Lavandula austroapennina: Assessment of the Antiviral Activity of Lipophilic Extracts from Its Organs.

In a framework aimed at the recovery and enhancement of medicinal plants endemic to the territory of the Cilento and Vallo di Diano National Park, Lavandula austroapennina N.G. Passal., Tundis and Upson has aroused interest. An insight into the chemical composition of the corolla, calyx, leaf, stem, and root organs was carried out following ultrasound-assisted maceration in n-hexane. The obtained lipophilic extracts were explored using ultra-high-performance chromatography coupled to high-resolution mass spectrometry (UHPLC-ESI-QqTOF-MS/MS). The extracts from the different organs varied in their relative content of fatty acids, ursanes, and oleanane-type triterpenes. In particular, the oleanolic acid content appeared to increase in the order of corolla < leaf < stem. An MTT assay was performed to verify the possible cytotoxicity of the organ extracts of L. austroapennina at a concentration ranging from 12.5 to 400 µg/mL on the Vero CCL-81 cell line. Antiviral activity against herpes simplex virus type 1 (HSV-1), alpha human coronavirus 229E (HCoV-229E), and poliovirus type 1 (PV-1) was evaluated via a plaque reduction assay in the same cellular model. All the extracts did not show cytotoxic effects after 2 and 24 h exposure times, and the antiviral efficacy was particularly important for the stem extract, capable of completely inhibiting the tested viruses at low doses. The antiviral activity in a non-enveloped virus PV-1 allowed the assertion that the extracts from the organs of L. austroapennina, and especially the stem extract, interfered directly with the viral envelope. This study underlines how much knowledge of a territory's medicinal plant heritage is a harbinger of promising discoveries in the health field.

Investigation of Volatile Components and Assessment of Antioxidant Potential in Seven Lamiaceae Plant Hydrosols.

Aromatic plants of the family Lamiaceae, especially of the genus Thymus, have promising antioxidant applications in pharmacology, medicine, food, cosmetology, and aromatherapy. Hydrosols (HDs) were extracted by hydrodistillation from seven species of Lamiaceae, including Thymus vulgaris, Thymus mongolicus, Mentha × piperita, Melissa officinalis, Rosmarinus officinali, Salvia elegans, and Leonurus artemisia. In total, 369 volatile components were determined and analyzed by gas chromatography-mass spectrometry (GC-MS). Among them, alcohols (2.86-28.48%), ethers (2.46-10.69%), and phenols (0.11-21.78%) constituted a large proportion, mainly linalool (0.28-19.27%), eucalyptol (0.16-6.97%), thymol (0-19.54%), and carvacrol (0-26.82%). Multivariate statistical analyses were performed and 27 differential metabolites were screened. Three different methods (ABTS+•, DPPH•, and FRAP) were used to determine the in vitro antioxidant activity of seven HDs. Thymus vulgaris hydrosols (Tv HDs) and Thymus mongolicus hydrosols (Tm HDs) had the strongest antioxidant activity and their stronger antioxidant capacity was related to their high levels of phenolic constituents, mainly thymol. The antioxidant activity of the other five Lamiaceae HDs was associated with their high alcohol (mainly linalool and eucalyptol) content, and the alcohol constituents may synergistically affect their antioxidant capacity. Therefore, the present study suggests that Lamiaceae plants can be utilized as antioxidant products or antioxidants in different industrial sectors including pharmaceuticals, food, cosmetics, and agrochemicals.

Phytochemical Study and Antiglioblastoma Activity Assessment of Plectranthus hadiensis (Forssk.) Schweinf. ex Sprenger var. hadiensis Stems.

Glioblastoma (GB) is the most malignant form of primary astrocytoma, accounting for more than 60% of all brain tumors in adults. Nowadays, due to the development of multidrug resistance causing relapses to the current treatments and the development of severe side effects resulting in reduced survival rates, new therapeutic approaches are needed. The genus Plectranthus belongs to the Lamiaceae family and is known to be rich in abietane-type diterpenes, which possess antitumor activity. Specifically, P. hadiensis (Forssk.) Schweinf. ex Sprenger has been documented for the use against brain tumors. Therefore, the aim of this work was to perform the bioguided isolation of compounds from the acetonic extract of P. hadiensis stems and to investigate the in vitro antiglioblastoma activity of the extract and its isolated constituents. After extraction, six fractions were obtained from the acetonic extract of P. hadiensis stems. In a preliminary biological screening, the fractions V and III showed the highest antioxidant and antimicrobial activities. None of the fractions were toxic in the Artemia salina assay. We obtained different abietane-type diterpenes such as 7α-acetoxy-6ß-hydroxyroyleanone (Roy) and 6ß,7ß-dihydroxyroyleanone (DiRoy), which was also in agreement with the HPLC-DAD profile of the extract. Furthermore, the antiproliferative activity was assessed in a glioma tumor cell line panel by the Alamar blue assay. After 48 h treatment, Roy exerted strong antiproliferative/cytotoxic effects against tumor cells with low IC50 values among the different cell lines. Finally, we synthesized a new fluorescence derivative in this study to evaluate the biodistribution of Roy. The uptake of BODIPY-7α-acetoxy-6ß-hydroxyroyleanone by GB cells was associated with increased intracellular fluorescence, supporting the antiproliferative effects of Roy. In conclusion, Roy is a promising natural compound that may serve as a lead compound for further derivatization to develop future therapeutic strategies against GB.

Morpho-palynological assessment of some species of family Asteraceae and Lamiaceae of District Bannu, Pakistan on the bases of light microscope & scanning electron microscopy.

Pollen micro-morphological features have proven to be helpful for the plant taxonomists in the identification and classification of plants. The aim of this study was to evaluate the palynological features of family Asteraceae and Lamiaceae from flora of District Bannu, Khyber Pakhtunkhwa, Pakistan using both scanning electron microscopy (SEM) and light microscope (LM) for their taxonomic importance. Pollen of seven Asteraceous species belonging to four genera and four Lamiaceae species categorized into four genera were collected from different localities of research area. The present research work provides detailed information of diverse morpho-palynological characters both qualitatively and quantitatively including pollen shape, type, diameter, P/E ratio, exine sculpturing and thickness. Type of pollen in Asteraceae and Lamiaceae was ranged from tricolporate, tricolpate, trizonocolpate and hexazonocolpate. The maximum polar diameter (40.05 µm) and equatorial diameter (37.66 µm) was observed in the Ajuga bracteoosa while minimum polar and equatorial diameter was noted in Isodon rugosus (11.10 µm) and Erigeron canadensis (13.20 µm) respectively. Sculpturing of exine include; echinate, reticulate scabrate, aerolate, reticulate-verrucate, reticulate-scabrate, perforate and reticulate to perforate. Exine thickness was examined maximum 1.50 µm in Helianthus tuberosus, whereas minimum in Conyza Canadensis (0.16 µm). The pollen fertility was found highest in C Canadensis (83.33%) and lowest in Ajuga bracteosa (58.06%). The observed pollen morphology has many valuable qualitative and quantitative attributes for the better understanding of their taxonomy and play significant role in correct identification.

A high-throughput method for dereplication and assessment of metabolite distribution in Salvia species using LC-MS/MS.

Dereplication of crude plant extracts through liquid chromatography-mass spectrometry is a powerful technique for the discovery of novel natural products. Unfortunately, this technique is often plagued by a low level of confidence in natural product identification. This is mainly due to the lack of extensive chromatographic and mass spectrometric optimizations that result in improper and incomplete MS/MS fragmentation data. This study proposes a solution to this problem by the optimization of chromatographic separation and mass spectrometry parameters. We report herein a direct and high-throughput strategy for natural product dereplication in five Salvia species using high-resolution ESI-QTOF-MS/MS data. In the present study, we were able to identify a total of forty-seven natural products in crude extracts of five Salvia species using MS/MS fragmentation data. In addition to dereplication of Salvia species, quantitative profiling of twenty-one bioactive constituents of the genus was also performed on an ion trap mass spectrometer. For the quantitation study, method development focused on chromatographic optimizations to achieve maximum sensitivity. The developed dereplication and quantitation strategy can be extended to develop comprehensive metabolic profiles of other plant genera and species and thus can prove useful in the field of drug discovery from plants.

A chemometric assessment of essential oil variation of three Salvia species indigenous to South Africa.

Indigenous Salvia species from southern Africa are popular traditional medicines for the treatment of a variety of conditions. They produce fragrant volatiles that can be isolated as essential oils. Some of these volatile organic compounds may play a role in the biological activities of the extracts. Three indigenous Salvia species, Salvia africana-lutea, S. lanceolata and S. chamelaeagnea, were selected for this study as they are commonly used in traditional medicine in South Africa, and the essential oils from these species have potential for commercialisation. Although some studies have described the essential oil compositions and some biological activities, only single composite samples were used. The aim of this study was to investigate the intra- and interspecies variation of the essential oils, sampled over a wide geographical area and using a representative sample size, to encourage commercialisation of the essential oil. Essential oils were isolated from individual plants using conventional hydrodistillation of the aerial parts, harvested from several localities. Gas chromatography coupled simultaneously to mass spectrometry/flame ionisation detection (GC-MS/FID) was used to identify and quantify the volatile constituents. The essential oils of S. africana-lutea consisted mainly of terpinene-4-ol + ß-caryophyllene (1.4 - 29.0%), T-cadinol (1.2 - 20.0%), α-eudesmol (trace - 23.0%) and ß-eudesmol (trace - 26.0%), those of S. lanceolata comprised mainly terpinene-4-ol + ß-caryophyllene (4.3 - 31.0%), α-humulene (2.3 - 15.0%), bicyclogermacrene (trace - 37.0%) and spathulenol (trace - 25.0%), while the essential oils of S. chamelaeagnea were characterised by δ-3-carene (trace - 18.0%), limonene (1.6 - 36.0%), viridiflorol (9.8 - 61.0%) and 1,8-cineole (not detected - 11.0%). The compounds identified in the essential oils of the three selected Salvia species have been identified in other Salvia essential oils. To add to the novelty of this study, the superior resolving power of two-dimensional gas chromatography was demonstrated through analysis of selected essential oils. Many additional compounds were identified, and previously co-eluting compounds were clearly separated. Chemometric modelling of the GC-MS data using SIMCA P+ 14 software allowed distinct clustering patterns to be discerned. The unsupervised principal component analysis model revealed separate clusters for the three species, confirming substantial chemical differences between their essential oils. Quantitative, rather than qualitative differences were evident when individual essential oil samples representing the same species, were compared. For each species, two chemically distinct groups were observed and unique marker compounds could be identified. This study has contributed detailed information on the major and minor volatile compounds present in the essential oils of the three Salvia species investigated.

Polyphenolic Profiling, Quantitative Assessment and Biological Activities of Tunisian Native Mentha rotundifolia (L.) Huds.

Phenolic profiling of ten plant samples of Mentha rotundifolia (L.) Huds. collected from different bioclimatic areas of Tunisia, was for the first time carried out by using a fast ultra-high-performance liquid chromatography (UHPLC)-high resolution tandem mass spectrometry (HRMS/MS) method on a Q Exactive platform equipped with an electrospray ionization (ESI) source. An intraspecific, interpopulation variability was evidenced and a total of 17 polyphenolic metabolites were identified and quantified by using the UHPLC-HRESIMS/MS method, here validated for specificity, linearity, limit of detection, limit of quantitation, accuracy and precision. The quantitative method resulted sensitive at the nM level and reliable for rapid polyphenol quantification in vegetal matrices. The metabolomic study allowed us to identify a new compound, named salvianolic acid W, which was isolated and characterized mainly by NMR and MS analysis. A statistical correlation of the phenolic composition with antioxidant and anti-acetylcholinesterase activities was provided.

Assessment of Antidepressant Effect of the Aerial Parts of Micromeria myrtifolia Boiss. & Hohen on Mice.

The currently available antidepressant agents necessitate the development of newer alternatives because of their serious adverse effects and costs. Traditional medicinal knowledge is likely the key that opens the door to discover new medicines. In Turkish folk medicine, the infusion prepared from aerial parts of Micromeria myrtifolia Boiss. & Hohen is used as pleasure and medicinal tea for its relaxing action. The present research was conceived to confirm the antidepressant's potential of this traditional medicinal plant. In this process, first of all, the collected and shade-dried aerial parts of M. myrtifolia were powdered and then, extracted using solvents with different polarity as follows; n-hexane, ethyl acetate (EtOAc), and methanol (MeOH). The antidepressant activity of the extracts was evaluated by using several in vivo and in vitro experimental models of depression. When the data obtained from the control and experimental groups were compared, it was determined that the MeOH extract was the most active. The active components of this extract were isolated and identified utilizing various chromatographic separation techniques. The MeOH extract was applied to reversed phase (RP-18) column chromatography to obtain five main fractions and they were tested on antidepressant activity models. The isolated compounds from the obtained fractions were elucidated as rosmarinic acid (1), myricetin (2), apigenin (3), and naringenin (4) which were assumed to be responsible for the antidepressant activity of the aerial parts. According to the results, rosmarinic acid, myricetin, apigenin, and naringenin showed statistically significant activity on forced swimming test and tetrabenazine-induced ptosis models, whereas only rosmarinic acid showed statistically significant activity on the tail suspension test. Apigenin displayed the highest inhibitory activity on MAO A and B enzymes. Studies in the future should be performed to investigate the antidepressant activity mechanism of these natural compounds. The current research could be an important step in the development of the new agents that can be used in the treatment of depression.

Chemical composition and assessment of larvicidal and repellent capacity of 14 Lamiaceae essential oils against Aedes albopictus.

In the current laboratory study, 14 essential oils (EOs) derived from 12 Lamiaceae plant species and their major components were screened for their larvicidal and repellent properties against Aedes albopictus, an invasive mosquito species of great medical importance. The results of toxicity bioassays revealed that the EOs from Thymus vulgaris, Ocimum basilicum, Origanum dictamnus, Origanum majorana, and Origanum vulgare, as well as their major components (terpenes), namely thymol, carvacrol, p-cymene, and γ-terpinene exerted the highest larvicidal effect. Essential oils from Mellisa officinalis, Origanum dictamus, Mentha spicata (chem. piperitenone epoxide), Origanum majorana, and Satureja thymbra were the most potent repellents, with the last two assigned as the best ones. Among the terpenes tested, piperitenone epoxide, carvacrol, thymol, and piperitenone provided the highest level of protection against Ae. albopictus adults. Chemical analysis revealed the presence of a high number of terpenes in the EOs, while in most cases, the biological action of the tested EOs and their major components was in consistency. The most effective EOs and terpenes that were identified through the current laboratory bioassays could be used as alternative agents to control larvae and repel adults of Ae. albopictus.

Genetic toxicological assessment and anti-arthritic effects of an ethanolic extract obtained from Salvia lachnostachys Benth leaves in mice.

In the present study, mice were subjected to prolonged treatment with ethanolic extract of Salvia lachnostachys Benth leaves (SLEE), and the inflammatory and arthritic parameters were evaluated using the Complete Freund's Adjuvant (CFA) model. The genotoxicity of SLEE were also assayed using genetic toxicological tests. For the CFA model, 28 male C57BL/6 mice were distributed randomly into four groups (control, 50 mg/kg of SLEE, 100 mg/kg of SLEE and dexamethasone) for the evaluation of hyperalgesia and paw edema for 21 days after injection of CFA into the paw. To conduct the toxicogenetic assessments (comet assay and micronuclei assay), apoptosis and splenic phagocytosis were evaluated in male Swiss mice after the administration of saline (control group), cyclophosphamide (positive control group) and SLEE (10, 100 and 1000 mg/kg). SLEE significantly reduced the mechanical hyperalgesia and edema caused by CFA injection. The results of the toxicogenetic assessment revealed no toxicogenetic potential in the mice, and the evaluation of apoptosis showed an increase in apoptotic cells in the spleen after 72 h of treatment with SLEE (1000 mg/kg). SLEE exhibited anti-arthritic activity with no toxicogenetic damage. These toxicogenic results support the safety of SLEE.