Morpho-anatomical features of the leaves and stems of Baccharis notosergila (Asteraceae) and their relationship with the environment and chemical control

Background and aims: Baccharis notosergila is an aggressive weed inhabiting the Salado river basin, Buenos Aires province, Argentina. The aims of this work were: to analyze the morpho-anatomy and histochemistry of aerial vegetative organs in order to understand the adaptation strategies that ensure its survival, as well as to expand knowledge on traits determining resistance to the control methods applied. M&M: The material collected was prepared and examined with conventional techniques of microscopy. Histochemical tests to identify starch, resins, polyphenols, and lipophilic substances were performed. Results: The major features found were small and deciduous leaves; uniseriate epidermis with massive and striate cuticle; stomata at level or slightly above the other epidermal cells and glandular trichomes secreting oily substances; stomata on both surfaces and isobilateral mesophyll. Tannins, starch and lipophilic substances were identified in leaves and stems; polyphenols, resins and lipids in ducts, and calcium oxalate crystals in leaves, stems and capitate trichomes. Conclusions: The aerial vegetative organs features of B. notosergila explain its tolerance to the unfavorable conditions of the Salado river basin area, as well as its high competitive ability over others species of the natural prairie. The reduced and deciduous leaves, the epidermal traits, and chemical substances found constitute a physical and chemical barrier reducing dehydration as well as the penetration of the herbicides applied for its control. Botanical knowledge of B. notosergila is the basis for the design and development of new and appropriate management methods for this species.

Intraperitoneal oil application causes local inflammation with depletion of resident peritoneal macrophages

Oil is frequently used as a solvent to inject lipophilic substances into the peritoneum of laboratory animals. Although mineral oil causes chronic peritoneal inflammation, little is known whether other oils are better suited. Here we show that olive, peanut, corn or mineral oil causes xanthogranulomatous inflammation with depletion of resident peritoneal macrophages. However, there were striking differences in the severity of the inflammatory response. Peanut and mineral oil caused severe chronic inflammation with persistent neutrophil and monocyte recruitment, expansion of the vasculature and fibrosis. Corn and olive oil provoked no or only mild signs of chronic inflammation. Mechanistically, the vegetal oils were taken up by macrophages leading to foam cell formation and induction of cell death. Olive oil triggered caspase-3 cleavage and apoptosis, which facilitates the resolution of inflammation. Peanut oil and, to a lesser degree, corn oil triggered caspase-1 activation and macrophage pyroptosis, which impairs the resolution of inflammation. As such, intraperitoneal oil administration can interfere with the outcome of subsequent experiments. As a proof-of-principle, intraperitoneal peanut oil injection was compared to its oral delivery in a thioglycolate-induced peritonitis model. The chronic peritoneal inflammation due to peanut oil injection impeded the proper recruitment of macrophages and the resolution of inflammation in this peritonitis model. In summary, the data indicate that it is advisable to deliver lipophilic substances like tamoxifen by oral gavage instead of intraperitoneal injection.

Investigation of lipophilic substances of the leaves and flowers of spindle tree (Euonymus Europaea L.)

Spindle Tree (Euonymus europaea L.) has long been used in folk medicine for the treatment of dermatomycoces and as an antiparasitic and anthelmintic agent. However, with what substances it is associate this activity, it is not known for certain. In a previous work, a was developed extracting scheme from the bark of Spindle Tree (Euonymus europaea L.) to obtain water-soluble and lipophilic substances and the composition of the lipophilic fraction was studied using the GC-MS method. The purpose of this work was to obtain lipophilic substances from the flowers and leaves of Spindle Tree and to compare their chemical composition with the composition of substances obtained from the bark of the plant. Flowers and leaves were dried in a drying oven at a temperature of 52 °C, after which a fraction of lipophilic substances was obtained by extraction with petroleum ether. This fraction was analyzed using gas chromatography-mass spectrometry (GC-MS). For this, a chromatographic column was used, which was washed with methanol (model NR-5ms, length 30 m, diameter 0.25 mm, filler: 95% dimethylpolysiloxane + 5% diphenylpolysiloxane; carrier gas ‒ helium with a constant flow of 1.5 ml/min) and 6C / MS Agilent Technologies 6890 N / 5975 B mass spectrometer (USA). The etxtraction by petroleum ether was obtained fraction of lipophilic substances in the amount of 0.83% of the taken weight of dried flowers and 0.62% by weight of dried leaves. In total, 28 compounds were detected in flowers, and 19 in leaves. It was established that flowers and leaves of Spindle Tree, like bark, are characterized by a high content of paraffinic carbohydrates (from C20H42 to C31H64). Their content is higher in flowers (more than 50% of the composition of the fraction), and in leaves and bark ≈ 35%. Steroid and terpene compounds in the lipophilic fractions of the leaves and bark of the plant are respectively 28.26% and 24.32%. Among them quantitatively dominated by freedeen. A rather high content of squalene (2.06%) was found in the leaves of the plant. Among other compounds, a significant proportion of fatty acids and their esters, of which 24.59% by weight of the lipophilic fraction of flowers and 14.74% by weight of the lipophilic fraction of leaves. An analysis of the results indicates a greater promise for the use in medicine of lipophilic substances of the bark and leaves of Spindle Tree than of flowers.

THE STUDY OF PHYSICO-TECHNOLOGICAL FACTORS’ INFLUENCE ON THE OUTPUT OF LIPOPHILIC SUB-STANCES FROM MEDICINAL PLANT MATERIALS

Currently, the lipophilic fractions of well-known medicinal plants are still understudied despite of contains of unique groups of BAS (chlorophylls, carotenoids, tocopherols, sterols, unsaturated fatty acids, phospholipids, etc.), what is the actual task of modern pharmaceutical science. The aim of our work was to perform experimental studies for the choice of optimal conditions of the extraction of lipophilic compounds from the meal of pumpkin pulp. Extraction of lipophilic substances from the meal of pumpkin pulp was carried out in laboratory conditions in the Soxhlet apparatus by the method of circulation extraction. The weight loss on drying, the content of extractive lipophilic substances and the amount of carotenoids (in terms of β-carotene) were determined by compendial procedures. According to the results of the experimental work, the optimum humidity (not more than 7%) and drying temperature (+ 60 °C) of pumpkin pulp meal containing carotenoids were established. The obtained data are necessary for the development of technology of lipophilic pumpkin extract.

Validation Cytotoxicity Assay for Lipophilic Substances

It is challenging to disperse lipophilic substances in a validated cytotoxicity assay, especially for compounds with log Kow greater than or equal to 5 that may show false negative results. The purpose of this study was to explain the challenges in conducting a cytotoxicity validated test of lipophilic substances: Minthostachys setosa, Pimenta pseudocaryophyllus, and Drimysbrasiliensis essential oils. Additionally, we compared the equivalence of Neutral Red (NR) and 3- (4,5-dimethylthiazol-2-yl) -5- (3- carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H -tetrazolium, inner salt (MTS) in detecting cell viability. The Hydrophile-Lipophile Balance (HLB) technique was used to evaluate the dispersion of essential oils and cytotoxicity in accordance to the guidelines of the OECD / GD 129 validated cytotoxicity assay. We compared the equivalence of vital dyes by TOST equivalence test. According to the results, we demonstrated the possibility of using other ways to disperse the lipophilic substances. Based on the HLB theory, we selected polysorbate 20 as the best solubilizing agent of the essential oils studied in D10 culture medium.