In vitro antioxidant and cytotoxic activities of polyherbal extracts from Vetiveria zizanioides,Trichosanthes cucumerina,and Mollugo cerviana on HeLa and MCF-7 cell lines

Various metabolites exist in the medicinal plants have lot of potential to cure various diseases and disorders. Plants such as, Vetiveria zizanioides, Trichosanthes cucumerina, and Mollugo cerviana were collected from Western Ghats, Tamilnadu, India. Phytochemicals were extracted from these plants using various organic solvents and tested against Gram-positive and Gram-negative bacteria. The phytochemicals such as, carbohydrate, alkaloids, steroids, saponins, flavonoids and tannin were detected from these medicinal plants. Among the extracts, methanol showed potent activity and this solvent was used to extract polyherbal medicinal plants. Methanol extract of V. zizanioides was found to be highly active against E. coli (27 ± 2 mm), P. mirabilis (19 ± 3 mm) and B. subtilis (18 ± 2 mm). Ethyl acetate extract showed high activity against E. coli (24 ± 2 mm), P. mirabilis (22 ± 3 mm) and B. subtilis (20 ± 1 mm). These three plants were taken at 1:1:1 ratio and extracted with methanol at 1:10 ratio and synergistic activity was tested against bacterial pathogens. Synergistic activity of polyherbal extract was analyzed. The extracted crude herbal medicine was found to be effective against Staphylococcus aureus, E. coli, Enterbacter sp., Pseudomonas aeruginosa, Bacillus subtilis and Proteus mirabilis. The zone of inhibition was 33 ± 3 mm, 17 ± 2 mm, 22 ± 2 mm, 40 ± 2 mm, 33 ± 1 mm and 38 ± 2 mm zone of inhibition against E. coli, S. aureus, P. aeruginosa, P. mirabilis, B. subtilis and Enterobacter sp. Polyherbal extract was found to be highly effective against P. mirabilis and Enterobacter sp. MIC values of polyherbal extract ranged from 29 ± 2.5 µg/ml to 34 ± 2.5 µg/ml. MIC value was found to be less against P. mirabilis and was high against S. aureus. Antioxidant property varied between 49 ± 3% and 95.3 ± 2%. At 20 µg/ml antioxidant activity was reported as 49 ± 3% and it was increased at higher concentrations of polyherbal extract. Two cell lines (HeLa and MCF cell lines) were selected to analyze cytotoxic activity of polyherbal extract. The methanol extract of polyherbal fraction showed cytotoxicity against these two cell lines. The LC50 value was 467 ± 2.9 µg/ml against HeLa cell line and >800 µg/ml against MCF-7 cell lines. The polyherbal extract showed antibacterial, antioxidant and anticancer activities.     

Cardioprotective properties of natural medicine in isoproterenol induced myocardial damage in the male Albino rats

The main aim of this study is to investigate cardioprotective properties of natural medicine inmyocardial damage induced male Albino rats. The aqueous extractof Allium sativumwas used for the determination of phenolic compounds and flavonoids. The amount of phenol (1.39 ± 0.37 GAE/g dry weight) and flavonoids (49.1 ± 2.79 QE/g dry weight) were high in aqueous extract. A. sativumextract and showed 68.39 ± 3.6% DPPHscavenging activity. Isoproterenol was used to induce myocardial injury in Albino rats in vivo by subcutaneous injection (100 mg/kg body weight). To achieve this, experimental animals were categorized into six groups (n = 4), namely, positive, negative control, only isoproterenol administered groups, and garlic extract administered group at 100–300 mg extract/kg body weight. Oxidative stress marker and cardiac markers were assayed to analyze the cardioprotective properties of garlic extract. At 300 mg/kg doseof garlic extract, rat was recovered from various altered factors such as, aspartate aminotransferase, alkaline transminase and alkaline phosphatase. The rats treated with 300 mggarlic extract/kg body weight decreased the level of asparate aminotransferase (126 ± 6.4 IU/L) than other lower doses (100 mg extract/kg and 200 mg extract/kg). Alkaline transaminase level of rat serum level was 81 ± 4.34 IU/L. In the isoproterenol treated rats elevated level was observed (152 ± 4.42 IU/L enzyme activity). Pre-treatment of Albino rat with A. sativum extract reduced cardiac damage. Isoproterenol exposed animal showed 207.6 ± 1.2 mg/dL triglyceride and the garlic administered rat (300 mgextract/kg) reduced LDL-cholesterol level (61.3 ± 1.3 mg/dL) significantly (p < 0.05). Creatinine kinase -MB level was 269.5 ± 12.5 IU/L in the control animal and stress induced animal showed elevated level (572.3 ± 19.4 IU/L). Garlic treated experimental animal (300 µg/kg bw) decreased CK-MB level. To conclude, the aqueous extract of A. sativumshowed cardio protective properties against myocardial injury.     

A Study on Phytochemicals from Medicinal Plants Against Multidrug Resistant Streptococcus mutans

Microbial drug resistance is creating severe problems worldwide. Medicinal important plants are a rich source of phytochemicals. These active compounds have antimicrobial and anticancer properties. Over a period of years, various plants based active compounds and its active principles have been analyzed for phytochemicals with antibacterial activity against Streptococcus mutans but this study was conducted on phytochemicals against multi-drug resistant S. mutans from dental plaque samples. Identification and isolation of S. mutans from dental plaque was done by using standard tests like Gram staining, phenol red test and blood agar hemolysis. Sensitivity/resistance pattern was done by antimicrobial sensitivity test by Kirby–Bauer disc diffusion test. Phytochemical extraction from Myristica fragrans (nutmeg) seed (MFS) and leaves (MFL), cloves (Syzigium aromaticam), Punica granatum fruit peel (PGP), Morus alba L. plant type I (MLP I) and M. alba L. plant type II leaves (MLP II) were done using solvent maceration with methanol, ethanol and water. Extracts were tested for antimicrobial properties against multidrug resistant S. mutans using antibiotic sensitivity test by agar well diffusion method and then were subjected for screening and identification of active phytochemical groups by chemical tests and high performance thin layer chromatography. The confirmatory analysis of the active compounds was done by using chromatography techniques: HPLC and GCMS. In the screening of medicinal plants clove bud, nutmeg seed and pomegranate peel showed effective antimicrobial activity against MDR S. mutans. The minimum inhibitory concentration for pomegranate peel was found to be 20 mg/ml, for clove 10 mg/ml and for Myristica seed 15 mg/ml.

     

Preliminary screening of antioxidant and cytotoxic potential of green seaweed,Halimeda opuntia (Linnaeus) Lamouroux

Marine natural products have displayed numerous advantageous effects on biological activities, including antioxidants and cytotoxicity. The total lipids, carotenoids, chlorophyll a and b content, total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity of methanolic crude extract of the green seaweed Halimeda opuntia were all measured in this study. The TPC of the extracts was determined according to the Folin-Ciocalteu method, yielding a result of 55.04 ± 0.98 mg GAE/g of extract. As determined by the aluminium chloride colorimetric method, the TFC of the extract was 40.02 ± 0.02 mg QE/g of extract. Antioxidant activity was determined by using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay with different concentrations that ranged between 200 and 1000 µg/mL, noted H. opuntia as the highest in DPPH reduction (63.61 %) at 1000 µg/mL concentration. Total antioxidant capacity (TAC) of the extract was 57.36 ± 0.004 mg AAE/g extract at concentration of 1.0 mg/mL. The cytotoxic activity of this seaweed was pre-screened against a panel of cell lines including estrogen receptor-positive human breast adenocarcinoma (MCF-7), estrogen negative human breast adenocarcinoma (MDA-MB-231), human colorectal adenocarcinoma (HT-29), human hepatocellular carcinoma (HepG2), and mouse embryonic fibroblast (3T3) using the MTT assay. The content of total lipids in H. opuntia was 1.60 ± 0.002 %. Total carotenoids were 115.57 ± 0.98 µg/g, while chlorophyll a and b were 148.73 ± 2.60 µg/g and 290.83 ± 9.46 µg/g, respectively. In terms of cytotoxicity activity, methanolic extract of H. opuntia was found to be highly cytotoxic to MCF-7 cells, with an IC₅₀ of 25.14 ± 1.02 g/mL, and slightly less so to 3T3 cells (IC₅₀ 65.23 ± 0.25 µg/mL). This study's findings suggest that natural pigments (carotenoids and chlorophyll), phytochemicals like phenolic and flavonoid compounds found in this species may play an important role and could be used as a natural cancer treatment.     

In-vitro antibacterial,antioxidant potentials and cytotoxic activity of the leaves of Tridax procumbens

The present study explored the phytochemicals, antibacterial, antioxidant and cytotoxic effect of Tridax procumbens leaves. The leaves were dried and extracted with various organic solvents. The leaves contained the phytochemicals such as alkaloids, carbohydrates, polyphenols and tannins respectively. Antimicrobial potentials of the extracts were determined by performing the disc diffusion techniques. Results revealed that different organic solvents extracts namely methanol, ethanol and ethyl acetate extracts documented comparatively good activity against the studied microbial strains. The methanol extract of leaves of T. procumbens showed combatively better antioxidant potential. The tested plant leaf extract showed high activity against human lung cancer cells than breast cancer cell lines. 250 µg/ml plants extract showed 84 ± 2.8% toxicity against human lung cancer cells.     

Improving the Efficiency of the Modified Hodge Test in KPC-Producing Klebsiella pneumoniae Isolates by Incorporating an EDTA Disk

Streptococcus mutans (S. mutans) is the main etiological agent of dental caries, and adheres to the tooth surface through the sortase A (SrtA)-mediated cell wall-anchored protein Pac. Inhibition of SrtA activity results in a marked reduction in the adhesion potential of S. mutans, and the frequency of dental caries. Morin is a natural plant extract that was previously reported to inhibit Staphylococcus aureus SrtA activity. Here, we demonstrate that morin has an inhibitory effect against S. mutans UA159 SrtA, with an IC₅₀ of 27.2 ± 2.6 μM. Western blotting demonstrated that 30 μM morin induced the partial release of the Pac protein into the supernatant. The biofilm mass of S. mutans was reduced in the presence of 30 μM morin, which was not caused by a decrease in S. mutans viability. These results indicate that morin might be important as a new agent to prevent caries.     

Comparative assessment of biological activities of different parts of halophytic plant Tamarix articulata (T. articulata) growing in Saudi Arabia

Owing to extremely high salinity and harsh environmental conditions, T. articulata is one of the most abundant wild plants growing in the deserts of Saudi Arabia. Such plants may contain novel compounds to display promising biological activities. Here, in this study, we evaluate the biological activities of methanolic extracts of fresh leaves, dry leaves, stem, and roots of T. articulata. The antioxidant activity was determined by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and total phenolic and flavonoid content were determined using standard colorimetric methods. Whereas antimicrobial and ant-proliferative activities were determined by standard well-diffusion and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) methods, respectively. Our results demonstrate that all methanolic extracts of T. articulata showed antioxidant activity, however, the methanolic extract of dry leaves exhibits promising antioxidant effect with IC₅₀ value 49.08 ± 1.98, which was strongly supported by total phenolic (409.92 ± 6.03 mg GAE/g DW) and flavonoid (177.71 mg QE/g DW) content. Although, antimicrobial activity was also exhibited by all the methanolic extracts, however, methanolic extract of dry leaves exhibits promising antimicrobial activity in Gram-positive bacteria Staphylococcus epidemidis. Furthermore, MTT assay revealed that all methanolic extracts exhibit antiproliferative activity in MCF-7 (breast cancer) and RKO (colorectal cancer) cells with IC₅₀ values ranges from 219 ± 5.112 µg/ml to 253 ± 5.231 µg/ml and 220 ± 4.330 µg/ml to 325 ± 6.213 µg/ml, respectively. However, the most promising antiproliferative effect was displayed by methanolic extract of dry leaves with IC₅₀ values 219 ± 5.112 µg/ml and 220 ± 4.330 µg/ml, respectively. In summary, these findings provide evidence that T. articulata has promising biological activities and can be used for many pharmaceutical activities in the future.     

Pharmacological properties and preliminary phytochemical analysis of Pseudocaryopteris foetida (D.Don) P.D. Cantino leaves

Medicinal plants have significant contribution in pharmaceutical industries being producers of compounds utilized as precursors for drug development. A plant of Lamiaceae family; Pseudocaryopteris foetida had not been investigated for its biomedical potential. Current research was aimed to investigate phytochemical analysis, cytotoxic potential and antioxidant activity of crude methanolic extract and fractions of Pseudocaryopteris foetida (leaves). The preliminary phytochemical analysis of crude methanolic extracts and fractions of Pseudocaryopteris foetida revealed that plant is rich in phenolic and flavonoid classes of secondary metabolites while presence of tannin was observed only in crude methanolic extract. The cytotoxicity was determined using brine shrimp lethality test. Different concentrations (25, 50, 100, 150, 200 and 250 µg/mL) of crude methanolic extract and fractions exhibited dose dependent cytotoxicity. However, The LD₅₀ for all the extracts was more than 200 µg/mL indicating weak cytotoxic potential of Pseudocaryopteris foetida. The antioxidant capabilities of crude methanolic extract and fraction of Pseudocaryopteris foetida were analyzed by in vitro bio assays including DPPH, ABTS, Reducing power and phosphomolybdate antioxidant assays using ascorbic acid as standard. The crude methanolic extract showed IC₅₀ (256.38 ± 0.6 and 314.95 ± 1.1 µg/mL) for DPPH and ABTS respectively, while total antioxidant capacity was calculated as 55.79 ± 0.5 µg/mL for crude methanolic extract of Pseudocaryopteris foetida while ascorbic acid indicated total antioxidant capacity of 71.89 ± 2.3 µg/mL. Study concluded that leaves of Pseudocaryopteris foetida were the rich source of antioxidant phytochemicals. Based on preliminary investigations further research should be focused to isolate bioactive phytochemicals as leading source of clinical medicines in future.     

Synergistic antibacterial activity of carvacrol loaded chitosan nanoparticles with Topoisomerase inhibitors and genotoxicity evaluation

The increasing prevalence of antibiotic resistant bacteria is a significant healthcare crisis with substantial socioeconomic impact on global community. The development of new antibiotics is both costly and time-consuming prompting the exploration of alternative solutions such as nanotechnology which represents opportunities for targeted drug delivery and reduced MIC. However, concerns have arisen regarding genotoxic effects of nanoparticles on human health necessitating an evaluation of nanoparticle induced DNA damage.This study aimed to investigate the antibacterial potential of already prepared, characterized chitosan nanoparticles loaded with carvacrol and their potential synergism with Topoisomerase II inhibitors against S. aureus, E. coli and S. typhi using agar well diffusion, microdilution and checkerboard method. Genotoxicity was assessed through comet assay.Results showed that both alone and drug combinations of varying concentrations exhibited greater zones of inhibition at higher concentrations. Carvacrol nanoparticles combined with ciprofloxacin and doxorubicin significantly reduced MIC compared to the drugs used alone. The MIC₅₀ values for ciprofloxacin were 35.8 µg/ml, 48.74 µg/ml, 35.57 µg/ml while doxorubicin showed MIC₅₀ values of 20.79 µg/ml, 34.35 µg/ml, 25.32 µg/ml against S. aureus, E. coli and S. typhi respectively. The FICI of ciprofloxacin and doxorubicin with carvacrol nanoparticles found ≤ 0.5 Such as 0.44, 0.44,0.48 for ciprofloxacin and 0.45, 0.45, 0.46 for doxorubicin against S. aureus, E. coli and S. typhi respectively revealed the synergistic effect. The analysis of comet assay output images showed alteration of DNA at high concentrations.Our results suggested that carvacrol nanoparticles in combination with Topoisomerase inhibitors may prevent and control the emergence of resistant bacteria with reduced dose.     

Anti-quorum sensing and anti-biofilm activity of Amomum tsaoko (Amommum tsao-ko Crevost et Lemarie) on foodborne pathogens

Cell-to-cell communication or quorum sensing (QS) leads to biofilm formation and causing other virulence factors which are extreme problems for food safety, biofilm related infectious diseases etc. This study evaluated the anti-QS activity of the Amomum tsaoko extract (0.5–4 mg/ml) by using Chromobacterium violaceum a biosensor strain and biofilm formation by crystal violate assay. Experimental results demonstrated that the overall yield of Amomum tsao-ko extract was 11.33 ± 0.3% (w/w). MIC for Staphylococcus aureus (Gram positive), Salmonella Typhimurium and Pseudomonas aeruginosa (Gram negative) was 1, 2 and 2 mg/ml, respectively. A concentration of 4 mg/ml extract showed highest biofilm inhibition 51.96% on S. Typhimurium when 47.06%, 45.28% were shown by S. aureus, P. aeruginosa respectively. The damage of biofilm architecture was observed by Confocal Laser Scanning Microscopy (CLSM). A level of 44.59% inhibition of violacein production was demonstrated when the dose was 4 mg/ml. Swarming motility inhibition was observed in a dose dependent manner. Taken together, the treatment of A. tsaoko extract can deliver value to food product and medicine by controlling pathogenesis.     

Exopolysaccharide production of <Emphasis Type="Italic">Lactobacillus salivarius</Emphasis> BCRC 14759 and <Emphasis Type="Italic">Bifidobacterium bifidum</Emphasis> BCRC 14615

In the present study, the production of exopolysaccharides (EPS) by 13 strains of Lactobacillus and 6 strains of Bifidobacterium in a chemical defined medium (CDM) supplemented with 30 g lactose/l was first compared. The highest EPS production of the Lactobacillus strains was found in L. salivarius BCRC 14759 while among the Bifidobacterium strains examined, B. bifidum BCRC 14615 showed the highest EPS production. Analyzes of the effect of lactose concentration and cultivation temperature on EPS production revealed that L. salivarius produced the highest amount of EPS (45.3 mg/l) in CDM supplemented with 5 g lactose/l at 40°C while B. bifidum produced the highest EPS (17.0 mg/l) in CDM supplemented with 40 g lactose/l at 35°C. α-Phosphoglucomutase, UDP-glucose pyrophosphorylase and UDP-galactose-4-epimerase exhibited a markedly notable activity compared with other enzymes examined in the cell extract of both test organisms. This indicates their possible involvement in the biosynthesis of EPS.     

The antimicrobial activity of compounds from the leaf and stem of Vitis amurensis against two oral pathogens

Nine compounds isolated from the leaf and stem of Vitis amurensis Rupr. (Vitaceae) were evaluated for their antimicrobial activity against two oral pathogens, Streptococcus mutans and Streptococcus sanguis, which are associated with caries and periodontal disease, respectively. The results of several antimicrobial tests, including MIC, MBC, and TBAI, showed that three compounds inhibited the growth of the test bacteria at concentrations ranging from 12.5 to 50 μg/mL. Among these compounds, compound 5, trans-ε-viniferin, displayed the strongest activity against S. mutans and S. sanguis with MIC values of 25 and 12.5 μg/mL, respectively. This is the first report on the antimicrobial activity of stilbenes and oligostilbenes isolated from the leaf and stem of V. amurensis. Thus, this result suggests that natural antimicrobial compounds derived from V. amurensis may benefit oral health as plaque-control agents for the prevention of dental caries and periodontal disease.     

Refinement of an adherence assay for the measurement of bacterial attachment to silica beads

An in vitro adherence assay [4] which utilised radiolabelled bacteria and silica beads was used to investigate firm and loose attachment of bacteria. A systematic evaluation of the effects of the assay conditions, including incubation vessel, bacteria to bead interaction, bead washing and bead weight, was carried out in an attempt to improve the reproducibility of adherence measurements. Four oral streptococcal strains (S. mutans NCTC 10449, S. sanguis NCTC 7863 and 10904, S. salivarius NCTC 8618) were studied with saliva coated and uncoated silica beads.Using the improved system, total bacterial adherence to uncoated beads was high (45–60% of bacteria present). While saliva coating of beads reduced total and firm adherence of all strains, these reductions were less marked (P > 0.005) for S. sanguis NCTC 10904.     

Toxic copper level increases erythrocyte glycolytic rate,glutathione production and alters electrolyte balance in male Wistar rats

BackgroundCopper is a micronutrient vital to several cellular energy metabolic processes and drives erythropoiesis. However, it disrupts cellular biological activities and causes oxidative damage when in excess of cellular needs. This study investigated the effects of copper toxicity on erythrocyte energy metabolism in male Wistar rats.MethodsTen Wistar rats (150–170 g) were randomly divided into 2 groups: control (given 0.1 ml distilled water) and copper toxic (given 100 mg/kg copper sulphate). Rats were orally treated for 30 days. Blood, collected retro-orbitally after sodium thiopentone anaesthesia (50 mg/kg i.p.) into fluoride oxalate and EDTA bottles, was subjected to blood lactate assay and extraction of red blood cell respectively. Red blood cell nitric oxide (RBC NO), glutathione (RBC GSH), adenosine triphosphate (RBC ATP) levels, RBC hexokinase, glucose-6-phosphate (RBC G6P), glucose-6-phosphate dehydrogenase (RBC G6PDH), and lactate dehydrogenase (RBC LDH) activity was estimated spectrophotometrically. Values (Mean±SEM, n = 5) were compared by Student’s unpaired T-test at p < 0.05.Results and conclusionCopper toxicity significantly increased RBC hexokinase (23.41 ± 2.80 µM), G6P (0.48 ± 0.03 µM), G6PDH (71.03 ± 4.76nmol/min/ml) activities, ATP (624.70 ± 57.36 µmol/gHb) and GSH (3.08 ± 0.37 µM) level compared to control (15.28 ± 1.37 µM, 0.35 ± 0.02 µM, 330.30 ± 49.58 µmol/gHb, 54.41 ± 3.01nmol/min/ml and 2.05 ± 0.14 µM respectively, p < 0.05). Also, RBC LDH activity (145.00 ± 19.88mU/ml), NO (3.45 ± 0.25 µM) and blood lactate (31.64 ± 0.91 mg/dl) level were lowered significantly compared to control (467.90 ± 94.23mU/ml, 4.48 ± 0.18 µM and 36.12 ± 1.06 mg/dl respectively). This study shows that copper toxicity increases erythrocyte glycolytic rate and glutathione production. This increase could be connected to a compensatory mechanism for cellular hypoxia and increased free radical generation.     

Metabolic profiling and biological activities of the aerial parts of Micromeria imbricata Forssk. growing in Saudi Arabia

The hydroalcoholic extract (MIT) of Micromeria imbricata (Forssk.) growing in Saudi Arabia in addition to the chloroform (MIC) and n-butanol (MIB) fractions were investigated for the first time using UPLC-ESI-MS/MS. The analysis revealed the tentative identification of fifty-eight compounds including three organic acids, twenty-five phenolic compounds, three coumarins, two anthocyanins, twenty-one flavonoids, three terpenes, and one miscellaneous. Moreover, the therapeutic potential of M. imbricata (MIT) and its fractions (MIC and MIB) were determined by in vitro evaluation of their cytotoxic, antioxidant, and anti-obesity characteristics. The MIT extract showed the highest phenolic (125.23 ± 0.87 mg gallic acid equivalent/100 gm extract) and flavonoid (112.24 ± 2.45 mg quercetin equivalent/100 gm extract) contents followed by n-butanol and chloroform fractions. The MIT extract revealed a potent cytotoxic activity against HepG-2 (Hepatocellular carcinoma) and MCF-7 (Breast carcinoma) with IC₅₀ 28.5 ± 2.0 and 35.2 ± 1.2 µg/mL, respectively. Additionally, the tested hydroalcoholic extract exhibited a significant DPPH scavenging activity with SC₅₀ 28.4 ± 1.2 µg/mL and a remarkable lipase inhibitory activity with IC₅₀ 54.2 ± 1.2 µg/mL. In conclusion, the current study presents the first insights into the phytochemical constituents and pharmacological properties of M. imbricata extract and its chloroform and n-butanol fractions. The results revealed that M. imbricata hydroalcoholic extract might be a prolific source of bioactive constituents with potent antioxidant, cytotoxic and anti-obesity potential. It might be a natural alternative therapy and nutritional strategy for obesity treatment.     

In-vitro antioxidant,antimutagenic and cancer cell growth inhibition activities of Rhododendron arboreum leaves and flowers

In the current investigation, the active principles of the methanol extracts of Rhododendron arboreum leaves (MEL) and flowers (MEF) were investigated with the help of ultra-high performance liquid chromatography (UHPLC), amino acid analyzer and gas chromatography mass spectrometry (GC-MS). UHPLC revealed different polyphenols present in the extracts. GC-MS identified 20 phytochemicals in leaves and 17 in the flowers, whereas, amino acid analyzer confirmed 11 amino acids in leaves and 10 in the flowers. The extracts were subjected to the investigation of biological activity through analysis of antioxidant activity in different in vitro assays, antimutagenic activity in Ames assay and cancer cell growth inhibition activity by MTT (3-4,5 dimethylthiazol-2,5 diphenyltetrazolium bromide) assay. MEL showed higher antioxidant activity in lipid peroxidation inhibition assay (95.32 ± 0.37%) than MEF (77.09 ± 4.17%) with IC₅₀103.6 µg/ml for MEL and 271.17 µg/ml for MEF. In nitric oxide scavenging assay, an activity of 94.46 ± 0.32% (IC₅₀ 150.13) was observed in MEF followed by 83.71 ± 0.74% (IC₅₀ 179.52) in MEL. The antimutagenic activity of both the extracts was evaluated against sodium azide, 4-nitro-O-phenylenediamine and 2-aminofluorene mutagens in TA-98 and TA-100 strains of Salmonella typhimurium. The analysis was carried out using pre- and co-incubation modes. However, both extracts were observed to possess considerable antimutagenic activity against different known mutagens, flowers came out to be more effective than the leaves in terms of % inhibition. The extracts also exhibited significant cancer cell growth inhibition activity, when tested against 3 cancer cell lines namely, Human cervical cancer cell line (HeLa), Breast cancer cell line (MCF7) and Lung cancer cell line (A549). In case of HeLa and A549, MEL showed higher activity of 64.62 ± 2.65 and 75.08 ± 1.68% as compared to 53.11 ± 2.84 and 45.92 ± 2.43% in MEL, respectively. The EC₅₀ values for MEL in HeLa and A549 were noted to be 232.76 and 155.38 µg/ml, respectively, whereas, MEF had EC₅₀ of 395.50 µg/ml in HeLa and 660.26 µg/ml in A549. Further, MEF showed higher cytotoxicity in MCF7 cell line (84.93 ± 1.17%) followed by the MEL (73.57 ± 1.27%) with EC₅₀ value of 95.16 µg/ml for MEF followed by 172.19 µg/ml for MEL. The biological activities of the extracts can be attributed to the phyto-constituents identified by sophisticated instruments.The biological activities of the extracts can be attributed to the active principles identified by sophisticated instruments.     

Control of Staphylococcus aureus methicillin resistant isolated from auricular infections using aqueous and methanolic extracts of Ephedra alata

In the current study the potential use of aqueous and methanolic extracts of Ephedra alata aerial parts as biological control agent against pathogenic bacteria and especially Staphylococcus aureus methicillin resistant isolated from auricular infections was evaluated. Chemical tests and spectrophotometric methods were used for screening and quantification of phytochemicals. The assessment of the antioxidant activity was accomplished by DPPH and ABTS radicals scavenging assays. Extracts were evaluated for their antibacterial efficacy by diffusion and microdilution methods. Biofilm inhibition was tested using XTT assay and the cytotoxicity of extracts was carried out on Vero cell line. The GC-FID analysis revealed that E. alata was rich in unsatured fatty acids. In addition, the aqueous extract had the highest flavonoid and protein contents (30.82 mg QE /g dry extract and 98.92 mg BSAE/g dry extract respectively). However, the methanolic extract had the highest phenolic, sugars and tannins. The antioxidant activity demonstrated that the aqueous extract exhibited the strong potency (IC50 ranged between 0.001 and 0.002 mg/mL).Both extracts displayed antimicrobial activity on Gram negative and positive strains. They were effective against S. aureus isolated from auricular infections. The tested extracts were able to inhibit biofilm formation with concentration-dependent manner.Moreover, no cytotoxic effect on Vero cells line was demonstrated for the extracts. Overall, our findings highlight the potential use of E. alata extract as a novel source of bioactive molecules with antioxidant, antibacterial and antiobiofilm effects for the control of infectious disease especially those associated to S. aureus methicillin resistant.     

Analysis of bioactive composites and antiviral activity of Iresine herbstii extracts against Newcastle disease virus in ovo

The study was implemented to actuate the qualitative and quantitative phyto constituents of Iresine herbstii extracts and its antiviral efficacy against avian ND virus. Among four tested solvents, the ethanolic extract of Iresine herbstii revealed the presence of highest quantity of all tested phytochemicals while petroleum ether extract showed the least. Folin-Ciocalteu method assessed the range of TPC extended from 81.01 ± 0.67 to 126.35 ± 0.45 µg GAE/mg. Acetonic extract showed the highest amount among all extracts and petroleum ether possessed the lower quantity. TFC ranged from 54.37 ± 0.45 to 88.12 ± 0.26 µg QE/mg followed by colorimetric method. From all extract ethanolic extract showed highest quantity and petroleum ether revealed the lower. HPLC analysis of ethanolic extract of I. herbstii confesses the presence six bioactive components by using the HP5-MS column. To check the antiviral potential of plants, different prepared treatments of plant extract and live virus were inoculated at 9 days old SPF embryonated chicken eggs. Results exposed that all plant extracts produce antiviral activity against NDV in ovo according to their potential and phytochemical profile. The highest survival rate was observed in the ethanolic extract at 400 µg/mL and acetonic extract at 300 µg/mL as it controls the NDV activity completely, evidence of absence of embryo death and HA titre. Dichloromethane and petroleum ether could not inhibit the virus completely. 600 µg/mL concentration was proved as toxic in all extracts except petroleum ether extract which showed a dose dependent pattern.     

Antimicrobial effects of herbal extracts on Streptococcus mutans and normal oral streptococci

Streptococcus mutans is associated with dental caries. A cariogenic biofilm, in particular, has been studied extensively for its role in the formation of dental caries. Herbal extracts such as Cudrania tricuspidata, Sophora flavescens, Ginkgo biloba, and Betula Schmidtii have been used as a folk remedy for treating diseases. The purpose of this study was to evaluate and compare the antibacterial activity of herbal extracts against normal oral streptococci, planktonic and biofilm of S. mutans. Streptococcus gordonii, Streptococcus oralis, Streptococcus salivarius, Streptococcus sanguinis, and S. mutans were cultivated with brain heart infusion broth and susceptibility assay for the herbal extracts was performed according to the protocol of Clinical and Laboratory Standard Institute. Also, S. mutans biofilm was formed on a polystyrene 12-well plate and 8-well chamber glass slip using BHI broth containing 2% sucrose and 1% mannose after conditioning the plate and the glass slip with unstimulated saliva. The biofilm was treated with the herbal extracts in various concentrations and inoculated on Mitis-Salivarius bacitracin agar plate for enumeration of viable S. mutans by counting colony forming units. Planktonic S. mutans showed susceptibility to all of the extracts and S. mutans biofilm exhibited the highest level of sensitivity for the extracts of S. flavescens. The normal oral streptococci exhibited a weak susceptibility in comparison to S. mutans. S. oralis, however, was resistant to all of the extracts. In conclusion, the extract of S. flavescens may be a potential candidate for prevention and management of dental caries.     

Pharmacognostic evaluation of Artemisia maritima L. a highly medicinal specie of genus Artemisia

The light and scanning electron microscopic observations were carried out for anatomical features of leaf, pollens and powder.Microscopic studies provide useful information for identification and authentication of adulteration in A. maritima. Nutritional analysis of A. maritima revealed that life fundamental macromolecules such as carbohydrates (49.63 %) crude proteins (13.17 %) and crude fibers (21.06 %) were present in sufficient quantity while crude fats (4.11 %) reported in low quantity. The life essential elements such as Mg (9.472 ± 0.011), Ca (4.152 ± 0.135) and Fe (4.112 ± 0.002) were found in high concentration while heavy metals reported under the safety threshold of WHO. These observations favored A. maritima an alternative of food.Appreciable quantity of phenolics (17.64 ± 0.574) and flavonoids (7.67 ± 0.069) were found while qualitatively active phytochemicals were reported.The FTIR characterization of A. maritima crude powder revealed chromatogram in 3328.61 to 408.68 frequency range and 24 characteristic peaks on the basis of which different compounds of biological importance were classified. HPLC-UV technique quantifiedand identified six phenolic compounds morin,epigallocatechin gallate, catechin hydrate,ellagic acid, pyrogallol andrutin. Identification of compounds through GC–MS chromatogram revealed the presence of 46 compounds in methanolic fraction however 17 compounds of biological importance were selected.In-vitro biological evaluation of A. maritima for antioxidant, antimicrobial, antidiabetic (12.61 ± 0.113 %) and cytotoxic activities (LC₅₀ = 20 μg/ml) suggested that methanolic fractions exhibited the highest activity as compared to chloroform and ethyl acetate fractions. The MIC values of 10 or 15 mg/ml were recorded for most of the fungal pathogens. Antibacterial activity revealed 3.75 mg/ml of MIC values against B. subtilis and 1.87 mg/ml against S. aureus, E. coli and P. aeruginosa. In-vivo biological evaluation revealed thatmaximum inhibition was observed for crude extract at 250 mg/kg body weight. The mechanism underlined in-vivo analgesic responses was carried out which revealed that naloxone (morphine and tramadol antagonist) showed no prominent effect while Glibenclamide pretreatment minutely modified the analgesic action. These observations clearly indicted the absence of opiod receptors and involvement of ATP sensitive potassium channels.