Exploring the Pharmacognostical and Phytochemical Profiles of Aqueous Extracts of Kalanchoe.

The use of plants for medicinal purposes has a long history, however it is desirable a continuous evaluation seeking for complementary scientific evidences for their safe application. Species within the Kalanchoe genus are often referred to as "miracle leaf" due to their remarkable healing properties. Traditionally, these plants have been used to treat infections, inflammation, and cancer. Despite their widespread use, the identification of their active components remains incomplete. This study aimed to differentiate K. crenata (KC), K. marmorata (KM), and K. pinnata (KP) by conducting detailed histochemical and phytochemical analyses, and to assess their antioxidant capabilities. The investigation revealed significant differences between the species, highlighting the variability in phenolic (PC) and flavonoid contents (FC) and their distinct antioxidant effects. The KM demonstrated the greatest results (PC: 59.26±1.53 mgEqGA/g; FC: 12.63±0.91 mgEqCQ/g; DPPH⋅ (IC50): 110.66 ug/mL; ABTS⋅+ (IC50): 26.81 ug/mL; ORAC: 9.65±0.75 mmolTE) when compared to KC and KP. These findings underscore a new reference for research within the Kalanchoe genus.

Lemongrass essential oil micro- and nanoencapsulation for industrial application: Production techniques and potential applications.

Due to its characteristic aroma and diverse therapeutic properties, lemongrass essential oil (LEO) has garnered increased attention in the pharmaceutical, food, and cosmetic industries. However, LEO's volatile nature, low chemical stability, and limited solubility in water limits its applications in the industry. Micro- and nanoencapsulation technologies emerge as a promising solution to overcome these challenges. A systematic methodology involving keyword searches in databases was employed to gather relevant literature on LEO micro- and nanoencapsulation, providing an extensive overview of techniques, processes, encapsulating materials, and possible applications. Beyond established methods, emerging techniques were explored. This review highlights the critical role of encapsulation in enhancing the thermal and chemical stability, applicability, bioavailability, and controlled release of LEO.

Preliminary Evaluation of Wound Healing Potential of Leonurus japonicus Houtt. Extracts.

Leonurus japonicus Houtt. is a medicinal plant popular in Brazil as "rubim", used in local folk medicine for several applications as an anti-inflammatory, antioxidant, analgesic, and antimicrobial phytomedicine. The traditional use for wound healing is related; however, few studies have evaluated the wound healing activity. Thus, this study aimed to analyse the popular indication of the hydroalcoholic and aqueous extracts of L. japonicus aerial parts in a rat wound healing model. The initial chemical characterization was performed using flavonoid quantification and complemented with mass spectroscopy/chemometrics analysis. The wound's lesion contraction and tissue regeneration (histological study stained with hematoxylin-eosin and picrosirius) were determined. Hydroalcoholic and aqueous extracts presented high flavonoid content, and mass spectrometry analysis of the extracts demonstrated the presence of compounds with a mass between 100-650, reinforcing the presence of polyphenolic constituents. The extracts of L. japonicus improve various wound healing phases, like inflammatory modulation, wound contraction, and collagen synthesis, resulting in faster healing in rats. These effects could be related to the extracts' polyphenolic compounds.

Study of the Antioxidant, Antimicrobial, and Wound Healing Properties of Raw Hydrolyzed Extract from Nile Tilapia Skin (Oreochromis niloticus).

Oreochromis niloticus (Nile tilapia) skin is a by-product of Brazilian fish farming, rich in collagen. The present study aims to evaluate the wound healing, antioxidant, and antimicrobial potential of the raw hydrolyzed extract of Nile tilapia skin, as well as the identification of the main compounds. The in vitro activity was performed using antioxidant, antimicrobial and scratch wound healing assays. An in vivo experiment was performed to evaluate the wound healing potential. On days 1, 7, 14 and 21, the lesions were photographed to assess wound retraction and on the 7th , 14th and 21st  days the skins were removed for histological evaluation and the blood of the animals was collected for glutamic oxaloacetic transaminase and glutamic pyruvic transaminase determination. The chemical study was carried out through liquid chromatography-tandem mass spectrometry and de novo sequencing of peptides. The in vitro assays showed a reduction of the gap area in 24 h, dose-dependent antimicrobial activity for both bacteria, and antioxidant activity. The chemical analysis highlighted the presence of active biopeptides. The histological evaluation showed that the raw hydrolyzed extract of Nile tilapia skin has a healing potential, and does not present toxicological effects; therefore, is promising for the treatment of wounds.

A Review of the Popular Uses, Anatomical, Chemical, and Biological Aspects of Kalanchoe (Crassulaceae): A Genus of Plants Known as "Miracle Leaf".

Species of the genus Kalanchoe have a long history of therapeutic use in ethnomedicine linked to their remarkable healing properties. Several species have chemical and anatomical similarities, often leading to confusion when they are used in folk medicine. This review aims to provide an overview and discussion of the reported traditional uses, botanical aspects, chemical constituents, and pharmacological potential of the Kalanchoe species. Published scientific materials were collected from the PubMed and SciFinder databases without restriction regarding the year of publication through April 2023. Ethnopharmacological knowledge suggests that these species have been used to treat infections, inflammation, injuries, and other disorders. Typically, all parts of the plant are used for medicinal purposes either as crude extract or juice. Botanical evaluation can clarify species differentiation and can enable correct identification and validation of the scientific data. Flavonoids are the most common classes of secondary metabolites identified from Kalanchoe species and can be correlated with some biological studies (antioxidant, anti-inflammatory, and antimicrobial potential). This review summarizes several topics related to the Kalanchoe genus, supporting future studies regarding other unexplored research areas. The need to conduct further studies to confirm the popular uses and biological activities of bioactive compounds is also highlighted.

Bioactive Secondary Plant Metabolites from Euphorbia umbellata (PAX) BRUYNS (Euphorbiaceae).

The species Euphorbia umbellata has been used to treat inflammatory diseases, cancer, and ulcers. Biological activities reported in the literature, including antiproliferative, cytotoxic and anti-inflammatory, are attributed to the chemical constituents present in its composition as terpenes and polyphenolic compounds. The most recurrently verified metabolites in the Euphorbiaceae family plant species are terpenes, of which euphol is a major constituent with broadly reported cytotoxic, antinociceptive and anti-inflammatory effects; it frequently appears in various extracts obtained from the plant. Euphol has a documented inhibitory effect on neutrophil chemotaxis and can modulate the complement system. Since complement system activation is intimately intertwined with autoimmune and inflammatory diseases, tumor growth promotion and metastasis, plant metabolites from Euphorbia umbellata might influence the outcomes of inflammatory processes. We believe that this is the first review presenting the current knowledge on Euphorbia umbellata secondary metabolites and their biological activities.

Polyphenolic compounds of Euphorbia umbellata (Pax) Bruyns (Euphorbiaceae) improved endothelial dysfunction through arginase inhibition.

Euphorbia umbellata is used for its anti-inflammatory properties; however, there are limited data available regarding its effects on vascular function. Its bark is rich in polyphenolic compounds, which potentially improve endothelial dysfunction (ED). This study proposes to investigate the effects of E. umbellata bark extracts and its polyphenolic compounds on arginase (ARG) activity and nitric oxide (NO)-related targets. Chromatographic procedures were used for the chemical characterisation of the extracts. Furthermore, in silico (molecular docking), in vitro (ARG inhibition), in vivo (streptozotocin-induced hyperglycemia model), and ex vivo (l-arginine metabolism, vascular reactivity, western blot, and biochemical) techniques were carried out. Quercetin, gallic acid, and ellagic acid were identified in the extracts. In silico screening predicted that gallic acid and quercetin would have the most promising interactions with ARG -identified cavities. This was confirmed in vitro as both compounds had a direct inhibitory effect on ARG, as was the case regarding the extracts. Oral treatment preserved endothelium-dependent vasodilation through ARG inhibition together with an increase in l-arginine bioavailability and endothelial NO synthase expression. Biochemical parameters determined the lack of toxicity for sub-chronic treatment. E. umbellata bark extracts and its compounds can contribute to ED treatment, at least partly, through the inhibition of vascular ARG.

Enriched Terpenes Fractions of the Latex of Euphorbia umbellata Promote Apoptosis in Leukemic Cells.

The current study was carried out by a bioguided fractionation of a hexane extract of the latex of Euphorbia umbellata against leukemic cells. Samples were analyzed by NMR, GC/MS, triterpenes quantification, and MTT reduction assay. Morphological, cell cycle, mitochondrial membrane potential and caspases 3/7 analyses were performed for the dichloromethane and ethanol fractions, and selectivity index for the dichloromethane fraction. NMR analysis presented characteristic signals of terpenes and steroids, data were confirmed by the quantification of triterpenes and GC/MS analysis. MTT reduction assay demonstrated that HL-60 was the most sensitive cell lineage against dichloromethane and ethanol fractions. Compounds of these matrices caused morphological changes compatible with apoptosis induction, altered cell cycle, increment of depolarized population cells and activation of caspases 3/7. Selectivity indices were higher than 22.44. Bioguided-fractionation study showed that samples of the latex of E. umbellata raised the activity of the phytocomplex against leukemic cells, and the cytotoxicity can be associated with an apoptosis pathway.

Phenolic Compounds as Arginase Inhibitors: New Insights Regarding Endothelial Dysfunction Treatment.

Endothelial dysfunction is characterised by the low bioavailability of nitric oxide with a relevant negative impact on the nitric oxide/cGMP pathway. The loss of nitric oxide/cGMP signaling may be caused by an increased arginase activity. Plant-derived substances, especially polyphenols, are compounds that have the potential to inhibit arginase activity and they may represent an attractive therapeutic option to combat clinical outcomes related to endothelial dysfunction. An extensive review was carried out using all available data published in English in the Pubmed database, and without restriction regarding the year of publication. Despite the increased number of new substances that have been tested as arginase inhibitors, it is rare to find a compound that satisfies all the toxicological criteria to be used in the development of a new drug. On the other hand, recent data have shown that substances from plants have great potential to be applied as arginase inhibitors, most of which are polyphenols. Of the relevant mechanisms in this process, the inhibition of arginase by natural products seems to act against endothelial dysfunction by reestablishing the vascular function and elevating nitric oxide levels (by increasing the amounts of substrate (L-arginine, and endothelial nitric oxide synthase activation and stabilisation) as well as decreasing the generation of reactive species (formed by uncoupledendothelial nitric oxide synthase). This review summarises several topics regarding arginase inhibition by natural substances as well as indicating this pathway as an emergent strategy to elevate nitric oxide levels in disorders involving endothelial dysfunction. In addition, some aspects regarding structural activity and future perspectives are discussed.

Cytotoxic biomonitored study of Euphorbia umbellata (Pax) Bruyns.

ETHNOPHARMACOLOGICAL RELEVANCE: Euphorbia umbellata latex (sap) has normally been used in folk medicine in southern Brazil to treat different types of cancers. AIM OF STUDY: To carry out a biomonitored investigation of partitioned latex using in vitro assay, to identify the main mechanisms related with the action of the most active fraction as well as to develop a phytochemical study with this material. MATERIALS AND METHODS: Biological screening was performed with hexane, chloroform, ethyl acetate and methanol fractions from the latex of E. umbellata using MTT, trypan blue, and neutral red assays to determine the cytotoxicity against HRT-18, HeLa and Jurkat cells and flow cytometry, DNA quantification, acridine orange and Hoechst 33342 staining to investigate mechanisms of action for the hexane extract. The phytochemical study of the hexane fraction was performed by chromatographic procedures and the substances were identified by NMR analysis. The isolated terpenes were evaluated using MTT to determine the cytotoxicity against Jurkat cells. RESULTS: All the fractions presented concentration and time dependent cytotoxicity. The hexane fraction showed the highest cytotoxicity; whereas the Jurkat cell was the lineage with the highest sensitivity (IC50 1.87µg/mL). Fragmentation of DNA and apoptosis are two mechanisms related with the toxicity of hexane fraction. The hexane fraction arrested the cell cycle in the G0/G1 phase, and the selectivity index was 4.30. Phytochemical study of the hexane fraction led to isolation of euphol (main compound) and germanicol acetate. Both substances demonstrated some slight cytotoxic activity against Jurkat cells after 72h; however the activity was minimal compared to vincristine (anticancer standard drug). CONCLUSION: The current research proves that the fractions of the latex from E. umbellata have a cytotoxic effect against three different cancer cells lines. The hexane fraction showed high in vitro cytotoxic effects against Jurkat cells demonstrating that the effect may be due to non-polar constituents. The two isolated terpenes (euphol and germanicol acetate) showed poor cytotoxic activity indicating that the anticancer properties of the extract may be caused by other substances present in the hexane fraction.

Avaliação preliminar da associação entre anfotericina e hesperina com oxidantes e células huvec / Preliminary assessment of the association between amphotericin and hesperetin with oxididants and huvec cells

A anfotericina B (AmB) é fármaco o “padrão ouro” para o tratamento de infecções fúngicas invasivas desde 1960, Entretanto, a anfotericina B apresenta elevada toxicidade, a qual manifesta-se mais frequentemente nos rins e no fígado, Sabe-se, desde 1985, que a auto-oxidação da AmB origina diferentes formas de espécies reativas oxidativas e estas, por serem tóxicas para a célula, seriam responsáveis, em parte, pela toxicidade, Diferentes estudos indicam que a hesperidina contribui por meio do decréscimo do estresse oxidativo, para a proteção renal e contra a injúria gerada pela isquemia, Tal fato e o envolvimento da AmB na geração de radicais livres tornam interessante a avaliação preliminar do efeito da hesperidina e AmB (isoladamente ou associadas) frente a espécies reativas do oxigênio e radicais livres, bem como o estudo das mesmas em modelos de citoxicidade, Frente ao ABTS•+, a AmB apresentou IC50 igual a 0,0124mg/mL, mas quando associada à hesperidina este valor caiu para 0,0003mg/mL, Frente ao HOCl, a Amb apresentou IC50 igual a 0,0056, mas quando associada à hesperidina este valor caiu para 0,0023mg/mL, No ensaio com DPPH• e radical ânion superóxido as amostras não foram efetivas, No ensaio com células endoteliais em cultivo (HUVEC), as associações reduziram a viabilidade celular, Estes resultados preliminares evidenciam a interação dos compostos com espécies reativas bem como indicam possibilidade de exacerbação do dano pela AmB na presença dos antioxidantes em um modelo in vitro...

Amphotericin B (AmB) is drug “gold standard” for the treatment of invasive fungal infections since 1960. However, amphotericin B has high toxicity, which manifests itself most often in the kidneys and in the liver. It is known, since 1985, that self-oxidation of AmB gives different forms of reactive oxidative species and these, being toxic to the cell, would be responsible, in part, by its toxicity. Different studies indicate that hesperidin contributes, through the reduction of oxidative stress, to protect against renal injury generated by ischemia. This fact and the involvement of AmB in the generation of free radicals make it interesting the preliminary evaluation of the effect of hesperidin and AmB (alone or associated) against reactive oxygen species and free radicals, as well as the study on models of cytotoxicity. Front ABTS•+, AmB presented IC50 equal to 0.0124 mg/mL, but when it was associated to hesperidin this value has decreased to 0.0003 mg/mL. Front HOCl, Amb presented IC50 equal to 0.0056, but when it was associated to hesperidin this value has decreased to 0.0023 mg/mL. In the trials with DPPH• and the superoxide anion radical samples were not effective. In the assay with endotelial cell culture (HUVEC cells), the association has decreased cell viability. These preliminary results demonstrate the interaction of the compounds with reactive species as well as indicate the possibility of damage exacerbation by AmB in the presence of antioxidants in an in vitro model...

Antitumoural effect of Synadenium grantii Hook f. (Euphorbiaceae) latex.

ETHNOPHARMACOLOGICAL RELEVANCE: Synadenium grantii Hook f. has traditionally been used to treat various neoplastic diseases in southern Brazil. AIM OF STUDY: Evaluation of the antitumoural potential of Synadenium grantii latex against B16F10 melanoma cell line using in vitro and in vivo models, as well as a phytochemical study of the latex. MATERIALS AND METHODS: The in vitro antitumoural activity was performed using MTT and trypan blue assays with different latex concentrations (1.7 µg-7.0 µg/well and 1.22 mg-4.88 mg/well). Flow cytometry was used to determine the progression of the cell cycle. The in vivo activity was performed by subcutaneously injecting melanoma cells in the dorsum of C57BL6 mice, followed by treating the mice with a popular form of use of the latex (garrafada) administered orally. After sacrificing the animals, histological analysis of the organs was performed by hematoxylin-eosin staining. The phytochemical study of the latex was performed by NMR and chromatographic procedures and the extracts and isolated substances were evaluated by IR, 1D and 2D NMR analysis. RESULTS: The Synadenium grantii latex exhibited decreased cell viability of the melanoma line in a concentration and time-dependent manner, and also cell cycle arrest in the S-G2/M phase. The latex caused a 40% reduction in the volume of tumours of the mice with melanomas. Histological examination of the organs of these animals showed no differences between groups. The phytochemical investigation resulted in the isolation and identification of triterpene euphol and the steroid citrostadienol, which were tested against the strain of melanoma. Euphol showed no antitumoural activity, while the steroid citrostadienol showed reduced cytotoxic activity. CONCLUSION: The Synadenium grantii latex presented in vitro and in vivo cytotoxic effects with antitumoural activity against B16F10 melanoma cells.

Determinação de ferro (III) em produtos farmacêuticos por titulação fotométrica / Determination of iron (III) in pharmaceutical products by photometric titration

Este trabalho descreve a montagem de um sistema de titulação fotométrica simples e de baixo custo para a determinação de ferro (III) em produtos farmacêuticos. O sistema de titulação fotométrica foi construído utilizando-se a bomba peristáltica de um espectrofotômetro convencional. O procedimento é baseado na titulação de ferro (III) com EDTA e ácido salicílico como indicador. A absorção do complexo ferro (III)-ácido salicílico foi monitorada espectrofotometricamente em 525 nm. O limite de quantificação foi de 5 µg de ferro (III). O procedimento de titulação fotométrica foi aplicado para a determinação de ferro (III) em amostras contendo sulfato ferroso e hidróxido férrico polimaltosado. O procedimento mostrou sensibilidade, reprodutibilidade e precisão para a utilização em análise rotineira de ferro (III) em produtos farmacêuticos.

This paper describes a simple, precise and low-cost photometric titration method for iron (III) determination in pharmaceutical preparations. The photometric titration system was constructed using the peristaltic pump of a conventional spectrophotometer. The method is based on titration of iron (III) with EDTA using salicylic acid as indicator. The absorption of the iron (III)-salicylic acid complex was monitored spectrophotometrically at 525 nm. The limit of quantification was 5 µg of iron (III). The photometric titration procedure was applied for the determination of iron (III) in samples of ferrous sulfate and ferric hydroxide polymaltose complex. The procedure showed sensibility, reproducibility and accuracy for use as a method for the routine analysis of iron (III) in pharmaceutical formulations.

Determinação de compostos fenólicos em amostras comerciais de chás verde e preto - Camellia sinensis (L. ) Kuntze, Theaceae / Determination of phenolic componds in commercial samples of green and black tea - Camellia sinensis (L. ) Kuntze, Theaceae

O chá de Camellia sinensis é rico em compostos fenólicos antioxidantes, responsáveis por efeitos benéficos à saúde humana. Neste trabalho, foram realizadas as determinações da concentração de fenóis totais e flavonoides de sete amostras comerciais dos chás preto e verde. Os teores de fenóis totais variaram de 4,80 a 26,60 mg de pirogalol g-1 de amostra e os de flavonoides, de 0,46 a 1,10 mg de quercetina g-1 de amostra. A análise anatômica do conteúdo das amostras permitiu caracterizar a autenticidade dos produtos, observando-se os marcadores anatômicos típicos da espécie (estômatos anomocíticos, tricomas tectores unicelulares, mesofilo dorsiventral, drusas de oxalato de cálcio e esclereides).

The tea from Camellia sinensis (green tea and black tea) is rich in antioxidant phenolic compounds, responsible for beneficial effects to human health. In this work, determinations were carried out on the concentration of total phenols and flavonoids in seven commercial samples of black tea and green tea. The samples presented concentrations of total phenols varying between 4.80 and26.60 mg of pyrogallol g-1 and concentrations of flavonoids from 0.46 to 1.10 mg of quercetin g-1. With the purpose of characterizing the authenticity of the product contained in the sachets, the samples were fixed for histological studies. The samples presented the typical anatomical markers of the species (anomocytic stomata, unicellular non-glandulartrichomes, dorsiventral mesophyll, calcium oxalate druses and sclereids).

Avaliação da qualidade das amostras comercias de Baccharis trimera L. (Carqueja) vendidas no Estado do Paraná / Quality evaluation of commercial samples of Baccharis trimera L. (Carqueja) sold in Paraná State

O presente trabalho descreve a avaliação da qualidade de 12 amostrascomerciais de "carqueja" (Baccharis trimera L.) que são empregadas na medicina tradicional como estomáquicas e diuréticas e são vendidas em estabelecimentos comerciaisfarmacêuticos no Estado do Paraná, visando ao preparo de chás. Foram avaliados parâmetros de qualidade preconizados na Farmacopéia Brasileira e na literatura específica: característicasdos rótulos e bulas, análise sensorial, autenticidade das amostras, material estranho e doseamento de marcadores químicos. Todos os produtos avaliados apresentaram algum tipo de irregularidade segundo os códigos oficiais, sendo necessária maior intensificação na vigilância de produtos à base de plantas medicinais.

This study aimed to evaluate the quality of 12 samples prepared with "carqueja" (Baccharis trimera L.) used in traditional medicine as stomachic anddiuretic medicines, and sold in commercial pharmacies in Paraná State. The analyses used parameters from the Brazilian Pharmacopoeia and specific literature. The methodology consisted in evaluating the product labels and instructions of the samples to verify theiraccordance to the specific legislation, sensorial analysis, authenticity of the samples, purity and dosage of chemical markers. All products showed some type of mistake or irregularity according to the official codes. These results showed that is necessary to intensify theinspection of medicinal plants in Brazil.

Quantitative determination of 20-hydroxyecdysone in methanolic extract of twigs from Vitex polygama Cham.

20-Hydroxyecdysone (20E) is effective in stimulating protein synthesis, therefore, it has been largely used as anabolic agent in several commercial formulas. Phytochemical study of methanolic extract of twigs from Vitex polygama, used in traditional Brazilian medicine as emenagogue, yielded a large quantity of 20E. This finding led us to developing and validating a simple and reliable method to determine 20E in the surveyed extract. Chromatographic separation of 20E was achieved on a phenyl-hexyl-based column using reversed elution mode. Extract was cleaned-up by solid phase extraction employing C(18) cartridge, and an absolute recovery of 97% was acquired. External standard and standard addition calibration graphs were obtained and good linearity was accomplished (r>0.999 for both curves). The limit of quantification and detection were determined. The results for accuracy fell within the -5 to +7% range.