Evaluation of the antidiarrheal activity of 80% methanol extract and solvent fractions of the leaf of Bersama abyssinica fresen (Melianthaceae) in mice

Introduction The use of traditional medicinal plants in the management of diarrhea has long been practiced in Ethiopia. B. abyssinica fresen is one of the plants traditionally used to treat diarrhea whereas an in vivo study had not yet been conducted. Thus, this study aimed to evaluate the antidiarrheal activity of crude extract and solvent fractions of the leaf of B. abyssinica in mice. Methods Cold maceration within 80% methanol was used to extract the leaf powder and extract of the leaf was fractionated using n-hexane, chloroform, and distilled water. The in vivo antidiarrheal activity of crude extracts and solvent fractions were tested in experimental models of castor oil-induced diarrhea, enteropooling, and antimotility test. Five groups each with 6 mice were used under the three antidiarrheal models. Positive controls were treated with loperamide 3 mg/kg and atropine 5 mg/kg and 2% tween 80 was used in the treatment of negative controls. The extract and solvent fractions were administered at doses of 100, 200, and 400 mg/kg. Time of onset of diarrhea, number and weight of total and wet feces, the percent reduction in the number of wet feces, weight and volume of intestinal contents, and percent inhibition of intestinal motility were recorded. Data were analyzed using SPSS version 20. Result Defecation of castor oil-induced diarrheal or loose stools was inhibited (p < 0.01 to p < 0.001) at 200 mg/kg and 400 mg/kg of crude extract and aqueous fraction. The crude extract and the aqueous fraction at three doses (p < 0.01 to p < 0.001), the chloroform fraction at 200 mg/kg and 400 mg/kg (p < 0.01 to p < 0.001), and the n-hexane fraction at 400 mg/kg (p < 0.05) reduced intraluminal fluid accumulation compared with the negative control. Castor oil-induced intestinal motility was significantly suppressed with the three-doses of aqueous fraction (p < 0.05 to p < 0.001), 200 mg/kg and 400 mg/kg of crude extract (p < 0.05 to p < 0.01), 400 mg/kg of chloroform and n-hexane (p < 0.01 to p < 0.001) compared with negative control. Conclusion The crude extract, aqueous, and chloroform fractions of B. abyyssinica leaves have promising anti-diarrheal effects, supporting the plant's traditional use to treat diarrhea.

Antimicrobial activity by solvents extracted from Ocimum basilicum herb against multidrug resistant gram negative rods.

ABSTRACT: Background: Failure of treatment with antibiotics occurs due to increase in number of Multidrug resistant gram negative bacteria, worldwide. The objective of this study was to find out the antimicrobial activity of crude ethanolic extract and its further three fractions by Ocimum basilicum leaves against multi drug resistant gram negative rods. Material and Methods: This descriptive study was conducted in the Department of Microbiology, University of Health Sciences, Lahore from 1st july 2016 to 30th june 2017. Total 80 multidrug resistant gram negative rods were included in this study. Agar dilution method was performed to determine MIC of crude ethanolic extract and different fractions i-e n-hexane, chloroform and ethyl acetate of Ocimum basilicum leaves against multidrug resistant gram negative rods i-e ESBLs and carbapenemase producers. Muti-inoculater was used for inoculation. Results: The mean MICs of crude ethanolic extract, n-hexane fraction, chloroform fraction, and ethyl acetate fraction of Ocimum basilicum against ESBLs were 100.0±8.00, 168.13±8.00, 176.88±8.00 41.75±8.00 respectively. Similarly, the mean MICs of crude ethanolic extract, n-hexane fraction, chloroform fraction, and ethyl acetate fraction of Ocimum basilicum against carbapenemase producers were 77.50±8.00, 113.75±8.00, 132.50±8.00 and 29.50±8.00 respectively. Conclusions: Ethyl acetate fraction and crude ethanolic extract from leaves of Ocimum basilicum showed good antibacterial effectiveness against ESBLs and carbapenem resistant organisms than other fractions. This finding may also promote the effective use of O. basilicum herb and its components in modern medicine.

Isolation of Alkaloids from Papaver rhoeas (Papaveraceae) Wildly Grown in Iraq

The plant Papaver rhoeas ,which belongs to family Papaveraceae and known as common poppy is wildly grown in Iraq .It was used in traditional medicine in wide range of diseases including inflammation, diarrhea, sleep disorders, treatment of cough, analgesia, and also to reduce the withdrawal signs of opioid addiction. The project provide the first comprehensive research done in Iraq to study the phytochemical and the methods of extraction and separation of alkaloids from Papaver rhoeas wildly grown in Iraq .The plant was harvested in April 2019 from Zurbatiya is an Iraqi town located at the northeast of Waist province in Iraq.The collected plant was washed thoroughly, dries under shade, and grounding in a mechanical grinder to fine powder. The plant was extracted by hot extraction method using Methanol then fractionation was done to separate alkaloids from chloroform Fraction by TLC and PTLC .The alkaloids were isolated and purified by PTLC then subjected to various analytical techniques for alkaloids identification such as UV, LC mass and IR .The result was indicated of three alkaloids (dihydrocodien, chelidonine and papaverrubine C) in Papaver rhoeas plant.

Isolation, characterization and quantification of a pentacyclic triterpinoid compound ursolic acid in Scabiosa palaestina L. distributed in the north of Iraq

Ursolic acid (UA, 3 ?-hydroxy-urs-12-en-28-oic acid) are isomeric triterpenic acids. The high quantities of pentacyclic triterpenoids in Scabiosa species seems to be obvious and there is an evidence that most of pentacyclic triterpenoids that have been isolated are saponins. This is one of the most important characteristic of the genus Scabiosa, the main aglycones are ursolic acid and oleanolic acid. In the current study, isolation from the aerial part and roots of Scabiosa palaestina L. was performed using Preparative HPLC. Furthermore, detection and quantitation of ursolic acid was performed by high performance thin layer chromatography (HPTLC). The identification of isolated triterpenoid involves two methods including FT-IR coupled with LC-MS/MS that have been used for the simultaneous determination of the isolated UA. Quantitative analysis of Ursolic acid content in chloroform fractions revealed that both of the aerial parts and roots contain comparable concentration of 0.052 and 0.054 mg/ml respectively. The FT-IR and LC-MS/MS spectra of the isolated compound shows good agreement with those reported in literatures of Ursolic acid. Quantitative concentration of UA in chloroform fraction revealed that aerial parts and roots contain comparable concentrations and the spectral data for the isolated unknown were in good agreement with those reported in literature of UA.

Evaluation of The Antioxidant, Antimicrobial, and Anticancer Activities of Dicliptera bupleuroides Isolated Compounds Using In Vitro and In Silico Studies

Phytochemical investigation of chloroform fraction (DBC) and ethyl acetate fraction (DBE) of D. bupleuroides (Acanthaceae) resulted in the isolation of β-sitosterol (1) from DBC and vanillic acid (2) from DBE, which were first to be isolated from D. bupleuroides. β-Sitosterol (1) exhibited substantial antioxidant activity (IC50 = 198.87 µg/mL), whereas vanillic acid (2) showed significant antioxidant power (IC50 = 92.68 µg/mL) employing 1,1-diphenyl-2-picrylhydrazyl (DPPH*) radical scavenging capacity assay. Both compounds showed pronounced antimicrobial activity using the agar disc diffusion method, particularly against fungi showing MIC values of 0.182 and 0.02 concerning Candida albicans, respectively, and 0.001 mg/mL regarding Penicillium notatum. They revealed considerable antibacterial activity with MIC values ranging between 0.467 and 0.809 mg/mL. Vanillic acid (2) exhibited substantial anticancer potential displaying 48.67% cell viability at a concentration of 100 μg/mL using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-Diphenyl-2H-Tetrazolium Bromide) assay concerning HepG2 cell lines. These results were further consolidated by in silico studies on different enzymes, where vanillic acid displayed a high fitting score in the active pockets of DNA-gyrase, dihydrofolate reductase, aminoglycoside nucleotidyltransferase, and β-lactamase. It also inhibited human cyclin-dependent kinase 2 (CDK-2) and DNA topoisomerase II, as revealed by the in silico studies. ADME/TOPKAT (absorption, distribution, metabolism, excretion, and toxicity) prediction showed that vanillic acid exhibited reasonable pharmacodynamic, pharmacokinetic, and toxicity properties and, thus, could perfectly together with D. bupleuroides crude extract be incorporated in pharmaceutical preparations to counteract cancer and microbial invasion, as well as oxidative stress. Thus, it is concluded that D. bupleroides could be a potential source of therapeutically active compounds, which would be helpful for the discovery of clinically effective and safe drugs.

Evaluation of Anti-Psychotic potential of Phytocompound rich Fractions of Methanolic extract of Lagenaria siceraria (Bottle Gourd) fruits in Murine Models of Schizophrenia

Context: Psychosis is a mental disorder, which is characterized as the reducing association with certainty and conviction. Around 1-2% of the world's population suffers from psychosis. The people affected by psychosis have hallucinations, delusions, forgetfulness, abolition, and anhedonia. Currently existing most of the anti- psychotic medications express the adverse drug reactions. Thus for the management of psychosis, satisfactory and adequate remedies are not available. Lagenaria siceraria (Molina) Standley (LS) (Cucurbitaceae) fruits, popularly known as “bottle gourd” (English) and “Lauki” (Hindi), exhibit numerous intrinsic therapeutic properties and actions. However limited scientific investigations have been performed to determine the neuroprotective potential of LS fruits. With this background, the present study was intended to investigate LS fruits for the assessment of anti-psychotic activity. Aims and Objectives: The rationale of this study was to evaluate the anti-psychotic activity of the phytocompound rich fractions of the methanolic extract of LS i.e., chloroform fraction of the methanolic extract (CFME), and acetone fraction of the methanolic extract (AFME) by using experimental models of cataleptic and stereotypic behavior in Swiss Albino mice. Materials and Methods: Haloperidol-induced catalepsy and apomorphine-induced stereotypy were the experimental screening tests performed for the assessment of anti-psychotic activity of the phytocompound rich fractions of methanolic extract of LS fruits at the doses of 100, 200, and 400mg/kg, p.o. in mice. Results: CFME and AFME, 100-400 mg/kg, significantly reduced cataleptic score and stereotypic behavior in mice, in dose-dependent manner. At doses of 200 and 400mg/ kg, AFME (P<0.001) exhibited more significant effect than CFME. In addition CFME and AFME did not generate motor dysfunction in mice. Preliminary phytochemical screening on CFME revealed the presence of saponins, phytosterols, and terpenoids, while phytochemical investigations on AFME indicated the presence of saponins, phenolic compounds, flavonoids and tannins. This was the first scientific report, which indicates that the LS fruits exhibit anti-psychotic action, confirming the folklore claims. Future investigations and studies should emphasize separation, characterization, and identification of the bioactive phytoconstituents and also on evaluating the precise mechanism involved in the anti-psychotic activity of both the fractions.

Antiplasmodial diterpenoid alkaloid from Aconitum heterophyllum: Isolation, characterization, and UHPLC-DAD based quantification

Aconitum heterophyllum is a traditionally important medicinal plant having numerous therapeutic actions as documented in Ayurveda. This plant has been used alone as well as in combination with other plants for the preparation of different anti-malarial formulations. However, there is no report on the assessment of its anti-plasmodial activity, and the compound responsible for this activity. The main aim of this study was to conduct phytochemical investigation of A. heterophyllum roots for the preparation of extract, fractions and isolation of pure molecules to identify active fractions/molecules responsible for the anti-plasmodial activity, and development of UHPLC-DAD based analytical method which can be used for the quantification of marker compounds in the extracts and fractions. Hydro-alcoholic extract (1:1 v/v) and fractions (n-hexane, chloroform, ethyl acetate, n-butanol and water) were prepared from the dried powdered roots of A. heterophyllum. Fractions were further subjected to silica gel-based column chromatography to isolate pure specialized secondary metabolites from this plant. All extracts, fractions and pure molecules were evaluated against the chloroquine resistant Pf INDO and chloroquine sensitive Pf3D7 strains in culture for calculating their IC50 values. UHPLC-DAD based analytical method was also developed for the first time for the quantification and quality assessment of this commercially important Himalayan medicinal plant. Phytochemical investigation of A. heterophyllum root led to the isolation of six specialized metabolites named as 2-O-cinnamoyl hetisine (1), atisinium (2), 4-oxabicyclo [3.2.2] nona-1(7),5,8-triene (3), atisinium cinnamate (4), aconitic acid (5), and atisinium formate (6). Compound 1 is a new hetisine type diterpenoid alkaloid, compounds 4 and 6 are new counter ionic forms observed with atisinium ion, and compound 3 is being reported for the first time from this genus. Chloroform fraction was found to be the most active with IC50 (µg/mL) 1.01 (Pf INDO) and 1.32 (Pf3D7). The isolated molecule 2-O-cinnamoyl hetisine (1), a new diterpenoid alkaloid isolated from chloroform fraction, showed promising antiplasmodial activities with IC50 (µM) 1.92 (Pf INDO) and 10.8 (Pf 3D7). Activity of chloroform fraction was further validated by the developed UHPLC-DAD based method as the quantity of 2-O-cinnamoyl hetisine (1) was higher in the chloroform fraction (≅200 µg/mL) than in all other fractions (< 7µg/mL). Atisinium (2) and 2-O-cinnamoyl hetisine (1) were found to be the main marker compounds of this plant based on quantity and antiplasmodial activity, respectively. This study provides the scientific rational for the traditional use of this plant in treating malaria. Further, this study revealed that anti-malarial potential of this plant might be due to the presence of diterpenoid alkaloids.

Antimalarial Activity of Hydromethanolic Crude Extract and Chloroform Fraction of Brucea antidysenterica Leaves in Plasmodium berghei-Infected Mice

Background. Different parts of Brucea antidysenterica are used in traditional and alternative medicine in Ethiopia for the treatment of different health problems including malaria and have good in vitro antimalarial activity. However, no in vivo study was conducted to substantiate the claim. Our study planned to determine the antimalarial effect of B. antidysenterica extract. Methods. Swiss albino mice (6–8 weeks old, 20–28 g) were inoculated with Plasmodium berghei. Different doses of both hydromethanolic extract and chloroform fraction were orally given at 100, 200, and 400 mg/kg/day. Results. The parasitemia suppression percent of hydromethanolic crude extract and chloroform fraction in chemosuppressive tests ranged between 33.48 and 75.93% and 38.32 and 76.64%, respectively. The hydromethanolic crude extract and chloroform fraction exhibited the curative effect of 46.75–70.91% and 50.30–80.06% parasitemia suppression, respectively ( p < 0.001), compared with negative control. Conclusion. From our study, it is concluded that the hydromethanolic crude extract and chloroform fraction of B. antidysenterica leaves showed promising antiplasmodial effects against Plasmodium berghei. This upholds the folkloric use of B. antidysenterica leaves and the thought of as a possible source to develop new antimalarial agents.

ANTIMICROBIAL, ANTIOXIDANT AND CYTOTOXIC EVALUATION OF VARIOUS EXTRACTS OF IFLOGA SPICATA (FORSSK.) SCH.BIP

The current study reveals the pharmacological and phytochemical potential of different solvent fractions of Ifloga spicata. Antimicrobial and free radical scavenging activities, DPPH (1, 1-diphenyl-2-picrylhydrazyl), hydrogen peroxide scavenging assay, and cytotoxic effects of plant samples on lymphocytes stressed with hydrogen were performed. Ifloga spicata ethyl acetate fraction markedly showed higher antibacterial potential against P. aeruginosa (ATCC 27853) and E. Coli (ATCC 25922) bacteria. While higher antifungal potential was shown by the chloroform fraction against all three different a strain A. fumigates (ATCC 204305), A. flavus (ATCC 9643) and A. niger (ATCC 16404) as compared to others fractions of sample. DPPH maximum antioxidant potential was noted for methanolic extract followed by ethyl acetate and chloroform. Similarly, hydrogen peroxide effect was significantly decreased by ethyl acetate fraction. In blood lymphocytes the raise in ROS and TBARS level due to stress given by H2O2 was also recovered by different extracts of I. spicata. The decrease level of CAT, SOD and POD was also restored