Metabolic Composition of Methanolic Extract of the Balkan Endemic Species Micromeria frivaldszkyana (Degen) Velen and Its Anti-Inflammatory Effect on Male Wistar Rats.

Extracts from medicinal plants are widely used in the treatment and prevention of different diseases. Micromeria frivaldszkyana is a Balkan endemic species with reported antioxidant and antimicrobial characteristics; however, its phytochemical composition is not well defined. Here, we examined the metabolome of M. frivaldszkyana by chromatography-mass spectrometry (GC-MS), ultra-performance liquid chromatography-mass spectrometry (UPLC-MS-MS), and inductively coupled plasma mass spectrometry (ICP-MS). Amino acids, organic acids, sugars, and sugar alcohols were the primary metabolites with the highest levels in the plant extract. Detailed analysis of the sugar content identified high levels of sucrose, glucose, mannose, and fructose. Lipids are primary plant metabolites, and the analysis revealed triacylglycerols as the most abundant lipid group. Potassium (K), magnesium (Mg), zinc (Zn), and calcium (Ca) were the elements with the highest content. The results showed linarin, 3-caffeoil-quinic acid, and rosmarinic acid, as well as a number of polyphenols, as the most abundant secondary metabolites. Among the flavonoids and polyphenols with a high presence were eupatorin, kaempferol, and apigenin-compounds widely known for their bioactive properties. Further, the acute toxicity and potential anti-inflammatory activity of the methanolic extract were evaluated in Wistar rats. No toxic effects were registered after a single oral application of the extract in doses of between 200 and 5000 mg/kg bw. A fourteen-day pre-treatment with methanolic extract of M. frivaldszkyana in doses of 250, 400, and 500 mg/kg bw induced anti-inflammatory activity in the 1st, 2nd, and 3rd hours after carrageenan injection in a model of rat paw edema. This effect was also present in the 4th hour only in the group treated with a dose of 500 mg/kg. In conclusion, M. frivaldszkyana extract is particularly rich in linarin, rosmarinic acid, and flavonoids (eupatorin, kaempferol, and apigenin). Its methanolic extract induced no toxicity in male Wistar rats after oral application in doses of up to 5000 mg/kg bw. Additionally, treatment with the methanolic extract for 14 days revealed anti-inflammatory potential in a model of rat paw edema on the 1st, 2nd, and 3rd hours after the carrageenan injection. These results show the anti-inflammatory potential of the plant, which might be considered for further exploration and eventual application as a phytotherapeutic agent.

Chemical Composition Antioxidant and Anti-Inflammatory Activities of Myrtus communis L. Leaf Extract: Forecasting ADMET Profiling and Anti-Inflammatory Targets Using Molecular Docking Tools.

Compounds derived from natural sources continue to serve as chemical scaffolds for designing prophylactic/therapeutic options for human healthcare. In this study, we aimed to systematically unravel the chemical profile and antioxidant and anti-inflammatory activities of myrtle methanolic extract (MMEx) using in vitro, in vivo, and in silico approaches. High levels of TPC (415.85 ± 15.52 mg GAE/g) and TFC (285.80 ± 1.64 mg QE/g) were observed. Mass spectrophotometry (GC-MS) analysis revealed the presence of 1,8-cineole (33.80%), α-pinene (10.06%), linalool (4.83%), p-dimethylaminobenzophenone (4.21%), thunbergol (4%), terpineol (3.60%), cis-geranyl acetate (3.25%), and totarol (3.30%) as major compounds. MMEx induced pronounced dose-dependent inhibition in all assays, and the best antioxidant activity was found with H2O2, with an IC50 of 17.81 ± 3.67 µg.mL-1. MMEx showed a good anti-inflammatory effect in vivo by limiting the development of carrageenan-induced paw edema. The pharmacokinetic profiles of the active molecules were determined using the SwissADME website, followed by virtual screening against anti-inflammatory targets including phospholipase A2 (PLA-2), cyclooxygenase-2 (COX-2), tumor necrosis factor alpha (TNF-α), interleukin-1ß (IL-1ß), and NF-κB. A pharmacokinetic study revealed that the molecules have good absorption, distribution, and metabolism profiles, with negative organ toxicity. Among the compounds identified by GC-MS analysis, pinostrobin chalcone, cinnamyl cinnamate, hedycaryol, totarol, and p-dimethylaminobenzophenone were observed to have good binding scores, thus appreciable anti-inflammatory potential. Our study reveals that MMEx from Algerian Myrtus communis L. can be considered to be a promising candidate for alleviating many health complaints associated with oxidative stress and inflammation.

In vitro antibiofilm, antibacterial, antioxidant, and antitumor activities of the brown alga Padina pavonica biomass extract.

The antibiofilm, antibacterial, antioxidant, and anticancer activities of the methanolic extract of Padina pavonica L. were determined. Results deduced that the algal extract had a high biofilm formation inhibitory action done via crystal violet (CV) assay, to 88-99%. The results showed a strong antibacterial against the identified bacteria species. Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, Klebsiella pneumonia, Bacillus subtilis, and the extract had moderate antibacterial activity against Escherichia coli, Pseudomonas fluorescens and Streptococcus agalactiae. The algal extract has a concentration-dependent DPPH radical scavenging activity (84.59%, with IC50 = 170.31 µg/ml). The inhibitory percent of P. pavonica methanolic extract in vitro antiproliferative activity was 1.79-98.25% with IC50 = 15.14 µg/ml against lung carcinoma. Phenols, terpenes, amino acids, alkaloids, flavones, alcohols, and fatty acids were among the metabolites whose biological actions were evaluated. In conclusion, for the first time, P. pavonica methanolic extract exhibited effective antibiofilm, antibacterial, antioxidant, and anticancer activities. .

Acaricidal activity of a 2-methoxy-clovan-9-ol rich fraction from Eugenia copacabanensis (Myrtaceae) extract on Rhipicephalus microplus (Acari: Ixodidae).

Rhipicephalus microplus (Ixodidae, canestrini, 1888) is an invasive ectoparasite of cattle which causes high economic losses in emerging countries such as Brazil. Phytochemical compounds have been tested as an alternative to synthetic acaricides due to potentially lower mammalian toxicity. This study evaluated the acaricidal activity against R. microplus of the 2-methoxy-clovan-9-ol rich fraction obtained from Eugenia copacabanensis Kiaersk.leaves, a little known Myrtaceae species from the Brazilian Atlantic Forest. This fraction was obtained by maceration with methanol, partition with n-hexane and purification by normal-phase column chromatography. GC-MS, 1H and 13C NMR and IR analysis contributed to the identification of a major compound as the sesquiterpene 2-methoxy-clovan-9-ol, reported for the first time for the Myrtaceae family. The fraction was tested against R. microplus unfed larvae and engorged females and a 93% larval mortality was observed at the concentration of 50 mg mL-1. Lower concentrations of the solution tested demonstrated a significant difference in egg mass weight, hatching and control percentage. Experiments with 50.0 mg mL-1 showed significative results, with lower concentration and maximum efficacy for both assays. The IC50 values for unfed larvae and engorged females were 21.76 and 11.13 mg mL-1, respectively. These results were similar to those obtained in other studies with isolated botanical compounds and essential oils. The lower IC50 for engorged females than for unfed R. microplus larvae had also been described for other plant materials, including plants from the same Myrtaceae family. The present result suggested different mechanisms of action of the compound on the reproductive biology of engorged females, improving its effect against egg viability. These results are important for tick control, suggesting that 2-methoxy-clovan-9-ol could be a potential natural acaricidal product against both R. microplus unfed larvae and engorged females.

Leaf-buds of Pistacia atlantica: a novel source of bioactive molecules with high anti-inflammatory, antioxidant, anti-tyrosinase and antimicrobial properties.

In this study, for the first time, the anti-inflammatory, antioxidant, anti-tyrosinase and antimicrobial property of P. atlantica Desf. subsp. atlantica leaf-bud extract have been investigated. The anti-inflammatory activity was determined in vivo by reducing carrageenan-induced hind paw edema in mice, while the antiradical function was evaluated using DPPH, total antioxidant capacity (TAC) and reduction power assays. The extract induced a significant reduction of the edema, from 1 to 6 h in a dose-dependent manner (150, 200 and 300 mg/kg). Histological observations of the inflamed tissues also confirmed this. An effective antioxidant activity of the plant samples was demonstrated, showing an EC50 = 0.183 ± 0.005 mg/mL for the DPPH test, a value of 28.776 ± 2.541 mg AAE/g for the TAC and an EC50 = 0.136 ± 0.003 mg/mL for reducing power. The leaf-bud extract also revealed a good antimicrobial activity against S. aureus and L. monocytogenes (mean diameter of inhibition zones of 13.2 and 17.0 mm, respectively), while a slight antifungal effect was observed. The plant preparation was then documented to inhibit tyrosinase activity, with an EC50 value of 0.098 ± 0.00 mg/mL in a dose-dependent manner. HPLC-DAD analysis revealed that dimethyl-allyl caffeic acid and rutin were the most abundant molecules. The current data documented that P. atlantica leaf-bud extract has strong biological properties and constitutes a potential source of pharmacological molecules.

Methanolic extract of Hemidesmus indicus root augments the antibacterial and antibiofilm activity of amoxicillin and clindamycin against methicillin-resistant Staphylococcus aureus of bovine origin.

The present study evaluated the antibacterial and antibiofilm activity of methanolic extract of Hemidesmus indicus root (MHIR) in combination with amoxicillin and clindamycin against biofilm-forming methicillin-resistant Staphylococcus aureus (MRSA) isolated from milk samples of mastitic cows. Microdilution susceptibility testing and microtitre plate assays were used to evaluate the in vitro efficacy of MHIR and antibiotic combinations against MRSA (n = 12). Furthermore, in vitro findings were validated in a murine model. Minimum inhibitory concentration and minimum biofilm inhibitory concentration of amoxicillin and clindamycin in combination with MHIR were significantly (P < 0·05) lower than when used alone against MRSA. In terms of antibacterial activity, MHIR showed additive interaction (fractional inhibitory concentrationindex >0·5-4) with amoxicillin and clindamycin against all the MRSA isolates, whereas MHIR synergizes (fractional biofilm inhibitory concentrationindex ≤0·5) the antibiofilm activity of amoxicillin and clindamycin against 58·33% and 83·33% of the MRSA isolates respectively. Amoxicillin/clindamycin in combination with MHIR significantly (P < 0·05) reduced disease activity score, and bacterial load and Gram-positive spots in kidney and liver of MRSA-infected mice. The combined efficacy of MHIR and amoxicillin/clindamycin was comparable to clindamycin alone but superior to amoxicillin alone. Hence, the combination of MHIR with amoxicillin/clindamycin is advocated in the treatment of MRSA-associated infections.

Trichoderma reesei as an elicitor triggers defense responses in tea plant and delays gray blight symptoms.

Gray blight caused by Pestalotiopsis-like species is a major disease of tea crop worldwide including India, causes significant losses in tea production. Management of disease using fungal biocontrol agents is considered an alternative eco-friendly approach to synthetic fungicides. The present study explores the efficacy of Trichoderma reesei in the gray blight management in tea crop and activation of defense related enzymes against gray blight pathogen by developing a tri-trophic interaction system. Out of 16 isolates of Trichoderma species screened in laboratory against Pseudopestalotiopsis theae, a gray blight pathogen, isolate TRPATH01 had highest antagonistic activity (81.2%) against Ps. theae and was found to produce inhibitory volatile and non-volatile metabolites. Based on ITS and TEF-1 alpha sequencing, the isolate TRPATH01 was recognised as T. reesei. The methanolic extract of T. reesei was also found effective against Ps. theae at 200 µg/mL also confirmed presence of highest volatile compounds. The isolate also produced hydrolytic enzymes such as chitinase, cellulase, protease, and lipase. Under nursery conditions, 2% and 5% concentrations with 2 × 106 conidia/ml of T. reesei were able to reduce 67.5% to 75.0% of disease severity over pathogen inoculated controls. Moreover, compared with positive and negative controls, T. reesei -treated tea plants showed increased shoot height, stem diameter, shoot and root fresh weight at 45 days after inoculation. Principal component analysis capturing 97.1% phenotypic variations, which revealed that the tea plants co-inoculated with Ps. theae and T. reesei exhibited significantly upregulated accumulation of defensive enzymes viz., polyphenol oxidase, peroxidase, phenylalanine ammonia lyase, phenolics, ß-1, 3-glucanase, and chitinase when compared to both controls. Hence, T. reesei could provide an eco-friendly and viable mitigation option for gray blight in tea gardens by inducing defense-related enzymes.

Phytochemical Profiling, In Vitro Biological Activities, and In Silico Molecular Docking Studies of Dracaena reflexa.

Dracaena reflexa, a traditionally significant medicinal plant, has not been extensively explored before for its phytochemical and biological potential. The present study was conducted to evaluate the bioactive phytochemicals and in vitro biological activities of D. reflexa, and perform in silico molecular docking validation of D. reflexa. The bioactive phytochemicals were assessed by preliminary phytochemical testing, total bioactive contents, and GC-MS analysis. For biological evaluation, the antioxidant (DPPH, ABTS, CUPRAC, and ABTS), antibacterial, thrombolytic, and enzyme inhibition (tyrosinase and cholinesterase enzymes) potential were determined. The highest level of total phenolic contents (92.72 ± 0.79 mg GAE/g extract) was found in the n-butanol fraction while the maximum total flavonoid content (110 ± 0.83 mg QE/g extract) was observed in methanolic extract. The results showed that n-butanol fraction exhibited very significant tyrosinase inhibition activity (73.46 ± 0.80) and acetylcholinesterase inhibition activity (64.06 ± 2.65%) as compared to other fractions and comparable to the standard compounds (kojic acid and galantamine). The methanolic extract was considered to have moderate butyrylcholinesterase inhibition activity (50.97 ± 063) as compared to the standard compound galantamine (53.671 ± 0.97%). The GC-MS analysis of the n-hexane fraction resulted in the tentative identification of 120 bioactive phytochemicals. Furthermore, the major compounds as identified by GC-MS were analyzed using in silico molecular docking studies to determine the binding affinity between the ligands and the enzymes (tyrosinase, acetylcholinesterase, and butyrylcholinesterase enzymes). The results of this study suggest that Dracaena reflexa has unquestionable pharmaceutical importance and it should be further explored for the isolation of secondary metabolites that can be employed for the treatment of different diseases.

Antioxidant and Toxic Activity of Helichrysum arenarium (L.) Moench and Helichrysum italicum (Roth) G. Don Essential Oils and Extracts.

Helichrysum arenarium (L.) Moench (sandy everlasting) is the only species from genus Helichrysum Mill that grows spontaneously in Lithuania. The chemical composition of the essential oils (EOs) from inflorescences and leaves of H. arenarium wild plants was analysed by GC-MS. Palmitic (≤23.8%), myristic (≤14.9%) and lauric (6.1%) acids, n-nonanal (10.4%), and trans-ß-caryophyllene (≤6.5%) were the major constituents in the EOs. For comparison, the main components in EO from flowers (commercial herb material) of H.italicum were γ-curcumene (21.5%), ß-selinene (13.6%), α-selinene (8.1%), ß-eudesmol (8.3%), and α-pinene (6.5%). Composition of H. arenarium methanolic extracts was investigated by HPLC-DAD-TOF. The main compounds were the following: luteolin-7-O-glucoside, naringenin and its glucoside, apigenin, chlorogenic acid, arenol, and arzanol. Antioxidant activity of EOs and extracts was tested by DPPH● and ABTS●+ assays. Sandy everlasting extracts exhibited significantly higher radical scavenging activities (for leaves 11.18 to 19.13 and for inflorescences 1.96 to 6.13 mmol/L TROLOX equivalent) compared to those of all tested EOs (0.25 to 0.46 mmol/L TROLOX equivalent). Antioxidant activity, assayed electrochemically by cyclic and square wave voltammetry correlated with total polyphenolic content in extracts and radical scavenging properties of EOs and extracts. The toxic activity of EOs of both Helichrysum species was evaluated using a brine shrimp (Artemia salina) bioassay. H. italicum inflorescence EO was found to be toxic (LC50 = 15.99 µg/mL) as well as that of H. arenarium (LC50 ≤ 23.42 µg/mL) oils.

HPLC-Analysis, Biological Activities and Characterization of Action Mode of Saudi Marrubium vulgare against Foodborne Diseases Bacteria.

The present study aims to evaluate the chemical composition, metabolites secondary and pharmacology activities of methanolic extract of Marrubium vulgare collected from King Saudi Arabia. Moreover, the primary mode of action of the tested extract was studied here for the first time against E. coli and L. monocytogenes. HPLC analysis shows that the major components in the tested extract are luteolin-7-O-d-glucoside, ferulic acid and premarrubiin. Obtained data demonstrated that the investigated extract was richer in phenol (26.8 ± 0.01 mg/GAE g) than in flavonoids (0.61 ± 0.05 mg EC/mL). In addition, the methanolic extract showed an important antioxidant capacity against the DPPH (IC50 = 35 ± 0.01 µg/mL) and ABTS (IC50 = 25 ± 0.2 µg/mL) radical scavenging and a strong inhibition of acetylcholinesterase enzyme with an IC50 value corresponding to 0.4 mg/mL. The antibacterial activity demonstrated that the evaluated extract had significant activity against both Gram-positive and Gram-negative bacteria. The effect of time on cell integrity on E. coli and L. monocytogenes determined by time-kill and bacteriolysis tests showed that the M. vulgare extract reduced the viability of both strains after 8 and 10 h and had a bacteriolytic effect against two different categories of bacteria, Gram-positive and negative, which are not of the same potency. Based on obtained data, it can be concluded that Saudi M. vulgare has a high pharmacological importance and can be used in preparation of food or drugs.

Characterization of Citrusnobilis Peel Methanolic Extract for Antioxidant, Antimicrobial, and Anti-Inflammatory Activity.

Currently, the potential utilization of fruits and vegetable waste as a source of micronutrients and antioxidants has increased. The present study, therefore, aimed to determine the antimicrobial and anti-inflammatory activities of Citrus nobilis peel extract. A modified solvent evaporation technique was employed for peel extract preparation. For effective utilization of the natural product, quantitative analysis of phenolic compounds was carried out using liquid chromatography and mass spectroscopy technique. Phenolic and flavonoids were present in high amounts, while ß-carotene and lycopene were present in vestigial amounts. The antimicrobial efficiency of peel extract was evaluated against four bacterial strains including Staphylococcus aureus (MTCC 3160), Klebsiella pneumoniae (MTCC 3384), Pseudomonas aeruginosa (MTCC 2295), and Salmonella typhimurium (MTCC 1254), and one fungal strain Candida albicans (MTCC 183), and zone of inhibition was comparable to the positive control streptomycin and amphotericin B, respectively. The extract of Citrus nobilis peels showed effective anti-inflammatory activity during human red blood cell membrane stabilization (HRBC) and albumin denaturation assay. The extracts also exhibited 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity ranging from 53.46 to 81.13%. Therefore, the obtained results suggest that Citrus nobilis peel could be used as an excellent source of polyphenols and transformed into value-added products.

Vitex negundo Linn.: phytochemical composition, nutritional analysis, and antioxidant and antimicrobial activity.

Vitex negundo (VN) is a widely used plant in folk medicine, namely for the treatment of jaundice, wounds, body ache, toothache, asthma, eye pain, and migraine. These effects have been increasingly attributed to its chemical composition. Here, we assessed the VN chemical and nutritional composition and biological activity, with particular emphasis on antioxidant and antimicrobial effects. VN methanol and hexane extracts revealed the presence of important phytochemical, such as terpenoids, polyphenol, steroids, saponins, phenolic compound and flavonoids. Total phenolic content of VN methanol extract from bark was 1082.473 mg/g GAE and that of leaves was 1382.984 mg/g GAE. The total flavonoids content in VN methanol extract from VN bark was 127.744 mg/g QE and that of leaves was 123.776 mg/g QE. VN methanol extract from bark exhibited high antioxidant effects (IC50=38.47 ppm). The content (%) of ash, moisture, crude fiber, crude protein and fat in VN leaves was, respectively, 7.86%, 18.35%, 6.52%, 9.687% and 6.19%. VN leaves methanol extract revealed antimicrobial activity against Bacillus subtilis and Staphylococcus aureus, with inhibition halos being, respectively, 13 mm and 14 mm, and the MBC values were found to be 1.562 mg/mL and 6.25 mg/mL, respectively. Similarly, VN bark methanol extract led to inhibition halos of 18 mm and 15 mm for B. subtilis and S. aureus, respectively, and the MBC values were found to be 2.372 mg/mL and 0.245 mg/mL. GC-MS analysis of the VN bark methanol extract revealed that monoolein was the major compounds, with an area of 0.57%. Thus, our results encourage the potential use of VN as a medicinal product, with high protein contents, and prominent antioxidant and anti-bacterial effects.

Nephroprotective effects of Datura metel extract in gentamicin induced mice model: biochemical and histological evidences.

Datura metel is traditionally used as a remedy for renal toxicity. However, the nephroprotection has not been scientifically validated yet. To evaluate the nephroprotective like effect of methanolic extract of D. metel in gentamicin induced mice model, mice of either sex were divided into groups. One group received normal saline as negative control. The 2nd group received gentamicin 100mg/kg for 8 days as positive control, 3rd group received 50mg/kg silymarin as standard, while the reaming groups received 100, 200 and 300 mg/kg of MEDM and gentamicin 100mg/kg, for 8 days. The blood and urine samples were collected on 9th day, animals were then dissected and whole kidneys were removed and preserved in formalin for later histological examinations. The level of serum creatinine, blood urea nitrogen, urine creatinine and urine urea were significantly (P<0.05) elevated and the renal MDA level was also elevated significantly (P<0.05) by gentamicin in mice. After the treatment of test animals with MEDM, the elevated level of serum and urine biomarkers by gentamicin were reversed by MEDM. The nephroprotective effect was found in dose dependent manner. As the MEDM significantly protected the nephrotoxicity via its antioxidant effect. The findings of our study thus proved the scientific background for the nephroprotective effect of MEDM.

Improvement of endothelial function by Gunnera tinctoria extract with antioxidant properties.

BACKGROUND: Gunnera tinctoria has been collected by Mapuche-Pewenche people for food and medicinal purposes. The high polyphenol content of methanolic extract from G. tinctoria leaves with chemical constituents such as ellagic acid and quercetin derivatives suggests its application to prevent endothelial dysfunction and oxidative stress. The aim of this study was to provide evidence of the protective effect of this extract on endothelial function by reducing oxidative stress induced by high D-glucose and H2O2, as well as by stimulating nitric oxide (NO) levels in human umbilical vein endothelial cells (HUVECs). RESULTS: A methanolic extract with a high content of polyphenols (520 ± 30 mg gallic acid equivalents/g dry extract) was obtained from G. tinctoria leaves. Its main constituent was ellagic acid. The results of Ferric reducing antioxidant power and 2,2-diphenyl-1-picrylhydrazyl radical scavenging assays of the extract confirmed its antioxidant activity by inhibition pathway of radical species. The incubation of HUVECs with the extract decreased the apoptosis and reactive oxygen species (ROS) synthesis induced by high extracellular concentration of D-glucose or hydrogen peroxide. The extract increased endothelial NO levels and reduced vasoconstriction in human placental vessels. CONCLUSIONS: This study provides evidence about the antioxidant and endothelial protective properties of methanolic G. tinctoria leaf extract. The extract improves the availability of NO in HUVECs, inhibiting the production of ROS and vasoconstriction.

Anastatica hierochuntica (L.) methanolic and aqueous extracts exert antiproliferative effects through the induction of apoptosis in MCF-7 breast cancer cells.

Breast cancer therapy using anticancer bioactive compounds derived from natural products as adjuvant treatment has gained recognition due to expensive and toxic conventional chemotherapeutic drugs. The whole plant of Anastatica hierochuntica (L.) (A. hierochuntica) has been investigated for its pharmacologically important anticancer properties but without categorizing the biological activities of the plant parts. We assessed the anticancer potential of different parts of A. hierochuntica (seeds, stems and leaves) and explored their mechanisms of action using the human breast cancer cell line, MCF-7. Currently, we investigated the antiproliferative effects of methanolic (MSD, MST, ML) and aqueous (ASD, AST, AL) extracts of A. hierochuntica plant parts on the MCF-7 cells using cell viability assays. Flow cytometry, Western Blot, DNA fragmentation, and gene expression assays were performed to evaluate apoptosis and cell cycle regulatory proteins. The results indicate that the methanolic and aqueous extracts decreased MCF-7 cell viability in a dose-dependent manner. The induction of apoptosis was observed in all the methanolic and aqueous-treated MCF-7 cells. The cell death process was confirmed by the visualization of DNA fragmentation and cleavage of the intrinsic apoptotic pathways, caspase-9 and caspase-3, the key enzyme causing apoptosis hallmarks. In addition, the most pro-apoptotic extracts, ASD and ML, up-regulated the expression of pro-apoptotic Bax, tumor suppressor TP53 genes and the cyclin inhibitor CDKN1A gene. In conclusion, of the aqueous and methanolic extracts of A. hierochuntica plant parts exerting antiproliferative effects through the induction of apoptosis in breast cancer MCF-7 cells, ASD and ML extracts were the most promising natural-based drugs for the treatment of breast cancer.

In Vivo Validation of Folkloric Use of Costus pictus D. Don as Antidiabetic Plant in Assam, Using Streptozotocin Induced Wister Rat Model.

Costus pictus D. Don belonging to the family Costaceae is one of the most commonly used plant among traditional healers in the upper Assam region in India, specifically used in the treatment of diabetes. Aerial parts of the plant are said to have potent anti diabetic property. The present study was aimed to evaluate the traditionally claimed antidiabetic activity of aerial parts of C. pictus in animal models. Healthy male Wister rats (120 ± 30 gm) were used in the study and diabetes was induced by Streptozotocin (STZ) i.p. prepared by dissolving in citrate buffer (pH 4.5), along with nicotinamide (120 mg/kg/b wt). Diabetic rats were treated for 14 days with daily doses of methanolic extract of C. pictus (MECP) in three different scheduled amounts (50, 100 and 200 mg/kg/b wt; p.o.). Control rats were treated with 0.3% CMC (Carboxy Methyl Cellulose) suspension (10 ml/kg/b wt; p.o.). Blood glucose level and plasma lipid profile was examined on 1st, 7th, 14th and 21st day 1 h after treatment. On the last day, 1 h after the treatment, animals were sacrificed followed by isolation of pancreas and liver for histopathological examination. The results were compared with that of the standard group treated with Glibenclamide (10 mg/kg/day; p.o). Comparison of the experimental data of different cohorts demonstrated the potential antidiabetic activity of C. pictus however the highest dose of 200 mg/kg/b wt; p.o. of MECP significantly (P < 0.05) reversed the STZ induced diabetic parameters (increased blood glucose level, altered plasma profile and histoarchitecture of the pancreatic and hepatic cells) that is comparable with that of the standard. The observed results suggest anti diabetic efficacy of C. pictus thereby uphelding the folkloric usage.

Assessment of Garcinia pedunculata Roxb. Role on Biological Markers in Carbon Tetra Chloride Induced Rat Model.

Garcinia pedunculata Roxb. is an important medicinal plant of North Eastern (NE) region of India, having number of medicinal properties and used against various diseases in folk medicine. An empirical research was designed to carry out evaluation of hepatoprotective activity of the leaves of G. pedunculata with special reference to its putative protective role. Methanolic extract from the dried leaf powder of G. pedunculata Roxb. was prepared by hot continuous extraction method. The prepared extract was investigated at different dose levels for its hepatoprotective nature and further histopathological study was carried out to ascertain the degree of reversing the hepatotoxic manifestation induced by CCl4 (Carbon tetrachloride). LD50 values of the G. pedunculata Roxb. extract was found to be safe up to 2000 mg. The in vivo biological studies on serum and tissues of male Wister rats at the doses of 100 mg, 300 mg and 600 mg/kg body weight respectively was carried out taking Silymarin as standard. The methanolic extract of G. pedunculata Roxb, improved the cholesterol level along with significant improvement of SGPT (Serum Glutamate Pyruvate Transaminase), SGOT (Serum Glutamate Oxaloacetate Transaminase), ALP (Alkaline Phosphatase) and total protein in respect to Silymarin group. The test extract at the dose 600 mg/kg body weight was found to significantly reverse the elevated marker enzymes i.e. SGOT, SGPT, ALP indicating its hepatoprotective role. The higher dose extracts were also found to have pronounced effect on oxidative stress parameters such as GSH and catalase on CCl4 induced rats. The histopathological studies further augmented the protective activity of G. pedunculata leaf extract thereby endorsing the traditional usage of the plant.

Investigation of common chemical components and inhibitory effect on GES-type ß-lactamase (GES22) in methanolic extracts of Algerian seaweeds.

This study aimed to investigate the total phenolic content (TPC), the identification of the common compounds by HPLC-ESI-MS and HPLC-ESI-MS-TOF and the inhibitory effects against class A-type ß-lactamase (GES-22 variant, produced recombinantly) in methanolic extracts (MEs) of four Algerian seaweeds [Ulva intestinalis, Codium tomentosum, Dictyota dichotoma and Halopteris scoparia]. The TPC varied among the four species, ranging between 0.93 ±â€¯0.65 and 2.66 ±â€¯1.33 mg GAEs/g DW. C.tomentosum had higher total phenol content than other seaweeds while, all of them inhibited uncompetitively GES-22 activity in a dose-dependent manner. Nitrocefin was used as chromogenic substrate to evaluate the inhibitory effect on GES-22. The methanolic extract of D.dichotoma exhibited significant inhibitory effect on GES-22 (IC50 = 13.01 ±â€¯0.046 µg/mL) more than clavulanate, sulbactam and tazobactam (classical ß-lactam inhibitors) (IC50 = 68.38 ±â€¯0.17 µg/mL, 52.68 ±â€¯0.64 µg/mL, and 29.94 ±â€¯0.01 µg/mL, respectively). IC50 of the other ME of U.intestinalis, C.tomentosum, and H.scoparia were 16.87 ±â€¯0.10 µg/mL, 16.54 ±â€¯0.048 µg/mL, and 25.72 ±â€¯0.15 µg/mL, respectively. Except H. scoparia, other three seaweed extracts showed almost two times or more inhibition on GES-22. Furthermore, four common compounds in these MEs were identified, α-linolenic acid (C18:3ω3), linoleic acid (C18:2ω6), oleic acid (C18:1ω9), the eicosanoid precursors ''arachidonic acid'' (C20:4ω6). Baicalein (C15H10O5) was identified in U.intestinalis and D.dichotoma seaweeds. The fact that all seaweed extracts inhibited the GES-22 better than commercial samples makes these seaweeds candidate for discovering new inhibitors against ß-lactamases. Besides that, they contain important components with potential health benefits.

Wound healing activity of Pimpinella anisum methanolic extract in streptozotocin-induced diabetic rats.

OBJECTIVE: To assess the wound healing potential of Pimpinella anisum on cutaneous wounds in diabetic rats. METHOD: Full-thickness excisional wounds were made on the back of male, Sprague-Dawley rats with diabetes. The rats were randomly allocated into four treatment groups: 1ml basal cream; tetracycline (3%); Pimpinella anisum 10% for 14 days; and a control group. At days seven, 14 and 21 post-injury, five animals of each group were euthanised, and wounds were assessed through gross, histopathological and oxidant/antioxidant evaluations. Additionally, the dry matter and hydroxyproline contents of the skin samples were measured. RESULTS: A total of 60 rats were used in the study. A significant decrease in the wound size was observed in treated animals with Pimpinella anisum compared with other groups during the experiment. Additionally, treatment with Pimpinella anisum decreased the number of lymphocytes and improved the number of fibroblasts at the earlier stages and increased a number of fibrocytes at the later stages of wound healing. Other parameters such as re-epithelialisation, tissue alignment, greater maturity of collagen fibres and large capillary-sized blood vessels revealed significant changes when compared with the control. Pimpinella anisum significantly reverted oxidative changes of total antioxidant capacity, malondialdehyde and glutathione peroxidase induced by diabetic wounds (p<0.05). Furthermore, it significantly increased the dry matter and hydroxyproline contents at various stages of wound healing (p<0.05). CONCLUSION: The present study showed that application of Pimpinella anisum extract promotes wound healing activity in diabetic rats. The wound-healing property of Pimpinella anisum can be attributed to the phytoconstituents present in the plant.

Anticoccidial activity of fruit peel of Punica granatum L.

In the interests of food safety and public health, plants and their compounds are now re-emerging as an alternative approach to treat parasitic diseases. Here, we studied the anticoccidial effect of different solvent extracts of the fruit peel of Punica granatum-a commercial waste from pomegranate juice industries. The hope underlying these experiments was to find a sustainable natural product for controlling coccidiosis. The plant extracts were prepared using solvents of different polarity. Acute oral toxicity study was first carried out to see the safety of crude extracts. A high dose of crude extracts (300 mg/kg body weight) was tested for possession of anticoccidial activity against experimentally induced coccidial infection in broiler chicken. Activity was measured in comparison to the reference drug amprolium on the basis of oocyst output reduction, mean weight gain of birds and feed conversion ratio. Oocyst output was measured using Mc-Masters counting technique. Acute oral toxicity study showed that crude extracts of P. granatum are safe up to dosage of 2000 mg/kg body weight. LD50 was not determined as mortalities were not recorded in any of the five groups of chicken. For anticoccidial activity crude methanolic extract (CME) of the fruit peel of P. granatum showed the maximum effect as evident by oocyst output reduction (92.8 ±â€¯15.3), weight gain of birds (1403.0 ±â€¯11.9 g) and feed conversion ratio (1.66 ±â€¯0.04), thereby affirming the presence of alcohol soluble active ingredients in the plant. We also tested different doses (100-400 mg/kg body weight) of the CME of the fruit peel of P. granatum, the most active extract on E. tenella and observed a dose dependent effect. From the present study it can be concluded that alcoholic extract of the fruit peel of P. granatum has significant potential to contribute to the control of coccidian parasites of chicken.