Bioactivity-guided isolation of the antidiabetic principle in Pterocarpus Santalinoides leaf extract

Abstract Pterocarpus santalinoides is used in Nigerian ethnomedicine to treat diabetes mellitus. This study aimed to establish the antidiabetic property of the plant, and isolate and characterize its active principle. Dried and pulverized leaves (500 g) of P. santalinoides were extracted with 1.8 L of 80 % hydromethanol by cold maceration. The dried extract (10 g) was partitioned into n-hexane, ethyl acetate (EtOAc), n-butanol, and water. Antidiabetic activitiy-guided isolation by column chromatographic separation of the EtOAc soluble and purification of the sub-fractions by repeated preparative thin layer chromatography (pTLC) yielded a C-glycosyl flavonoid, identified as isovitexin. The chemical structure was elucidated based on high-resolution mass spectroscopy, 1D, and 2D nuclear magnetic resonance spectroscopic analyses. Alloxan-induced diabetic rat model was adopted for antidiabetic screening. The extract of P. santalinoides (100-200 mg/kg), fraction F4 (50 mg/kg), sub-fraction F4.3 (10 mg/kg), and the semi-purified compound F4.3.2 (5 mg/kg) significantly (p<0.05) reduced the fasting blood glucose of alloxan-induced diabetic rats, causing 48.4, 69.4, 57.7 and 64.5 % antidiabetic activity respectively, compared with > 68 % recorded in glibenclamide (2 mg/kg) control. These results reveal that isovitexin is the antidiabetic principle in P. santalinoides

In vitro antibacterial-antibiofilm effect of Hypericum atomarium Boiss and chemical composition

Abstract Treatment with plant is considered an effective option against increased antibiotic resistance. In this study antibiofilm activity of methanol (CH3OH), chloroform (CHCl3), ethyl acetate (EtOAc) and water (H2O) extracts of Hypericum atomarium Boiss. which is member of Hypericum genus was evaluated in Pseudomonas aeruginosa PAO1 and antibacterial performance against Gram (+) and Gram (-) strains and also bioactive compounds of extract were analysed using by HPLC and GC-MS. According to antibacterial activity test results the extracts were effective all Gram (+) bacteria and Gram (-) Chromobacterium violaceum (MICs ranging from 0.42 µg/ml to 4.3 mg). Inhibition effect of biofilm formation was found to be different rate in extracts (methanol-63%, chloroform-52%). The major flavonoids were detected (−)-epicatechin (2388.93 µg/ml) and (+)-catechin (788.94 µg/ml). The main phenolic acids were appeared as caffeic acid 277.34 µg/ml and chlorogenic acid 261.79 µg/ml. And according to GC results α-pinene was found main compound for three solvent extracts methanol, chloroform and ethyl acetate 67.05, 62.69, 49.28% rate respectively

Influência do metil-jasmonato na formação de compostos voláteis do aroma em Bananas (Musa acuminata cv. Nanicão) e Laranjas (Citrus sinensis cv. Pera) / Influence of methyljasmonate on the formation of aroma volatile compounds in Bananas (Musa acuminata cv. Nanicão) and Oranges (Citrus sinensis cv. Pera)

O principal hormônio associado aos processos do amadurecimento é o etileno, porém, na formação de compostos voláteis nos frutos, observa-se que as auxinas, o ácido abscísico e os jasmonatos também podem atuar como reguladores. Estudos indicam que em frutos climatéricos deve haver uma interação entre o metil jasmonato (MeJA) e o etileno na formação de compostos voláteis, mas em frutos não-climatéricos tal interação não é tão evidente. Há evidências de que o MeJA atue na regulação de algumas vias metabólicas relacionadas ao amadurecimento em frutos, sendo capaz de induzir aumento na produção de várias classes de compostos voláteis, através da expressão de genes que codificam as enzimas relacionadas às suas vias biossintéticas. Neste sentido, o objetivo deste projeto foi avaliar o efeito do metil jasmonato sobre o padrão de produção de compostos voláteis do aroma em frutos climatéricos e não-climatéricos. Precedentes do laboratório de Química, Bioq. e Biol. Molecular de Alimentos indicam que o MeJA apresentou padrões diferentes de comportamento em frutos climatéricos e não-climatéricos no que tange a formação do aroma. Assim, o presente projeto tem por hipótese a diferença de influência que o MeJA exerce sobre a produção de compostos voláteis em frutos climatéricos e não-climatéricos. Para testar esta hipótese foi avaliado o efeito do tratamento com MeJA na produção de compostos voláteis do aroma durante o amadurecimento de banana (Musa acuminata, cv. Nanicão), como exemplo de fruto climatérico e laranja (Citrus sinensis cv Pêra) para não-climatéricos. Os frutos foram divididos em grupo controle e tratado com MeJA (10 ppm/24h), armazenados em caixas plásticas tampadas e lacradas. Após tratamento foram submetidos a análises diárias da produção de etileno por cromatrogafia gasosa (CG), cor da casca e pesagem. Baseado em escalas de cor e a polpa foi congelada em N2 líq. e armazenada a -80°C para posterior análise dos compostos voláteis por cromatografia gasosa acoplada à espectrometria de massas (GC-MS). Ésteres, álcoois, cetonas e aldeídos foram compostos majoritariamente identificados na banana e terpenos, aldeídos, ésteres na laranja. As Bananas sofreram influência no perfil de acetato de isoamila, butonoato de butila, isobutirato de isoamila e isolvalerato de isoamila do começo ao fim do tratamento com MeJA, e as laranjas o tratamento influenciou os compostos Cis-muirola-3-5-diene, gamageraniol, alfa-copaeno, valenceno, alfa-pineno, carvone, geranial, entre outros terpenos, aldeídos como 3-hexanal e 2-hexenal (E) e ésteres como butirato de etila, nerol e tiglato de etilo. Os ésteres em frutos são produzidos por várias isoformas das álcool acil transferases (AATs). Estudos explicam que, ao menos 31 transcritos de AATs foram identificados em bananas, sendo 8 com altos níveis de expressão. Assim, é plausível supor que tal variedade de transcritos, e por conseguinte as AATs que codificam, sejam reguladas por múltiplos fatores, o que pode incluir o MeJa dentre outros sinais hormonais. Os terpenos são formados a partir de duas rotas, a do ácido mevalônico (MVA) e a rota do metileritritol fosfato (MEP). Compostos como, D-limoneno (51) e beta-selineno (62) tiveram níveis relativos maiores nos frutos do grupo controle, enquanto compostos terpênicos como geranial (59), valenceno (79) e o-cimeno (128), apresentaram maiores níveis nos frutos tratados com MeJa, no primeiro dia após o tratamento. Os resultados mostraram que o tratamento hormonal com MeJA causou mudanças do início ao fim do amadurecimento na composição do aroma de bananas (Musa acuminata cv Nanicão) e laranjas (Citrus sinensis cv Pera)

The main hormone associated with ripening processes is ethylene, but in the formation of volatile compounds in fruits, auxins, abscisic acid and jasmonates can also act as regulators. Studies indicate that in climacteric fruits there should be an interaction between methyl jasmonate (MeJA) and ethylene in the formation of volatile compounds, but in nonclimacteric fruits such interaction is not so evident. There is evidence that MeJA acts in the regulation of some metabolic pathways related to fruit ripening, being able to induce an increase in the production of several classes of volatile compounds, through the expression of genes that encode enzymes related to their biosynthetic pathways. In this sense, the objective of this project was to evaluate the effect of methyl jasmonate on the production pattern of aroma volatile compounds in climacteric and non-climacteric fruits. Precedents from the Laboratory of Food Chemistry, Bioq. and Molecular Biol. Molecular Chemistry, Bioq. and Molecular Biol. of Foods laboratory indicate that MeJA showed different behavior patterns in climacteric and non-climacteric fruits regarding aroma formation. Thus, the present project hypothesizes the different influence that MeJA has on the production of volatile compounds in climacteric and non-climacteric fruits. To test this hypothesis the effect of MeJA treatment on the production of volatile aroma compounds during ripening of banana (Musa acuminata, cv. Nanicão) as an example for climacteric fruit and orange (Citrus sinensis cv Pêra) for non-climacteric fruit was evaluated. Fruits were divided into control and MeJA treated group (10 ppm/24h), stored in capped and sealed plastic boxes. After treatment they were subjected to daily analysis of ethylene production by gas chromatography (GC), peel color and weighing. Based on color scales and the pulp was frozen in liquid N2 and stored at -80°C for subsequent analysis of volatile compounds by gas chromatography coupled to mass spectrometry (GC-MS). Esters, alcohols, ketones and aldehydes were compounds mostly identified in banana and terpenes, aldehydes, esters in orange. Bananas were influenced in the profile of isoamyl acetate, butyl butonoate, isoamyl isobutyrate and isoamyl isolvalerate from the beginning to the end of the MeJA treatment, and oranges the treatment influenced the compounds Cis-myrola-3-5-diene, gamma-geraniol, alpha-copaene, valencene, alpha-pinene, carvone, geranial, among other terpenes, aldehydes like 3-hexanal and 2-hexenal (E), and esters like ethyl butyrate, nerol, and ethyl tiglate. Esters in fruits are produced by various isoforms of the alcohol acyl transferases (AATs). Studies explain that at least 31 AAT transcripts have been identified in bananas, 8 of which have high expression levels. Thus, it is plausible to assume that such a variety of transcripts, and therefore the AATs they encode, are regulated by multiple factors, which may include MeJa among other hormonal signals. Terpenes are formed from two routes, the mevalonic acid (MVA) route and the methylerythritol phosphate (MEP) route. Compounds such as, D-limonene (51) and beta-selinene (62) had higher relative levels in the fruits of the control group, while terpenic compounds such as geranial (59), valencene (79) and o-cymene (128), showed higher levels in the MeJa treated fruits on the first day after treatment. The results showed that the hormonal treatment with MeJA caused changes from the beginning to the end of ripening in the aroma composition of bananas (Musa acuminata cv Nanicão) and oranges (Citrus sinensis cv Pera)

Dissolution profile of different brands of low solubility drugs available in the Nigerian and Brazilian pharmaceutical markets / Dissolution profile of different brands of low solubility drugs available in the Nigerian and Brazilian pharmaceutical markets

The purpose of this work was to elaborate a diagnosis of the dissolution test in Africa in comparison with Brazil, evaluating the dissolution profile of low solubility drugs such as albendazole, ibuprofen, furosemide, glibenclamide, hydrochlorothiazide and carvedilol to ascertain their quality. The dissolution profiles were evaluated by utilizing the United States Pharmacopeia (USP). The glibenclamide medicine was evaluated according to the Food and Drug Administration (FDA), while a dissolution method was developed for the carvedilol medicine. A filter selection test for all the drugs showed that cannula is suitable for all, except for carvedilol, which is centrifuged. The various brands of Nigerian and Brazilian medicines tested showed some statistical differences. The suitable conditions that allowed the dissolution of carvedilol to be determined were the USP type II apparatus at 75 rpm containing 900 mL of acetate buffer, pH 4.5. The results of the dissolution test showed that out of the 17 different brands of Brazilian medicines and 17 different products from Nigeria, 94.12% and 58.82% passed respectively

O objetivo deste trabalho foi elaborar um diagnóstico do teste de dissolução na África em comparação ao Brasil, avaliando o perfil de dissolução de medicamentos de baixa solubilidade como albendazol, ibuprofeno, furosemida, glibenclamida, hidroclorotiazida e carvedilol para verificar sua qualidade.Os perfis de dissolução foram avaliados utilizando a Farmacopeia dos Estados Unidos (USP). O medicamento glibenclamida foi avaliado de acordo com a Food and Drug Administration (FDA), enquanto um método de dissolução foi desenvolvido para o medicamento carvedilol.Um teste de seleção de filtro para todos os medicamentos mostrou que a cânula é adequada para todos, exceto para o carvedilol, que é centrifugado. As diversas marcas de medicamentos Nigerianos e Brasileiros testadas apresentaram algumas diferenças estatísticas. As condições adequadas que permitiram a determinação da dissolução do carvedilol foram o aparelho USP tipo II a 75 rpm contendo 900 mL de tampão acetato, pH 4,5. Os resultados do teste de dissolução mostraram que das 17 diferentes marcas de medicamentos brasileiros e 17 diferentes produtos da Nigéria, 94,12% e 58,82% foram aprovados, respectivamente

Spectrophotometric determination of acid dissociation constants of some arylpropionic acids and arylacetic acids in acetonitrile-water binary mixtures at 25ºC

Abstract The acid dissociation constant of drug active compounds (arylpropionic and aryl acetic acids) were determined in acetonitrile and water binary mixtures (corresponding volume fractions of 0.40, 0.45, 0.50, and 0.55) by using a multi-wavelength spectrophotometric method. Drug active compounds, which were slightly soluble in water, were studied in these binary mixtures. The dissociation constants of drug active compounds are important in drug design studies and in any research of the biopharmaceutical and physicochemical properties of drugs. The STAR program was used for the determination of dissociation constants. The acidity constants of arylpropionic and aryl acetic acids were correlated with the Kamlet and Taft solvaatochromic parameters. Aqueous pKa values of these arylpropionic and aryl acetic acids were determined from pKa values obtained from acetonitrile and water binary mixtures with varying volume fractions. The studied drugs had a pKa value corresponding to base functional group. Results showed that the acid dissociation constant values of the drug active compounds increased with an increase in acetonitrile content in the medium.

Development and validation of a reversed-phase HPLC method for quantification of 1'-acetoxychavicol acetate content in a nanostructured lipid carrier formulation

Abstract 1'-acetoxychavicol acetate (ACA)-loaded nanostructured lipid carriers (NLCs) were formulated for prostate cancer therapy and to determine the optimal therapeutic dose, we developed a rapid, specific, and accurate reversed-phase high-performance liquid chromatography (RP-HPLC) method to quantify the ACA content in NLCs. The method was validated according to International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines. Chromatographic separation of ACA from the lipid components was performed with an Agilent 1220 Infinity LC system and ultraviolet detector using an Agilent Poroshell C18 column (4.6 x 250.0 mm). The mobile phase consisted of acetonitrile and water (80:20 [v/v]) with a flow rate of 0.8 mL/min in isocratic mode. Linearity of the standard curve was assessed at an ACA concentration range of 5-200 µg/mL, and a 1/x weighted linear regression was adopted for the calibration curve. The calculated limits of detection and quantification were 0.59 µg/mL and 1.79 µg/mL, respectively. The mean percent recovery of ACA was 100.02% (relative SD, 2%), and the coefficients of variation for intraday and interday assays were within the values required by the ICH. We also demonstrated robustness of the method by altering the mobile phase ratio and flow rate. Furthermore, we proved specificity of the method for ACA by comparing chromatograms of the blank NLC and ACA-NLC. Hence, we effectively used this validated method to determine the drug-loading capacity and entrapment efficiency of the NLCs.

Effects of reducing postprandial hyperglycemia and metabolism of acetate wheat starch on healthy mice

Abstract Recently, the acetate wheat starch (AWS) has been prepared by acetylation with an acetyl content of 2.42%, containing of rapidly digestible starch (RDS), slowly digestible starch (SDS) and resistant starch (RS) with 25.0%; 22.9% and 34.5%, respectively. In this study, this kind of starch was continuously evaluated with the postprandial blood glucose response and determined short-chain fatty acids (SCFAs) metabolized from AWS in the gastrointestinal tract of healthy mice by HPLC. The result showed that the mice fed with AWS exhibited a very limited increase in blood glucose level and remained stable for 2 hours after meals efficiently comparing with the control group fed with natural wheat starch (NWS). Simultaneously, the content of SCFAs produced in the caecum of the mice fed with AWS was significantly higher than mice fed with NWS, especially with acetic and propionic acids by 28% and 26%, respectively. Thus, AWS has shown to limit the postprandial hyperglycemia in mice effectively through the resistance to amylase hydrolysis in the small intestine. When going into the caecum, it is fermented to form SCFAs providing a part of energy for the body's activities, avoiding rotten fermentation causing digestive disorders which are inherent restrictions of normal high cellulose and fiber food.

Solid Phase Extraction and Simultaneous Chromatographic Quantification of some Non-steroidal Anti-inflammatory Drug Residues; an Application in Pharmaceutical Industrial Wastewater Effluent

Abstract Two sensitive and selective methods were developed for the simultaneous determination of four commonly used non-steroidal anti-inflammatory drugs (NSAIDs), namely; paracetamol (PCM), diclofenac sodium (DCF), ibuprofen (IBP), and indomethacin (IND) in wastewater effluents. The first method used HPLC for the determination of the studied drugs using a mobile phase consisting of phosphate buffer (pH 3.0) and acetonitrile at a flow rate of 1 mL/min. in gradient elution mode and detection at 220 nm. The separation process was performed on BDS Hypersil Cyano column (250 x 4.6 mm, 5 µm). The second method was a TLC-densitometric one which was performed using n-Hexane: ethyl acetate: acetic acid in the ratio (6:3.5:0.5) as a developing system. The proposed chromatographic methods were successfully applied for the selective determination of the four studied drugs in simulated and real pharmaceutical wastewater samples after their solid-phase extraction

Acetate wheat starch improving blood glucose response and bilan lipid on obesity dyslipidemia mice

Abstract Resistant starch is particularly concerned with beneficial effects in regulating blood glucose concentration and lipid metabolism, reducing the risk of diabetes and cardiovascular diseases. This study aimed to validate the effects of wheat starch acetate containing 32.1% resistant starch on postprandial blood glucose response and lipid profile on obesity, dyslipidemia Swiss mice induced by a high-fat diet. The result showed that there was a restriction on postprandial hyperglycemia and remained stable for 2 hours after meal efficiently comparing with the control group fed natural wheat starch. Simultaneously, when maintaining the dose of 5g/kg once or twice a day for 8 weeks, wheat starch acetate to be able to reduce body weight and blood glucose, triglyceride, cholesterol levels compared to the control group (p<0.05)

Cytotoxic activity of extracts from Tecoma species and isolated lignans

Abstract A phytochemical study of Tecoma genus (Bignoniaceae) was accomplished by antitumor activity of ethanolic extracts. Species of this genus are composed of small shrubs often used as ornamental plants. The Tecoma stans species is used in folk medicine for different purposes. Recent work shows in vitro anticancer activity against human breast cancer. The ethanolic extracts from leaves and trunks of Tecoma casneifolia, T. garrocha, T. stans var. angustata and T. stans var. stans were tested in vitro. The assays used were against line tumor cells by the MTT method and the most active extracts were further studied. In this way, the ethanolic extract from T. stans var. stans trunks presented the higher cytotoxicity against the tumor cell lines studied (CC50 0.02 to 0.55 µg/ml) when compared to the other extracts tested (CC50 0.08 to 200.0 µg/ml). Accordingly, this extract was selected for chromatographic fractionation from which five known lignans were isolated. Further, paulownin, paulownin acetate, sesamin, olivil and cycloolivil were identified using 13C and 1H NMR, IR, UV and spectroscopy and spectrometric MS techniques. These isolated compounds were tested and exhibited CC50 ranging from 13.01 to100.0 µg/ml which is superior to the ethanolic extract of trunk of T. stans

Antioxidant and antimicrobial properties of dihydroquercetin esters

Abstract Flavonoids display various beneficial biological properties, such as antioxidant activity and low cytotoxicity, which make them useful ingredients in foods, pharmaceuticals, and functional cosmetics. In particular, dihydroquercetin (DHQ) is found in various forms, and its derivatives exhibit interesting biological properties. Herein, we report the synthesis of acetylated and butyrylated dihydroquercetin derivatives and their antimicrobial and antioxidant properties. The DHQ derivatives were identified using 1H and 13C NMR spectroscopies and high-performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry. The chemical stabilities of the acetylated dihydroquercetin derivatives were found to depend on the number of acetate groups, with 3,3',4',4,7-pentaacetyldihydroquercetin found to be the most stable acetylated dihydroquercetin. Furthermore, 7,3',4'-triacetyl- dihydroquercetin exhibited potent antioxidant activity, with an IC50 of 56.67 ± 4.79 µg/mL in the 1,1-diphenyl-2-picrylhydrazyl assay, with DHQ exhibiting a value of 32.41 ± 3.35 µg/mL. The reactive-oxygen-species-scavenging activity of 7,3',4'-triacetyldihydroquercetin was highest among the esters in the ferric reducing ability of plasma assay, but lower than that of DHQ. Overall, both DHQ and 7,3',4'-triacetyldihydroquercetin exhibited antimicrobial behavior against S. aureus and P. acnes using the paper disc assay. DHQ displayed a higher antimicrobial activity, with minimum inhibitory concentrations of 625 µg/mL (P. acnes), 2,500 µg/mL (S. aureus), and 5,000 µg/mL (E. coli). DHQ and acetylated dihydroquercetins are potentially useful as complex antioxidant and antimicrobial materials

Cytotoxic evaluation and LC-MS/MS analysis of aerial parts of Eryngium kotschyi Boiss. grown in Turkey

Abstract Increasing biological activity and phytochemical investigations on Eryngium species showed its potential as pharmaceutical approach. Eryngium kotschyi Boiss. is one of the species of Eryngium genus and is endemic to Turkey. It is known that this plant is traditionally used in the South-western part of Turkey for the treatment of various diseases. This study focuses on cytotoxic activities of methanol extract and ethyl acetate, n-butanol and water sub-extracts from E. kotschyi in A549, COLO 205 and MDA-MB-231 cell lines by Sulforhodamin B assay and qualitative and quantitative determination of phytochemical constituents in active extract by LC-MS/MS. From the result of the study, it was seen that E. kotschyi ethyl acetate (EKE) sub-extract showed the strongest cytotoxic effect with the low IC50 values (50.00; 31.96 and 22.26 µg/mL in A549; COLO 205 and MDA-MB-231 cells at 48 h, respectively). Preliminary examination of the mass spectrums revealed the presence of 15 phytochemical compounds in active sub-extract and 7 of them was quantified. According to quantitative analyses the main compounds of EKE sub-extract were rosmarinic acid (485.603 µg/mgextract), chlorogenic acid (62.355 µg/mgextract) and caffeic acid (59.266 µg/mgextract). Moreover, this preliminary study on inhibitory activity of EKE sub-extract suggests further toxicologic investigations and detailed investigation on cytotoxic effect of various combinations of determined compounds

Comparator product issues for biowaiver implementation: the case of Fluconazole

The aim of this work was to assess if the commercially available Fluconazole drug products (Reference, Generic and Similar) would meet the biowaiver criteria from Food and Drug Administration (FDA) and Brazilian Agency for Health Surveillance (ANVISA) agencies. All formulations were evaluated considering the dissolution profile carried out in Simulated Gastric Fluid (SGF) pH 1.2, Acetate Buffer (AB) pH 4.5 and Simulated Intestinal Fluid (SIF) pH 6.8. The results demonstrated that all formulations fulfilled the 85% of drug dissolved at 30 min criterion in SGF pH 1.2. However, in AB pH 4.5 and SIF pH 6.8, some formulations, including the comparator, did not achieve this dissolution percentage. The discrepant dissolution profiles also failed the ƒ2 similarity factor analysis, since none of the formulations showed values between 50 and 100 in the three dissolution media. Comparative dissolution profiles were not similar, considering that the main issues concerning the dissolution were evidenced for the comparator product. Hence, a revision in the regulatory norms in order to establish criteria to switch the comparator could result in an increased application of drugs based on biowaiver criteria

Mechanism of paeoniflorin inhibiting apoptosis of hippocampal neurons of rats induced by lead acetate / 中华劳动卫生职业病杂志

Objective: To investigate the effect and underlying mechanism of paeoniflorin on hippocampal neuron apoptosis induced by lead acetate. Methods: In September 2020, primary hippocampal neuronal cells were isolated and cultured from fetal rats, and identified using cellular immunofluorescent. MTT assay was used to measure the cell viability to determine the concentration and time of lead acetate-induced hippocampal neuron apoptosis. MTT was also used to evaluate the effect of paeoniflorin concentration on the apoptosis of hippocampal neurons induced by lead acetate. According to the results, different concentrations of paeoniflorin were selected to intervene hippocampal neuron cells, after 24 h, lead acetate was added to the cells, meanwhile, blank and model groups were set up, the content of reactive oxygen species (ROS) , superoxide dismutase (SOD) , lactate dehydrogenase (LDH) , malondialdehyde (MDA) and Caspase-3 were measured. Extracellular signal regulated kinase (ERK) , phosphorylated ERK (p-ERK) , p38 mitogen -activated protein kinases (p38MAPK) , phosphorylated p38MAPK (p-p38MAPK) , c-Jun N-terminal kinase (JNK) and phosphorylated JNK (p-JNK) protein expression in hippocampal neuronal cells were determined by Western blotting. Results: The isolated and cultured hippocampal neurons were identified by immunofluorescence chemical staining and then treated with lead acetate, MTT results showed that lead acetate had the best toxicity effect when treated for 24 h at a concentration of 25 μmol/L. Paeoniflorin showed no cytotoxic effect on hippocampal neuronal cells when the concentrations below 80 μmol/L. Compared with the model group, the activity of hippocampal neuronal cells was significantly increased after treating with 20, 40 or 80 μmol/L paeoniflorin (P<0.05) . Compared with the blank group, the ROS activity, LDH release level, MDA content and caspase-3 content were significantly increased (P<0.01) , and the SOD activity was significantly decreased (P< 0.01) in the hippocampal neuronal cells of the model group. Compared with the model group, the ROS activity, LDH release level, MDA content and caspase-3 content were obviously decreased (P<0.05) , SOD activity was significantly increased (P <0.01) after hippocampal neuronal cells were treated with 40 or 80 μmol/L paeoniflorin. Relative to the model group, the ratio of p-ERK/ERK were significantly up-regulated (P<0.01) , while the ratios of p-p38MAPK/p38MAPK and p-JNK/JNK were significantly down-regulated after hippocampal neuronal cells were treated with 40 or 80 μmol/L paeoniflorin (P<0.05) . Conclusion: Paeoniflorin may down-regulate the expression of p-p38MAPK and p-JNK protein, up-regulate the expression of p-ERK protein, and inhibit the apoptosis of hippocampal neurons induced by lead acetate through the MAPK signaling pathway.

Ethyl Acetate Fraction of Dicliptera chinensis (L.) Juss. Ameliorates Liver Fibrosis by Inducing Autophagy via PI3K/AKT/mTOR/p70S6K Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To investigate the molecular mechanism underlying the anti-hepatic fibrosis activity of ethyl acetate fraction Dicliptera chinensis (L.) Juss. (EDC) in human hepatic stellate cells (HSCs) in vitro and in a carbon tetrachloride (CCl4)-induced hepatic fibrosis mouse model in vivo.@*METHODS@#For in vitro study, HSCs were pre-treated with platelet-derived growth factor (10 ng/mL) for 2 h to ensure activation and treated with EDC for 24 h and 48 h, respectively. The effect of EDC on HSCs was assessed using cell counting kit-8 assay, EdU staining, transmission electron microscopy, immunofluorescence staining, and Western blot, respectively. For in vivo experiments, mice were intraperitoneally injected with CCl4 (2 ° L/g, adjusted to a 25% concentration in olive oil), 3 times per week for 6 weeks, to develop a hepatic fibrosis model. Forty 8-week-old male C57BL/6 mice were divided into 4 groups using a random number table (n=10), including control, model, positive control and EDC treatment groups. Mice in the EDC and colchicine groups were intragastrically administered EDC (0.5 g/kg) or colchicine (0.2 mg/kg) once per day for 6 weeks. Mice in the control and model groups received an equal volume of saline. Biochemical assays and histological examinations were used to assess liver damage. Protein expression levels of α -smooth muscle actin (α -SMA) and microtubule-associated protein light chain 3B (LC3B) were measured by Western blot.@*RESULTS@#EDC reduced pathological damage associated with liver fibrosis, downregulated the expression of α -SMA and upregulated the expression of LC3B (P<0.05), both in HSCs and the CCl4-induced liver fibrosis mouse model. The intervention of bafilomycin A1 and rapamycin in HSCs strongly supported the notion that inhibition of autophagy enhanced α -SMA protein expression levels (P<0.01). The results also found that the levels of phosphoinositide (PI3K), p-PI3K, AKT, p-AKT, mammalian target of rapamycin (mTOR), p-mTOR, and p-p70S6K all decreased after EDC treatment (P<0.05).@*CONCLUSIONS@#EDC has anti-hepatic fibrosis activity by inducing autophagy and might be a potential drug to be further developed for human liver fibrosis therapy.

Analysis of chemical constituents in ethyl acetate extract of Taxilli Herba by UPLC-Q-Exactive-MS and screening of potential xanthine oxidase inhibitors / 中国中药杂志

The present study analyzed and identified the chemical constituents from ethyl acetate(EA) extract of Taxilli Herba with UPLC-Q-Exactive-MS and screened active xanthine oxidase(XO) inhibitors with HPLC. The analysis was performed on an Hypersil GOLD C_(18) reversed-phase column(2.1 mm×50 mm, 1.9 μm), with the mobile phase of water containing 1% formic acid(A) and methanol(B) under gradient elution, the flow rate of 0.3 mL·min~(-1), and the injection volume of 5 μL. ESI source was used for MS and the compounds were collected in positive and negative ion modes. Xcalibur 4.1 was used to analyze the retention time, accurate relative molecular weight, and fragmentation of the compounds. The inhibitory activity of some known compounds on XO was screened by HPLC. Thirty chemical constituents were identified, including phenolic acids and flavonoids by experimental data combined with information of standards, data reported previously, and databases, such as MzCloud and ChemSpider. The activities of 10 chemical components were screened. Gallic acid and naringenin chalcone had strong inhibitory activities on XO with IC_(50) of 57 μg·mL~(-1) and 108 μg·mL~(-1). UPLC-Q-Exactive-MS allows the accurate, rapid, and comprehensive identification of main chemical constituents from Taxilli Herba. Gallic acid and naringenin chalcone may be the active components of XO inhibitors.

The relationship between methane production metabolic flux and microorganisms in a microbial electrolytic cell coupled anaerobic digestion / 生物工程学报

In this study, voltage was used as a disturbance factor to investigate the relationship between microbial community and methane (CH4) production flux in a microbial electrolytic cell coupled anaerobic digestion (MEC-AD). Metabolic flux analysis (MFA) was used to explore the relationship between the CH4 metabolic flux produced and the microbes. The results showed that both methane production flux and hydrogen production flux changed significantly upon voltage disturbance, while the voltage disturbance had little effect on acetic acid production flux. The maximum CH4 production flux under 0.6 V disturbance was 0.522±0.051, which increased by 77% and 32%, respectively, compared with that of the control group under 1.0 V (0.295±0.013) and under 1.4 V (0.395±0.029). In addition, an average of 15.7%±2.9% of H2 (flux) was used to reduce CO2 to produce CH4 and acetic acid, and an average of 27.7%±6.9% of acetic acid (flux) was converted to CH4. Moreover, the abundance of Lachnospiraceae significantly affected the flux of acetic acid. The flux of CH4 production is positively correlated with the abundances of Petrimonas, Syntrophomonas, Blvii28, and Acinetobacter, and negatively correlated with the abundances of Tuzzerella and Sphaerochaeta. The species that affected the flux of H2 and CH4 were similar, mostly belonging to Bacteroides, Clostridium, Pseudomonas and Firmicutes. Furthermore, the interspecies interaction is also an important factor affecting the MEC-AD methanogenesis flux.

Reacciones adversas a montelukast: de la teoría a la práctica. Serie de casos / Adverse drug reactions of montelukast: from theory to practice. Case report

El montelukast se utiliza ampliamente en el tratamiento de sibilancias recurrentes y/o asma. Están descritas numerosas reacciones adversas medicamentosas (RAM) en niños relacionadas con montelukast; se destacan las neuropsiquiátricas. Realizamos un estudio observacional, retrospectivo, descriptivo, sobre RAM relacionadas con montelukast. Entre enero de 2012 y diciembre de 2017, en la Unidad de Neumonología Pediátrica se trataron con Montelukast 348 pacientes; de ellos, 20 presentaron RAM. Los síntomas más frecuentes fueron insomnio (n = 7), hiperactividad (n = 4), pesadillas (n = 3), dolor abdominal (n = 2) y parestesias en extremidades (n = 2). Se presentaron desde días hasta meses tras iniciar el tratamiento, y desaparecieron tras su suspensión. Se destacan dos pacientes con parestesias en extremidades, síntoma no descrito antes en niños. El 5,7 % de los pacientes tratados con montelukast presentaron RAM que requirieron suspender el tratamiento. Los trastornos del sueño fueron los más frecuentes.

Montelukast is widely used in recurrent wheezing and/or asthma treatment. Several adverse drug reactions (ADRs) have been described in children related to montelukast. Neuropsychiatric reactions are one of the most important. We designed an observational, retrospective, descriptive study on ADRs related to montelukast in the Pediatric Pulmonology Unit, Hospital Universitario Miguel Servet, Zaragoza, Spain. Between January 2012 and December 2017, in the Pediatric Pulmonology Unit, 348 patients were treated with Montelukast; of them, 20 presented RAM. The main symptoms described were insomnia (n = 7), hyperactivity (n = 4), nightmares (n = 3), abdominal pain (n = 2) and paraesthesia in extremities (n = 2). They appeared from the first days to months after the start of treatment and disappeared after stopping it. Two patients presented limb paresthesia, not described previously in children. The 5.7 % of our patients treated with montelukast had ADRs that required treatment discontinuation. Sleep disorders were the most frequent.

Antioxidant activity and hypoglycemic effect assessment of the leaves from Syzygium cumini (L. ) Skeels in Wistar rats

Syzygiumcumini(L.) Skeels wasadaptedto the climatic conditionsandsoil typesin Brazil. Its fruits, leaves andinner barkare usedin folk medicinedue to their highantioxidant, anti-inflammatory, anticarcinogenicandantidiabeticactivities mainlyassociated with the presenceof phenolic compounds. It is estimated thatat least300million peopleworldwide developdiabetesand approximately 11million peopleare carriersof the disease in Brazil.The objectiveof this workwas to evaluate thein vitro antioxidant activity, as well as thehypoglycemic actionofhydroethanolic extract(HEE), the ethyl acetate(EAF) andhydromethanolic(HMF) fractions from leavesofS.cumini(L.) Skeels in rats. All assays werecarried out in three replications. Data wereexpressed as meanSDand significance was evaluated by ANOVAand Bonferronitest (p < 0.05). The results indicatea significant(p < 0.05) total phenolcontent (207 ± 2.3GAE mg g-1) andantioxidant activity(EC50=9.05±0.170 µg mL-1) for EAF. HEE and its fractions showed no significant (p > 0.05) actionto modulateglucosebytheOGTT assayinnondiabetic micecompared to control. Thus the use of the plant against diabetes in individuals is not proven.

Chemical evaluation and anticholinesterase activity of Hippeastrum puniceum (Lam. ) Kuntz bulbs (Amaryllidaceae)

Hippeastrum puniceum is a species that belongs to the Amaryllidaceae family. A particular characteristic of this family is the consistent and very specific presence of isoquinoline alkaloids, which have demonstrated a wide range of biological activities such as antioxidant, antiviral, antifungal, antiparasitic, and acetylcholinesterase inhibitory activity, among others. In the present work, fifteen alkaloids were identified from the bulbs of Hippeastrum puniceum (Lam.) Kuntz using a GC-MS approach. The alkaloids 9-O-demethyllycoramine, 9-demethyl-2α-hydroxyhomolycorine, lycorine and tazettine were isolated through chromatographic techniques. The typical Amaryllidaceae alkaloids lycorine and tazettine, along with the crude and ethyl acetate extract from bulbs of the species were evaluated for their inhibitory potential on α-amylase, α-glucosidase, tyrosinase and acetylcholinesterase activity. Although no significant inhibition activity was observed against α-amylase, α-glucosidase and tyrosinase from the tested samples, the crude and ethyl acetate extracts showed remarkable acetylcholinesterase inhibitory activity. The biological activity results that correlated to the alkaloid chemical profile by GC-MS are discussed herein. Therefore, this study contributed to the knowledge of the chemical and biological properties of Hippeastrum puniceum (Lam.) and can subsidize future studies of this species