater radical-generating capacity and distinct cytotoxic effects compared to aged ESD, influenced by factors beyond CS content. ROS-independent mechanisms appear crucial for acute cytotoxicity. These findings indicate particle aging as a critical factor in ESD toxicological assessment.

This study explores the green synthesis of silver nanoparticles (AgNPs) using Cymbopogon citratus (lemongrass) extract as a reducing agent. Synthesis was confirmed by a color change (light yellow to dark brown) under optimal conditions: 1.50 mM silver nitrate, 3.5% v/v extract, at 100 °C, with a pH of 9, and for 60 min. The AgNPs exhibited spherical morphology, a hydrodynamic diameter of 135.41 ± 49.30 nm, a zeta potential of -29.9 ± 1.4 mV, crystalline structure, and minimal aggregation. AgNPs showed significant antibacterial activity, particularly at >20 µg/well, with the zones of inhibition varying by bacterial strain. In vitro studies demonstrated anti-inflammatory, antidiabetic (α-glucosidase and α-amylase inhibition), and antioxidant activities, with AgNPs outperforming plant extract and nearing standard efficacy at higher concentrations. Cyto-toxicity studies indicated that AgNPs and plant extract were less toxic than doxorubicin but exhibited concentration-dependent effects on cancerous and non-cancerous cells. Eco-toxicity assays revealed that AgNPs were less acutely toxic than controls but posed risks with prolonged exposure. This work highlights the eco-friendly synthesis of AgNPs and their potential in biomedical applications, demonstrating efficacy in antibacterial and antioxidant activities.

For paediatric patients suffering from neurofibromatosis, Selumetinib (SEL) is the only approved drug. Here an original ecofriendly and high pace method is introduced using 96- microwell spectrophotometric assay (MW-SPA) to measure SEL content in bulk and commercial pharmaceutical formulation (Koselugo[®] capsules). This assay was relied on in-microwell formation of a coloured charge transfer complex (CTC) upon interaction of SEL with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ). The complex was fully characterized by spectrophotometric and computational studies. The CTC exhibited an absorbance maximum (λmax) at 440 nm. The ease of reaction occurrence, complex stability and its high absorptivity were proved by measuring its association constant (0.63 × 10[2] L/ mol), standard free energy change (-10.31 KJ/mol), molar absorptivity (ε) (3.78 × 10[3] L/mol/cm), and the SEL: DDQ stoichiometric ratio (1:1). Establishments of the optimum values of the applied conditions in 96-well assay plate were refined regarding DDQ concentration, reaction time, temperature, and solvents. Validation of the assay was according to the ICH guidelines. The assay was linear in SEL' concentrations ranged from 10 to 200 µg/well, with limits of detection and quantitation of 4.1 and 12.5 µg/well, respectively. Then, the assay was efficaciously adapted to accurately and precisely determine SEL content in bulk form and Koselugo[®] capsules. The assay environmental safety was documented by three different comprehensive metric tools. Additionally, assessment of the assay's rate demonstrated its high throughput, enabling the processing of large number of samples in pharmaceutical quality control laboratories. The successful development of this assay provides a valuable fast and green analytical tool for ensuring the quality control of SEL's bulk form and capsules.

Background and Objectives: Ceritinib (CER) is a potent drug of the third-generation tyrosine kinase inhibitor class. CER has been approved for the treatment of patients with non-small-cell lung cancer (NSCLC) harboring the anaplastic lymphoma kinase (ALK) mutation gene. In the literature, there is no green and high-throughput analytical method for the quantitation of CER in its dosage form (Zykadia[®] capsules). This study describes, for the first time, the development and validation of two novel one-step and green microwell spectrophotometric methods (MW-SPMs) for the high-throughput quantitation of CER in Zykadia[®] capsules. Materials and Methods: These two methods were based on an in microwell formation of colored derivatives upon the reaction of CER with two different benzoquinone reagents via two different mechanisms. These reagents were ortho-benzoquinone (OBQ) and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), and their reactions proceeded via condensation and charge transfer reactions, respectively. The reactions were carried out in 96-well transparent plates, and the absorbances of the colored reaction products were measured with an absorbance microplate reader at 540 and 460 nm for reactions with OBQ and DDQ, respectively. The optimum conditions of reactions were established, their molar ratios were determined, and reaction mechanisms were postulated. Under the refined optimum reaction conditions, procedures of MW-SPMs were established and validated according to the guidelines of the International Council on Harmonization. Results: The limits of quantitation were 6.5 and 10.2 µg/well for methods involving reactions with OBQ and DDQ, respectively. Both methods were applied with great reliability to the determination of CER content in Zykadia[®] capsules and their drug uniformity. Greenness of the MW-SPMs was evaluated using three different metric tools, and the results proved that the two methods fulfil the requirements of green analytical approaches. In addition, the simultaneous handling of a large number of samples with microvolumes in the proposed methods gave them the advantage of a high-throughput analysis. Conclusions: The two methods are valuable tools for rapid routine application in pharmaceutical quality control units for the quantitation of CER.

Airborne micro- and nanoplastics are widely spread and pose a risk to human health. The third polymer plastic most commonly produced and present in atmospheric fallout is polystyrene (PS). For these reasons and for a more realistic assessment of biological effects, we examined in-home oxidised (ox-, simulating photoaging) nPS/mPS (0.1 and 1 μm), comparing the effects with virgin ones (v-). On human alveolar cells (A549), we quantified the cellular uptake, using FITC-functionalised nPS/mPS, while cytotoxicity, changes in the acidic compartment, ROS production, mitochondrial function, and DNA damage were assessed to study the effects of internalised v- and ox-nPS/mPS. The results showed that the uptake was dose-dependent and very fast (1 h), since, at the lowest dose (1.25 µg/well), it was 20.8% and 21.8% of nPS and mPS, respectively. Compared to v-, significant ROS increases, DNA damage, and mitochondrial impairment were observed after exposure to ox-nPS/mPS. The enhancement of effects due to environmental aging processes highlighted the true potential impact on human health of these airborne pollutants.

Royal jelly (RJ) is a multifunctional bee product with a unique composition and wide-ranging biological properties, including antioxidant, anti-inflammatory and antiproliferative activities. Still, little is known about the possible myocardial protective properties of RJ. Considering that sonication could enhance RJ bioactivity, this study aimed to assess the effects of non-sonicated (NS) and sonicated (S) RJ on fibrotic signaling, cell proliferation, and collagen production in cardiac fibroblasts. S-RJ was produced by ultrasonication at 20 kHz. Ventricular fibroblasts isolated from neonatal rats were cultured and treated with different concentrations of NS-RJ or S-RJ (0, 50, 100, 150, 200, and 250 µg/well). S-RJ significantly depressed the expression levels of transglutaminase 2 (TG2) mRNA across all the concentrations tested and was inversely associated with the expression of this profibrotic marker. S-RJ and NS-RJ displayed distinct dose-dependent effects on mRNA expression of several other profibrotic, proliferation, and apoptotic markers. Unlike NS-RJ, S-RJ elicited strong negative dose-dependent relationships with the expression of profibrotic markers (TG2, COL1A1, COL3A1, FN1, CTGF, MMP-2, α-SMA, TGF-β1, CX43, periostin), as well as proliferation (CCND1) and apoptotic (BAX, BAX/BCL-2) markers, indicating that RJ dose-response effects were significantly modified by sonification. NS-RJ and S-RJ increased the content of soluble collagen, while decreasing collagen cross-linking. Collectively, these findings show that S-RJ has a greater range of action than NS-RJ for downregulating the expression of biomarkers associated with cardiac fibrosis. Reduced biomarker expression and collagen cross-linkages upon cardiac fibroblast treatment with specific concentrations of S-RJ or NS-RJ suggests putative roles and mechanisms by which RJ may confer some protection against cardiac fibrosis.

Lorlatinib (LRL) is the first drug of the third generation of anaplastic lymphoma kinase (ALK) inhibitors used a first-line treatment of non-small cell lung cancer (NSCLC). This study describes, for the first time, the investigations for the formation of a charge transfer complex (CTC) between LRL, as electron donor, with chloranilic acid (CLA), as a π-electron acceptor. The CTC was characterized by ultraviolet (UV)-visible spectrophotometry and computational calculations. The UV-visible spectrophotometry ascertained the formation of the CTC in methanol via formation of a new broad absorption band with maximum absorption peak (λmax) at 530 nm. The molar absorptivity (ε) of the complex was 0.55 × 10[3] L mol[-1] cm[-1] and its band gap energy was 2.3465 eV. The stoichiometric ratio of LRL/CLA was found to be 1:2. The association constant of the complex was 0.40 × 10[3] L mol[-1], and its standard free energy was -0.15 × 10[2] J mole[-1]. The computational calculation for the atomic charges of an energy minimized LRL molecule was conducted, the sites of interaction on the LRL molecule were assigned, and the mechanism of the reaction was postulated. The reaction was adopted as a basis for developing a novel 96-microwell spectrophotometric method (MW-SPA) for LRL. The assay limits of detection and quantitation were 2.1 and 6.5 µg/well, respectively. The assay was validated, and all validation parameters were acceptable. The assay was implemented successfully with great precision and accuracy to the determination of LRL in its bulk form and pharmaceutical formulation (tablets). This assay is simple, economic, and more importantly has a high-throughput property. Therefore, the assay can be valuable for routine in quality control laboratories for analysis of LRL's bulk form and pharmaceutical tablets.

Background and Objective: Tyrosine kinase inhibitors (TKIs) are used for the treatment of different types of cancers. The current study describes, for the first time, the ultraviolet-visible spectrophotometric investigation of charge transfer complexes (CTCs) of seven TKIs, as electron donors, and iodine, as σ-electron. Materials and Methods: The formation of CTCs was promoted in dichloromethane, among the other solvents used in the investigation. The molar absorptivity values, association constants, and free energy changes of the CTCs were determined. Stoichiometric ratio of TKI: iodine as well as TKIs site(s) of interaction were addressed. Reaction was the basis for constructing a novel simple and accurate 96-microwell spectrophotometric assay (MW-SPA) with high-throughput property for the quantitative determination of TKIs in their pharmaceutical formulations. Results: Beer's law, which relates CTC absorbances to TKI concentrations, was followed within the optimal range of 2 to 100 µg/well (r ranged from 0.9991 to 0.9998). Detection and quantification limits ranged from 0.91 to 3.60 and 2.76 to 10.92 g µmL[-1], respectively. Relative standard deviations values for the intra- and inter-assay precisions of the proposed MW-SPA did not exceed 2.13 and 2.34%, respectively. Studies of recovery demonstrated MW-SPA accuracy, with results ranging from 98.9% to 102.4%. All TKIs, both in bulk form and in pharmaceutical formulations (tablets), were effectively determined using the suggested MW-SPA. Conclusions: The current MW-SPA involved a simple procedure and it was convenient as it could analyse all proposed TKIs utilizing a single assay system at once measuring wavelengths for all TKIs. In addition, the proposed MW-SPA has high throughput which enables the processing of a batch of huge samples' number in very short reasonable time period. In conclusion, TKIs can be routinely analysed in their dosage forms in quality control laboratories, and the assay can be highly valuable and helpful in this regard.