A Descriptive Chemical Composition of Concentrated Bud Macerates through an Optimized SPE-HPLC-UV-MS2 Method—Application to Alnus glutinosa, Ribes nigrum, Rosa canina, Rosmarinus officinalis and Tilia tomentosa

Concentrated bud macerates (CBMs) are obtained from meristematic tissues such as buds and young shoots by maceration in a solvent composed of glycerin, water and ethanol (1/1/1/, v/v). Their traditional utilization in gemmotherapy has gained interest in the past years, and the knowledge of their chemical characterization can provide commercial arguments, particularly to secure their quality control. Therefore, an optimized method for phytochemical analysis including glycerol removal by a preliminary solid phase extraction (SPE) followed by compound identification using high performance liquid chromatography coupled with ultra-violet and tandem mass detectors (HPLC-UV-MS2) was developed. This method was applied on 5 CBMs obtained from Alnus glutinosa, Ribesnigrum, Rosmarinus officinalis, Rosa canina and Tilia tomentosa in order to determinate their chemical composition. Their antioxidant effects were also investigated by radical scavenging activity assays (DPPH and ORAC). Glycerol removal improved the resolution of HPLC chemical profiles and allowed us to perform TLC antioxidant screening. Our approach permitted the identification of 57 compounds distributed in eight major classes, three of them being common to all macerates including nucleosides, phenolic acids and glycosylated flavonoids. Quantification of the later class as a rutin equivalent (RE) showed a great disparity between Rosa canina macerate (809 mg RE/L), and the other ones (from 175 to 470 mg RE/L). DPPH and ORAC assays confirmed the great activity of Rosa canina (4857 and 6479 μmol TE/g of dry matter, respectively). Finally, phytochemical and antioxidant analysis of CBMs strengthened their phytomedicinal interest in the gemmotherapy field.

Change of Ba concentration by species and organ in several fruits grown in city centers

Heavy metals are elements that are very harmful to human and environmental health. Heavy metal concentration in the fruits grown in city centers can reach very high levels and consuming these fruits as food causes a direct intake of heavy metals, which these fruits contain, into a human body and can pose a significant health risk. All the compounds of barium (Ba), which is one of the most dangerous heavy metals, are toxic. Thus, determining the Ba concentration in plants, which are grown in areas with high pollution and consumed as food, is very important. In the present study, it was aimed to determine the change of Ba concentration by species and organ in several fruits grown in areas with high traffic density. Within the scope of this study, Ba concentrations in leaf, branch, bark, seed, and fruits of Prunus ceresifera, Tilia tomentosa, Prunus avium, and Prunus cerasus were compared. As a result, it was determined that the change of Ba concentrations by species and the change by organ were statistically significant in all organs and in all species, respectively. In general, the lowest values were found in Prunus cerasus or Prunus avium, whereas the highest values were observed in Tilia tomentosa. Considering the organs, the lowest values were found in seed and the highest ones in bark and branch.

Thermal resistance and high-performance microwave decontamination assessment of Bacillus endospores isolated from food-grade herbal extracts

Generally, endospore contamination can occur from different sources during product manufacturing in many industries and therefore lower its quality by affecting physicochemical properties and shelf-life. Bacterial endospores can germinate inside the product and produce several enzymes, which can cause several undesirable changes. This study assessed the spores thermal resistance and applied a microwave decontamination technique toward herbal extracts (Tilia tomentosa and Centella asiatica) containing ethanol or glycerol. Based on 16S rRNA analysis, the detected contaminant endospores belonged to different Bacillus species, namely B. subtilis, B. zhangzhouensis, and B. pumilus. The thermal resistance assessment using inoculated endospores in the actual products revealed B. pumilus T2 as the most resistant endospore to the heat treatments tested in both T. tomentosa and C. asiatica extracts. Finally, a high-performance microwave technique was used to decontaminate T. tomentosa extract against the mixture of Bacillus spores. Results from the microwave technique indicate that the increase of temperature from 100°C to 105°C not only decontaminated the product but also could dramatically decrease the effective thermal treatment time (10 times), which can benefit the product quality. The results provided in this study considerably contribute to improving an original decontamination method for products containing glycerol and ethanol with the most negligible effect on product quality.

Flowers Characteristics of Selected Species of Lime-Tree (Tilia spp.) in Terms of miRNA-Based Markers Activity, Mannose Expression and Biological Compounds Content

The significant healing effect of Tilia platyphyllos Scop. and Tilia cordata Mill. flowers are well known. However, the flowers of Tilia tomentosa Moench. are not suitable for harvest due to their toxic effects. To investigate the diverse background of this effect, we applied a functional miRNA-based marker, mannose expression analysis and determined the content of bioactive compounds. Out of nine tested markers, three (miR160, miR167 and miR408) provided reproducible miRNA-based loci and two of them (miR160 and miR398) enabled the acquisition of fingerprinting specific to flower and leaf samples of T. platyphyllos and T. tomentosa. The most pronounced profiling was specific for miR408 marker, the function of which is connected to plant defense and adaptation mechanisms. We confirmed the suitability of microRNA-based markers for polymorphism determination of flowers of selected species of lime-tree. The highest values of antioxidant activity, flavonoids, total polyphenols and phenolic acids content have been reached in silver linden flowers. When comparing the transcription activity of mannose in flowers, more than 30 times higher levels of mannose transcripts for the silver linden flowers was observed.

Influence of Tilia tomentosa Moench Extract on Mouse Small Intestine Neuromuscular Contractility

Functional gastrointestinal disorders (FGIDs) are characterized by abdominal pain, bloating and bowel disturbances. FGID therapy is primarily symptomatic, including treatment with herbal remedies. Flower extract of Tilia tomentosa Moench (TtM) is occasionally used as an anti-spasmodic in popular medicine. Since its effect on intestinal response is unknown, we evaluated the influence of TtM extract on small intestine contractility. Ileal preparations from C57BL/6J mice were mounted in organ baths to assess changes in muscle tension, following addition of TtM extract (0.5–36 μg/mL) or a vehicle (ethanol). Changes in contractile response to receptor- and non-receptor-mediated stimuli were assessed in ileal preparations pretreated with 12 μg/mL TtM. Alterations in the enteric nervous system neuroglial network were analyzed by confocal immunofluorescence. Increasing addition of TtM induced a marked relaxation in ileal specimens compared to the vehicle. Pretreatment with TtM affected cholinergic and tachykininergic neuromuscular contractions as well as K+-induced smooth muscle depolarization. Following incubation with TtM, a significant reduction in non-adrenergic non-cholinergic-mediated relaxation sensitive to Nω-Nitro-L-arginine methyl ester hydrochloride (pan-nitric oxide synthase inhibitor) was found. In vitro incubation of intestinal specimens with TtM did not affect the myenteric plexus neuroglial network. Our findings show that TtM-induced intestinal relaxation is mediated by nitric oxide pathways, providing a pharmacological basis for the use of TtM in FGIDs.

The Influence of Tilia tomentosa Moench on Plant Species Diversity and Composition in Mesophilic Forests of Western Romania–A Potential Tree Species for Warming Forests in Central Europe?

Climate change challenges important native timber species in Central Europe. The introduction of non-native tree species originating from warmer climates is one option to make Central European forests compatible to global warming. This, however, requires an assessment of the species’ growth requirements, and of its impact on biodiversity in its native ranges. Silver lime (Tilia tomentosa), a moderately drought-tolerant, thermophilous tree species of South-eastern Europe is considered suitable for the future. Along three elevational transects in western Romania, we assessed the impact of changing climate and local site conditions on the abundance of this tree species and contrasted plant species diversity and composition of lime-dominated forests with mesophytic oak and beech forests. Local site conditions and disturbance histories shaped the distribution pattern of silver lime. When dominant, it reduced plant species diversity within stands due to its dense canopy. For shade-tolerant, mesophytic species, though, lime forests provided an additional habitat and extended their range into warmer environments. Thus, silver lime may have the potential as an admixed tree species forming a transitory meso-thermophilous habitat in the future. At the same time, silver lime may be limited under increasing drought frequency.

Old Forests from Dobrogea’s Plateau

From the oldest times, the forest has been the main food, shelter and energy source, playing an essential role in the development of human societies. Old stands are characterized by their long life, large heights and diameters and by their help in conserving biodiversity. The surfaces occupied by old stands have decreased lately because of their wood mass high value which leads to exploitation. This is furthermore coupled with climate changes, urbanization and human influences. These stands occupy a surface of 128.500 ha in the forests from Dobrogea’s Plateau, where broad-leaved species are preponderant (Quercus sp., Fraxinus ornus, Tilia tomentosa, Acer sp.). As such, the present paper intends to characterize the oldest stands from Dobrogea’s Plateau with the help of forest management plans realized during 1993-2007. The main characteristics analyzed in this study for stands older than 120 years were: location, surface, height, diameter, structure, forest type, soil type, altitude, exposition, slope and consistency. Stand older than 100 years occupy only 4% of the total surface and are composed of linden, brown oak and pedunculate oak which grow and develop on common luvisols.

Bazı Peyzaj Bitkilerinde Krom Konsantrasyonunun Tür, Organ ve Trafik Yoğunluğuna Bağlı Değişimi

Today, air pollution is one of the most important problems that threaten human and environmental health. Although there are many components of air pollution, heavy metals are of particular importance. The reason for this is that some of the heavy metals are pure toxic at low concentrations, many heavy metals that are nutrients for living beings are harmful at high costs in bile, live biodegradation process, they are not easily degraded in nature and are used in addition to them. For this reason, it is of great importance to monitor the heavy metal concentrations in the air, especially in risky conditions such as traffic processing, industrial activities and populated areas. In this study, it is grown in Samsun city center and frequently used in landscape studies; Tilia tomentosa, Aesculus hippocastanum, Ligustrum vulgare and Catalpa bignoides species have been tried to determine the changes of Cr concentrations in individuals grown in areas where traffic is heavy, less dense and almost none. Within the scope of the study, leaf, bark and wood samples were collected from the plants, and washing operations were also carried out on the bark and leaves. As a result of the study, Cr concentrations in the species subject to the study changed significantly on the basis of species, organs and traffic density.

Use of an Animal Model to Evaluate Anxiolytic Effects of Dietary Supplementation with Tilia tomentosa Moench Bud Extracts

Anxiety disorders are common and complex psychiatric syndromes affecting a broad spectrum of patients. On top of that, we know that aging produces an increase in anxiety vulnerability and sedative consumption. Moreover, stress disorders frequently show a clear gender susceptibility. Currently, the approved pharmacological strategies have severe side effects such as hallucinations, addiction, suicide, insomnia, and loss of motor coordination. Dietary integration with supplements represents an intriguing strategy for improving the efficacy and the safety of synthetic anxiolytics. Accordingly, a recent article demonstrated that glyceric bud extracts from Tilia tomentosa Moench (TTBEs) exert effects that are consistent with anxiolytic activity. However, the effects of these compounds in vivo are unknown. To examine this question, we conducted behavioral analysis in mice. A total of 21 days of oral supplements (vehicle and TTBEs) were assessed by Light Dark and Hole Board tests in male and female mice (young, 3 months; old, 24 months). Interestingly, the principal component analysis revealed gender and age-specific behavioral modulations. Moreover, the diet integration with the botanicals did not modify the body weight gain and the daily intake of water. Our results support the use of TTBEs as dietary supplements for anxiolytic purposes and unveil age and gender-dependent responses.