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Introduction:Kalanchoe pinnataextracts have been shown to possess beneficial cardiovascular effects, analgesic and myorelaxant activities. This study aimed at evaluating the haematological and anti-fatigue effects of its aqueous extract. Materials and methods: This was anin vivopharmacological experiment, carried out in the Laboratory of Zoology, University of Buea, from January to May 2018. A total of 48 mice were subjected to a 90mins/day-free endurance swimming test for 14 days. Two groups of mice received distilled water and 12% NaCl solution, respectively. Five groups received the plant extract (25, 50, 100 and 200 mg/kg/day) and 200mg/kg/day+NaCl. One positive control group received 5% Vitamin C (1.97mg/kg/day). All administrations were by gavage. Maximum swimming time, glycaemia, lactatemia, uremia, triglyceridemia, haematological indices, tissue glutathion peroxidase, malondialdehyde, glucose and creatine kinase-MB in the heart or gastrocnemius muscle were measured. Results:K. pinnata(50 and 100mg/kg/day) induced a non-significant increase of the swimming duration, compared to neutral control. However, the 12%NaCl group recorded a significant (PK. pinnata(200mg/kg/day). Conclusions: Significant biochemical variations support the hypothesis thatK. pinnatacould be used to delay exercise induced fatigue.&nbsp

The aim of this study was to analyze the chemical components and evaluate the biological activity of the extracts from the leaves Kalanchoe pinnata and Kalanchoe daigremontiana, which are cultivated in the province of Chiriqui, Republic of Panama. Phytochemicals components, antioxidant and anti-inflammatory activities were studied. The composition of the obtained petroleum ether, ethanol and aqueous extracts was analyzed by phytochemical screening. The antioxidant activity of the extracts was studied using three in vitro model systems (DPPH radical scavenging assay, nitric oxide radical scavenging assay, and superoxide radical scavenging activity). The anti-inflammatory activity of these species was studied using an in vivo model (ʎ-carrageenan-induced paw edema in rats). Phytochemical analysis of the extracts showed the presence of alkaloids, steroids, triterpenes, flavonoids, phenolic compounds, saponins and glycosides. The greatest radical inhibitory effect was observed in the DPPH model where the ethanolic extracts of both species developed a concentration-dependent inhibitory effect, the K. pinnata extract reached a maximum inhibitory effect of 49.5 ± 5.6% (2000

Bryophyllum pinnatum (Lam) Pers. (Crassulaceae) is widely used in folk medicine as leaf juice, aqueous, or hydro-ethanolic extracts. It is also listed as a medicinal plant in several countries such as France and Brazil. The main reported constituents are flavone glycosides, especially those with the rare 3-O-α-l-arabinopyranosyl-(1 → 2)-α-l-rhamnopyranoside moiety. Despite several phytochemical screenings indicating the presence of cyanide derivatives or alkaloids, there are no reports of nitrogenous metabolite characterization from this plant species. Nevertheless, the occurrence and the type of such compounds are of particular interest, as they may account for some of the numerous biological activities and ethnomedicinal uses described for B. pinnatum and could be regarded as chemical/taxonomic markers. Consequently, a hydro-ethanolic extract of B. pinnatum was investigated by using UHPLC-HRMS/MS and the nitrile glucoside sarmentosin was detected for the first time within the genus Bryophyllum/Kalanchoe. Considering the wide use of B. pinnatum and its closely related species for health purposes, the target metabolite was isolated by a combination of centrifugal partition chromatography in elution/extrusion mode and MPLC in order to confirm its structure. A linear, selective, precise, fast, and reliable 1H NMR quantitation method was then developed and validated and may become a tool for easy quality assessment of the plant species. The amount of sarmentosin was determined as 2.07% of the examined sample. Sarmentosin was also detected in Kalanchoe laciniata, confirming the occurrence of this compound within the genus.

Latterly, the green synthesis of metallic nanoparticles has stock pile and terminal interest over the last decade due to their idiosyncratic properties that make them felicitous in various fields of science and technology. In this study the silver nanoparticles can be manufacture by using the leaf extract of Kalanchoe pinnata plant using microwave radiation method. The nanoparticles that are synthesized by plants are non-toxic and eco-friendly and it is characterized by using UV-Visible, FT-IR , X-ray diffraction (XRD) and TEM.The crystalline natures of the synthesized silver nanoparticles were identified by XRD and its shape was confirmed by TEM analysis. Anti-urolithiatic activity of synthesized silver nanoparticles was accomplished which reduces the growth of kidney stone which was appraisal by using the In-Vitro growth of urinary calculi.

A large population is suffering from multifactorial urolithiasis worldwide with a reoccurrence rate of almost 70%–80% in males and 47%–60% in females. In the present study, the nephroprotective effect of silver nanoparticles (AgNPs) synthesised by Bryophyllum pinnatum was evaluated in ethylene glycol‐induced urolithiasis in rat. B. pinnatum‐mediated AgNPs which were found to be spherical and polydispersed particles with an average size of 32.65 nm determined by transmission electron microscopy analysis, and showing an absorption peak at 432 nm by the UV‐Vis spectrophotometric analysis, revealing the role of hydroxyl group in the synthesis by Fourier Transformed Infrared Spectroscopy analysis, with a zeta potential value of −15.7 mV. The crystalline nature and fcc structure was demonstrated based on X‐ray diffraction analysis. Animal study was performed on 36 male Wistar rats divided into six equal groups, which demonstrated significant increase in serum total protein, albumin and globulin and significant decrease in AST, ALT, creatinine, BUN, calcium and phosphorus in group V and VI when compared with group II and IV. No crystalluria was observed in rats given B. pinnatum AgNPs. Histopathological observations in group V and VI showed mild degenerative changes and restoration or maintenance of kidney parenchyma when compared with group II and IV rats. Thus, the authors conclude with the beneficial preventive and therapeutic nephroprotective effect of B. pinnatum‐mediated AgNPs against ethylene glycol‐induced urolithiasis in rats.

The evaluation of toxic effects of nanoparticles (NPs) has become an important aspect of Nanotechnology research in the 21st century. The present investigation deals with the green synthesis of biogenic zinc oxide nanoparticles (ZnO-NPs) using Bryophyllum pinnatum leaves, their characterization and evaluation of acute oral toxicity in Wistar rats. The characterization of synthesized ZnO-NPs revealed maximum absorbance at 307 nm on UV-Vis spectrophotometric analysis, NTA showed mean size of particles and mode of the particles distribution as 128.2 nm and 12.6 nm, respectively. Zeta potential was found to be −0.369 mV. The absorbance shown by FTIR at 3469, 1644, 1355 and 887 cm-1 indicates the involvement of biomolecules that are accountable for capping and stabilization of ZnO-NPs. The XRD assessment further demonstrated the crystalline nature of the ZnO-NP. The TEM analysis of the synthesized ZnO-NPs revealed the presence of spherical NPs with the mean size of 3.7 nm. The acute oral toxicity evaluation in rat showed an approximate median lethal dose to be more than 2000 mg/kg body weight. It is thus concluded that biogenic ZnO-NPs showed absence of acute oral toxicity symptoms at the doses employed in the present study.

Leaves of Kalanchoe pinnata are used worldwide for healing skin wounds. This study aimed to develop and compare two creams containing a leaf aqueous extract of K. pinnata (KP; 6%) and its major flavonoid [quercetin 3-O-α-L-arabinopyranosyl-(1→2)-α-L-rhamnopyranoside] (0.15%). Both creams were topically evaluated in a rat excision model for 15 days. On the 12th day, groups treated with KP leaf-extract and KP major flavonoid creams exhibited 95.3 ± 1.2% and 97.5 ± 0.8% of healing, respectively (positive control = 96.7 ± 0.8%; negative control = 76.1 ± 3.8%). Both resulted in better re-epithelialization and denser collagen fibres. Flavonol glycosides are the main phenolics in KP leaf-extract according to HPLC-ESI-MS/MS analysis. KP major flavonoid plays a fundamental role in the wound healing. The similar results found for both creams indicate that the use of KP crude extract should be more profitable than the isolated compound.

Kalanchoe delagoensis is adapted to intense solar irradiation, drought, and heat, partially due to the presence of phenols, important photo-protective compounds and antioxidants. This study aimed to evaluate the distribution of flavonoids and phenolic acid derivatives throughout the erect-tubular leaves of K. delagoensis. Specimens grown under sunny conditions were used for histochemical and high-performance liquid chromatography coupled with diode array detection (liquid HPLC-DAD) analysis. The NP (2-aminoethyl diphenylborinate) test suggested the presence of phenolic acids throughout the leaf blade below the epidermis and in chloroplasts, mainly in the leaf base. Flavonoids were detected specifically in chloroplasts, on the adaxial side of the middle third and at the leaf apex, near the meristematic cells. There was a tendency of flavonoid accumulation from the middle third to the apex, especially surrounding the gem, while phenolic acids were observed mainly in the base. This can be explained by the more exposed leaf apex and to the presence of apical buds (high production and regulation sites of ROS). The HPLC-DAD analysis showed different classes of flavonoids and phenolic acid derivatives in the leaf extracts, agreeing with the NP test results. This is the first time that the substitution of phenolic acids by flavonoids from the leaf base to the apex has been described.

Diabetes mellitus (DM) is one of the major health problems. One way to deal with diabetes mellitus is by inhibiting the work of enzymes that hydrolyze carbohydrates, thereby reducing glucose absorption. The enzyme that plays a role in the breakdown of starch into simple sugars is the α-amylase. The genus Kalanchoe or Panda plant has been widely reported to contain a variety of secondary metabolites that have several activities such as antimalarial, antibacterial, antidiabetic, and anticancer potential. This study aims to isolate, determine the chemical structure, and test the inhibitory activity of α-amylase from the ethyl acetate fraction of K. tomentosa leaves. The isolation stage began with the maceration process with n-hexane and fractionated with ethyl acetate. Ethyl acetate extract was refracted using a gradient liquid vacuum chromatography (KCV) method (n-hexane: ethyl acetate: methanol). Repeated gravity columns separated the fraction of KCV results and preparative thin-layer chromatography (TLC-P) to obtain isolate pure. Isolate pure was characterized using several spectroscopic methods, including UV-Vis, IR, NMR. Determination of the structure of the compound against isolate pure shows that this isolate is a 3,7,4’-trihydroxyflavonol or kaempferol compound. The results of the test activity of 3,7,4’-trihydroxyflavonol compounds in inhibiting the enzyme α-amylase in vitro showed an IC50 value of 346 µU/mL. The results obtained indicate that the IC50 value is higher than that of the Acarbose, which was 39.3 µU/mL.

This is the first investigation on the in vitro cytotoxicological and genotoxicological effects of Kalanchoe pinnata (Lam.) Pers. fresh leaf juice, for establishing a safe and effective quantity fo...

Species of kalanchoe are rich in bioactive compounds and are widely used in folk medicine; however, these plants are not well known from the point of view of aroma. Two species, Kalanchoe pinnata and Kalanchoe daigremontiana, were examined after six months and two years of growth and their vitamin C content, succulence, and aroma composition were determined. The efficiency of juice extraction was highest (72%) for the leaves of K. daigremontiana after six months of growth. The concentration of vitamin C was highest in juices from two-year-old plants and much higher in the juice of K. pinnata (81 mg/100 g). SPME/GC/MS analysis identified 32 aroma components, considering those with the spectrum similarity over 75%. The main components were furan-2-ethyl, hexanal, 2-hexenal, 2,4-hexadienal, 1-octen-3-ol, nonanal. The quantitative relations of these compounds were somewhat different in the two species. The most dominant component, 2-hexenal, is responsible for the green-like aroma noted by the sensory panel.

RESUMEN Objetivo: Evaluar la actividad antioxidante y antiinflamatoria en extractos hidroalcohólicos de las partes aéreas de Kalanchoe daigremontiana Raym. -Hamet & H. Perrier. Materiales y métodos: Se obtuvieron extractos hidroalcohólicos utilizando como solventes etanol (EtOH) y metanol (MeOH) en diferentes concentraciones. Los fenoles totales y los flavonoides se cuantificaron con el método de Folin Ciocalteu y el de complejación de aluminio respectivamente. Metodología: Para determinar la actividad antioxidante en los extractos se utilizó la técnica de decoloración del radical libre DPPH» (2,2-difenil-1-picrilhidracilo) y para la actividad antiinflamatoria se evaluó la capacidad inhibitoria de la enzima lipoxigenasa (LOX). Resultados: El extracto etanólico al 70% presentó mayor capacidad antioxidante frente a los radicales DPPH» y mayor inhibición sobre la enzima lipooxigenasa como respuesta antiinflamatoria, lo cual estaría relacionado con el contenido de fenoles y flavonoides. Conclusiones: Esto sugiere que el K. daigremontiana posee propiedades antioxidantes y antiinflamatorias. SUMMARY Aim: To evaluate the antioxidant and anti-inflammatory activity in hydroalcoholic extracts of the aerial parts of Kalanchoe daigremontiana Raym. -Hamet & H. Perrier. Materials and methods: Extracts were obtained using ethanol (EtOH) and methanol (MeOH) as solvents at different concentrations. Total phenols and flavonoids were quantified using the Folin-Ciocalteu method and the aluminum complexation reaction, respectively. Methodology: The antioxidant activity of the extracts was determined using the DPPH» (2,2-diphenyl-1-picrylhydracil) free radical discoloration technique, while lipoxygenase (LOX) inhibition assays were used to measure the anti-inflammatory activity. Results: The 70% ethanolic extract showed a greater antioxidant capacity against DPPH» and a greater inhibition of the lipoxygenase enzyme, which should be related to the overall phenols and flavonoids content, suggesting that K. daigremontiana exhibits antioxidant and anti-inflammatory properties. RESUMO Objetivo: Avaliar a atividade antioxidante e anti-inflamatória em extratos hidroalcoólicos da parte aérea de Kalanchoe daigremontiana Raym. -Hamet & H. Perrier. Materiais e métodos: Os extratos foram obtidos usando etanol (EtOH) e metanol (MeOH) como solventes em diferentes concentrações. Fenóis e flavonóides totais foram quantificados pelo método de Folin-Ciocalteu e pela reação de complexação com alumínio, respectivamente. Metodologia: A atividade antioxidante dos extratos foi determinada pela técnica de descoloração do radical livre DPPH» (2,2-difenil-1-picrilidracila), enquanto ensaios de inibição da lipoxigenase (LOX) foram utilizados para medir a atividade anti-inflamatória. Resultados: O extrato etanólico 70% apresentou maior capacidade antioxidante contra DPPH» e maior inibição da enzima lipoxigenase, que deve estar relacionada ao teor geral de fenóis e flavonoides, sugerindo que K. daigremontiana apresenta propriedades antioxidantes e anti-inflamatórias.

The responses of stomatal aperture to light intensity and CO2 concentration were studied in both Vicia faba (C3) and Kalanchoë fedtschenkoi (Crassulacean acid metabolism; CAM), in material sampled from both light and dark periods. Direct comparison was made between intact leaf segments, epidermises grafted onto exposed mesophyll, and isolated epidermal peels, including transplantations between species and between diel periods. We reported the stomatal opening in response to darkness in isolated CAM peels from the light period, but not from the dark. Furthermore, we showed that C3 mesophyll has stimulated CAM stomata in transplanted peels to behave as C3 in response to light and CO2. By using peels and mesophyll from plants sampled in the dark and the light period, we provided clear evidence that CAM stomata behaved differently from C3. This might be linked to stored metabolites/ions and signalling pathway components within the guard cells, and/or a mesophyll-derived signal. Overall, our results provided evidence for both the involvement of guard cell metabolism and mesophyll signals in stomatal responses in both C3 and CAM species.

Background: Parasitic infections represent one of the main public health problems in humans according to the WHO. Therefore, the need has arisen to find new treatments that can be used as an alternative cure to parasitosis. We aimed to investigate the in-vitro effects of the methanolic extract of Kalanchoe daigremontiana as well as its main component, quercetin against Entamoeba histolytica and Trichomonas vaginalis. Methods: For this purpose, the in-vitro activity of the methanol extract of K. daigremontiana also its main component, quercetin, against trophozoites of E. histolytica and T. vaginalis was evaluated, using the microassay technique. Furthermore, the antioxidant activity was determined. Finally, the cytotoxic and cytoprotective capacity was determined using the hemolysis technique. Results: The IC50 indicated that quercetin significantly (P < 0.05) inhibited the growth rate of the trophozoite stage of E. histolytica and T. vaginalis in comparison to the methanolic extract of K. daigremontiana (KalL). Also, quercetin significantly (P < 0.05) was a better antioxidant as compared with the positive control. In the evaluation of cytotoxicity effects, it could be observed that KalL as compared with quercetin exhibited more cytotoxicity against human erythrocytes. Quercetin significantly (P < 0.001) exhibited better cytoprotective activity compared to KalL. Conclusion: Both K. daigremontiana methanolic extract and quercetin alone demonstrated high antiparasitic activity against E. histolytica and T. vaginalis. However, the in-vivo efficacy of K. daigremontiana and quercetin also requires to be evaluated using an animal model.

The objective of this investigation was to study the effects of cycocel (CCC) and paclobutrazol (PBZ) on Kalanchoe blossfeldiana, L. to produce plants more compact, shorter, with a plurality of branches and inflorescences, flowering earlier, and to be more suitable as a pot plant. Different concentrations of CCC at 1000, 2000, and 3000 ppm and PBZ 50, 100, and 150 ppm were applied as foliar spray for once (after 45 days) or twice (after 45 and 60 days from planting). All treated plants were shorter with profuse branches specially the ones which have been treated with PBZ at 150 ppm twice (6.00 branches/ plant), and by the same treatment achieved the maximum number of inflorescences as it recorded 5.00 inflorescences/ plant. Sprayed plants with PBZ at 150 ppm twice gave the minimum number (125.33) of days to flowering. The total chlorophyll and carbohydrates content were increased in all sprayed plants, as it was 0.679 mg/ g FW for chlorophyll (PBZ 100 ppm twice) and 207.07 mg/ g DW for carbohydrates (PBZ 150 ppm twice). For compressed Kalanchoe plants with dark green leaves, abundant branches, abundant blooms and early blooms, paclobutrazol may be recommended at 150 ppm twice.

Objective: The main purpose of the present study was to analyze the antidiabetic activity of methanolic extract of the leaf of Kalanchoe pinnata in alloxan-induced diabetic rats. Methods: Diabetes mellitus (DM) is a chronic metabolic and endocrine disease regarded as a serious global public health problem. Albino Wistar rats were divided into four groups. Group I (normal) received drinking water throughout the course till 20 days. Groups II-IV received alloxan (120 mg/kg b.w) i.p on the 1st day of the study period. Group III animals received glibenclamide (10 mg/kg p.o) and Group IV received K. pinnata (200 mg/kg b.w p.o) for 20 days. The body weight, blood glucose level, serum urea, blood urea nitrogen (BUN), creatinine, cholesterol, total protein (TP), uric acid, and tissue (liver) antioxidant parameters: Malondialdehyde (MDA) and glutathione (GSH) were measured. Results: K. pinnata treated rats showed the percentage increase in the body weight, decrease in the blood sugar level, creatinine, TP level, urea, uric acid, and BUN, MDA, and GSH when compared to alloxan-induced diabetic mellitus control rats. Thus, K. pinnata could be possibly employed to treat DM. Conclusion: This preliminary analysis tested the most promising profile. It seems that methanolic extract of the leaf of K. pinnata improved general, blood serum, and liver antidiabetic agent. However, further studies confirming its potential is certainly warranted.

Kalanchoe pinnata (Lam.) Pers. has many pharmacological properties such as anticancer, antidiabetic, insecticidal, antimicrobial, anti-urolithiatic, etc. The plant leaves are used as ethno medicine traditionally. Many diseases are associated with oxidative stress caused by free radicals. The aim of the present study was to evaluate physicochemical, qualitative and quantitative phytochemical analysis and in vitro antioxidant activities of leaf of Kalanchoe pinnata collected from Bhopal region of Madhya Pradesh.The physicochemical evaluations carried out in terms of loss on drying, ash value, extractive values and acid insoluble ash value ect. Qualitative analysis of various phytochemical constituents and quantitative analysis of total phenolics and flavonoids were determined by the well-known test protocol available in the literature. Quantitative analysis of phenolic and flavonoids was carried out by FolinsCiocalteau reagent method and Aluminium chloride method respectively. The In vitro antioxidant activity of ethanolic extract of the leaf was assessed against DPPH assay method using standard protocols. Phytochemical analysis revealed the presence of phenols, flavonoids, tannins, saponins, alkaloids. The total phenolics content of leaves ethanolic extract was (4.63mg/100mg), followed by flavonoids (2.03mg/100mg). The activities of ethanolic leaves extract against DPPH assay method were concentration dependent with IC 50 values of ascorbic acid and extracts 24.44 and 49.81μg/ml respectively. These studies provided information for standardization and correct identification of this plant material. The diverse array of phytochemicals present in the plant thus suggests its therapeutic potentials which may be explored in drug manufacturing industry as well as in traditional medicine. Keywords: Kalanchoe pinnata, Physicochemical, Qualitative, Quantitative phytochemical, Antioxidant

This study provides a very first report on the use of a sprayable formulation of 1-MCP, HarvistaTM, on ornamental plant species. Individual florets of Phalaenopsis and inflorescences of Kalanchoe b...

Benign Prostatic Hyperplasia (BPH) affects mainly older men. It is estimated to affect 50% of 51-60-year-old men and 70% of 61-70-year-old men. BPH is a nonmalignant proliferation of epithelial and stromal cells of the prostate gland regions. Despite the use of conventional pharmacological therapy, herbal medicines are used in BPH therapy, and several mechanisms of action have been suggested based on their complex chemical composition. Considering the ethnomedicinal uses ofKalanchoe gastonis-bonnieri(KGB), we evaluated the inhibitory effects on the proliferation of stromal cells from primary benign prostatic hyperplasia (BPH) of four different aqueous extracts from this plant: underground parts from specimens in flower (T1 treatment), leaves from specimens in flower (T2 treatment), and flowers (T3 treatment) and leaves from specimens not in flower (T4 treatment). T1, T2, T3, and T4 treatments at 250μg/ml for 72 hours inhibited BPH cells by 56.7%, 29.2%, 39.4%, and 13.5%, respectively, showing that the KGB underground parts extract (T1 treatment) was the most active. Our findings show that the extract of the KGB underground parts (150 and 250μg/ml) stimulates important changes in the BPH cells, modulating crucial processes such as proliferation, viability, and apoptosis. HPLC-DAD-MS/MS analysis provided a tentative identification of glycosylated syringic acid derivatives, glycosylated forms of volatile compounds, and lignans in this extract. Finally, these results suggest that there is a potential therapeutic use for KGB in BPH, which could improve the clinical management of the disease.

Anti-inflammatory activity possessing bio-compounds were identified from Kalanchoe pinnata methanolic leaf extract using Gas chromatography-mass spectrometry (GC-MS) analysis and their anti-inflammatory potential was checked using inhibition of albumin denaturation assay, anti-proteinase activity assay, and heat and hypotonicity induced hemolysis assay. GC-MS analysis revealed the presence of aldehydes, ketones, carboxylic acid in the crude. Antioxidant activity analysis using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay revealed the half maximal inhibitory concentration (IC50) value of methanol crude leaf extract as 100 μg/mL. The biocompatibility of the methanol extract was determined using 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay.The anti-inflammatory property of identified bio-compounds was also validated using a computational approach. Drug likelihood properties of phyto-compounds were analyzed by predicting the absorption, distribution, metabolism, and excretion (ADMET) score and the bioactivity score. Six chemical compounds with excellent drug relevant properties and availability were selected for a molecular docking study versus a common inflammatory molecular target COX-2 protein. The present study showed that anti-inflammatory properties of phyto-compounds from methanol crude leaf extract of Kalanchoe pinnata can be exploited for novel drug design to treat inflammatory disorders. Keywords: Inflammation, Kalanchoe pinnata, In-silico, GCMS, Cytokines, Anti-oxidant

Bryophyllum pinnatum has been used since the 1970s to prevent premature labour, first in anthroposophic hospitals and, more recently, also in the main Swiss perinatal centres. However, it is not known which compounds in B. pinnatum leaves contribute to the tocolytic effect. Here we studied the effects of a flavonoid-enriched fraction, the corresponding flavonoid aglycon mixture, a bufadienolide-enriched fraction, and B. pinnatum leaf press juice on human myometrial contractility in vitro. The strength (area under the curve and amplitude) and frequency of contractions were recorded using strips of human myometrium mounted in an organ bath system. Cell viability assays were performed with the human myometrium hTERT-C3 and PHM1 – 41 cell lines. Repeated addition of the flavonoid-enriched fraction, flavonoid aglycon mixture, bufadienolide-enriched fraction, or B. pinnatum leaf press juice led to a progressive decrease of contraction strength, without jeopardising the vitality of myometrium strips. The bufadienolide-enriched fraction was the most active, since 1 µg/mL of the bufadienolide-enriched fraction lowered the area under the curve to 40.1 ± 11.8% of the initial value, whereas 150 µg/mL of the flavonoid-enriched fraction, 6.2 µg/mL of the flavonoid aglycon mixture, and 10 mg/mL of the B. pinnatum leaf press juice were required to achieve comparable inhibition. A progressive increase of contraction frequency was observed, except in the case of the flavonoid aglycon mixture, which did not affect frequency. None of the test substances decreased myometrial cell viability, even at concentrations of 500 µg/mL of the flavonoid-enriched fraction, 40 µg/mL of the flavonoid aglycon mixture, 3.8 µg/mL of the bufadienolide-enriched fraction, and 75 mg/mL of the B. pinnatum leaf press juice, i.e., higher than those used in the myometrium experiments. Given the concentrations of flavonoids in the flavonoid-enriched fraction and B. pinnatum leaf press juice, and of bufadienolides in the bufadienolide-enriched fraction and B. pinnatum leaf press juice, it appears that bufadienolides may be mainly responsible for the relaxant effect.

Massive blood loss is still a great challenge for modern medicine. To stop the hemorrhage during the surgery or after injury apart from suturing or electrocoagulation, the most efficient method of hemostasis restoration is the use of hemostatic agents. Although there are numerous products on the market, there is still a need for biomaterials that are capable of fast and efficient bleeding management without affecting wound closure or embolism. Chitosan is known for its hemostatic activity, however, its quite poor mechanical properties and heterogenous chemical composition still needs some improvements to become superior compared to biological adhesives. The following study deals with the preparation and evaluation of chitosan-derived natural biomaterials containing Kalanchoe pinnata extract with the potential application as a blood-clotting agent. The materials were obtained under microwave-assisted conditions in two different forms (granules/dressing), whose chemical structure and morphology were studied. Their antioxidant properties have been proven. The chitosan-derived hemostatic agents exhibited superior blood sorption abilities and lack of cytotoxicity to L929 mouse fibroblasts. The study also showed the differences in biological properties depending on their preparation method. The potential mechanism of action was proposed as well as their potential in hemostasis revival.

Bryophyllum pinnatum (Lam.) Oken (Crassulaceae) is widely used as leaf juice or extracts in traditional medicine all over tropical areas, especially in Brazil, to relieve inflammation-associated symptoms. Flavonol glycosides with unusual sugar moiety are among the major metabolites. Nevertheless, there are not enough quality control studies that can contribute to authentication of B. pinnatum and determination of their markers. As it is also used as medicinal plant in several countries, it is necessary to provide data related to safety, efficacy and quality. In this context, this work aims to isolate the major flavonoids from B. pinnatum hydroethanolic extract, to validate a method to quantify the content of chemical markers and to evaluate their xanthine oxidase inhibition and antioxidant activity. The extract was submitted to centrifugal partition chromatography (CPC). The solvents system CyHex-EtOAc-EtOH-H2O, 0.5:9:3:5.5, v/v/v/v was selected by shake-flask method. Four flavonoids (quercetin 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (1), kaempferol 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (2), quercetin 3-O-α-L-rhamnopyranoside (3) and kaempferol 3-O-α-L-rhamnopyranoside (4)) were isolated in a single and fast CPC run and their structures were confirmed by NMR analysis. An UPLC-DAD quantification method was established for the first time with validation of required parameters, according to RDC 166/2017. The calibration curves were linear with correlation coefficient ranging from 0.9996 to 0.9997 while the values of LOD (0.0077–1.984 ng.mL−1), LOQ (0.0263–6.012 ng.mL−1), recovery (≥ 80.7 %) and inter-day (%RSD ≤ 3.581) and intra-day precision (%RSD ≤ 2.628) were satisfactory. Quantitative analysis of these compounds showed that the proportion of 1, 2 and 3 were 2.43, 0.25 and 0.33 % (24.3 mg.g−1, 0.25 mg.g−1 and 0.33 mg.g−1 of extract), respectively. Moreover, in vitro xanthine oxidase (XO), DPPH and ABTS inhibition were evaluated for the extract and the major flavonoids. Compounds 2 (168 μM) and 3 (124 μM) moderately inhibited XO, while compounds 1 and 3 displayed average radical scavenging activity. In conclusion, our results suggest the flavonoid 1 as a specific marker which may be used for quality control of B. pinnatum hydroethanolic leaves extract.

Context Bufadienolide compounds occur in many plants and animal species and have strong cardiac and anti-inflammatory properties. The compounds have been recently investigated for cytotoxic and antitumor activity. Objective The cytotoxic effect of bersaldegenin-1,3,5-orthoacetate – a bufadienolide steroid occuring in plants from Kalanchoe genus (Crassulaceae), was evaluated with cervical cancer HeLa cells in vitro. Materials and methods The cytotoxic activity of the compound (at 0.1–20.0 μg/mL) on the cells was determined by Real-Time Cell Analysis (RTCA) system for 24 h. The estimation of cell cycle arrest, reactive oxygen species (ROS) production, reduction of mitochondrial membrane potential (MMP), and caspases-3/7/9 activity in the HeLa cells treated with the compound was done by flow cytometry and luminometric technique. DNA damage in the cells was estimated by immunofluorescence staining and the comet assay with etoposide as a positive control. Results The compound had strong effect on the cells (IC50 = 0.55 μg/mL) by the suppression of HeLa cells proliferation in G2/M phase of cell cycle and induction of cell death through double-stranded DNA damage and reactive oxygen species overproduction. Furthermore, we did not observe an increase in the activity of caspase-3/7/9 in the treated cells as well as a decrease in cellular mitochondrial membrane potential. Gene expression analysis revealed the overexpression of NF-Kappa-B inhibitors genes (>2-fold higher than control) in the treated cells. Conclusions Bersaldegenin-1,3,5-orthoacetate induces cell cycle arrest and caspase-independent cell death through double-stranded DNA damage. These results are an important step in further studies on cell death signalling pathways induced by bufadienolides.

In this work, the combination of elicited plant suspension cultured cells (PSCCs) with untargeted metabolomics establishes a powerful, cutting-edge strategy to unravel the effects of elicitors on the biosynthetic potential of medicinal plants, as Bryophyllum sp. The PSCC technology constitutes a successful biotechnological system for the study and production of bioactive compounds throughout the inclusion of elicitors with the ability to modulate secondary metabolism. The use of methyl jasmonate (MJ) and salicylic acid (SA) as abiotic elicitors on bryophyllum PSCCs, resulted in differential effects on cell growth and secondary metabolism, depending on the species, including synergistic and antagonistic effects. This fact suggests that both elicitors play a pleiotropic effect on plant secondary metabolism, showing complex interactions, according to the UHPLC-QTOF mass spectrometry profiling. Thus, the combination of both elicitors induced a strong synergistic on B. daigremontianum PSCCs, with 2272 putatively annotated compounds, whereas it caused a negative effect on the secondary metabolism of B. × houghtonii PSCCs, being MJ the only elicitor driving a positive effect, presenting 2972 annotated compounds. Meanwhile, B. tubiflorum PSCCs did not show a significant modulation of secondary metabolism, with 1521 annotated compounds. The metabolite annotation indicated that three families of secondary metabolites were mainly affected by elicitation: phenolic compounds constituted the most affected family by elicitation, mainly represented by flavonoids and lignans; N-containing compounds included glucosinolates, amines, and alkaloids, reported to Bryophyllum sp. for the first time; and terpenoids included mainly phytoalexins and saponins. The results depict a deep genotype-dependent metabolomic reprogramming of secondary metabolism in response to elicitors, thanks to the application of untargeted metabolomics. This knowledge will allow the consideration of Bryophyllum sp. as a valuable source of bioactive compounds, with the potential associated to PSCCs, for being included in food, cosmetic, and pharmaceutical applications.

Phenolic compounds constitute an important family of natural bioactive compounds responsible for the medicinal properties attributed to Bryophyllum plants (genus Kalanchoe, Crassulaceae), but their production by these medicinal plants has not been characterized to date. In this work, a combinatorial approach including plant tissue culture, untargeted metabolomics, and machine learning is proposed to unravel the critical factors behind the biosynthesis of phenolic compounds in these species. The untargeted metabolomics revealed 485 annotated compounds that were produced by three Bryophyllum species cultured in vitro in a genotype and organ-dependent manner. Neurofuzzy logic (NFL) predictive models assessed the significant influence of genotypes and organs and identified the key nutrients from culture media formulations involved in phenolic compound biosynthesis. Sulfate played a critical role in tyrosol and lignan biosynthesis, copper in phenolic acid biosynthesis, calcium in stilbene biosynthesis, and magnesium in flavanol biosynthesis. Flavonol and anthocyanin biosynthesis was not significantly affected by mineral components. As a result, a predictive biosynthetic model for all the Bryophyllum genotypes was proposed. The combination of untargeted metabolomics with machine learning provided a robust approach to achieve the phytochemical characterization of the previously unexplored species belonging to the Bryophyllum subgenus, facilitating their biotechnological exploitation as a promising source of bioactive compounds. Xunta de Galicia | Ref. ED431E 2018/07 Xunta de Galicia | Ref. ED431D 2017/18 Ministerio de Educación | Ref. FPU15 / 04849 European Molecular Biology Organization | Ref. 8659 Ministerio de Ciencia e Innovación | Ref. EQC2019-006178-P

Garugapinnata Roxb. (Burseraceae) is a medium-sized tree widely available all over the tropical regions of Asia. Bryophyllum pinnatum (Lam) Oken. (Crassulaceae) is an indigenous and exotic plant grown in tropical regions. Both plants have been used for their anti-inflammatory, antioxidant, anticancer, wound healing, antidiabetic activities, etc. This investigation was designed to explore the result shown by methanolic extract of Garuga pinnata bark and Bryophyllum pinnatum leaves, on cognitive power and retention of the memory in experimental mice along with quantification of phenolic compounds and DPPH radicals neutralizing capacity. The memory-enhancing activity was determined by the elevated plus-maze method in Scopolamine-induced amnesic mice, using Piracetam as allopathic and Shankhpushpi as ayurvedic standard drugs. Two doses (200 and 400 mg/kg p.o.) of both extracts were administered to mice up to 8 consecutive days; transfer latency of individual group was recorded after 45 minutes and memory of the experienced things was examined after 1 day. DPPH assay method and the Folin–Ciocalteu method were employed to determine antioxidant potency and total phenol amount, respectively. 400 mg/kg of the methanolic B. pinnatum bark extract significantly improved memory and learning of mice with transfer latency (TL) of 32.75 s, which is comparable to that of standard Piracetam (21.78 s) and Shankhpushpi (27.83 s). Greater phenolic content was quantified in B. pinnatum bark extract (156.80 ± 0.33 µg GAE/mg dry extract) as well as the antioxidant potency (69.77% of free radical inhibition at the 100 µg/mL concentration). Our study proclaimed the scientific evidence for the memory-boosting effect of both plants.

* Plants utilizing crassulacean acid metabolism (CAM) concentrate CO2 around RuBisCO while reducing transpirational water loss associated with photosynthesis. Unlike stomata of C3 and C4 species, CAM stomata open at night for the mesophyll to fix CO2 into malate (Mal) and store it in the vacuole. CAM plants decarboxylate Mal in the light, generating high CO2 concentrations within the leaf behind closed stomata for refixation by RuBisCO.\ud \ud * CO2 may contribute to stomatal closure but additional mechanisms, plausibly including Mal activation of anion channels, ensure closure in the light.\ud \ud * In the CAM species Kalanchoë fedtschenkoi, we found that guard cell anion channel activity, recorded under voltage clamp, follows KfSLAC1 and KfALMT12 transcript abundance, declining to near‐zero by the end of the light period. Unexpectedly, however, we found that extracellular Mal inhibited the anion current of Kalanchoë guard cells, both in wild‐type and RNAi mutants with impaired Mal metabolism.\ud \ud * We conclude that the diurnal cycle of anion channel gene transcription, rather than the physiological signal of Mal release, is a key factor in the inverted CAM stomatal cycle.

The current industrial requirements for food naturalness are forcing the development of new strategies to achieve the production of healthier foods by replacing the use of synthetic additives with bioactive compounds from natural sources. Here, we investigate the use of plant tissue culture as a biotechnological solution to produce plant-derived bioactive compounds with antioxidant activity and their application to protect fish oil-in-water emulsions against lipid peroxidation. The total phenolic content of Bryophyllum plant extracts ranges from 3.4 to 5.9 mM, expressed as gallic acid equivalents (GAE). The addition of Bryophyllum extracts to 4:6 fish oil-in-water emulsions results in a sharp (eight-fold) increase in the antioxidant efficiency due to the incorporation of polyphenols to the interfacial region. In the emulsions, the antioxidant efficiency of extracts increased linearly with concentration and levelled off at 500 &mu, M GAE, reaching a plateau region. The antioxidant efficiency increases modestly (12%) upon increasing the pH from 3.0 to 5.0, while an increase in temperature from 10 to 30 &deg, C causes a six-fold decrease in the antioxidant efficiency. Overall, results show that Bryophyllum plant-derived extracts are promising sources of bioactive compounds with antioxidant activity that can be eventually be used to control lipid oxidation in food emulsions containing (poly)unsaturated fatty acids.

The effect of drying on the phytoconstituents, antioxidative, and anti-inflammatory properties of Bryophyllum pinnatum leaves was investigated. The phenolic constituents were characterized using HPLC-DAD. The aqueous extraction was done and various assays (Inhibition of membrane stabilization, albumin Denaturation and heat-induced hemolysis, malondialdehyde (MDA), and reduced glutathione (GSH) contents, as well as superoxide dismutase (SOD) activity), were carried out on human erythrocytes. The fresh portion (89.12 µg/ml) exhibited the highest potential to inhibit heat-induced hemolysis compared to the standard drug-Diclofenac (91.51 µg/ml). Freeze-dried sample showed the highest inhibitory potential on albumin denaturation ([Freeze-dried-330.72 µg/ml], [Diclofenac-318.63 µg/ml]) and membrane destabilization ([Freeze-dried-331.93 µg/ml], [Diclofenac-289.57 µg/ml]) when compared with Diclofenac. Similarly, the freeze-dried sample showed the highest GSH and SOD level and lowest MDA level when human erythrocytes challenged with tertiary butyl hydroperoxide (tBHP) were treated with the extract. This study confirms the retention of a considerable quantity of bioactive constituents of plants when freeze-dried. PRACTICAL APPLICATIONS: The ideal method of drying Bryophyllum pinnatum and possible anti-inflammatory potential was investigated. This work may apply to the development of anti-inflammatory agents from a natural source with little or no side effect in managing inflammation.

Organogenesis constitutes the biological feature driving plant in vitro regeneration, in which the role of plant hormones is crucial. The use of machine learning (ML) technology stands out as a novel approach to characterize the combined role of two phytohormones, the auxin indoleacetic acid (IAA) and the cytokinin 6-benzylaminopurine (BAP), on the in vitro organogenesis of unexploited medicinal plants from the Bryophyllum subgenus. The predictive model generated by neurofuzzy logic, a combination of artificial neural networks (ANNs) and fuzzy logic algorithms, was able to reveal the critical factors affecting such multifactorial process over the experimental dataset collected. The rules obtained along with the model allowed to decipher that BAP had a pleiotropic effect on the Bryophyllum spp., as it caused different organogenetic responses depending on its concentration and the genotype, including direct and indirect shoot organogenesis and callus formation. On the contrary, IAA showed an inhibiting role, restricted to indirect shoot regeneration. In this work, neurofuzzy logic emerged as a cutting-edge method to characterize the mechanism of action of two phytohormones, leading to the optimization of plant tissue culture protocols with high large-scale biotechnological applicability The authors acknowledge the FPU grant awarded to Pascual García-Pérez from the Spanish Ministry of Education (grant number FPU15/04849) SI

The interaction of methyl jasmonate (JA-Me) and indole-3-acetic acid (IAA) to induce the formation of the secondary abscission zone in the middle of internode segments of Bryophyllum calycinum was investigated in relation to auxin status and histology. When IAA at 0.1% (w/w, in lanolin) was applied to the segments, the formation of the secondary abscission zone at a few mm above the treatment in the apical direction was observed. On the contrary, IAA at 0.5% (w/w, in lanolin) did not induce the formation of the secondary abscission zone. JA-Me at 0.5% (w/w, in lanolin) applied to the middle of internode segments kept in the normal (natural) or inverted positions also induced the formation of the secondary abscission zone below and above parts of the treatment. IAA at 0.5% applied to the cut surface of the upper part of the segments completely prevented the formation of the secondary abscission zone induced by JA-Me. Simultaneous application of IAA 0.5% with JA-Me 0.5% in the middle part of the internode segments induced the formation of the secondary abscission zone at 10 mm to 12 mm above the treatment. Histological analyses indicated that the formation of the secondary abscission zone was characterized by the presence of newly synthesized cell plates that resulted from periclinal cell division within one layer of mother cells in stems. The effects of IAA (0.1%) and JA-Me (0.5%) on the formation of the secondary abscission zone were histologically similar. Comprehensive analyses of plant hormones revealed that the balance of the endogenous levels of IAA in both sides adjacent to the abscission zone was significantly disturbed when the secondary abscission formation was induced by the application of IAA. These results strongly suggest that an auxin gradient is important in the formation of the secondary abscission zone in the internode segments of B. calycinum, and IAA gradient results from polar IAA transport from the application site. IAA is important in the regulation of formation of the secondary abscission zone induced by JA-Me. Further possible mechanisms of the formation of the secondary abscission zone in the internode segments of B. calycinum are also discussed in the interaction of JA-Me and IAA.

Kalanchoe species are well-known medicinal plants used in traditional medicine as anti-inflammatory and analgesic remedies. Recently, it has been reported that Kalanchoe plants have cytotoxic properties; however, data on traditional use of these plants in tumor treatment are extremely limited. Kalanchoe daigremontiana is one of the most popular species cultivated in Europe, and it is used, among other things, as a remedy in treating skin injuries and wounds. Studies on the biological activity of this species are scarce, and there is a lack of data on the cytotoxic activity of K. daigremontiana extracts on epithelial cancer cells in the literature. In our present study, we analyzed the phytochemical composition of K. daigremontiana ethanol extract and fractions–water and dichloromethane–by the HPLC-DAD-ESI-MS method and estimated cytotoxic activity of the extracts on human adenocarcinoma (HeLa), ovarian (SKOV-3), breast (MCF-7), and melanoma (A375) cell lines by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, real-time cell analyzer (RTCA), and flow cytometry. We identified 6 bufadienolide compounds and 19 flavonoids, mostly kaempferol, quercetin, isorhamnetin, and myricetin glycosides, of which only 3 flavonoids have been identified in K. daigremontiana to date. Other flavonoids that were characterized in our study have not yet been found in this plant. The ethanol extract and water fraction of K. daigremontiana did not show significant cytotoxic activity on the tested cell lines. In contrast, the dichloromethane fraction showed the strongest activity against all cell lines with IC50 values of ≤ 10 µg/mL. The results indicated that this activity is mainly due to the presence of bersaldegenin-1,3,5-orthoacetate.

We combined machine learning and plant in vitro culture methodologies as a novel approach for unraveling the phytochemical potential of unexploited medicinal plants. In order to induce phenolic compound biosynthesis, the in vitro culture of three different species of Bryophyllum under nutritional stress was established. To optimize phenolic extraction, four solvents with different MeOH proportions were used, and total phenolic content (TPC), flavonoid content (FC) and radical-scavenging activity (RSA) were determined. All results were subjected to data modeling with the application of artificial neural networks to provide insight into the significant factors that influence such multifactorial processes. Our findings suggest that aerial parts accumulate a higher proportion of phenolic compounds and flavonoids in comparison to roots. TPC was increased under ammonium concentrations below 15 mM, and their extraction was maximum when using solvents with intermediate methanol proportions (55&ndash, 85%). The same behavior was reported for RSA, and, conversely, FC was independent of culture media composition, and their extraction was enhanced using solvents with high methanol proportions (&gt, 85%). These findings confer a wide perspective about the relationship between abiotic stress and secondary metabolism and could serve as the starting point for the optimization of bioactive compound production at a biotechnological scale.

Kalanchoe pinnata from the stonecrop family (Crassulaceae) is a medicinal plant with high content of bioactive compounds. The plant is known for its anti-allergic, antioxidant, anti-inflammatory, antimicrobial and antibacterial activities, which are mainly attributed to flavonoids. Low yields of bioactive compounds in medicinal plants have led to new strategies for enhancing their biosynthetic capacity in order to increase the content of specialized metabolites. Herein, the accumulation of specialized metabolites in plants can be triggered by elicitation methods. In the current study, different elicitation strategies were conducted towards the enhancement of bioactive compounds in K. pinnata leaves. We investigated the effect of natural transformation with the Agrobacterium rhizogenes strain A4 as means of biological elicitation on the total content of flavonoids in the leaves of K. pinnata. Furthermore, the effect of supplemental UV-B radiation, as physical elicitor, was assessed on the total flavonoid content of both wild-type (WT) and rol-transformed plants. The combined effect of the two mentioned elicitation methods was also examined. The data showed that presence of rol genes resulted in an increase of 24% in the total flavonoid content compared to WT plants. The supplemental UV-B radiation increased the total content of flavonoids with 95% and 89% in the WT and the rol+ plants, respectively. Collectively, a synergistic effect was shown as the combination of the two factors dramatically increased (133%) the total flavonoid content in K. pinnata leaves.

Introduction : Kalanchoe laxiflora (Family Crassulaceae) contains different bioactive phytochemicals, which possess several biological activities. The present study aimed at assessing the antioxidant, cytotoxic, antidiabetic, and antibiofilm activities of Kalanchoe laxiflora flower extracts, as well as identification of their phytochemicals. Methods : The total phenolic and flavonoid contents of the tested extracts were established. The antioxidant, cytotoxic, antidiabetic and antibiofilm activities were also evaluated using different methods. The phytoconstituents of K. laxiflora flower extracts were profiled using LC-ESI(-ve)-MS analysis. Results : The phytochemical investigations showed that the n-BuOH fraction had the highest total phenolic content (397.25 ± 1.18 mg GAE / g ext.) and flavonoid content (99.71 ± 0.24 mg RE / g ext.). The n-BuOH fraction also possessed the most potent antioxidant activities (DPPH; SC50=18.72±1.81 µg/mL, RPA; 78.26±0.49 mg AAE /g ext.), and TAC; 446.37±0.23 mg AAE /g ext.) and cytotoxic potential in HepG2 cells with IC50= 9.15±0.33 μg/mL. Furthermore, it exhibited the strongest α-amylase and α-glucosidase inhibitory activities (IC50= 54.72±0.96 µg/mL and 89.02±0.99 µg/mL, respectively). Whereas, the 85% methanolic extract exhibited the highest antibiofilm activity against Escherichia coli (MBIC100= 15.63 µg/ml). Additionally, the LC-ESI(-ve)-MS analysis of K. laxiflora extracts led to the characterization of a total of 35 compounds classified to flavonoid glycosides, gallotannins, alkaloids and fatty acids, which are considered to be the major components. Conclusions : K. laxiflora extracts reduce ROS, improve antidiabetic responses and inhibit bacterial growth. The identification of K. laxiflora specific phytoconstituents will provide a starting point to carry out further in vivo studies on these extracts.

ETHNOPHARMACOLOGICAL RELEVANCE Advancement in cancer therapy has improved survival among patients. However, use of anticancer drugs like anthracyclines (e.g., doxorubicin) is not without adverse effects. Notable among adverse effects of doxorubicin (DOX) is cardiotoxicity, which ranges from mild transient blood pressure changes to potentially serious heart failure. Anecdotal reports suggest that Kalanchoe integra (KI) may have cardio-protective potential. AIMS OF THE STUDY This study sought to determine the cardio-protective potential of KI against doxorubicin-induced cardiotoxicity and also examined any possible genotoxic potential of KI in selected organs. Additionally, the nitric oxide modulatory potential of KI was assessed. MATERIALS AND METHODS The leaves of KI were collected, air-dried, pulverised and extracted using 70% ethanol. High-performance liquid chromatography (HPLC) fingerprinting was done for KI. Also, the single-cell gel electrophoresis assay (Comet assay) was employed to ascertain the genotoxic potential of KI. In assessment of cardio-protective potential of KI against doxorubicin-induced cardiotoxicity, a total of 42 female Sprague-Dawley rats were put into 7 groups (n = 6). Group I: vehicle control, received normal saline (1 mL/kg p.o) for 30 days. Group II: toxic control, received DOX (20 mg/kg i.p.) once on the 29th day. Group III: KI control, received KI (300 mg/kg p.o) for 30 days. Group IV: vitamin E control, received vitamin E (100 mg/kg p.o) for 30 days. Group V: KI treated-1, received KI (300 mg/kg p.o) for 30 days and DOX (20 mg/kg i.p) on the 29th day. Group VI: KI treated-2, received KI (600 mg/kg p.o) for 30 days and DOX (20 mg/kg i.p) on the 29th day. Group VII: vitamin E treated, received vitamin E (100 mg/kg p.o) for 30 days and DOX (20 mg/kg i.p) on the 29th day. Thirty-six (36) hours after last administration, rats were sacrificed. Blood samples were taken via cardiac puncture to determine levels of aspartate aminotransferase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), creatine kinase (CK), lactate dehydrogenase (LDH), enzymatic antioxidants such as glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT). Nitric oxide level was also determined. Hearts of rats in each group were excised and taken through histopathological examination. RESULTS In the HPLC fingerprint analysis, 13 peaks were identified, and peak with retention time of 24.0 min had the highest peak area (3.223 x104 mAU). Comet assay showed that the KI extract was non-genotoxic. Pretreatment with KI protected rats against doxorubicin-induced cardiotoxicity as evidenced by the low levels of AST, ALT, ALP, CK and LDH compared with the controls (p

It has long been recognized that plant invasions may alter carbon (C) and nitrogen (N) cycles, but the direction and magnitude of such alterations have been rarely quantified. In this study, we quantified the effects caused by the invasion of a noxious exotic plant, Kalanchoe daigremontiana (Crassulaceae), on C and N mineralization and enzymatic and microbial activities in the soil of a semiarid locality in Venezuela. We compared soil parameters associated with these processes (C and N mineralization time and the cumulative values, fluorescein diacetate hydrolytic activity, and activities of dehydrogenase, β-glucosidase, glucosaminidase, and urease) between invaded and adjacent non-invaded sites. In addition, correlations among these parameters and the soil physical-chemical properties were also examined to determine if a positive feedback exists between nutrient availability and K. daigremontiana invasion. Overall, our results showed that C mineralization and transformation of organic compounds to NH4+ were favored at sites colonized by K. daigremontiana. With this species, we found the highest cumulative amounts of NH4+-N and C and the lowest mineralization time. These results could be explained by higher activities of urease and glucosaminidase in soils under the influence of K. daigremontiana. In addition, higher amounts of organic matter and moisture content in invaded soils might favor C and N mineralization. In conclusion, invasion of Neotropical semiarid zones by K. daigremontiana may influence the chemical and biological properties of the soils covered by this species, increasing nutrient bioavailability, which, in time, can facilitate the invasion process.

Ethnopharmacological importance Kalanchoe flammea Stapf (Crassulaceae) is a medicinal plant grown in the South of Mexico (State of Tabasco), which is commonly used in traditional medicine for the treatment of fever, wounds, inflammation, and cancer. Aim of the study To establish the potential of K. flammea for the treatment of prostate cancer, evaluating its cytotoxic activity, its probable mechanism of action, and carrying out some toxicological safety studies. Materials and methods The cytotoxic activity of the ethyl acetate extract of K. flammea (Kf-EtOAc) was evaluated in several cell lines of prostate cancer by MTT viability assay. The cellular death mechanism was studied by evaluating the translocation of phosphatidylserine (Annexin V); overproduction of reactive oxygen species [2'-7'-Dichlorodihydrofluorescein diacetate (DCFH-DA) assay]; release of Cytochrome C; activation of caspase-3 and -9, and regulation of Bcl-2, XIAP, and PKCe proteins by Western Blot analysis. For the evaluation of the safety of Kf-EtOAc, the Ames test, Micronucleus assay, and acute toxicity study were determined. Results Kf-EtOAc exhibited selective cytotoxic activity against prostate cell lines as follows: PC-3, LNCaP, and PrEC (IC50 = 1.36 ± 0.05; 2.06 ± 0.02, and 127.05 ± 0.07 μg/mL, respectively). The F82-P2 fraction (rich in coumaric acid and palmitic acid) obtained by bioassay-guided fractionation of Kf-EtOAc also demonstrated selective cytotoxic activity against PC-3 cells (IC50 = 1.05 ± 0.06 μg/mL). Kf-EtOAc induces apoptosis by the intrinsic pathway; this mechanism of cell death was confirmed after observing that the extract produces phosphatidylserine translocation, overproduction of reactive oxygen species, release of Cytochrome C at mitochondrial level, and activation of caspase-3 and -9. It was also observed that Kf-EtOAc produces significant downregulation of apoptosis-related proteins Bcl-2, XIAP, and PKCe and induces DNA fragmentation and cell cycle arrest. In addition, Kf-EtOAc is non-genotoxic in vitro by Ames test and non-genotoxic in vivo by Micronucleus assay, and no signs of toxicity or death were reported after the administration of a single acute exposure of 2000 mg/kg. Conclusion K. flammea is a potential candidate for the development of new drugs for the treatment of prostate cancer. However, to propose their use in clinical trials, additional studies are required to understand their pharmacokinetic behavior, as well as the development of a suitable pharmaceutical form.

Objective Kalanchoe hybrida (Crassulaceae) is naturalized throughout all the island of Taiwan in China. The preliminary bioassay-guided fractionation of the crude extract of K. hybrida exhibited that the chloroform and n-butanol fractions possessed potent cytotoxicity against MCF-7, NCI-H460, and SF-268 tumor cell lines at 50 µg/mL concentration. Therefore, K. hybrida was selected as a target and the chemical constituents from the chloroform and n-butanol fractions of the crude extracts of K. hybrida were identified. The potential constituents were examined for their cytotoxicity against the tumor cell lines. Methods A combination of conventional chromatographic techniques was performed on the crude extract of K. hybrida. The chemical structures of the purified constituents were identified on the basis of spectroscopic and spectrometric analysis. Results The purification results had led to the characterization of totally 37 compounds. The isolated compounds 1, 2, and 4–12 were examined for their cytotoxicity in vitro, and bufadienolides 4–8 and flavonol glycoside 11 displayed significant cytotoxicity towards all the tested tumor cell lines among these tested compounds. Conclusion The results indicated that these principles should be responsible for the bioactivity of corresponding partial fractions. The potential constituents could be further investigated to explore the new natural lead drugs.

Kalanchoë is an economically important genus comprising numerous potted plants and recently is also emerging as cut flowers. However, the lack of information about flower-inducing factors limits the number of species that can be used in commercial production and breeding programs. Therefore, the aim of this study was to investigate the effects of exogenous application of gibberellic acid (GA3) on flower induction and flowering quality of Kalanchoë longiflora and Kalanchoë pinnata. The experiment was conducted under a short day (SD) photoperiod with a day temperature of 22 °C and a night temperature of 15 °C for 8 weeks. The treatments consisted of four applications of either 0.25 or 0.50 μg of GA3 per plant per week, providing a total of 100 μg or 200 μg/plant and 0 μg/plant for the control. The volume of 100 μL of GA3 solution containing 1% agarose was applied to the shoot apex using a pipette. For both species, flowering was enhanced by the GA3 treatments compared with the control plants. Gibberellin-treated plants flowered earlier, produced more inflorescences, and exhibited an increased number of flowers compared with the control plants. Moreover, the GA3 treatments in K. longiflora delayed the appearance of wilted flowers. Plant height increased in plants that received GA3, but the number of nodes did not differ from the control plants. Thus, we conclude that the application of GA3 improves flowering of Kalanchoë species and can be a useful tool for the production of cut flower cultivars.

Alien invasive species have strategies that can maintain fitness in a variety of environments. This flexibility is associated with environmental tolerance in several traits, such as allocation of resources to shoots versus roots, clonal versus sexual reproduction, and survival of seedlings. These traits were explored in the chandelier plant (Kalanchoe delagoensis Eckl. & Zeyh.), which has invasive populations in several countries. Light and water tolerance and herbicide treatments were tested on plantlet survival. Plantlet survival in the most extreme cases (full sunlight and no watering) was close to 30%, whereas in less severe conditions (water and shaded), it was close to 100%. Stress conditions triggered the onset of plantlet production from the margin of leaves, which increased clonality. Biomass was allocated primarily to aboveground structures. Although all herbicides resulted in high plantlet mortality (>85%), only 2,4-D and glyphosate+2,4-D amine achieved the maximum recorded mortality a few days after the chemical application. The high tolerance of K. delagoensis plantlets to varying conditions shows that under stress, plantlet production is enhanced as survival of established individuals decreases. Biomass is primarily aboveground, which can potentially alter nitrogen and carbon in poor arid environments, and the proportion of the biomass assigned to belowground roots increased with an increase in sunlight received. Even though the chemical treatments 2,4-D and glyphosate+2,4-D amine have been shown to be the only effective treatments, the 2,4-D treatment may be the most viable (cost+quantity) to reduce the propagation of K. delagoensis. Plantlets have become the main reason for population persistence, partially due to the plant’s environmental tolerance and ability to reproduce asexually in short time periods. Susceptibility of plantlets to the two herbicides presents a means to adequately manage invasions of K. delagoensis in Mexico.

Introduction The medicinal plant Kalanchoe pinnata is a phenolic-rich species used worldwide. The reports on its pharmacological uses have increased by 70% in the last 10 years. The leaves of this plant are the main source of an unusual quercetin-diglycosyl flavonoid (QAR, quercetin arabinopyranosyl rhamnopyranoside), which can be easily extracted using water. QAR possess a strong in vivo anti-inflammatory activity. Objective To optimize the aqueous extraction of QAR from K. pinnata leaves using a three-level full factorial design. Material and methods After a previous screening design, time (x1 ) and temperature (x2 ) were chosen as the two independent variables for optimization. Freeze-dried leaves were extracted with water (20% w/v), at 30°C, 40°C or 50°C for 5, 18 or 30 min. QAR content (determined by HPLC-DAD) and yield of extracts were analyzed. The optimized extracts were also evaluated for cytotoxicity. Results The optimal heating times for extract yield and QAR content were similar in two-dimensional (2D) surface responses (between 12.8 and 30 min), but their optimal extraction temperatures were ranged between 40°C and 50°C for QAR content and 30°C and 38°C for extract yield. A compromise region for both parameters was at the mean points that were 40°C for the extraction temperature and 18 min for the total time. Conclusion The optimized process is faster and spends less energy than the previous one (water; 30 min at 55°C); therefore is greener and more attractive for industrial purposes. This is the first report of extraction optimization of this bioactive flavonoid. Copyright © 2018 John Wiley & Sons, Ltd.

The method of producing of water extract from transgenic Kalanchoe pinnata L. plants expressing cecropin P1 gene has been developed and adapted for pharmaceutical production. Chlorophorm has been shown to be an efficient preservative for this extract. The antibiotic activity of the extract has kept well even after 1-h boiling. In addition, the prolonged antimicrobial activity of the extract has been confirmed by the “accelerated aging” method. Being added to culture medium, the leaf extract from transgenic K. pinnata plants has provided more intensive callus growth and improved rhizogenesis in ice plant (Mesembryanthemum crystallinum). The obtained results demonstrate an increased biological activity of the extract from kalanchoe plants producing antimicrobial cecropin P1 peptide and good prospects for its use in pharmacology.