Background: Periodontitis is an infectious disease caused by microorganisms naturally present in the oral cavity that, under certain conditions, can disrupt oral homeostasis and trigger inflammatory responses that lead to tissue damage. Periodontal inflammation may release inflammatory mediators into the systemic circulation and activate immune cells. Among these cells, monocytes and macrophages play important roles in tissue homeostasis, inflammation, and repair during the development of periodontal disease. Objective: This study evaluated the effects of sera from patients with stage III or IV periodontitis on THP-1 cell activation in vitro. Methods: The human monocytic cell line THP-1 was stimulated for 24 h with 10% serum from 10 patients with periodontitis (stage III, n = 7; stage IV, n = 3) or 10 healthy controls. THP-1 cell adhesion to plastic, an indicator of cellular activation, was assessed by spectrofluorometry, and surface CD69 expression was assessed by flow cytometry. LPS was used as a positive control in both assays. Results: Sera from patients with stage IV periodontitis significantly increased THP-1 cell adhesion to plastic compared with sera from healthy controls, whereas sera from patients with stage III periodontitis did not. CD69 expression was higher in THP-1 cells treated with sera from patients with stage III or IV periodontitis than in cells treated with sera from healthy controls. Conclusion: Sera from patients with advanced periodontitis induced THP-1 cell activation in vitro. The response differed by outcome: increased adhesion was observed with sera from patients with stage IV periodontitis, whereas increased CD69 expression was observed with sera from patients with stage III or IV periodontitis. These findings suggest that circulating factors associated with advanced periodontitis may contribute to monocyte activation; further studies are needed to identify the responsible serum components and determine their clinical relevance.
BACKGROUND: A rapid and cost-effective HPLC-UV method was developed and validated for the quantification of acetaminophen in human plasma. OBJECTIVE: To develop and validate a simple and rapid assay for acetaminophen quantification in human plasma, aimed at strengthening clinical monitoring, particularly in overdose scenarios. METHODS: The method utilizes isocratic elution with ultraviolet detection and a simple one-step protein precipitation using acetonitrile. This approach yields a short run time of approximately 8 minutes and requires a minimal injection volume of 20 μL. The assay provides a broad dynamic range (2–250 μg/mL), covering both therapeutic and toxic concentrations. RESULTS: The calibration curve exhibited excellent linearity (R² = 0.999), and method validation demonstrated acceptable precision (CV ≤ 15% for quality control samples and ≤ 20% at the lower limit of quantification) and accuracy (within ±15%, or ±20% at the LLOQ). Its clinical applicability was confirmed by successfully quantifying acetaminophen in 55 plasma samples from healthy volunteers receiving a therapeutic oral dose of 1000 mg. Although no internal standard was used, the method’s rigorous validation and minimal matrix effects support its omission, in accordance with FDA, EMA, and ICH guidelines. CONCLUSION: This method offers a reliable and accessible alternative to more complex analytical techniques, with significant potential to support patient care in both therapeutic drug monitoring and overdose management. Further validation in overdose settings is required and will be pursued as part of future work.
BACKGROUND: Currently available hemostatic agents have important limitations, including high cost, adverse effects, and limited accessibility. Vernonanthura tweedieana (Baker) H.Rob. is traditionally used in Paraguayan folk medicine as a topical hemostatic agent; however, its hemostatic activity has not been experimentally validated. OBJECTIVE: To evaluate the hemostatic activity, preliminary phytochemical composition, and safety of the ethanolic leaf extract of V. tweedieana (EEVt) in a murine model. METHODS: Acute oral toxicity was evaluated in accordance with OECD Guideline 423. Mice were orally treated with EEVt (50, 100, or 200 mg/kg) for 14 days. Hemostatic activity was assessed by measuring bleeding time, coagulation time, platelet count, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen levels, erythrocyte morphology, and body weight. A preliminary phytochemical screening of the extract was also performed. RESULTS: Oral administration of V. tweedieana extract in mice promoted primary hemostasis by reducing bleeding time by 48–56% and coagulation time by 29–36% compared with the control group. In contrast, no significant effects were observed on platelet count, prothrombin time, activated partial thromboplastin time, fibrinogen levels, erythrocyte morphology, or body weight, suggesting no detectable effect on secondary hemostasis. EEVt exhibited low acute oral toxicity and produced no behavioral alterations or changes in body weight. Preliminary phytochemical screening revealed the presence of steroids/triterpenoids and terpene lactones. CONCLUSIONS: This study provides the first experimental evidence supporting the traditional hemostatic use of V. tweedieana. Oral administration of EEVt to mice promoted primary hemostasis without affecting coagulation parameters associated with secondary hemostasis. Although preliminary phytochemical analysis identified steroids/triterpenoids and terpene lactones, further bioassay-guided fractionation and mechanistic studies are required to identify the active(s) constituent(s) and elucidate the mechanism underlying its hemostatic activity.
Background: Amazonian fruits stand out as promising resources. Beyond their nutritional properties, which classify them as superfoods, they contain bioactive compounds with significant added-value potential for various industries. However, these biocompounds should be extracted using sustainable and user-friendly alternatives for local communities. Objectives: A proposed extraction approach combining simple methods such as maceration with advanced user-friendly techniques like ultrasound and centrifugation—tested under different fruit-to-solvent and temperature–time conditions for Colombian Amazonian fruits—enables the analysis and selection of an useful alternative for the bioactive compound recovery. Methods: Extracts of annatto (Bixa orellana), Amazonian chili (Capsicum chinense var. A. charapita), and açaí (Euterpe oleracea) from the Colombian Amazon region were obtained through a process combining ultrasound, maceration, and centrifugation. For each fruit, three fruit-to-solvent ratios (0.01, 0.05, and 0.1 g mL⁻¹) and two extraction temperature–time conditions were tested, resulting in a total of 54 trials. The parameters measured included extraction efficiency, antioxidant activity, total phenols, total flavonoids, and UV-Vis spectral scanning. Results: The highest extraction efficiency (19.34 ± 2.02 g extract 100 g⁻¹ dry fruit) was obtained from açaí at a fruit-to-solvent ratio of 0.01 g mL⁻¹. The annatto extract under conditions of 0.01 g mL⁻¹ at 50°C showed promissory antioxidant activity (572.14 ± 17.62 µmol TE g⁻¹ extract DW), total phenols (121.06 ± 3.86 mg GAE g⁻¹ extract DW), and Area Under the Curve index (AUC) (AU · nm) by UV-VIS scanning for flavonoids (250–430 nm) and carotenoids (410–510 nm), at a concentration of 100 µg mL⁻¹ of extract. The outstanding total flavonoid content was recorded for the açaí extract at a ratio of 0.1 g mL⁻¹ (196.08 ± 4.8 mg CE g⁻¹ extract). The relationship between the fruit-to-solvent ratio (x) and the extraction parameters was predominantly negative, showing a high efficiency of extraction at low fruit ratio. Conclusions: Considering consistency across more than three measured parameters and shorter extraction times, the order of optimal configuration for extraction was: annatto0.01-1h > chili0.05-3h > açaí0.01-3h.
Background: Mammea americana L. (“mamey”) is a tropical plant traditionally used in South America for its therapeutic, antimicrobial, and insect-repellent properties. Its ethnopharmacological relevance and reported in vitro antibacterial activity support its potential application in topical pharmaceutical formulations with anti-infective properties. However, additional studies are needed to characterize the physicochemical behavior of its extracts. Objective: This study aimed to evaluate the physicochemical and phytochemical properties of an ethanolic extract obtained from the endosperm of M. americana and assess its suitability for topical antimicrobial formulations. Methods: Two ethanolic extracts (EMa01 and EMa02) were prepared from dried and ground plant material with different particle size distributions determined by laser diffraction analysis to evaluate the effect of granulometry on the extraction process. The extracts were characterized by TLC, HPLC-PDA, and UV-Vis spectroscopy. Solubility studies for EMa01 were conducted according to OECD guidelines using 18 formulation excipients, including solvents of different polarities, essential oils, surfactants, and pH conditions, in order to identify suitable components for topical formulations. In addition, Shinoda and ferric chloride assays and accelerated stability studies were performed. Results: EMa01 showed a higher extraction yield (7.75%) than EMa02 (4.83%), indicating that particle size influenced extraction efficiency, although TLC profiles revealed similar chemical compositions. HPLC analysis confirmed the presence of the coumarins mammea B/BA and mammea B/BC, compounds previously associated with antibacterial activity, which were selected as markers of extract quality and biological activity. Flavonoids and tannins were also detected in the extract. Among the evaluated excipients, Span 80 and rosemary oil exhibited the best solubility and compatibility with EMa01. Conclusions: These findings demonstrate that particle size affects extraction yield but not the chromatographic profile of the extract. Although the antibacterial activity of the extract was not evaluated in the present study, the detection of the coumarin mammea B/BA may serve as an indicator of potential pharmacological activity. The preformulation studies provide preliminary information supporting the future development of topical antimicrobial or insect-repellent formulations based on M. americana extract.
