The alphabet of sea fennel: Comprehensive phytochemical characterisation of Croatian populations of Crithmum maritimum L.

Extreme environmental conditions affect the synthesis and accumulation of bioactive metabolites in halophytic plants. The aim of this study was to investigate the presence and quantity of key health-promoting phytochemicals in Croatian sea fennel, one of the most popular Mediterranean halophytes with a wide range of uses. The EOs were characterised by a high content of limonene (up to 93%), while the fatty acid profile shows a low content of oleic acid and the presence of valuable linoleic acid (ω-6) and linolenic acid (ω-3) in high percentages. The dominances of lutein and α-tocopherol were also confirmed in all samples. The results confirm the great variability in the chemistry of sea fennel populations in the Mediterranean region, with significant differences in the composition of the Croatian samples compared to the others, as well as the presence and high concentrations of the analysed bioactive compounds that contribute to the plant's health-promoting attributes.

Spontaneous fermentation of Marastina wines: The correlation between autochthonous mycobiota and phenolic compounds.

Understanding fungal community dynamics during fermentation is important for assessing their influence on wine's phenolic content. The present study represents the first effort to explore the correlation between the autochthonous mycobiota of Marastina grapes collected from Dalmatian winegrowing sub-regions in Croatia and the phenolic composition, as well as the physicochemical parameters of wines produced through spontaneous fermentation. The metataxonomic approach revealed Metschnikowia pulcherrima, Metschnikowia fructicola and Hanseniaspora uvarum as the core mycobiota detected at the initial phase of fermentation. By contrast, Saccharomyces cerevisiae took over the dominance starting from the middle stage of fermentation. The wine's phenolic compounds were revealed by high-performance liquid chromatography, with tyrosol being the most abundant. Rhodotorula babjevae and Botrytis cinerea showed a positive correlation with p-hydroxybenzoic acid, gentisic acid, caffeic acid and cinnamic acid, while demonstrating a negative correlation with protocatechuic acid and chlorogenic acid. Heterophoma novae-verbascicola exhibited the opposite behaviour regarding the same phenolic compounds. The concentration of lactic acid was positively correlated with B. cinerea and negatively correlated with Het. novae-verbascicola. These findings serve as a foundation for in-depth investigations into the role of autochthonous grape mycobiota in phenolic transformation during spontaneous fermentation, potentially leading to the production of high-quality wines with unique terroir characteristics. Future studies should aim to explore the specific role played by individual yeast isolates in the formation of phenolic compounds.

Conventional vs. Microwave-Assisted Hydrodistillation: Influence on the Chemistry of Sea Fennel Essential Oil and Its By-Products.

The main objectives of this study were to investigate the effects of the applied essential oil (EO) isolation method, conventional hydro-distillation (HD), and microwave-assisted hydro-distillation (MHD) on the chemical profile of sea fennel (Crithmum maritimum L.) essential oil and to investigate the main constituents present in the liquid by-products of EOs isolation (hydrolate and residual wastewater). Headspace-solid phase microextraction (HS-SPME) was used to isolate hydrolate components, while gas chromatography coupled with mass spectrometry (GC-MS) was used to detect and analyse the chemical constituents of the essential oils and hydrolates. The phenolic composition of the wastewater extracts was analysed by high performance liquid chromatography (HPLC). The EO obtained by MHD had a higher yield of limonene and sabinene. The chemical composition of the hydrolates differed from the EO compositions. The content of terpinen-4-ol in the MHD hydrolate was higher, while several compounds were detected in relatively high proportions only in the HD hydrolate. MHD also resulted in a higher phenolic content of the wastewater, where an increase in the concentration of chlorogenic acid was also observed. It can be concluded that the isolation method had a great influence on the profile of sea fennel EOs, especially on their corresponding hydrolates and residual wastewater extracts. Due to their valuable chemical composition, these by-products can be a cost-effective source of bioactive compounds that have great potential for use in various industries.

A Lifetime of a Dispenser-Release Rates of Olive Fruit Fly-Associated Yeast Volatile Compounds and Their Influence on Olive Fruit Fly (Bactrocera oleae Rossi) Attraction.

The objective of this study was to evaluate the release rate, duration, and biological efficiency of yeast volatile compounds associated with olive fruit flies in slow-release dispensers, polypropylene vials, and rubber septa attached to yellow sticky traps under different environmental conditions in order to protect the environment, humans, and nontarget organisms. Isoamyl alcohol, 2-octanone, and 2-phenethyl acetate were placed in dispensers and tested over a four-week experiment. The weight loss of the volatile compounds in both dispensers was measured, and a rapid, inexpensive, and simple HS-GC/FID method was developed to determine the residual amount of volatiles in the septa. 2-Phenethyl acetate stood out in the rubber septa and showed a statistically significant difference in the release ratio compared to the other volatiles under all conditions tested. Our results showed that the attraction of olive fruit flies increased with decreasing concentrations of the tested volatiles. Regarding the number of flies attracted by rubber septa containing 2-phenethyl acetate, significantly better results were obtained than for septa containing isoamyl alcohol and 2-octanone, in contrast to the attraction of olive fruit flies to polypropylene vials containing these compounds but without significant difference. Since the presence of all tested chemicals was detected during the experiment, this opens the possibility of using more environmentally friendly and cost-effective dispensers with a significantly lower amount of semiochemicals.

Golden Chanterelle or a Gold Mine? Metabolites from Aqueous Extracts of Golden Chanterelle (Cantharellus cibarius) and Their Antioxidant and Cytotoxic Activities.

Cantharellus cibarius, the golden chanterelle, is the second most-collected wild edible mushroom in Europe and very frequently harvested in Croatia. Wild mushrooms have been considered a healthy food since ancient times and are today highly valued for their beneficial nutritional as well as medicinal properties. Since golden chanterelle is added to different food products to improve their nutritive value, we studied the chemical profile of aqueous extracts of golden chanterelle (at 25 °C and 70 °C) and their antioxidant and cytotoxic activities. Malic acid, pyrogallol and oleic acid were some of the main compounds identified by GC-MS from derivatized extract. p-Hydroxybenzoic acid, protocatechuic acid and gallic acid were the most abundant phenolics quantitatively determined by HPLC, with somewhat higher amounts for samples extracted at 70 °C. Antioxidant activity was determined by ferric reducing antioxidant power assay and oxygen radical absorption method, and the highest results were recorded for golden chanterelle extracted at 70 °C, being 41.54 ± 1.54 and 38.72 ± 2.47 µM TE/L, respectively. Aqueous extract at 25 °C showed the better response against human breast adenocarcinoma MDA-MB-231 (IC50 = 375µg/mL). Our results confirm the beneficial effect of golden chanterelle even under aqueous extraction conditions and highlight its significance as a dietary supplement and in the development of new beverage products.

In the Beginning Was the Bud: Phytochemicals from Olive (Olea europaea L.) Vegetative Buds and Their Biological Properties.

Even though Olea europaea L. is one of the most important and well-studied crops in the world, embryonic parts of the plants remain largely understudied. In this study, comprehensive phytochemical profiling of olive vegetative buds of two Croatian cultivars, Lastovka and Oblica, was performed with an analysis of essential oils and methanol extracts as well as biological activities (antioxidant, antimicrobial, and cytotoxic activities). A total of 113 different volatiles were identified in essential oils with hydrocarbons accounting for up to 60.30% and (Z)-3-heptadecene being the most abundant compound. Oleacein, oleuropein, and 3-hydroxytyrosol had the highest concentrations of all phenolics in the bud extracts. Other major compounds belong to the chemical classes of sugars, fatty acids, and triterpenoid acids. Antioxidant, antimicrobial, and cytotoxic activities were determined for both cultivars. Apart from antioxidant activity, essential oils had a weak overall biological effect. The extract from cultivar Lastovka showed much better antioxidant activity than both isolates with both methods (with an oxygen radical absorbance capacity value of 1835.42 µM TE/g and DPPH IC50 of 0.274 mg/mL), as well as antimicrobial activity with the best results against Listeria monocytogenes. The human breast adenocarcinoma MDA-MB-231 cell line showed the best response for cultivar Lastovka bud extract (IC50 = 150 µg/mL) among three human cancer cell lines tested. These results demonstrate great chemical and biological potential that is hidden in olive buds and the need to increase research in the area of embryonic parts of plants.

Chemical Profiling of Sea Fennel (Crithmum maritimum L., Apiaceae) Essential Oils and Their Isolation Residual Waste-Waters.

Sea fennel (Crithmum maritimum L.) is a wild-growing halophyte used in cuisine, traditional medicine or cosmetic products for its beneficial nutritive value and pleasant sensory characteristics. This study aimed to investigate sea fennel essential oils (EOs) from different parts of the plant (flowers, leaves and stems) and the corresponding hydrodistillation by-products (residual water) to validate their potential use and application in different industries. EOs were analyzed by gas chromatography coupled with mass spectrometry (GC-MS), while the phenolic profile of the residual water was analyzed by high-performance liquid chromatography (HPLC) and spectrophotometric methods. The EO analysis confirmed the presence of 14 compounds, dominated by sabinene (from 42.55 to 51.47%) and limonene (from 36.28 to 43.58%), while among the 12 detected phenolics, chlorogenic acid and its isomers (cryptochlorogenic and neochlorogenic acid) were found in the highest concentrations. Total phenolic, flavonoid and tannin contents were concentrated in the order flowers > leaves > stems. Although the sea fennel samples showed differences in chemical profiles, overall they were rich in bioactive compounds with relatively high amounts of key compounds with already proved good biological properties, especially in waste-water, indicating great potential for re-use in accordance with green processing technology trends.

Physiological and Biochemical Response of Wild Olive (Olea europaea Subsp. europaea var. sylvestris) to Salinity.

In the face of climate change, water deficit and increasing soil salinity pose an even greater challenge to olive cultivation in the Mediterranean basin. Due to its tolerance to abiotic stresses, wild olive (Olea europaea subsp. europaea var. sylvestris) presents a good candidate in breeding climate-resilient olive varieties. In this study, the early response of the native Croatian wild olive genotype (WOG) to salinity was evaluated and compared with that of well-known cultivars (cv.) Leccino and Koroneiki. Potted olive plants were exposed either to 150 mM NaCl or 300 mM mannitol for 3 weeks to distinguish between the osmotic and ionic components of salt stress. To determine the impact of the plant age on salinity, 1-, 2-, and 3-year-old WOG plants were used in the study. The growth parameters of both the cultivars and WOG of different ages decreased in response to the mannitol treatment. In contrast to cv. Leccino, the NaCl treatment did not significantly affect the growth of cv. Koroneiki or WOG of any age. The contents of Na+ and Cl- were considerably higher in the salt-treated WOG, regardless of age, compared with the cultivars. However, while both treatments significantly reduced the K+ content of cv. Koroneiki, that nutrient was not significantly affected in either cv. Leccino or WOG. Unlike the cultivars and older WOG, the NaCl treatment caused a significant decline of photosynthetic pigments in the 1-year-old WOG. The cultivars and WOG of different ages experienced a similar drop in the chlorophyll a content under the isotonic mannitol treatment. The absence of lipid peroxidation, modulation of superoxide dismutase, and guaiacol peroxidase activity were noted in all WOG ages under both stressors. These data suggest that WOG resilience to salinity is associated with its large leaf capacity for Na+ and Cl- accumulation, K+ retention, and its adaptable antioxidative mechanisms. The results are promising with regard to obtaining a new olive cultivar with better resilience to soil salinity.

Essential Oil Volatile Fingerprint Differentiates Croatian cv. Oblica from Other Olea europaea L. Cultivars.

Olive leaves are a highly available by-product from table olive and olive oil production. They are nowadays strongly valuable for their major bioactive compounds and their beneficial effects. To determine the differences between two Croatian domestic (Lastovka, Oblica) and two introduced (Leccino, Frantoio) cultivars, physical and chemical analysis of olive leaves were performed: surface area, color variability, total phenolic amounts, and essential oil volatile profiles were analyzed at three harvest periods. All cultivars greatly differed in surface area, with cv. Lastovka being the smallest. Color variability resulted in an overall decrease in darkness and amounts of green and yellow that could be attributed to a decrease in photosynthetic demand and chlorophyll content. The highest amount of total phenolic content occurred in the summer months, followed by a reduction until October. Essential oils volatiles were determined by GC-MS and showed great diversity not only amongst cultivars but also between harvest periods, with overall 45 compounds identified. Principal component analysis distinguished domestic cultivar Oblica from the other observed cultivars, mainly due to its essential oil volatile fingerprint. Compounds that differentiated cv. Oblica were aldehydes ((E,Z)-2,4-heptadienal, (E,E)-2,4-heptadienal, decanal), ketones ((E)-ß-damascone, dihydrodehydro-ß-ionone), sesquiterpenes (cyclosativene, α-copaene, α-muurolene) and saturated hydrocarbons (tetradecane, hexadecane). Essential oil volatile fingerprint attributed the highest to the biodiversity of domestic cv. Oblica through all three harvest periods.

Examination of a New Delivery Approach for Oral Cannabidiol in Healthy Subjects: A Randomized, Double-Blinded, Placebo-Controlled Pharmacokinetics Study.

INTRODUCTION: Therapeutic effects of cannabidiol (CBD) in specialized populations continue to emerge. Despite supra-physiological dosing being shown to be tolerable in various pathologies, optimization of CBD absorption has obvious benefits for general health and recreational usage. Our objectives were to: (1) to investigate a joint pharmacokinetic-physiological time course of multiple recreational-equivalent (< 100 mg) dosages of oral CBD in young healthy adults and (2) evaluate a newly developed technology (TurboCBD™) for the enhanced delivery of CBD. METHODS: In a double-blinded, placebo-controlled, cross-over design, 12 participants received placebo, generic 45 or 90 mg of CBD, or TurboCBD™ delivery technology capsules on five separate occasions. RESULTS: Although there were no differences in the 45 mg conditions, circulating CBD levels were higher with the TurboCBD™ 90 mg group at both 90 (+ 86%) and 120 (+ 65%) min compared with the 90 mg control (p < 0.05). Total area under the curve tended to be higher with TurboCBD™ 90 mg compared with 90 mg (10,865 ± 6322 ng ml-1 vs. 7114 ± 2978 ng ml-1; p = 0.088). Only the TurboCBD™ 90 mg dose was elevated greater than placebo at 30 min (p = 0.017) and remained elevated at 4 h (p = 0.002). CONCLUSION: Consistent with higher bioavailability, TurboCBD™ 90 mg at the peak CBD concentration was associated with an increase in cerebral perfusion and slight reduction in blood pressure compared with baseline and the 90 mg control. Further studies are needed to establish the mechanisms of action of this technology and to explore the therapeutic potential of acute and chronic dosing on more at-risk populations. FUNDING: Lexaria Bioscience Corp. TRIAL REGISTRATION: ClinicalTrials.gov identifier, NCT03295903.

Liver damage indices as a tool for modifying methadone maintenance treatment: a cross-sectional study.

AIM: To assess the effect of liver damage on methadone metabolism in opiate addicts undergoing methadone maintenance treatment (MMT). METHODS: This cross-sectional study recruited 74 patients treated at the outpatient clinic of Public Health Institute of Split-Dalmatia County from 2013-2016. Concentrations of methadone and its main inactive metabolite were measured in participants' biological samples on regular check-ups. Urine samples obtained before oral methadone intake, and blood and urine samples obtained 90 minutes after methadone intake were analyzed using gas chromatography/mass spectrometry. Participants were divided into groups according to liver damage criteria: hepatitis C virus status (positive, negative, or clinical remission); aspartate aminotransferase to platelet ratio (APRI) index (<0.7 and ≥0.7); and fibrosis-4 score (<1.45, 1.45-3.25, >3.25). RESULTS: Metabolic ratio and methadone metabolite concentration in plasma decreased linearly with HCV infection status by the factor of 1.67 (P=0.001) and 2.2 (P=0.043), respectively. Metabolic ratio in plasma decreased in patients with APRI index ≥0.7 by the average factor of 2.12 (P=0.01) and methadone metabolite concentration in plasma decreased by the factor of 6.16 (P=0.009). Metabolic ratio in urine decreased with the severity of fibrosis-4 score by the average factor of 1.63 (P=0.008), whereas methadone metabolite concentration decreased by the factor of 3.53 (P=0.007). CONCLUSION: Liver damage decreases methadone metabolism. Indices of liver function should be calculated regularly during MMT for methadone dose titration.

Blood alcohol stability in postmortem blood samples.

The effect of long-term storage on alcohol stability in postmortem blood samples was investigated. Seventy-nine blood samples were used in this study. For the first time, blood alcohol concentrations were measured within 1 to 4 days after being taken at the Laboratory of Forensic Toxicology. After storage (ranging from 191 to 468 days), each sample was reanalyzed for blood alcohol concentration, and results were compared with the original analysis. Results showed good agreement between the experimental measurements. About 90% of the results lie within 95% limits, and 10% were outside. Despite these positive correlations, the results of some blood samples showed a high variation in ethanol concentration. Deviation in alcohol concentration between replicates of up to 10% (observed in 39% samples) is not acceptable as precise forensic evidence.

The role of alcohol in road traffic accidents with fatal outcome: 10-year period in Croatia Split-Dalmatia County.

OBJECTIVE: The aim of the study was to investigate the relationship between alcohol consumption and fatal road traffic accidents that took place in Split-Dalmatia County (Croatia) during a 10-year period (from August 2001 to August 2011). METHODS: A retrospective analysis of 474 autopsy reports was performed, with an emphasis on 337 toxicologal findings of blood alcohol concentration (BAC) and the mode of participation in traffic (driver, passenger, pedestrian, or cyclist). Relations between the BAC, the weekday on which death occurred, victim's age and gender, and differences according to 3 successive legislation periods within the observed time frame were analyzed. The BAC was measured by gas chromatography with headspace and flame ionization detection. RESULTS: Alcohol was regarded as a contributive death factor if BAC was positive; that is, higher than 0.5 g/kg. BAC was positive in 177 cases. Most often it was the car driver who had a positive BAC. Victims of traffic accidents were mostly male drivers, and those accidents were more often associated with alcohol consumption. Consumption of alcohol produced a significant increase in culpability. CONCLUSION: The results of our study show that alcohol remains one of the main contributing factors of traffic accidents in Split-Dalmatia County. Unfortunately, legislation changes regarding the allowed BAC did not cause any reduction in casualties among drivers driving under the influence of alcohol (DUIA). Chi-square test of the number of total driver victims and driver victims who were DUIA revealed no significant difference (P = .173).