Metabolite Profiling of Rosemary Cell Lines with Antiproliferative Potential against Human HT-29 Colon Cancer Cells.

Rosemary (Rosmarinus officinalis) is a culinary and medicinal plant used in food and pharmaceutical industry. The wide range of biological activities is mainly related to phenolic and terpenic compounds; like carnosic acid (CA), carnosol (CS) and rosmarinic acid (RA), mainly reported in rosemary leaf extracts, and recently described in rosemary callus extracts. The aim of this work was to investigate the chemical profile of rosemary cell lines and evaluate their antiproliferative potential against human HT-29 colorectal cancer cell lines. For this purpose, rosemary leaf explants were dedifferentiated on MS medium and added with 2, 4-D (2, 4-dichlorophenoxyacetic acid; 2 mg/L) and BAP (6-benzylaminopurine; 2 mg/L). Cell aggregates were separated according to colour and three rosemary cell lines cultures were established: green (RoG), yellow (RoY) and white (RoW). The chemical profile of rosemary cell lines extracts was characterized by combining HPLC and GC platforms coupled to HR-MS/MS. The antiproliferative activity against HT-29 cell line was analyzed with MTT assay. A total of 71 compounds, including hydroxycinnamic acid and hydroxybenzoic acid derivatives, flavonoids, phenolic di- and triterpenes, as well as relevant unsaturated fatty acids and their esters, phytosterols, and carotenoids were tentatively identified in the extract of the target cell lines. The antiproliferative activity test against HT-29 cell using the MTT assay revealed that the viability of HT-29 colon cancer cells was affected after treatment with the RoW extract (IC50 of 49.63 µg/mL) at 48 h. These results showed that rosemary cell lines can also accumulate other bioactive phytochemicals with demonstrated antiproliferative potential.

Improving Diversity in Pediatric Residency Selection: Using an Equity Framework to Implement Holistic Review.

BACKGROUND: Many programs struggle to recruit, select, and match a diverse class of residents, and the most effective strategies for holistic review of applications to enhance diversity are not clear. OBJECTIVE: We determined if holistic pediatric residency application review guided by frameworks that assess for bias along structural, interpersonal, and individual levels would increase the number of matched residents from racial and ethnic groups that are underrepresented in medicine (UiM). METHODS: Between 2017 and 2020, University of California San Francisco Pediatrics Department identified structural, interpersonal, and individual biases in existing selection processes and developed mitigation strategies in each area. Interventions included creating a shared mental model of desirable qualities in residents, employing a new scoring rubric, intentional inclusion of UiM faculty and trainees in the selection process, and requiring anti-bias training for everyone involved with recruitment and selection. RESULTS: Since implementing these changes, the percentage of entering interns who self-identify as UIM increased from 11% in 2015 to 45% (OR 6.8, P = .008) in 2019 and to 35% (OR 4.6, P = .035) in 2020. CONCLUSIONS: Using an equity framework to guide implementation of a pediatric residency program's holistic review of applications increased the numbers of matched UiM residents over a 3-year period.

Chemical characterization of leaves and calli extracts of Rosmarinus officinalis by UHPLC-MS.

Rosmarinus officinalis L. (Lamiaceae) is an aromatic plant widely popular mainly due to its uses in traditional medicine as an anti-inflammatory, diuretic and antimicrobial, as well as in the prevention and treatment of diseases. These biological activities are mainly related to the presence of phenolic and terpenic compounds. This work reports a chemical profile analysis of extracts from leaves and calli of rosemary obtained by both pressurized liquid extraction and maceration. Chemical profiles were determined on calli extracts of 3, 6, 9, and 15 days of culture; chemical characterization and quantification of compounds was carried out using ultrahigh performance liquid chromatography-mass spectrometry. A total of 53 metabolites were identified in callus and 47 compounds in leaf extracts, of which 25 correspond to phenolic compounds, mainly flavonoids and flavones, 13 terpenes that include phenolic terpenes and one diterpenolactone, two glycosides which correspond to 6-O-caffeoyl-ß-D-fructofuranosil-(2→1)-α-D-glucopyranoside and primulaverin, an aromatic compound identified as fenantrenone and a growth regulator 12-hydroxy jasmonic acid. These results showed that undifferentiated rosemary cells accumulate the same compounds identified mainly in highly specialized tissues such as leaves. The plant cell culture supply the possibility of developing biotechnological processes to obtain compounds of commercial interest.

Comparative Study of Phenolic Profile, Antioxidant Capacity, and Color-composition Relation of Roselle Cultivars with Contrasting Pigmentation.

Roselle is a plant that accumulates anthocyanins significantly, hence its importance as food coloring and as a source of antioxidant compounds for human health. This study was aimed to determine phenolic composition and antioxidant capacity of methanolic extracts, and beverages obtained from native roselle cultivars in Mexico (Negra, Sudan, Rosa and Blanca) with different degrees of pigmentation, and to establish the color-composition relationship. Chromatographic methods were used to determine phenolic compounds: flavanols, flavonols, benzoic, hibiscus and phenolic acids as well as two main anthocyanins (cyanidin 3-sambubioside and delphinidin 3-sambubioside). The antioxidant capacity was evaluated by ABTS and FRAP assays. Tristimulus colorimetry showed to be a useful technique to determine the color-composition relationship, leading to equations that allowed to predict anthocyanin content of roselle (R > 0.84). Also, a stepwise linear discriminant analysis (SLDA) was developed in order to classify roselle cultivars. The obtained mathematical model could be an important tool to be used in colorimetric characterization of functional compounds used in food processing.

Vimentin filament organization and stress sensing depend on its single cysteine residue and zinc binding.

The vimentin filament network plays a key role in cell architecture and signalling, as well as in epithelial-mesenchymal transition. Vimentin C328 is targeted by various oxidative modifications, but its role in vimentin organization is not known. Here we show that C328 is essential for vimentin network reorganization in response to oxidants and electrophiles, and is required for optimal vimentin performance in network expansion, lysosomal distribution and aggresome formation. C328 may fulfil these roles through interaction with zinc. In vitro, micromolar zinc protects vimentin from iodoacetamide modification and elicits vimentin polymerization into optically detectable structures; in cells, zinc closely associates with vimentin and its depletion causes reversible filament disassembly. Finally, zinc transport-deficient human fibroblasts show increased vimentin solubility and susceptibility to disruption, which are restored by zinc supplementation. These results unveil a critical role of C328 in vimentin organization and open new perspectives for the regulation of intermediate filaments by zinc.

Callus culture development of two varieties of Tagetes erecta and carotenoid production

Background The properties of natural pigments, such as antioxidants, functional, medical, and nutraceutical, have demonstrated the advantages of these natural compounds over synthetic ones. Some products are accepted only when they are pigmented with natural, food-quality colorants: for example poultry products (manly marigold flower extracts). Carotenoids such as β-carotene, β-criptoxanthin and lutein are very attractive as natural food colorants due to their antioxidant and pro-vitamin activities which provide additional value to the target products. Marigold (Tagetes erecta) is an Asteraceous ornamental plant native to Mexico, and it is also important as a carotenoid source for industrial and medicinal purposes but nowadays its production is destined mainly for ornamental purposes. Results Friable callus of T. erecta yellow flower (YF) and white flower (WF) varieties was induced from leaf explants on Murashige and Skoog (MS) medium supplemented with 9.0 µM 4-dichlorophenoxyacetic acid (2,4-D) and 8.8 µM benzyladenine (BA). Calluses developed from both varieties were different in pigmentation. Extract characterization from callus cultures was carried out by high-performance liquid chromatography (HPLC). This analytical process detected several carotenoids; the main pigments in extracts from YF callus were lutein and zeaxanthin, whereas in the extracts of the WF callus the main pigments were lutein, zeaxanthin, β-cryptoxanthin and β-carotene. Callus cultures of T. erecta accumulated pigments even after several rounds of subculture. Conclusions WF callus appeared to be a suitable candidate as a source of different carotenoids, and tested varieties could represent an alternative for further studies about in vitro pigment production.

Estudio del desarrollo, calidad de planta y acumulación de pigmentos en lígulas de tagetes erecta l: expuestas a dos intensidades de luz / Study of development, quality of plant and accumulation of pigments in ligules of tagetes erecta l: exposed to two light intensities

The carotenoids are photosensitive pigments during photosynthesis. The objective of this work was to study the effect on development and accumulation of carotenoids in ligules of Tagetes erecta exposed under two different lighting ambient (with mesh and without mesh of 50 percent). The plant development was evaluated measuring the height of the plant, number of floral buds, the ligules diameter. In adition, the quantification and identification of carotenoids from ligules was done by HPLC. The results showed significant differences (p<0.05) in the height of the plant, number of floral buds and ligules diameter of T. erecta. The group grown without mesh received greater UV radiation and different temperature, that under a mesh. The first conditions lead to a reduction of the ligules diameter and total content of xanthophylls (lutein and zeaxanthin). The plastids ultrastructure in the cells of T. erecta developed with mesh showed the greatest amount of thylakoid membranes and more conspicuous starch granules.

Los carotenoides son pigmentos fotosensibles frente a un exceso de intensidad luminosa durante el proceso de fotosíntesis. El objetivo de este trabajo fue el estudio del efecto en el desarrollo de la planta y la acumulación de carotenoides por la exposición a dos diferentes intensidades lumínicas (con y sin malla de sombra al 50 por ciento). Se evaluó el desarrollo de T. erecta en cuanto a la altura de la planta, número de botones florales y el diámetro de las lígulas. Adicionalmente, en las lígulas se cuantificaron e identificaron los carotenoides por HPLC. Los resultados mostraron diferencias significativas (p<0.05) en cuanto al desarrollo de las plantas expuestas a mayor radiación UV y temperatura, presentaron reducción del diámetro de las lígulas y disminución en el contenido de Xantófilas totales ( luteína y zeaxantina) con respecto a las cultivadas con malla,. La ultraestructura de los plastidios mostró mayor cantidad de membranas tilacoidales y gránulos de almidón más conspicuos en las células de las plantas de T erecta desarrolladas con malla.