Determination of the Cytotoxicity and Antibiofilm Potential Effect of Equisetum arvense Silver Nanoparticles.

This study aimed to synthesize and characterize silver nanoparticles (AgNPs) by green synthesis from Equisetum arvense (Ea) extracts and to investigate their cytotoxicity, antibiofilm activity, and α-glucosidase enzyme inhibition. Diverse characterization techniques were applied to verify the production of nanoparticles. SEM examination confirmed that the size of nanoparticles is in the range of 40-60 nm. Also, interactions between silver and natural compounds of plant extract were confirmed through FT-IR and EDX analyses. It was determined that Equisetum arvense silver nanoparticles had antibiofilm activity against three different clinical strains with high biofilm-forming ability. AgNPs reduced the biofilm-forming capacity of clinical A. baumannii isolate with strong biofilm-forming capacity by approximately twofold, while the capacity of clinical K.pneumonaie and E.coli isolates decreased by 1.5 and 1.2 fold, respectively. The α-glucosidase enzyme inhibition potential of the AgNPs, which is determined as 93.50%, was higher than the plant extract with, and the α- 30.37%. MTT was performed to assess whether incubation of nanoparticles with A549 and ARPE-19 cell lines affected their viability, and a dramatic reduction in cell growth inhibition of both A549 and ARPE-19 cells was observed. It has been shown that A549 cells treated with 200 and 150 µg/mL nanoparticles had less cell proliferation compared to control cells at 24-h and 48-h incubation time. According to these results, Ea-derived AgNPs appear to have potential anticancer activity against A549 cancer cells. Investigating the effects of green synthesis nanoparticles on microbial biofilm and various tumors may be important for developing new therapies. The outcomes of this study have showed that Ea-AgNPsmay have a high potential both in the treatment of pathogenic strains that form biofilms, as well as in anticancer therapy use.

Genus Equisetum L: Taxonomy, toxicology, phytochemistry and pharmacology.

INTRODUCTION: The genus Equisetum (Equisetaceae) is cosmopolitan in distribution, with 41 recognized species. Several species of Equisetum are widely used in treating genitourinary and related diseases, inflammatory and rheumatic problems, hypertension, and wound healing in traditional medicine practices worldwide. This review intends to present information on the traditional uses, phytochemical components, pharmacological activities, and toxicity of Equisetum spp. and to analyze the new insights for further study. METHODS: Relevant literature has been scanned and collected via various electronic repositories, including PubMed, Science Direct, Google Scholar, Springer Connect, and Science Online, from 1960 to 2022. RESULTS: Sixteen Equisetum spp. were documented as widely used in traditional medicine practices by different ethnic groups throughout the world. A total of 229 chemical compounds were identified from Equisetum spp. with the major group of constituents being flavonol glycosides and flavonoids. The crude extracts and phytochemicals of Equisetum spp. exhibited significant antioxidant, antimicrobial, anti-inflammatory, antiulcerogenic, antidiabetic, hepatoprotective, and diuretic properties. A wide range of studies have also demonstrated the safety of Equisetum spp. CONCLUSION: The reported pharmacological properties of Equisetum spp. support its use in traditional medicine, though there are gaps in understanding the traditional usage of these plants for clinical experiments. The documented information revealed that the genus is not only a great herbal remedy but also has several bioactives with the potential to be discovered as novel drugs. Detailed scientific investigation is still needed to fully understand the efficacy of this genus; hence, very few Equisetum spp. were studied in detail for phytochemical and pharmacological investigation. Moreover, its bioactives, structure-activity connection, in vivo activity, and associated mechanism of action ought to be explored further.

New substances of Equisetum hyemale L. extracts and their in vivo antitumoral effect against oral squamous cell carcinoma.

ETHNOBOTANICAL RELEVANCE: Equisetum hyemale is used in traditional medicine as an anti-inflammatory, antioxidant, diuretic and anticancer agent. Recent studies have observed antiproliferative activity of this species in some tumor cell lines. AIM OF THE STUDY: The aim of this study was to evaluate the antiproliferative activity of the ethanol extract of E. hyemale and its partitions in oral squamous carcinoma cell lines, the death pathways induced by the most active partition, the acute toxicity and therapeutic activity, and the identification of the main compounds. MATERIALS AND METHODS: The ethanol crude extract was prepared from the stems of E. hyemale and partitions were obtained from this extract with n-hexane, dichloromethane and ethyl acetate. Cytotoxicity assays were performed using MTT on human oral tumor lines SCC-9, SCC4 and SCC-25, and normal primary fibroblasts. The main pathways of programmed cell death were analyzed. Acute toxicity in mice was performed using the most active partition, ethyl acetate. Antitumor activity was accessed in xenotransplants grafts of SCC-9 cells in Balb/nude mice. Phytochemical analysis was performed using UHPLC-MS/MS and dereplication was done using Global Natural Product Social Molecular Networking (GNPS) analysis. RESULTS: Ethanol extract, n-hexane and ethyl acetate partitions showed dose-dependent activity and selectivity towards oral tumor cells, with the ethyl acetate being the most bioactive. This medium polarity partition was shown to induce tumor cell death through apoptosis due to the presence of activated caspase 3/7, DNA fragmentation, chromatin condensation and phosphatidylserine exposure. The ethyl acetate partition also produced low toxicity in mice, provoking mild hepatic changes, but without causing necrosis and significantly reduced tumors volume and weight in xenotransplants of SCC-9 cells. Phytochemical analysis allowed identification of kaempferol glycosides and cinnamic acid derivatives previously described for E. hyemale. In addition it was possible to identify 6 new non-glycolyzed flavonoids 5-Hydroxy-3',4',7,8-tetramethoxyflavone (14), 5,4'-Dihydroxy-7,8,3'-trimethoxyflavone (15), 5,7-Dihydroxy-3',4'-dimethoxyflavone (16), 3',4,5,7-Tretramethoxyflavone (17), 5-Hydroxy-3'4',7-trimethoxyflavone (18), and 5,4'-Dihydroxy-3'-7'-dimethoxyflavone (19); besides 5 compounds already determined to be cytotoxic in other species, Isoferulic acid (1), Ferulic acid (2), Atractylenolide III (6), Dihydroxy-3',4'-dimethoxyflavone (16), and 5-Hydroxy-3'4 ',7-trimethoxyflavone (18). CONCLUSION: The results indicate that the E. hyemale extract and partitions inhibited 3 different cell lines of OSCC in a highly selective nontoxic way by inducing apoptosis of the cells. We identified 6 new non-glycosylated flavonoids and 5 other substances in this species.

Heavy metal accumulation efficiency and subsequent of cytogenotoxicity evaluation in the medicinal plant Equisetum hyemale.

Heavy metals in soils represent a threat to the environment, food safety, as well as human and animal health. The bioaccumulation of these elements in plants might enhance medium- and long-term adverse health risk promoting genetic alterations that lead to dermal, gastrointestinal, circulatory, renal, and brain disorders. The present study aimed to determine the bioaccumulation potential and cytogenotoxic effect of Equisetum hyemale extracts. E. hyemale seedlings were divided into two groups: exposed group (plants cultivated in soil with heavy metals solution) and control (plants cultivated in soil with distilled water). Heavy metals were quantified in the cultivation soils (control and exposed) and extracts (ethanolic and infusion) of vegetative parts from E. hyemale cultivated in both soils. Root length and cytogenotoxic effect were determined utilizing Allium cepa bioassay. Data demonstrated that Equisetum hyemale present the ability to absorb and bioaccumulate different heavy metals including lead, copper, cobalt manganese, zinc, iron and chromium. Given this property E. hyemale may be considered a reliable bioindicator to assess cytogenotoxicity of certain substances that exert adverse risks to environment and human and animal health.

Four new glucosides from the aerial parts of Equisetum sylvaticum.

Two previously undescribed megastigmane glucosides, (3S)-3-hydroxy-4-oxo-7,8-dihydro-ß-ionone-3-O-ß-D-glucopyranoside (1), (3S)-3-hydroxy-4-oxo-ß-ionone-3-O-ß-D-glucopyranoside (2), an apocarotenoid glucoside named equiseoside A (3) and an unusual aromatic compound with a glucose-fused skeleton named equiseoside B (4), together with 35 known compounds (5-39) were isolated from the aerial parts of Equisetum sylvaticum. The structures of these compounds were elucidated by spectroscopic methods, including 1D and 2D NMR, IR, CD, and HR-MS.

Aqueous extract from Equisetum arvense stimulates the secretion of Tamm-Horsfall protein in human urine after oral intake.

BACKGROUND: Within European traditional phytotherapy, extracts from different herbal plants are used for prevention and therapy of uncomplicated urinary tract infections and for flushing out of kidney grits. Besides increased urine flow by slight diuretic effects, also stimulation of Tamm-Horsfall protein (syn. THP, uromodulin) in the distal part of the kidney could explain reduced kidney gravel and anti-virulent activity against uropathogenic E. coli. PURPOSES: Evaluation of THP-inducing activity of extracts from Equisetum arvense, Levisticum officinalis, Ilex paraguariensis, Juniperus communis, Urtica dioica, and Taraxacum officinale by quantification of THP in urine samples after oral application to humans. STUDY DESIGN: 7 days p.o. application of the test intervention to healthy volunteers (n = 10 per intervention group) and analysis of urine samples at day 1 (untreated control values), and days 3, 6 and 8 on THP content by validated ELISA. Antiadhesive activity of urine samples was monitored by flow cytometry using UPEC strain NU14 against human T24 bladder cells. RESULTS: An aqueous extract from E. arvense, fully characterized by a specific LC-MS method, induced THP concentration in urine samples significantly during a 7-day p.o. application up to 300%, related to the untreated controls. Ex vivo investigation of the individual and pooled urine samples with elevated THP concentrations showed good correlation to antiadhesive effects against UPEC NU14 to T24 cells. Urine samples of the Equisetum treated volunteers had no effect on the proliferation and on biofilm formation of UPEC NU14. Silica excretion in the urine samples had no correlation to the respective THP levels. Monitoring of electrolyte content in the urine samples indicat ed diuretic effects of the intervention with Equisetum extract. Detailed phytochemical analysis of the Equisetum extract by LC-MS and LC-UV revealed an analytical protocol, which identified > 80 compounds from the extract by MS evaluations and 18 compounds by UV detection. This protocol will provide a valuable tool for future quality control of Equisetum extract. CONCLUSION: Aqueous extract from E. arvense significantly stimulates THP secretion in urine samples after 7 days of oral intake and inhibits the interplay between UPEC and bladder host cells. This could explain the therapeutic use of this herbal material for urinary tract infections and kidney gravel. Detailed phytochemical analysis of the Equisetum extract by LC-MS and LC-UV revealed an analytical protocol, which identified > 82% of all eluted compounds. This protocol will provide a valuable tool for future quality control of Equisetum extract.

Dose-Dependent Variation in Anticancer Activity of Hexane and Chloroform Extracts of Field Horsetail Plant on Human Hepatocarcinoma Cells.

Horsetail fern plant is botanically known as Equisetum arvense L., and it is a good source of phenolic flavonoids, phenolic acids, and compounds. Anticancer properties of hexane and chloroform extracts of the horsetail fern plant and their mechanisms involved in the anticancer activity on human hepatocarcinoma (HuH-7) cells were examined. Cytotoxicity was evaluated by using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and NRU (neutral red uptake) assays. Other parameters such as oxidative stress and apoptosis in pretreated hexane and chloroform extracts of the horsetail fern plant were examined in HuH-7 cells. The observation showed that hexane and chloroform extract of the horsetail fern plant exhibited cytotoxicity against HuH-7 cells. The value of IC50-24 h of hexane and chloroform extract of the horsetail fern plant was determined as 199.0 µg/ml and 161.90 0 µg/ml for HuH-7 cells, respectively, and on the basis of IC50 value, three acute concentrations, viz., 75% of IC50, 50% of IC50, and 25% of IC50, were determined for further study. The lower dose of extracts hexane and chloroform extract of the horsetail fern plant did not show significant toxicity. Higher concentrations of extract induced significant antioxidant effects as well as apoptosis effects. However, exposure to hexane and chloroform extract of the horsetail fern plant upregulated the expression of Bax and p53 in HuH-7 cells. These data suggest that hexane and chloroform extract of the horsetail fern plant plays a significant role in the induction of toxicity via the regulation of oxidative stress in HuH-7 cells. This work may be useful for cancer chemotherapy.

Biomarker-Based Determination of Equiseti herba Contamination by Equisetum palustre Using HPLC-MS/MS.

Equiseti herba has been traditionally indicated in bacterial diseases of the efferent urinary tract or bad healing wounds in many regions worldwide. Most of the plant material used for medical purposes comes from collections of wild growing plants. The European Pharmacopoeia requires that Equiseti herba should consist of a minimum of 95% Equisetum arvense and a maximum of 5% foreign ingredients. This includes Equisetum palustre, which is known for its potentially toxic alkaloid palustrine. However, both Equisetum species are quite common, look morphologically very similar, and share similar habitats, hence, are therefore often confused. Recently, several structurally related Equisetum alkaloids have been identified in E. palustre but not in E. arvense. We have established a hydrophilic interaction liquid chromatography HPLC-ESI-MS/MS method for the detection of these E. palustre-specific Equisetum alkaloids in order to quantify the contamination of Equiseti herba (E. arvense) by E. palustre plant material. In a second, independent approach, the results of the HPLC-MS/MS analysis were confirmed by scanning electron microscopy, looking for the species-specific characteristics of the stoma apparatus of E. palustre. Thirty-four Equiseti herba products obtained from different pharmacies, drug stores, supermarkets, and web stores were analyzed. The majority of the products (26 out of 34) were Equisetum alkaloid positive, with contents ranging from 0.29 - 21.7 mg of Equisetum alkaloids/kg (d. w.). In addition, the transfer of Equisetum alkaloids into tea infusions was investigated, demonstrating a 42 to 60% transfer rate for cold and hot water extraction of Equisetum alkaloid-contaminated Equiseti herba, respectively.

Silicon Resorption from Equisetum arvense Tea - A Randomized, Three-Armed Pilot Study.

Equisetum arvense tea (TEA) contains high concentrations of silicon and has been used in folk medicine for the treatment of inflammatory ailments. We examined the resorption of silicon after TEA consumption. Safety and immunological effects were secondary outcomes. A monocentric, randomized, three-armed pilot study was conducted with 12 voluntary, healthy, male subjects. The study is registered in the German register for clinical trials (DRKS-ID: DRKS00016628). After a low silicon diet for 36 hours, 1000 mL TEA1 with approximately 200 000 µg silicon/L, TEA2 with approximately 750 000 µg silicon/L, or Si-low-Water (approximately 10 - 10 000 µg silicon/L as a control) were ingested on three consecutive days. Blood and urine samples were collected at baseline, day 1 examining silicon kinetics, day 3 examining silicon accumulation, and day 8 (safety, immunological parameters). Si-low-Water intake did not change silicon serum (Cmax 294 µg/L) or urine (19 000 µg/24 h) concentrations compared to baseline. Cmax was 2855 µg/L for TEA1 and 2498 µg/L for TEA2; tmax was 60 and 120 min, respectively. Silicon accumulation did not occur. Urine silica within 24 h (E24 h) was higher after TEA2 compared to TEA1 ingestion (142 000 vs. 109 000 µg/24 h). Serum silicon levels at t = 120 min differed significantly after intake of TEA2 or intake of Si-low-Water (p = 0.029). The immunological parameters did not show any significant changes indicating immunosuppressive effects in volunteers. TEA1 was well tolerated, while TEA2 caused diarrhoea in 4 subjects. Our investigations show that intake of TEA1 leads to significant rise in serum silicon concentration.

Relative absorption of silicon from different formulations of dietary supplements: a pilot randomized, double-blind, crossover post-prandial study.

The purpose of the present study was to compare the relative absorption of a new powder presentation of silicon (Si) as orthosilicic acid with maltodextrin (Orgono Powder) compared to usual Si liquid presentations as orthosilicic acid with Equisetum arvense and Rosmarinus officinalis (G5 Siliplant) and orthosilicic acid with aloe vera (G7 Aloe). All dietary supplements were administered at the same Si oral dose (21.6 mg) in a randomized, double-blind, crossover post-prandial study conducted in 5 healthy men. Urine was collected at baseline and over the 6-h post-dose period in 2 separate 3-h collections for the analysis of Si concentration, which was conducted by inductively coupled plasma optical emission spectrometry as the gold standard method. No significant differences in total urinary Si excretion were found after the intake of these 3 dietary supplements; 34.6%, 32.4% and 27.2% of the ingested Si from G7 Aloe, G5 Siliplant and Orgono Powder, respectively, was excreted in urine over the 6-h follow-up period. The 3 different oral Si formulations tested, in powder and liquid presentations, provide highly bioavailable Si and present an equivalent relative absorption in healthy humans.

Neuroprotective and Antioxidant Enhancing Properties of Selective Equisetum Extracts.

The sterile stems belonging to the Equisetum species are often used in traditional medicine of various nations, including Romanians. They are highly efficient in treating urinary tract infections, cardiovascular diseases, respiratory tract infections, and medical skin conditions due to their content of polyphenolic derivatives that have been isolated. In this regard, this study aimed to provide the chemical composition of the extracts obtained from the Equisetum species (E. pratense, E. sylvaticum, E. telmateia) and to investigate the biological action in vitro and in vivo. For the chemical characterization of the analyzed Equisetum species extracts, studies were performed by using ultra-high-performance liquid chromatography (UHPLC-DAD). In vitro evaluation of the antioxidant activity of the plant extracts obtained from these species of Equisetum genus was determined. The neuroprotective activity of these three ethanolic extracts from the Equisetum species using zebrafish tests was determined in vivo. All obtained results were statistically significant. The results indicate that E. sylvaticum extract has a significant antioxidant activity; whereas, E. pratense extract had anxiolytic and antidepressant effects significantly higher than the other two extracts used. All these determinations indicate promising results for the antioxidant in vitro tests and neuroprotective activity of in vivo tests, particularly mediated by their active principles.

Antioxidant and Antiproliferative Activity of The Ethanolic Extract of Equisetum Myriochaetum and Molecular Docking of Its Main Metabolites (Apigenin, Kaempferol, and Quercetin) on ß-Tubulin.

Equisetum myriochaetum is a semi-aquatic plant found on riverbanks that is commonly used in traditional medicine as a diuretic agent. Additionally, the genus Equisetum stands out for its content of the flavonoid kaempferol, a well-known antiproliferative agent. Therefore, in this study, E. myriochaetum ethanolic extract was tested in vitro against a cervical cancer cell line (SiHa). Additionally, the antioxidative activity was evaluated through a 2,2-diphenyl-1-picrilhidrazil (DPPH) assay. Finally, a molecular docking analysis of apigenin, kaempferol, and quercetin on the active site of ß-tubulin was performed to investigate their potential mechanism of action. All fractions of E. myriochaetum ethanolic extract showed antioxidative activity. Fraction 14 displayed an antiproliferative capacity with a half maximal inhibitory concentration (IC50) value of 6.78 µg/mL against SiHa cells.

Anti-inflammatory effect of glycyrrhizin with Equisetum arvense extract.

Periodontal disease is the most prevalent infectious disease, and inflammatory mediators play critical roles in its progression. Therefore, controlling pro-inflammatory cytokine production, especially at initial disease stages, is essential to maintaining gingival and periodontal health. Glycyrrhizin (GL) has an anti-inflammatory effect and has been added to toothpaste and mouth rinse to prevent periodontal disease. However, there is a maximum dose for the use of GL. The aim of the present study is to screen plant extracts which can effectively enhance the effects of GL. The effects of extracts from six different plants on GL-suppressed TNF-α expression in Aggregatibacter actinomycetemcomitans (A.a.)-LPS-stimulated human oral keratinocytes (RT7) were examined. Results demonstrated that Equisetum arvense (EA) extract had the strongest additive effect on the suppression of TNF-α by GL at both mRNA and protein levels. In addition, GL downregulated the production of TNF-α by suppressing NF-κB p65 phosphorylation, but not JNK or p38 phosphorylation. In contrast, EA decreased JNK phosphorylation but not NF-κB p65 or p38 phosphorylation. The combination of GL and EA effectively attenuated A.a.-LPS-induced phosphorylation of NF-κB p65 and JNK. Furthermore, an LPS-induced periodontitis rat model showed that GL with EA supplementation significantly downregulated TNF-α mRNA in the gingival tissue. These results indicate that EA can suppress A.a.-LPS-induced pro-inflammatory cytokine production by inhibiting JNK activation and can promote the anti-inflammatory effects of GL. Our findings suggest that a combination of GL and EA may improve the development of new oral hygiene products aimed at enhancing periodontal health.

SIRT1 Activation by Equisetum Arvense L. (Horsetail) Modulates Insulin Sensitivity in Streptozotocin Induced Diabetic Rats.

BACKGROUND: Equisetum arvense L., commonly known as field horsetail is a perennial fern of which extracts are rich sources of phenolic compounds, flavonoids, and phenolic acids. Activation of SIRT1 that was shown to be involved in well-known signal pathways of diabetic cardiomyopathy has a protective effect against oxidative stress, inflammatory processes, and apoptosis that are the basis of diseases such as obesity, diabetes mellitus, or cardiovascular diseases. The aim of our study was to evaluate the antidiabetic and cardioprotective effects of horsetail extract in streptozotocin induced diabetic rats. METHODS: Diabetes was induced by a single intraperitoneal injection of 45 mg/kg streptozotocin. In the control groups (healthy and diabetic), rats were administered with vehicle, whilst in the treated groups, animals were administered with 50, 100, or 200 mg/kg horsetail extract, respectively, for six weeks. Blood glucose levels, glucose tolerance, and insulin sensitivity were determined, and SIRT1 levels were measured from the cardiac muscle. RESULTS: The horsetail extract showed moderate beneficial changes in blood glucose levels and exhibited a tendency to elevate SIRT1 levels in cardiomyocytes, furthermore a 100 mg/kg dose also improved insulin sensitivity. CONCLUSIONS: Altogether our results suggest that horsetail extract might have potential in ameliorating manifested cardiomyopathy acting on SIRT1.

Antidiabetic effect of Equisetum giganteum L. extract on alloxan-diabetic rabbit.

ETHNOPHARMACOLOGICAL RELEVANCE: Equisetum giganteum has been traditionally used as an anti-diabetic herbal remedy to treat diabetes in the southern state of Rio Grande do Sul in Brazil. AIM OF THE STUDY: Considering the ethonopharmacology and historical importance of E. giganteum, its potential antidiabetic effect was evaluated in alloxan induced diabetic rabbits. MATERIAL AND METHODS: Samples of Equisetum giganteum were collected in the city of Ouro Preto, Minas Gerais, Brazil. Butanolic and aqueous extracts were prepared and subsequently evaluated for anti-diabetic properties in vivo using albino male rabbits. At the end of the treatment period, the animals were euthanized, and histopathological analysis were carried out. The following biochemical parameters were studied: glucose, triacylglycerol, cholesterol, albumin, creatinine, glycosylated hemoglobin and lipase. The phytochemical profile of the extracts was studied by liquid chromatography techniques coupled to a UV/VIS detector and high-resolution mass spectrometry. RESULTS: Both aqueous and butanolic extracts were capable of reducing significantly the levels of glucose, cholesterol and triacylglycerol and thus demonstrating their hypolipidemic and hypoglycemiant effects. Furthermore, the extracts prevented the occurrence of hepatic complications during treatment. The phytochemical profile of the extracts was investigated, and the natural products detected were in agreement with those that had been previously described in the literature. CONCLUSION: Based on the significant reductions in biochemical parameters and the histologic evidence for the absence of complications in the liver, pancreas of the treated animals, Equisetum giganteum can be a therapeutically relevant resource in the treatment of diabetes and hyperlipidemia.

Efficacy of application of Equisetum pyramidale Goldm. hydrogel for tissue restoration of induced skin lesions in Wistar rats / Eficácia da aplicação de Equisetum pyramidale Goldm. hidrogel na restauração tecidual em lesões cutâneas induzidas em ratos Wistar

Abstract Medicinal plants, such as E. pyramidale, are alternatives against infections and assist in the healing process of wounds. We evaluated the effects of Hydrogel of E. pyramidale on the healing of cutaneous wounds in animal models by morphological, morphometric and histological analyses, and elucidated major components. The ethanolic extract of E. pyramidale shoots was submitted to phytochemical analysis. For cicatrizing activity assay, Wistar rats were used, and the groups evaluated were hydrogel containing ethanolic extract of E. pyramidale at 2% compared to control groups. The treatment periods were 3, 7 and 21 days, and macroscopic and microscopic analyses were performed. Data were submitted to Analysis of Variance (p <0.05). Phytochemistry and quantification evidenced that flavonoids are main constituents and that they were evidenced by thin layer chromatography (TLC), high efficiency (HPLC) and infrared (IR) methods. In TLC, using flavonoid patterns, we observed pinocembrin. The hydrogel containing ethanolic extract of E. pyramidale at 2% was effective in wound regression. E. pyramidale can be used for the treatment of second intention wounds, and the effective healing may be due to a high flavonoid content.

Resumo As plantas medicinais, como E. pyramidale, são uma alternativa para combater infecções e ajudar o processo de cicatrização de feridas. Avaliar os efeitos do hidrogel de E. pyramidale na cicatrização de feridas cutâneas em modelos animais através de análises morfológicas, morfométricas e histológicas, bem como para elucidar os principais componentes. O extrato etanólico das partes aéreas de E. pyramidale foi submetido a análise fitoquímica. Para o ensaio de atividade cicatrizante, foram utilizados ratos Wistar e os grupos avaliados foram o hidrogel com extrato etanólico de E. pyramidale 2% em comparação com os grupos controle. Os períodos de tratamento foram 3, 7 e 21 dias e as análises macroscópicas e microscópicas foram realizadas e os dados submetidos à Análise de Variância (p <0,05). A fitoquímica e a quantificação indicaram que os flavonóides são os constituintes principais e que mesmos foram evidenciados na cromatografia de camada delgada (CCD), de alta eficiência (HPLC) e por infravermelho (IV). Em CCD, empregando padrões de flavonoides, foi observado a pinocembrina. O hidrogel com extrato etanólico de E. pyramidale 2% foi eficaz na regressão da ferida. E. pyramidale pode ser usado no tratamento de feridas de segunda intenção e a cura efetiva pode ser devido ao alto teor de flavonoides.

Efficacy of application of Equisetum pyramidale Goldm. hydrogel for tissue restoration of induced skin lesions in Wistar rats.

Medicinal plants, such as E. pyramidale, are alternatives against infections and assist in the healing process of wounds. We evaluated the effects of Hydrogel of E. pyramidale on the healing of cutaneous wounds in animal models by morphological, morphometric and histological analyses, and elucidated major components. The ethanolic extract of E. pyramidale shoots was submitted to phytochemical analysis. For cicatrizing activity assay, Wistar rats were used, and the groups evaluated were hydrogel containing ethanolic extract of E. pyramidale at 2% compared to control groups. The treatment periods were 3, 7 and 21 days, and macroscopic and microscopic analyses were performed. Data were submitted to Analysis of Variance (p <0.05). Phytochemistry and quantification evidenced that flavonoids are main constituents and that they were evidenced by thin layer chromatography (TLC), high efficiency (HPLC) and infrared (IR) methods. In TLC, using flavonoid patterns, we observed pinocembrin. The hydrogel containing ethanolic extract of E. pyramidale at 2% was effective in wound regression. E. pyramidale can be used for the treatment of second intention wounds, and the effective healing may be due to a high flavonoid content.

A randomised, double-blind, placebo-controlled clinical trial assessing the efficacy of bedtime buddy® for the treatment of nocturnal enuresis in children.

BACKGROUND: Nocturnal enuresis (NE), or 'bedwetting', is a form of night-time urinary incontinence occurring in younger children. A diagnosis of NE can be socially disruptive and psychologically stressful for a child. The most common strategies used by parents to manage NE are waking the child during the night to use the bathroom and limiting the child's water intake before going to bed. Behavioural or educational therapies for NE such as urotherapy or bladder retraining are widely accepted and considered as a mainstream treatment option for non-neurogenic lower urinary tract dysfunction in children. Pharmacotherapy also plays an ancillary role. However, there is no gold standard therapy or intervention to effectively manage NE. METHODS: This study aims to determine the efficacy of a herbal combination in the treatment of NE in children. The target population for this study is 80 children aged between 6 and 14 years old (males and females) who have primary nocturnal enuresis ≥3 per week (wet nights). The active group will receive one or two capsules per day containing 420 mg of a proprietary blend of Urox® (Seipel Group, Brisbane, Australia) containing Cratevox™ (Crataeva nurvala L; Capparidaceae; Varuna) stem bark extract standardised for 1.5% lupeol: non-standardised Equisetum arvense L. (Equisetaceae; Horsetail) stem extract; and, non-standardised Lindera aggregata Sims. The primary outcome for this study is the frequency of nocturia. Secondary outcomes include safety, quality of life, and daytime incontinence. Each participation will be involved in the trial for 32 weeks including contact with the research team every 2 weeks for the first 8 weeks and then every 8 weeks until trial completion. DISCUSSION: This study examines a novel treatment for an under-researched health condition affecting many children. Despite the availability of several therapies for NE, there is insufficient evidence to support the use of any one intervention and as such this randomised placebo-controlled phase II trial will be an important contribution to understanding potential new treatments for this condition. TRIAL REGISTRATION: Australian and New Zealand Clinical Trials Registration Number: 12618000288224. PROTOCOL: 23 February 2018, version 1.1.

Structural, rheological and antioxidant properties of pectins from Equisetum arvense L. and Equisetum sylvaticum L.

The pectins were isolated from sterile stems of E. arvense (EA, yield 5.9%) and E. sylvaticum (ES, yield 4.8%) (Equisetaceae) using ammonium oxalate extraction after preliminary treatment with dilute HCl (рH 4.0). The pectins possessed high molecular weight (Mw, 340-360 kDa), high GalA content (ca. 85%), low degrees of methyl-esterification (14-16%) and acetylation (3-8%). NMR analysis indicated extensive regions of partially methyl-etherified and 3-O-acetylated HG and minor regions of low branched RG in the fragment isolated after hydrolysis of pectin EA by pectinase. Pectin EA produced a higher viscosity solution, formed a stronger and more rigid ionotropic hydrogel than pectin ES. The pectins scavenged DPPH and hydroxyl radicals, but not the superoxide radical and hydrogen peroxide. Phenolic compounds (0.11 and 0.23%) associated with polysaccharide moieties were apparently responsible for the differences in the anti-DPPH scavenging activity of pectins EA and ES (63 and 49%). The findings suggested that pectin from E. arvense should be more perspective than pectin from E. sylvaticum on their use as components of wound healing remedies.

A randomized, open-label, multicenter, comparative study of therapeutic efficacy, safety and tolerability of BNO 1030 extract, containing marshmallow root, chamomile flowers, horsetail herb, walnut leaves, yarrow herb, oak bark, dandelion herb in the treatment of acute non-bacterial tonsillitis in children aged 6 to 18 years.

Seventy to 95% of acute tonsillitis episodes are caused by viral infection, therefore why antibiotic therapy is not indicated in majority of cases. In such cases, acetaminophen or ibuprofen are used to alleviate the symptoms. The objective of this study was assessment of efficacy of phytoneering extract BNO 1030 (Imupret®) in patients with acute non-bacterial tonsillitis. METHODS: This randomized, open-label, multicenter, comparative study randomised 238 outpatients aged 6-18 years to receive either BNO 1030 (Imupret®) as a supplement to standard symptomatic therapy, or standard therapy. Assessment criteria were as follows: sore throat dynamics at rest and at swallowing, throat irritation associated with cough, general condition, day of withdrawal of antipyretics, the share of treatment responders, as well as assessment of "therapeutic benefit" from the use of BNO 1030. RESULTS: Decreased intensity of acute tonsillitis symptoms to 1 point and lower, assessed using 4-point scale starting from the day 5 of treatment (p < 0.005), alleviation of local symptoms and general condition starting from day 2 of the disease (р < 0.001), withdrawal of antipyretics starting from day 4 of treatment (p < 0.005), increase of the number of treatment responders to 81.6% (p < 0.005) versus the control were reported. "Therapeutic benefit" was 4.2 days. All patients tolerated phytotherapy well, and no adverse reactions were seen. CONCLUSION: BNO 1030 (Imupret®) is a safe and effective product for treatment of acute non-bacterial tonsillitis in children aged 6-18 years, assuring therapeutic benefit when prescribed additionally to the standard symptomatic therapy.