Objectives: The aim of this work was to determine the phenolic composition of sweet wormwood (Artemisia annua L., Asteraceae) from controlled cultivation in Serbia and to assess the potential antioxidant effects and cytotoxicity. Methods: High-performance liquid chromatography was used to determine the phenolic composition of Artemisia annua ethanolic extract. The antioxidant activity was studied using in vitro tests of inhibition of the neutralization of 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (OH), and nitroso (NO) radicals, as well as the process of inhibiting lipid peroxidation and the ferric reducing antioxidant power (FRAP). The cytotoxicity was evaluated by the effect on three cell lines (the rat pancreatic insulinoma cell line (Rin-5F), the rat hepatoma cell line (H4IIE), and human hepatocellular carcinoma (Hep G2)) using the MTT test of viability. Results: Ethanol extract showed the highest potency in inhibiting the DPPH radical, and the half maximal inhibitory concentration (IC₅₀) was 5.17 μg/mL. Chlorogenic acid was the dominant phenolic compound with an amount of 651 μg/g of dry extract. The results of the MTT viability test showed that the extract has the potential to inhibit the growth of the Rin-5F and Hep G2 cell lines, while no growth inhibition was observed on the H4IIE cell line. Conclusions: Undoubtedly, Artemisia annua is a powerful plant and a rich source of phenolic compounds. Inhibitory activity on causes of oxidative stress shows that the plant has a good antioxidant effect. Also, the anticancer activity shown through the inhibition of cell growth is not negligible.

Background/Objectives: In previous studies, chloroquine and ivermectin separately exhibited similar anticancer effects on various known cancer modulatory targets. This study aimed (1) to identify a non-toxic synergistic combination of chloroquine and ivermectin that suppresses hamster fibrosarcoma; (2) to verify combined antitumor efficacy using dose–response analysis; and (3) to investigate potential synergistic mechanisms by restoring tumor progression with the reciprocal cancer-modulating agent deoxycholic acid. Methods: A BHK-21/C13 cell culture was subcutaneously inoculated into Syrian golden hamsters randomly divided into groups (6 animals per group): (1) untreated control; treated daily (17 days after inoculation) with (2) chloroquine 50 mg/kg; (3) ivermectin 5 mg/kg; (4) a combination of chloroquine 50 mg/kg and ivermectin 5 mg/kg; (5) a combination of chloroquine 50 mg/kg, ivermectin 5 mg/kg and deoxycholic acid 100 mg/kg; (6) a combination of chloroquine 25 mg/kg and ivermectin 2.5 mg/kg; (7) a combination of chloroquine 12.5 mg/kg and ivermectin 1.25 mg/kg. Dose–response curves were generated for chloroquine and ivermectin combinations. Characteristics of tumors (growth kinetics, biophysical, histological, immunohistochemical, pathological), hamster organs, biochemical and hematological blood tests were compared among the groups. Results: The synergistic, dose-dependent anticancer effects of two antiparasitic agents, similar tumor-regulation modulators chloroquine and ivermectin, in doses equivalent to human doses were observed in fibrosarcoma in hamsters (both drugs approximately 1/10 LD₅₀) without toxicity and in various cell lines of human lung, colon and cervical carcinomas and hamster fibrosarcoma in vitro. The addition of a reciprocal modulator of cancer regulation, NF-κB stimulator deoxycholic acid, caused a huge rescue effect on fibrosarcoma and a reversal of the successful anticancer therapy using the combination. Conclusions: The chloroquine and ivermectin combination may be recommended for comprehensive additional preclinical and clinical evaluation due to its synergistic anticancer effects. Further preclinical and clinical exploration will be crucial to thoroughly define the optimal role of the combination therapy in the treatment of fibrosarcoma and potentially other cancer types.

Volatile organic compounds (VOCs) emission from trees and other plants depends on various factors such as species, age, and biochemical characteristics, as well as the interaction with fungi and insects. VOCs derive from the tree native structure and/or are produced via oxidation and hydrolysis from wood components. The aim of this study was to determine VOCs in Pinus nigra Arnold bark extracts. Pinus nigra Arnold bark was sampled from the mountains Tara and Mokra Gora (Republic of Serbia). Dichloromethane and hexane were used as pure solvents and conventional liquid–liquid extraction (LLE) and ultrasound extraction were applied. Prepared extracts were analysed by gas chromatography coupled with a mass spectrometry detector. The NIST and Wiley libraries were used for the identification of VOCs. After analysis of the VOCs’ abundance and peak areas, dichloromethane was chosen as a more efficient solvent in comparison with hexane. The ultrasound technique was more effective for VOC extraction versus the conventional LLE. Additionally, bark extracts from Mokra Gora contained diverse VOCs in contrast to the ones obtained from the Tara locality. The main VOCs were monoterpenes, such as α- and β-pinene, camphene and limonene. In addition, sesquiterpenes: humulene, germacrene D, longipinene, longifolene and cadinene, as well as oxygenated terpenes such as verbenone and α-terpineol, were detected in bark extracts. The obtained results showed that location strongly affects the VOC profile in Pinus nigra.

Hyperlipidaemia is a common worldwide problem associated with an increased risk of coronary and extra-coronary atherosclerosis and cardiovascular disease. Besides changes in lifestyle habits that include controlling the diet with moderate alcohol consumption and maintaining normal weight, medication is usually prescribed in addition. The antioxidative potential of functional food in the treatment of hyperlipidaemia continuously attracts growing attention. Paraoxonase enzyme (PON1) prevents the oxidation of low- and high-density lipoprotein (LDL and HDL) and, hence, has an important role in acting against lipid peroxides. The aim of this study was to evaluate Spirulina platensis’s influence on blood and hepatic PON1 activity in an animal model. Male Wistar rats (approved by the Institutional Bioethics committee No. III-2011-01) were randomly divided into five groups based on the applied diet (I—normal diet; II—normal diet with spirulina; III—lipogenic diet; IV—lipogenic diet with concomitant spirulina supplementation; and V—lipogenic diet with spirulina treatment). The differences in PON1 activity were related to diet type. A lipogenic diet rich in saturated fats impaired the PON1 activity. Both blood and hepatic PON1 activity were significantly increased after the administration of a normal diet with spirulina supplementation. As expected, significantly reduced blood PON1 activity was measured in the lipogenic diet group. Blood PON1 activity was decreased in groups III, IV, and V, but PON1 in both the blood and liver had a tendency to increase in groups IV and V. Based on the obtained results, PON1 activity is affected by hyperlipidaemia, and spirulina supplementation may promote enzyme activity.

Since ancient times, many plant species within the Thymus genus have been used due to their numerous health benefits, such as antimicrobial, anti-inflammatory, antiseptic, or diuretic activity. While many of the species within this genus were well known and described, Thymus pannonicus All. or Pannonian thyme remains relatively unexplored despite its unique chemical composition and activity. T. pannonicus is a small shrub, spread over central and eastern Europe, with a diverse, location-dependent chemical composition. Within this study, the citral chemotype of T. pannonicus was used, which up to our knowledge was found and described only in Northern Serbia. Therefore, in order to explore the composition of T. pannonicus low-polar extracts and to present a preliminary insight into their bioactivity, supercritical carbon dioxide (ScCO₂) extraction was applied (with and without pre-treatments), followed by GC-MS analysis. Different ScCO₂ extraction pressures were tested (100, 150, 200, 250, and 300 bar) in order to determine the most suitable conditions for the extraction of T. pannonicus’s low-polar fraction. Additionally, in order to maximize the yield of the extraction, two types of ScCO₂ extraction pre-treatments were investigated (enzymatic and microwave). The highest extraction yield obtained from untreated plant material was 3.01% (w/w), and it was obtained at conditions of 150 bar and 40 °C, while the 4% (v/w) enzymatic pre-treatment, with the same conditions, provided a yield of 3.89%. For all of the obtained extracts, the GC-MS analysis showed that oxygenated monoterpenes and sesquiterpenes were the two most dominant groups with principal bioactive compounds such as (E)-citral (18.95–38.17%), (Z)-citral (6.68–14.66%), β-bisabolene (8.2–14.4%), and nerol (6.08–9.67%). The extracts that exhibited the highest concentration of principal bioactive compounds ((Z)-citral, and (E)-citral) were further analyzed for anticancer potential, using short- and long-term cell viability observations on liver cancer cells.