Phytochemical Analysis and Assessment of Antioxidant and Enzyme Inhibitory Activity of Alchemilla pseudocartalinica Juz.

This study aimed to evaluate and compare the antioxidant capacity and enzyme inhibitory activity of extracts, sub-extracts, and fractions prepared from the aerial parts and roots of A. pseudocartalinica. The phytochemical content of the active extracts was also analyzed. According to the results, ellagic acid (38.42 mg/g) was the major compound in the aerial part methanol extract and catechin (185.30 mg/g) in the root methanol extract. The DPPH inhibition activity of all fractions was monitored, with the most active one (Fr B) reaching an IC50 value of (4.92 ± 0.59 µg/mL). All the fractions prepared from the aerial parts' water sub-extract showed higher a-glucosidase inhibitory activity than the positive control acarbose. In the tyrosinase assay, Fr B (58.81 ± 7.50 µg/mL) exhibited the highest inhibitory actions among all fractions. The structure of the major substances of the most active fraction were elucidated as quercetin 7-O-ß-glucopyranosyl-3-O-ß-glucuronopyranoside- (1), and α-[(2-formyl-5-hydroxymethyl)pyrrol-1-yl]aspartic acid (2).

Identification of Flavonoids, Antioxidant and Antiproliferative Activity of Aqueous Infusions of Calendula officinalis L., Chelidonium majus L., Teucrium chamaedrys L. and Alchemilla vulgaris L.

Research background: The current changes in the global economy, characterised by the climate crisis and the economic and health impact of the COVID-19 pandemic, have led to a significant demand for medicinal herbs. This trend is expected to increase significantly by 2050. In this study, we investigated the biopotential of aqueous infusions of four medicinal plants: Calendula officinalis, Chelidonium majus, Teucrium chamaedrys and Alchemilla vulgaris. Experimental approach: The flavonoid analysis of the aqueous infusions of the selected plants was carried out using the RP-HPLC technique. The antiproliferative activity of the prepared aqueous plant infusions was analysed against three human cancer cell lines (MDA-MD-231, T24 and A549), while the antioxidant potential was measured using three antioxidant methods (DPPH, FRAP and Rancimat assay). Results and conclusions: T. chamaedrys had the highest total phenolics (expressed as GAE (2061±42) mg/L), free radical scavenging activity (IC50=1.9 mg/mL) and Fe(III) reducing antioxidant power (expressed as FeCl2 (9798±27) mg/L). At a concentration of 1 mg/mL, the antiproliferation of T24 by C. majus was 96 % and of MDA-MD-231 cells by A. vulgaris was 75 % after 72 h. After principal component analysis, T. chamaedrys and C. majus were grouped together. Quercetin glucoside and antioxidant capacity (DPPH) contributed the most to differentiate these infusions from the other two. Novelty and scientific contribution: This study represents a comparative analysis of the biopotential of four medicinal plants. A new RP-HPLC method was developed to separate the flavonoids in the herbal infusions. This is the first report on the presence of kaempferol-3-O-rutinoside in C. officinalis and isorhamnetin-3-O-rutinoside in A. vulgaris aqueous infusion. For the first time, C. majus has been shown to contribute to the oxidative stability of edible oil. Furthermore, this is the first comparative study on the antiproliferative activity of selected medicinal plants against the cell lines MDA-MD-231, T24 and A549.

The Aqueous Lyophilisate of Alchemilla Kiwuensis Engl. (Rosaceae) Displays Antiepileptogenic and Antiepileptic Effects on PTZ-induced Kindling in rats: Evidence of Modulation of Glutamatergic and GABAergic Pathways Coupled to Antioxidant Properties.

Alchemilla kiwuensis Engl. (Rosaceae) (A. kiwuensis) is an herbaceous plant traditionally used by Cameroonians to treat epilepsy and other central nervous system disorders. The present study evaluated the antiepileptogenic and antiepileptic effects of A. kiwuensis (40 mg/kg, 80 mg/kg) following Pentylenetetrazole (PTZ)-induced kindling as well as its sub-chronic toxicity. Following an initial i.p administration of a challenge dose (70 mg/kg), Wistar rats of both sexes received sub convulsive doses (35 mg/kg) of PTZ every other day, one hour after the oral gavage of animals with treatments, until two consecutive stage 4, in all animals of negative control. Seizure progression, latency, duration, and repetition were noted. Twenty-four hours later, animals were dissected to extract their hippocampi. The resulting homogenates were used to evaluate Malondialdehyde, reduced glutathione, catalase activity, GABA, GABA-Transaminase, glutamate, glutamate transporter 2, IL-1ß and TGF-1 ß. Sub-chronic toxicity was conducted according to OECD 407 guidelines. The lyophilisate of A. kiwuensis significantly increased the latency of seizure appearance, delayed seizure progression and decreased seizure repetition and duration. Biochemical analysis revealed that the lyophilisate significantly increased the catalase activity, reduced glutathione, GABA, glutamate transporter 2 and TGF-1B levels. The lyophilisate equally caused a significant decreased in the GABA-Transaminase activity, malondialdehyde, and IL-1 ß levels. There was no noticeable sign of toxicity. A. kiwuensis possesses antiepileptic and antiepiletogenic effects by enhancing GABAergic neurotransmission and antioxidant properties, coupled to modulation of glutamatergic and neuroinflammatory pathways and is innocuous in a sub-chronic model. These justifies its local use for the treatment of epilepsy.

Alchemilla viridiflora Rothm.: the potent natural inhibitor of angiotensin I-converting enzyme.

Alchemilla viridiflora Rothm., Rosaceae is a herbaceous plant widespread in central Greece, Bulgaria, North Macedonia and Serbia with Kosovo. Liquid chromatography-mass spectrometry analysis leads to the identification of 20 compounds in methanol extract, mainly ellagitannins and flavonoid glycosides. Given that various plant extracts have traditionally been used to treat hypertension and that some of the analyzed methanol extract constituents have beneficial cardiovascular effects, we hypothesized that some of these effects are achieved by inhibiting angiotensin I-converting enzyme (ACE). The dose-dependent ACE inhibitory activities of A. viridiflora and miquelianin were observed with an IC50 of 2.51 ± 0.00 µg/mL of A. viridiflora extract compared to the IC50 of 5.4139 ± 0.00 µM for miquelianin. The contribution of the single compounds to the tested activity was further analyzed through the in silico experimental approach. Computational docking results showed that tiliroside, ellagic acid pentose and galloyl-hexahydroxydiphenoyl-glucose exhibited even better binding affinity for the ACE active site than miquelianin, for which ACE activity was confirmed by an in vitro assay.

LC-ESI-MS/MS Identification of Biologically Active Phenolics in Different Extracts of Alchemilla acutiloba Opiz.

Liquid chromatography electrospray ionization tandem mass spectrometric (LC-ESI-MS/MS) qualitative and quantitative analysis of different extracts from the aerial parts and roots of Alchemilla acutiloba led to the identification of phenolic acids and flavonoids. To the best of our knowledge, isorhamnetin 3-glucoside, kaempferol 3-rutinoside, narcissoside, naringenin 7-glucoside, 3-O-methylquercetin, naringenin, eriodictyol, rhamnetin, and isorhamnetin were described for the first time in Alchemilla genus. In addition, the antioxidant, anti-inflammatory and cytotoxic activity of all extracts were evaluated. The results clearly showed that among analyzed extracts, the butanol extract of the aerial parts exhibited the highest biological activity comparable with the positive controls used.

Beyond Traditional Use of Alchemilla vulgaris: Genoprotective and Antitumor Activity In Vitro.

Alchemilla vulgaris L. (lady's mantle) was used for centuries in Europe and Balkan countries for treatments of numerous conditions and diseases of the reproductive system, yet some of the biological activities of lady's mantle have been poorly studied and neglected. The present study aimed to estimate the potential of A. vulgaris ethanolic extract from Southeast Serbia to prevent and suppress tumor development in vitro, validated by antioxidant, genoprotective, and cytotoxic properties. A total of 45 compounds were detected by UHPLC-HRMS analysis in A. vulgaris ethanolic extract. Measurement of antioxidant activity revealed the significant potential of the tested extract to scavenge free radicals. In addition, the analysis of micronuclei showed an in vitro protective effect on chromosome aberrations in peripheral human lymphocytes. A. vulgaris extract strongly suppressed the growth of human cell lines derived from different types of tumors (MCF-7, A375, A549, and HCT116). The observed antitumor effect is realized through the blockade of cell division, caspase-dependent apoptosis, and autophagic cell death. Our study has shown that Alchemilla vulgaris L. is a valuable source of bioactive compounds able to protect the subcellular structure from damage, thus preventing tumorigenesis as well as suppressing tumor cell growth.

In Silico and In Vitro Studies of Alchemilla viridiflora Rothm-Polyphenols' Potential for Inhibition of SARS-CoV-2 Internalization.

Since the outbreak of the COVID-19 pandemic, it has been obvious that virus infection poses a serious threat to human health on a global scale. Certain plants, particularly those rich in polyphenols, have been found to be effective antiviral agents. The effectiveness of Alchemilla viridiflora Rothm. (Rosaceae) methanol extract to prevent contact between virus spike (S)-glycoprotein and angiotensin-converting enzyme 2 (ACE2) and neuropilin-1 (NRP1) receptors was investigated. In vitro results revealed that the tested samples inhibited 50% of virus-receptor binding interactions in doses of 0.18 and 0.22 mg/mL for NRP1 and ACE2, respectively. Molecular docking studies revealed that the compounds from A. viridiflora ellagitannins class had a higher affinity for binding with S-glycoprotein whilst flavonoid compounds more significantly interacted with the NRP1 receptor. Quercetin 3-(6″-ferulylglucoside) and pentagalloylglucose were two compounds with the highest exhibited interfering potential for selected target receptors, with binding energies of -8.035 (S-glycoprotein) and -7.685 kcal/mol (NRP1), respectively. Furthermore, computational studies on other SARS-CoV-2 strains resulting from mutations in the original wild strain (V483A, N501Y-K417N-E484K, N501Y, N439K, L452R-T478K, K417N, G476S, F456L, E484K) revealed that virus internalization activity was maintained, but with different single compound contributions.

Phenolic Composition, Anti-Inflammatory, Antioxidant, and Antimicrobial Activities of Alchemilla mollis (Buser) Rothm.

The current study was designed to evaluate the antioxidant, anti-inflammatory and antimicrobial activities of Alchemilla mollis (Buser) Rothm. (Rosaceae) aerial parts extracts. Chemical composition was analyzed by spectrophotometric and chromatographic (HPLC) techniques. The antioxidant properties assessed included DPPH· and ABTS·+ radical scavenging, ß-carotene-linoleic acid co-oxidation assay. Antimicrobial activity was evaluated with disc diffusion and micro dilution method. In order to evaluate toxicity of the extracts, with the sulforhodamine B colorimetric assay L929 cell line (mouse fibroblast) was used. The anti-inflammatory activities of the potent antioxidant extracts (methanol, 70% methanol, and water extracts) were determined by measuring the inhibitory effects on NO production and pro-inflammatory cytokine TNF-α levels in lipopolysaccharide stimulated RAW 264.7 cells. 70% methanol and water extracts which were found to be rich in phenolic compounds (184.79 and 172.60 mg GAE/g extract) showed higher antioxidant activity. Luteolin-7-O-glucoside was the main compound in the extracts. Ethyl acetate and 70% methanol extracts showed higher antibacterial activity against Staphylococcus aureus and Salmonella enteritidis with MIC value of 125 µg/ml. 70% methanol extract potentially inhibited the NO and TNF-α production (18.43 µm and 1556.22 pg/ml, respectively, 6 h).

The evolution of dwarf shrubs in alpine environments: a case study of Alchemilla in Africa.

BACKGROUND AND AIMS: Alpine and arctic environments worldwide, including high mountains, are dominated by short-stature woody plants (dwarf shrubs). This conspicuous life form asserts considerable influence on local environmental conditions above the treeline, creating its own microhabitat. This study reconstructs the evolution of dwarf shrubs in Alchemilla in the African tropical alpine environment, where they represent one of the largest clades and are among the most common and abundant plants. METHODS: Different phylogenetic inference methods were used with plastid and nuclear DNA sequence markers, molecular dating (BEAST and RelTime), analyses of diversification rate shifts (MEDUSA and BAMM) and ancestral character and area reconstructions (Mesquite). KEY RESULTS: It is inferred that African Alchemilla species originated following long-distance dispersal to tropical East Africa, but that the evolution of dwarf shrubs occurred in Ethiopia and in tropical East Africa independently. Establishing a timeframe is challenging given inconsistencies in age estimates, but it seems likely that they originated in the Pleistocene, or at the earliest in the late Miocene. The adaptation to alpine-like environments in the form of dwarf shrubs has apparently not led to enhanced diversification rates. Ancestral reconstructions indicate reversals in Alchemilla from plants with a woody base to entirely herbaceous forms, a transition that is rarely reported in angiosperms. CONCLUSIONS: Alchemilla is a clear example of in situ tropical alpine speciation. The dwarf shrub life form typical of African Alchemilla has evolved twice independently, further indicating its selective advantage in these harsh environments. However, it has not influenced diversification, which, although recent, was not rapid.

Dual Role of Alchemilla vulgaris L. Extract in Breast Cancer Regression: Reestablishment of Effective Immune Response.

Ethnomedicinal records have long mentioned the historical usage of Alchemilla vulgaris L. in folk medicine, particularly for the treatment of gynecological issues. Building on this ethnomedicinal knowledge regarding female illnesses, the aim of this research was to evaluate the impact of ethanolic extract of A. vulgaris on mouse breast cancer cells (4T1) in vitro and in vivo, in addition to its effect on the immune compartment in the tumor microenvironment. Behind viability decrease of 4T1 cells induced by treatment with A. vulgaris extract was strong inhibition of cell proliferation accompanied by caspase-dependent apoptosis and autophagic cell death. Observed changes in 4T1 cell culture after treatment were well orchestrated and led to a reduction in metastatic potential through weakened adhesion, invasion, migration, and colony-forming abilities in vitro. Enhanced intracellular production of reactive oxygen and nitrogen species promoted by the treatment might interfere with all the observed effects. Apart from the direct effect on tumor cells, the A. vulgaris extract significantly reduced tumor growth in the solid orthotropic mammary carcinoma model through restitution of efficient local and systemic immune response reflected in enhanced antigen-presenting potential of dendritic cells (DCs) as well as the extent and activity of effector T cells.

Antimelanoma Effects of Alchemilla vulgaris: A Comprehensive In Vitro and In Vivo Study.

Due to the rich ethnobotanical and growing evidence-based medicine records, the Alchemillae herba, i.e., the upper parts of the Lady's mantle (Alchemilla vulgaris L.), was used for the assessment of antimelanoma activity. The ethanolic extract of A. vulgaris strongly suppressed the viability of B16F1, B16F10, 518A2, and Fem-X cell lines. In contrast to the in vitro study, where the B16F1 cells were more sensitive to the treatment than the more aggressive counterpart B16F10, the results obtained in vivo using the corresponding syngeneic murine model were quite the opposite. The higher sensitivity of B16F10 tumors in vivo may be attributed to a more complex response to the extract compared to one triggered in vitro. In addition, the strong immunosuppressive microenvironment in the B16F1 model is impaired by the treatment, as evidenced by enhanced antigen-presenting potential of dendritic cells, influx and activity of CD4+ T and CD8+ T lymphocytes, decreased presence of T regulatory lymphocytes, and attenuation of anti-inflammatory cytokine production. All these effects are supported by the absence of systemic toxicity. A. vulgaris extract treatment results in a sustained and enhanced ability to reduce melanoma growth, followed by the restoration of innate and adopted antitumor immunity without affecting the overall physiology of the host.

Alchemilla vulgaris modulates isoproterenol-induced cardiotoxicity: interplay of oxidative stress, inflammation, autophagy, and apoptosis.

Introduction: Isoproterenol (ISO) is regarded as an adrenergic non-selective ß agonist. It regulates myocardial contractility and may cause damage to cardiac tissues. Alchemilla vulgaris (AV) is an herbal plant that has garnered considerable attention due to its anti-inflammatory and antioxidant bioactive components. The present investigation assessed the cardioprotective potential of AV towards ISO-induced myocardial damage. Methods: Four groups of mice were utilized: control that received saline, an ISO group (85 mg/kg, S.C.), ISO + AV100, and ISO + AV200 groups (mice received 100 or 200 mg/kg AV orally along with ISO). Results and discussion: ISO induced notable cardiac damage demonstrated by clear histopathological disruption and alterations in biochemical parameters. Intriguingly, AV treatment mitigates ISO provoked oxidative stress elucidated by a substantial enhancement in superoxide dismutase (SOD) and catalase (CAT) activities and reduced glutathione (GSH) content, as well as a considerable reduction in malondialdehyde (MDA) concentrations. In addition, notable downregulation of inflammatory biomarkers (IL-1ß, TNF-α, and RAGE) and the NF-κB/p65 pathway was observed in ISO-exposed animals following AV treatment. Furthermore, the pro-apoptotic marker Bax was downregulated together with autophagy markers Beclin1 and LC3 with in ISO-exposed animals when treated with AV. Pre-treatment with AV significantly alleviated ISO-induced cardiac damage in a dose related manner, possibly due to their antioxidant and anti-inflammatory properties. Interestingly, when AV was given at higher doses, a remarkable restoration of ISO-induced cardiac injury was revealed.

Phenolic Composition and Antioxidant Activity of Alchemilla Species.

The genus Alchemilla, belonging to the Rosaceae family, is a rich source of interesting secondary metabolites, including mainly flavonoids, tannins, and phenolic acids, which display a variety of biological activities, such as anti-inflammatory, antimicrobial, and antioxidant. Alchemilla species are used in traditional medicine for treatment of acute diarrhea, wounds, dysmenorrhea, and menorrhagia. In this review, we focus on the phenolic compound composition and antioxidative activity of Alchemilla species. We can assume that phytomedicine and natural products chemistry are of significant importance due to the fact that extract combinations with various bioactive compounds possess the activity to protect the human body rather than disturb damaging factors.

Alchemilla monticola Opiz. Functional Traits Respond to Diverse Alpine Environmental Conditions in Karavanke, Slovenia.

Alpine plants are exposed to demanding environmental conditions, such as high ultraviolet (UV) and photosynthetic radiation, extreme temperatures, drought, and nutrient deficiencies. Alpine plants adapt and acclimate to harsh conditions, developing several strategies, including biochemical, physiological, and optical responses. However, alpine plants' survival strategies are hardly researched due to time-consuming and complex experimental conditions, which are supported by scarce studies. Our study focused on the functional traits of the alpine plant Alchemilla monticola Opiz (hairy lady's mantle) growing at two different altitudes (1500, 2000 m a.s.l.) and two different UV exposures per altitude. Near-ambient (UV) and reduced (UV-) UV radiations were provided by using two sorts of UV absorbing filters; temperatures were monitored hourly. The experimental plots were located at Tegoska Gora, Karavanke, Slovenia. Functional traits: physiological, biochemical, and optical characteristics were recorded three times during the growing season. A. monticola showed high maximum photochemical efficiency at both altitudes throughout the season, which confirms good adaptation and acclimatization of the plant. Furthermore, significantly higher maximum photochemical efficiency at the subalpine altitude coincided with significantly higher UV absorbing compounds (UV AC) contents at the subalpine compared to the montane altitude in August. A. monticola manifested high UV AC contents throughout the season, with significantly increased synthesis of UV AC contents in the subalpine conditions in August and September. The stomatal conductance rate increased with altitude and was correlated mostly to a lower temperature. A. monticola leaves did not transmit any UV spectrum, which corresponded to high total UV AC contents. The leaf transmittance of the photosynthetic spectrum increased at the subalpine altitude, while the transmittance of the green and yellow spectra increased under the reduced UV radiation in the autumn. A. monticola's high photosynthetic spectrum transmittance at the subalpine altitude in the autumn might therefore be due to subalpine harsh environmental conditions, as well as plant ontogenetical phase.

Optimization of Deep Eutectic Solvent Extraction of Phenolic Acids and Tannins from Alchemilla vulgaris L.

Alchemilla vulgaris L. is a good source of antioxidant components with an emphasis on phenolic acids and tannins. In this study, gallic acid, ellagic acid, and hydrolyzable tannins (HT) were extracted from this plant with different deep eutectic solvents (DESs), varying the amount of added H2O, temperature and extraction time. Seventeen DESs (n = 3) were used for the extraction, of which choline chloride:urea (1:2) proved to be the most suitable. The selection of the best solvent was followed by the examination of the influence of the extraction type and parameters using response surface methodology (RSM). Gallic acid content was in the range of 0.00-1.89 µg mg-1, ellagic acid content was 0.00-12.76 µg mg-1 and hydrolyzable tannin (HT) content was 3.06-181.26 µgTAE mg-1, depending on the used technique and the extraction conditions. According to the results, extraction by stirring and heating was the most suitable since the highest amounts of gallic acid, ellagic acid, and HT were extracted, and the obtained optimal values using response surface methodology (RSM) are confirmed by experimentally obtained values.

Physiological Performance, Antioxidant and Immune Status, Columnaris Resistance, and Growth of Nile Tilapia That Received Alchemilla vulgaris-Supplemented Diets.

The current perspective is a pioneering trial to assess the efficacy of the dietary supplementation of Alchemilla vulgaris powder (AVP) in the diet of Nile tilapia (Oreochromis niloticus) on growth performance, blood picture, hepatic and renal biomarkers, immune status, and serum and tissue antioxidant capacity and to investigate the resistance against Flavobacterium columnare challenge. Fish (n = 360) were distributed into six groups (three replicates each) and received increasing AVP supplementation levels (0, 2, 4, 6, 8, and 10 g kg-1) for 60 days. Furthermore, fish were exposed to the bacterial challenge of a virulent F. columnare strain and maintained under observation for 12 days. During the observation period, clinical signs and the cumulative mortality percentage were recorded. The results demonstrated that the growth performance, feed conversion ratio, and hematological profile were noticeably enhanced in the AVP-supplemented groups compared to the control. The most promising results of weight gain and feed conversion ratio were recorded in the groups with 6, 8, and 10 g AVP kg-1 diets in a linear regression trend. The levels of hepatorenal function indicators were maintained in a healthy range in the different dietary AVP-supplemented groups. In a dose-dependent manner, fish fed AVP dietary supplements displayed significant augmented serum levels of innate immune indicators (lysozyme, nitric oxide, and complement 3) and antioxidant biomarkers (Catalase (CAT), superoxide dismutase (SOD), total antioxidant (TAC), and reduced glutathione (GSH) with a marked decrease in myeloperoxidase (MPO) and malondialdehyde (MDA) levels). Likewise, hepatic CAT and SOD activities were significantly improved, and the opposite trend was recorded with hepatic MDA. The highest AVP-supplemented dose (10 g/kg) recorded the highest immune-antioxidant status. Based on the study findings, we highlight the efficacy of AVP as a nutraceutical dietary supplementation for aquaculture to enhance growth, physiological performance, and immune-antioxidant status and as a natural economic antibacterial agent in O. niloticus for sustaining aquaculture. It could be concluded that the dietary supplementation of 10 g AVP/kg enhanced O. niloticus growth, physiological performance, immune-antioxidant status, and resistance against F. columnare.

Climate and Land-Cover Change Impacts and Extinction Risk Assessment of Rare and Threatened Endemic Taxa of Chelmos-Vouraikos National Park (Peloponnese, Greece).

Chelmos-Vouraikos National Park is a floristic diversity and endemism hotspot in Greece and one of the main areas where Greek endemic taxa, preliminary assessed as critically endangered and threatened under the IUCN Criteria A and B, are mainly concentrated. The climate and land-cover change impacts on rare and endemic species distributions is more prominent in regional biodiversity hotspots. The main aims of the current study were: (a) to investigate how climate and land-cover change may alter the distribution of four single mountain endemics and three very rare Peloponnesian endemic taxa of the National Park via a species distribution modelling approach, and (b) to estimate the current and future extinction risk of the aforementioned taxa based on the IUCN Criteria A and B, in order to investigate the need for designing an effective plant micro-reserve network and to support decision making on spatial planning efforts and conservation research for a sustainable, integrated management. Most of the taxa analyzed are expected to continue to be considered as critically endangered based on both Criteria A and B under all land-cover/land-use scenarios, GCM/RCP and time-period combinations, while two, namely Alchemilla aroanica and Silene conglomeratica, are projected to become extinct in most future climate change scenarios. When land-cover/land-use data were included in the analyses, these negative effects were less pronounced. However, Silene conglomeratica, the rarest mountain endemic found in the study area, is still expected to face substantial range decline. Our results highlight the urgent need for the establishment of micro-reserves for these taxa.

Comparison of prophylactic effect of topical Alchemilla vulgaris in glycerine versus that of dexamethasone on postoperative sore throat after tracheal intubation using a double-lumen endobronchial tube: a randomized controlled study.

BACKGROUND: Sore throat and hoarseness frequently occur following general anesthesia with tracheal intubation and are effectively reduced when dexamethasone is used prophylactically. Alchemilla vulgaris in glycerine (Neo Mucosal Activator®) suppresses inflammatory response, possibly relieving sore throat. METHODS: We enrolled 94 patients (age ≥ 18 years) scheduled for thoracic surgery using double-lumen tube intubation. Before intubation, 0.2 mg/kg of dexamethasone was administered intravenously and 2 ml of normal saline was sprayed into the oropharyngeal cavity (Group D; n = 45), or 0.04 ml/kg normal saline was administered intravenously and 1 g of Neo Mucosal Activator® mixed with 1 ml of normal saline was sprayed into the oropharyngeal cavity (Group N; n = 43), in a double blind and prospectively randomized manner. Postoperative sore throat and hoarseness were recorded using a numeral rating scale and a 4-point scale to detect a change in voice quality following tracheal extubation (at 1, 6, and 24 h). The primary outcome was the incidence of sore throat at 24 h following surgery. The secondary outcomes were incidence and severity of sore throat and hoarseness. RESULTS: There were no significant differences in the incidence of sore throat at 24 h following surgery (57.8% vs. 46.5%; P = 0.290) or in the incidence and intensity of sore throat and hoarseness at 1, 6, and 24 h following surgery between the groups. CONCLUSIONS: A. vulgaris in glycerine did not significantly differ from dexamethasone for preventing sore throat and hoarseness owing to intubation.

Anti-Adipogenic Effect of Alchemilla monticola is Mediated Via PI3K/AKT Signaling Inhibition in Human Adipocytes.

Obesity is a persistent and continuously expanding social health concern. Excessive fat mass accumulation is associated with increased risk of chronic diseases including diabetes, atherosclerosis, non-alcoholic steatohepatitis, reproductive dysfunctions and certain types of cancer. Alchemilla monticola Opiz. is a perennial plant of the Rosaceae family traditionally used to treat inflammatory conditions and as a component of weight loss herbal mixtures. In the search for bioactive leads with potential anti-adipogenic effect from A. monticola extract (ALM), we have employed nuclear magnetic resonance (NMR) based metabolomics to obtain data for the phytochemical profile of the extract. Further, molecular docking simulation was performed against key adipogenic targets for selected pure compounds, present in the ALM extract. Evaluation of the biological activity was done in human adipocytes exposed to ALM (5, 10 and 25 µg/ml), pure astragalin (AST) or quercitrin (QUE) both at the concentrations of 5, 10 and 25 µM. Investigation of the molecular pathways involved was performed through real-time quantitative PCR and Western blot analyses. According to the docking predictions strong putative affinity was revealed for both AST and QUE towards peroxisome proliferator-activated receptor gamma (PPARγ) and phosphoinositide 3-kinase (PI3K). Assessment of the intracellular lipid accumulation revealed anti-adipogenic activity of ALM. Correspondingly, the expression of the adipogenic genes CCAAT/enhancer-binding protein alpha (CEBPA) and PPARG was downregulated upon ALM and AST treatment. The Western blotting results exposed protein kinase B (AKT), PI3K and PPARγ as targets for the inhibitory effect of ALM and AST on adipogenesis. Collectively, we provide a broader insight of the phytochemical composition of A. monticola. Additionally, we demonstrate the anti-adipogenic effect of ALM and its active compound AST in human adipocytes. Furthermore, PI3K/AKT signaling pathway is identified to mediate the ALM anti-adipogenic action. Hence, the ALM extract and its secondary metabolite AST are worth further exploration as potentially active agents in obesity management.

Initial Complete Chloroplast Genomes of Alchemilla (Rosaceae): Comparative Analysis and Phylogenetic Relationships.

The genus Alchemilla L., known for its medicinal and ornamental value, is widely distributed in the Holarctic regions with a few species found in Asia and Africa. Delimitation of species within Alchemilla is difficult due to hybridization, autonomous apomixes, and polyploidy, necessitating efficient molecular-based characterization. Herein, we report the initial complete chloroplast (cp) genomes of Alchemilla. The cp genomes of two African (Afromilla) species Alchemilla pedata and Alchemilla argyrophylla were sequenced, and phylogenetic and comparative analyses were conducted in the family Rosaceae. The cp genomes mapped a typical circular quadripartite structure of lengths 152,438 and 152,427 base pairs (bp) in A. pedata and A. argyrophylla, respectively. Alchemilla cp genomes were composed of a pair of inverted repeat regions (IRa/IRb) of length 25,923 and 25,915 bp, separating the small single copy (SSC) region of 17,980 and 17,981 bp and a large single copy (LSC) region of 82,612 and 82,616 bp in A. pedata and A. argyrophylla, respectively. The cp genomes encoded 114 unique genes including 88 protein-coding genes, 37 transfer RNA (tRNA) genes, and 4 ribosomal RNA (rRNA) genes. Additionally, 88 and 95 simple sequence repeats (SSRs) and 37 and 40 tandem repeats were identified in A. pedata and A. argyrophylla, respectively. Significantly, the loss of group II intron in atpF gene in Alchemilla species was detected. Phylogenetic analysis based on 26 whole cp genome sequences and 78 protein-coding gene sequences of 27 Rosaceae species revealed a monophyletic clustering of Alchemilla nested within subfamily Rosoideae. Based on a protein-coding region, negative selective pressure (Ka/Ks < 1) was detected with an average Ka/Ks value of 0.1322 in A. argyrophylla and 0.1418 in A. pedata. The availability of complete cp genome in the genus Alchemilla will contribute to species delineation and further phylogenetic and evolutionary studies in the family Rosaceae.