Chemical composition, free-radical-scavenging and insecticidal activities of the aerial parts of Stachys byzantina

Stachys byzantina K. Koch. is an Iranian endemic species of the genus Stachys L., which comprises about 300 species, and is one of the largest genera of the family Lamiaceae. A combination of solid phase extraction (SPE) and high pressure liquid chromatography (HPLC) of the methanolic extract of the aerial parts of S. byzantina afforded three phenylethanoids, 2'-O-arabinosyl verbascoside (1), verbascoside (2), aeschynanthoside C (3) and three flavones apigenin 7-O-glucoside (4), apigenin 7-O-(6-p-coumaroyl)-glucoside (5) and apigenin (6). The structures of these compounds were determined by spectroscopic methods. Free-radical-scavenging and insecticidal properties of the crude extracts, the fractions and the isolated compounds were assessed.

Quantitative multiplex immunohistochemistry reveals inter-patient lymphovascular and immune heterogeneity in primary cutaneous melanoma

IntroductionQuantitative, multiplexed imaging is revealing complex spatial relationships between phenotypically diverse tumor infiltrating leukocyte populations and their prognostic implications. The underlying mechanisms and tissue structures that determine leukocyte distribution within and around tumor nests, however, remain poorly understood. While presumed players in metastatic dissemination, new preclinical data demonstrates that blood and lymphatic vessels (lymphovasculature) also dictate leukocyte trafficking within tumor microenvironments and thereby impact anti-tumor immunity. Here we interrogate these relationships in primary human cutaneous melanoma. MethodsWe established a quantitative, multiplexed imaging platform to simultaneously detect immune infiltrates and tumor-associated vessels in formalin-fixed paraffin embedded patient samples. We performed a discovery, retrospective analysis of 28 treatment-naïve, primary cutaneous melanomas. ResultsHere we find that the lymphvasculature and immune infiltrate is heterogenous across patients in treatment naïve, primary melanoma. We categorized five lymphovascular subtypes that differ by functionality and morphology and mapped their localization in and around primary tumors. Interestingly, the localization of specific vessel subtypes, but not overall vessel density, significantly associated with the presence of lymphoid aggregates, regional progression, and intratumoral T cell infiltrates. DiscussionWe describe a quantitative platform to enable simultaneous lymphovascular and immune infiltrate analysis and map their spatial relationships in primary melanoma. Our data indicate that tumor-associated vessels exist in different states and that their localization may determine potential for metastasis or immune infiltration. This platform will support future efforts to map tumor-associated lymphovascular evolution across stage, assess its prognostic value, and stratify patients for adjuvant therapy

Phytochemical analysis of essential oil of Anthriscus nemorosa and evaluation of antioxidant and anti-malarial activity

Background and objectives: This investigation was performed in order to analyze the composition of the essential oil (EO) of Anthriscus nemorosa and evaluation of its anti-oxidant and anti-malarial activity of its extracts and determination of the total phenolics content (TPC) and total flavonoid content (TFC). Methods: One hundred g dried powder of Anthriscus nemorosa was submitted to hydro-distillation and also was extracted (with n-hexane, dichloromethane (DCM) and methanol (MeOH)), by using Clevenger and Soxhlet apparatus, respectively. Moreover, extracted essential oil (EO) was analyzed by GC-MS. Furthermore, the anti-oxidant, anti- malaria, Total phenolics content (TPC) and total flavonoid content (TFC) of EO and the extracts were investigated by DPPH, cell free -hematin formation, Folin- Ciocalteau and colorimetric methods, respectively. Results: Fifty nine compounds, representing 94% of total oil were identified High content of terpenoids (60.02%) were identified in the essential oil with isogeranol (28.86%), crystathenyl acetate  (13.86%) and farnesene (10.39%) as the most dominant compounds.. Methanol extract demonstrated free radical scavenging activity (RC50 0.192±0.133).Total phenol contents was (325.82±2.72 mg/g). Total flavonoid content was (140.4096±2.4 mg/g). None of the extracts showed anti-malaria effect. Conclusion: Main constituents of A. nemorosa were terpenoids. In comparison with other species of Anthriscus, antioxidant activity of A. nemorosa essential oil was less noticeable

Phytochemical analysis and biological activity evaluation of essential oil of Scrophularia atropatana

Background and objectives: Scrophularia atropatana (S. atropatana) is endemic to western and central regions of Iran. It belongs to the genus of Scrophularia (family: Scrophulariaceae). These species have been found to possess antibacterial, antitumor and anti-inflammatory properties. Most of these effects were proven to be attributed to the non-volatile components, which were known as secondary metabolites. Iridoids, phenylpropanoids, phenolic acids and flavonoids have been identified as their main secondary metabolites. This investigation was performed to analyze the composition of the essential oil (EO) of S. atropatana and evaluation of its anti-oxidant and total toxicity properties. Methods: About 150 g powder of the air dried parts of S. atropatana was submitted to hydro-distillation and also was successively extracted (with n-hexane, dichloromethane (DCM) and methanol (MeOH)), by using Clevenger and Soxhlet apparatus, respectively. In addition, Essential Oil (EO) was analyzed by GC-MS and GC-FID. Furthermore, the anti-oxidant activity of the extracts, fractions and EO as well as general toxicity were investigated by DPPH and brine shrimp larva assay, respectively. Results: Totally, sixty eight volatile compounds were identified, which constituted 91.78% of the total oil composition. Among the compounds, major constituents were identified as non-terpenoides (59.59%). Moreover, among the extracts and EO, methanol extract demonstrated high free radical scavenging and general toxic activities with RC50 equal 0.143±0.13 mg/mL and LD50 0.271 mcg/mL respectively. Conclusion: Overall, the main constituents of S. atropatana were hydrocarbons. In comparison to other genesis of Scrophularia, antioxidant activity of the essential oil of S. atropatana was less noticeable

Common ash (Fraxinus excelsior L.) seeds as a new vegetable oil source

In this study, common ash (Fraxinus excelsior L.) seed oil compositions were determined for the first time. Effect of the moisture content and microwave pretreatment of the seeds on the oil yield and its composition were also investigated. The optimum level of moisture content to obtain the maximum oil (18.18%) yield was 1% with 3 min pre-treatment. The main triacylglycerol composition of the oil was linoleic–linoleic–oleic, followed by oleic-oleic-linoleic and linoleic–linoleic-linoleic. The main fatty acid of the oil was linoleic acid (C18:2), followed by oleic acid (C18:1) and palmitic acid (C16:0). The main phytosterol of the oil was β-sitosterol, followed by Δ⁵-avenasterol and campesterol. The main tocopherol of the oil was α-tocopherol, followed by β and γ-tocopherols. The main essential oil of the oil was bicyclogermacrene, followed by delta-cadinene. These results suggest that this oil is a good source of valuable nutritional compounds and that microwave pretreatment is a promising and environmentally friendly method to extract it as a new vegetable oil source

In vitro bioactivity and phytochemical evaluation of extracts from aerial parts of Eremostachys macrophylla Montbr. & Auch. growing in Iran

Background and objectives: The aerial part extracts of Eremostachys macrophylla  from Labiatae family, which has been traditionally used in wound healing, snake bites, rheumatism and joint pains, were investigated for general toxicity, anti-proliferative, free radical scavenging and anti-bacterial effects.Moreover, preliminary phytochemical investigations were carried out on the extracts. Methods: Extracts were prepared using a soxhlet apparatus with n-hexane, dichloromethane (DCM) and methanol (MeOH), respectively. Brine shrimp lethality test (BSLT) and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay were performed to evaluate general toxicity and free radical scavenging properties, respectively. Anti-proliferative and anti-bacterial activities were assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and disc diffusion methods, respectively.  MTT assay was carried out on one normal and two cancer cell lines including human umbilical vein endothelial cells (HUVEC), human colorectal adenocarcinoma (HT-29) and human lung carcinoma (A-549), respectively. Additionally, all the extracts were tested for the presence of various phytoconstituents by different reagents. Results: The n-hexane extract was the most active fraction in BSLT whereas the MeOH extract showed significant free-radical-scavenging activity. The results indicated that the n-hexane and MeOH extracts possessed anti-proliferative effects against HT-29 cells while all the three extracts were effective against A-549 cell line. None of these three extracts showed any significant effect against HUVEC.The extracts didn’t show any antimicrobial effects against Gram positive, Gram negative and Candida albicans species. Conclusion: Considering the results, the species might be a good candidate for further phytochemical and biological studies for the isolation of active and pure ingredients and clarification of anti-neoplastic mechanism

Evaluation of chemical composition and biological activity of the aerial parts of Artemisia fragrans

Background and objectives: The genus Artemisia, an important member of the Asteraceae family, is mostly distributed in Europe, North America, Asia and South Africa, and represented in Iranian flora by 34 species. In the Iranian traditional medicine these plants were used as anthelmintic and anti-inflammatory. The presence of sesquiterpene lactones and germacranolides were reported in A. fragrans. Our objective in this investigation was to identify the chemical constituents and antioxidant activity of A. fragrans. Methods: The essential oil (EO) and different extracts (n-hexane, dichloromethane, and methanol) of the aerial parts of A. fragrans were obtained by hydro-distillation and Soxhlet apparatus. The essential oil was analyzed by GC-MS and GC-FID. For further investigation, the MeOH extract (2 g) was subjected to Sep-Pack fractionation using a step gradient of MeOH-water. Moreover, the free radical scavenging activity of the EO and extracts was evaluated by the 2, 2-diphenyl -1-picrylhydrazyl (DPPH) method. Results: The GC-MS analysis, led to the identification of six main monoterpenes accounting for the 88.7% of the total components present. Among the extracts, camphor was the main component. The essential oil of A. fragrans displayed weak antioxidant activity whereas the antioxidant activity of the MeOH extract was high. The analysis of the 20% and 40% methanolic Sep-Pack fractions resulted in the isolation of two flavonoid glycosides and one dipeptide. Conclusion: The results of the present study introduced A. fragrans as a new source of flavonoid glycosides and suggested it as an appropriate candidate for further radical scavenging studies