Ciprofloxacin loaded PEG coated ZnO nanoparticles with enhanced antibacterial and wound healing effects.

Antimicrobial resistance is a worldwide health problem that demands alternative antibacterial strategies. Modified nano-composites can be an effective strategy as compared to traditional medicine. The current study was designed to develop a biocompatible nano-drug delivery system with increased efficacy of current therapeutics for biomedical applications. Zinc oxide nanoparticles (ZnO-NPs) were synthesized by chemical and green methods by mediating with Moringa olifera root extract. The ZnO-NPs were further modified by drug conjugation and coating with PEG (CIP-PEG-ZnO-NPs) to enhance their therapeutic potential. PEGylated ZnO-ciprofloxacin nano-conjugates were characterized by Fourier Transform Infrared spectroscopy, X-ray diffractometry, and Scanning Electron Microscopy. During antibacterial screenings chemically and green synthesized CIP-PEG-ZnO-NPs revealed significant activity against clinically isolated Gram-positive and Gram-negative bacterial strains. The sustainable and prolonged release of antibiotics was noted from the CIP-PEG conjugated ZnO-NPs. The synthesized nanoparticles were found compatible with RBCs and Baby hamster kidney cell lines (BHK21) during hemolytic and MTT assays respectively. Based on initial findings a broad-spectrum nano-material was developed and tested for biomedical applications that eradicated Staphylococcus aureus from the infectious site and showed wound-healing effects during in vivo applications. ZnO-based nano-drug carrier can offer targeted drug delivery, and improved drug stability and efficacy resulting in better drug penetration.

Administration of vitamin E and C enhances immunological and biochemical responses against toxicity of silver nanoparticles in grass carp (Ctenopharyngodon idella).

The aim of the current study was to evaluate the toxic effect of silver nanoparticles (Ag-NPs) on biochemical biomarkers, immune responses, and the curative potential effects of vitamin C and E on grass carp. Fish (n = 420) with an average initial body weight of 8.045 ± 0.13 g were shifted to glass aquaria (36 x 18 x 18 inches, filled with 160-L tap water) in triplicates. Aquaria were randomly designated as A, B, C, D with alone Ag-NPs (Control (0), 0.25, 0.50, 0.75 mg/L) and E, F, G with Ag-NPs + Vit. C + Vit. E (0.25+0.25+0.25, 0.50+0.50+0.50, 0.75+0.75+0.75 mg/L). NPs particles were administrated viz, oral and intravenous routes for 7 days. The results indicated that both routes had non-significant effect, but levels of Ag-NPs had significant effect. Treatments C, D and G showed significant decrease in levels of RBC, HGB and HCT except for WBC and NEUT levels, which significantly increased. ALT, ALP, AST, urea, and creatinine showed significant increase in activity in the C, D, and G groups. CAT, SOD decreased significantly in all Ag-NPs alone groups, while significantly increased with vitamin E and C. LYZ, TP, ALB, GLB showed significant low activity in the B, C, and D groups while significantly high activity in the E, F, and G groups. Cortisol, glucose and triglycerides showed significant increase in the B, C, and D groups, while E, F, and G groups showed significant low levels of triglycerides, COR, and GLU. Cholesterol level was same across all treatment groups. In conclusion, vitamin E and C as powerful antioxidants protect the fish against Ag-NPs except high dose level of 0.75mg/L, while 0.25mg/L of Ag-NPs was presumably safe for C. idella.

Isolation, Characterization and Anticancer Activity of Two Bioactive Compounds from Arisaema flavum (Forssk.) Schott.

Medicinal plants play important role in the public health sector worldwide. Natural products from medicinal plants are sources of unlimited opportunities for new drug leads because of their unique chemical diversity. Researchers have focused on exploring herbal products as potential sources for the treatment of cancer, cardiac and infectious diseases. Arisaema flavum (Forssk.) is an important medicinal plant found in the northwest Himalayan regions of Pakistan. It is a poisonous plant and is used as a remedy against snake bites and scorpion stings. In this study, two bioactive compounds were isolated from Arisaema flavum (Forssk.) and their anticancer activity was evaluated against human breast cancer cell line MCF-7 using an MTT assay. The crude extract of Arisaema flavum (Forssk.) was subjected to fractionation using different organic solvents in increasing order of polarity. The fraction indicating maximum activity was then taken for isolation of bioactive compounds using various chromatographic and spectroscopic techniques such as column chromatography, thin-layer chromatography (TLC), gas chromatography−mass spectrometry (GC-MS), Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance spectroscopy (NMR). Crude extract of Arisaema flavum (Forssk.), as well as various fractions extracted in different solvents such as n-hexane, chloroform and ethyl acetate, were tested against human breast cancer cell line MCF-7 using an MTT assay. The crude extract exhibited significant dose-dependent anticancer activity with a maximum activity of 78.6% at 500 µg/mL concentration. Two compounds, hexadecanoic acid ethyl ester with molecular formula C18H36O7 and molar mass 284 and 5-Oxo-19 propyl-docosanoic acid methyl ester with molecular formula C26H50O3 and molecular mass 410, were isolated from chloroform fraction. These compounds were tested against the MCF-7cell line for cytotoxic activity and exhibited a significant (p < 0.00l) decrease in cell numbers for MCF-7 cells with IC50 of 25 µM after 48 h of treatment. Results indicated that Arisaema flavum (Forssk.) possesses compounds with cytotoxic activity that can further be exploited to develop anticancer formulations.

Isolation and Identification of Bioactive Compounds from Bidens spp. Using HPLC-DAD and GC-MS Analysis and Their Biological Activity as Anticancer Molecules.

The genus Bidens a member of family Compositae, is widely documented as an ethno-medicinally important genus of plants. In the present study, anticancer potential of three ethno-medicinally important species i.e., B. bipinnata, B. biternata and B. pilosa were tested. For in-vitro evaluation, an MTT (Thiazolyl blue tetrazolium bromide) assay was performed against cervical cancer cells (HeLa), hepatocellular carcinoma (HepG), and adenocarcinoma human alveolar basal epithelial cells (A549). For in vivo evaluation, Artemia salina, Danio rerio, and Caenorhabditis elegans were used. Among all the tested extracts, the ethanol extract of B. biternata appeared to have highest anticancer activity, and the compounds responsible for this activity were identified to be Tris (2,4-di-tert-butylphenyl), 4-hydroxy-2,4'-dimethoxychalcone, and 2,4-di-tert-butylphenol. This is the first report of the isolation of Tris (2,4-di-tert-butylphenyl) phosphate from the genus Bidens and the first report of 4-hydroxy-2,4'-dimethoxychalcone and 2,4-di-tert-butylphenol from B. biternata. Among the isolated compounds, 4-hydroxy-2,4'-dimethoxychalcone showed the highest anticancer activity with an LD50 value of 236.7 µg/mL. Therefore, this compound carries promising potential for being established as a pharmaceutical for chemoprevention and chemotherapy.

Therapeutic Potential of Selected Medicinal Plant Extracts against Multi-Drug Resistant Salmonella enterica serovar Typhi.

Salmonella enterica serovar Typhi is Gram negative, rod shaped, facultative anaerobic bacterium, belongs to enterobacteriaceae family that causes typhoid fever in humans. This bacterium has become a super bug due to acquisition of multi drug resistance. Bacteria is transmitted through food and water contaminated with human feaces. Present study reports the screening of Adhatoda vasica, Amaranthus hybridus and Aloe barbadensis and their evaluation against multi-drug resistant Salmonella enterica serovar Typhi. Qualitative analysis of ten phytochemicals was conducted using chemical method and Gas Chromatography-Mass Spectrometry (GCMS). Antibacterial activity of plants was carried out by agar well diffusion method on Mueller Hinton agar. Total tannins, total alkaloids and total flavonoids of different parts of three plants were estimated through spectrophotometer. Total tannins content in different parts of plants was present in the given order Amaranthus hybridus leaf > Aloe barbadensis leaf > Adhatoda vasica leaf > Adhatoda vasica flower > Adhatoda vasica stem. Whereas, the order of total flavonoid concentration was Amaranthus hybridus leaf > Aloe barbadensis leaf > Adhatoda vasica leaf > Amaranthus hybridus seed. Total alkaloids have order, Adhatoda vasica leaf > Amaranthus hybridus leaf > Adhatoda vasica flower > Amaranthus hybridus seed > Aloe barbadensis leaf. Results of phytochemical analysis suggested that plants have strong profile of antioxidants, total phenolic contents and various enzymes proposing them best alternate to cure bacterial infections. GC-MS analysis further confirmed stronger phytochemical profile that can be utilized as antagonists to Salmonella enterica serovar Typhi.

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.

Therapeutic prospects of endophytic Bacillus species from Berberis lycium against oxidative stress and microbial pathogens.

Endophytes are microorganisms residing within plant tissues. Bacterial endophytes are important sources for production of pharmaceutically important metabolites. Berberis lycium is an important medicinal plant and there exist no report regarding isolation and determination of bioactive potential of its bacterial endophytes. Therefore the present study was aimed to isolate and identify bacterial endophytes from Berberis lycium. The study resulted in isolation of 20 strains of bacterial endophytes. Based on their antibacterial activity three strains were identified as Bacillus cereus (LBL6), Bacillus thuringiensis (SBL3) and Bacillus anthracis (SBL4) on basis of 16SrRNA gene using universal primers. Crude ethyl acetate extracts of LBL6, SBL3 and SBL4 were further evaluated for antioxidant and antifungal activities. Moderate antioxidant activity (56 %) at a concentration of 1000 µg/mL was observed for LBL6 followed by 45 and 43 % activity by SBL4 and SBL3 respectively. Significant antifungal activity was observed against Aspergillus niger (60 %) and Aspergillus flavus (56 %) at concentration of 4 mg/mL of SBL3 and SBL4 respectively. GCMS analysis of extract (LBL6) exhibited presence of 12 bioactive secondary metabolites corresponding to antimicrobial, antifungal, antioxidant, antitumor and anticancer activities. In conclusion, present study highlighted the importance of Berberis lycium to host diverse bacterial endophytes of pharmaceutical importance.

Determination of ROS Scavenging, Antibacterial and Antifungal Potential of Methanolic Extract of Otostegia limbata (Benth.) Boiss.

Wide spectrum medicinal significance augments plant utilization as the primary source of significant pharmaceutical agents. In vitro investigation of antioxidant and antimicrobial activity highlights the therapeutic potential of Otostegia limbata. Methanol extract of the plant (MEP) shows considerable dose dependent antioxidant ability at six concentrations (7.81 µg/mL to 250 µg/mL) in 2.2-diphenyl-1-picrylhydrazyl (DPPH) assay, phosphomolybdate assay (PMA) and reducing power assay (RPA). The plant capability to scavenge free radicals in the mixture ranged from 37.89% to 63.50% in a concentration-dependent manner. MEP was active against five tested bacterial strains in the agar-well diffusion method. Staphylococcus aureus, gram-positive bacteria was found to be most susceptible followed by S. epidermidis with 18.80 mm and 17.47 mm mean zone of inhibition. The mean inhibition zone against gram-negative strains Klebsiella pneumonia, Pseudomonas spp. and Escherichia coli were 15.07 mm, 14.73 mm, and 12.17 mm. MEP revealed potential against Alternaria spp. and Aspergillus terreus fungal strains evaluated through agar-tube dilution assay. Aspergillus terreus was more sensitive than Alternaria spp. with an average 78.45% and 68.0% inhibition. These findings can serve as a benchmark for forthcoming scrutiny such as bioactive components discovery and drug development.

Using HPLC-DAD and GC-MS Analysis Isolation and Identification of Anticandida Compounds from Gui Zhen Cao Herbs (Genus Bidens): An Important Chinese Medicinal Formulation.

Gui Zhen Cao is an herbal formulation that has been documented in Chinese traditional medicine as a remedy for diarrhea, dysentery, inflammation, and toxicity. The sources of this formulation (Bidens pilosa L., Bidens biternata (Lour.) Merr. & Sherff, Bidens bipinnata L.) are also listed in ethnomedicinal reports all over the world. In this study, all these plants are tested for in vitro anticandida activity. A quantitative evaluation of the phytochemicals in all these plants indicated that their vegetative parts are rich in tannins, saponins, oxalates, cyanogenic glycoside and lipids; moreover, the roots have high percentages of alkaloids, flavonoids, and phenols. The results indicated significant anticandida activity, especially for the hexane extract of B. bipinnata leaves which inhibited C. albicans (42.54%), C. glabrata (46.98%), C. tropicalis (50.89%), C. krusei (40.56%), and C. orthopsilosis (50.24%). The extract was subjected to silica gel chromatography and 220 fractions were obtained. Purification by High Performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD) and Gas Chromatography tandem Mass Spectrometry (GC-MS/MS) analysis led to the identification of two anticandida compounds: dehydroabietic and linoleic acid having an inhibition of 85 and 92%, respectively.

Iriflophenone-3-C-ß-d Glucopyranoside from Dryopteris ramosa (Hope) C. Chr. with Promising Future as Natural Antibiotic for Gastrointestinal Tract Infections.

Ethnopharmacological approaches provide clues for the search of bioactive compounds. Dryopteris ramosa (Hope) C. Chr. (plant family: Dryopteridaceae) is an ethnomedicinal plant of the Galliyat region of Pakistan. The aqueous fraction (AqF) of D. ramosa is being used by inhabitants of the Galliyat region of Pakistan to treat their gastrointestinal tract ailments, especially those caused by bacteria. The aims of the present study were as follows: (i) to justify the ethnomedicinal uses of the AqF of D. ramosa; (ii) to isolate a bioactive compound from the AqF of D. ramosa; and (iii) to evaluate the antibacterial and cytotoxic potential of the isolated compound. Column chromatography (CC) techniques were used for the isolation studies. Spectroscopic techniques (UV-Vis, MS, 1&2D NMR) were used for structural elucidation. The agar-well diffusion method was used to evaluate the antibacterial potential of "i3CßDGP" against five bacterial strains, and compare it with the known antibiotic "Cefixime". The brine shrimp lethality test (BSLT) was used for cytotoxic studies. The AqF of D. ramosa afforded "iriflophenone-3-C-ß-D glucopyranoside (i3CßDGP)" when subjected to LH20 Sephadex, followed by MPLC silica gel60, and purified by preparative TLC. The "i3CßDGP" showed a strong potential (MIC = 31.1 ± 7.2, 62.5 ± 7.2, and 62.5 ± 7.2 µg/mL) against Klebsiella pneumoniae, Staphylococcus aureus, and Escherichia coli, respectively. On the other hand, the least antibacterial potential was shown by "i3CßDGP" (MIC = 125 ± 7.2 µg/mL), against Bacillus subtilis, in comparison to Cefixime (MIC = 62.5 ± 7.2 µg/mL). The cytotoxicity of "i3CßDGP" was significantly low (LD50 = 10.037 ± 2.8 µg/mL) against Artemia salina nauplii. This study not only justified the ethnomedicinal use of D. ramosa, but also highlighted the importance of ethnomedicinal knowledge. Further studies on AqF and other fractions of D. ramosa are in progress.

Antioxidative, protein kinase inhibition and antibacterial potential of seven mango varieties cultivated in Pakistan.

Peel, pulp and kernel extracts of seven mango fruit (varieties) were analyzedsubjected for antibacterial and antioxidative potential. Langra peel showed good activity against B. subtilis, S. aureus and E. aerogenes. Good zone of inhibition by chaunsa kernel (28mm) and pulp (22mm) against Streptomyces stipulate its potential as anticarcinogencancerous. Dosehri and almashil pulp and sindhri peel asserted free radical scavenger (upto 79%) determined through DPPH assay. The peel and kernel of almashil contained maximum (total) flavonoids contents (58 & 43µgQE/100mg, respectively) while; total phenolics were higher in kernel of sindhri, chaunsa, langra and hujra and almashil pulp. Reducing power potential demonstrated variation from 300 to 554µgAAE/100mg. Total antioxidant potential was maximum in hujra pulp (512µg AAE/100mg). Concluded This study concludes that mango has vast beneficial potential for prone to human health and may also be used for isolation of antioxidative and antimicrobial as well as a protein kinase inhibition agent.

Ethnophytotherapy Practices for Wound Healing among Populations of District Haripur, KPK, Pakistan.

Wounds, burns, cuts, and scarring may cause a serious problem for human health if left untreated, and medicinal plants are identified as potentially useful for wound healing. Therefore, the study focused on ethnophytotherapy practices for wound healing from an unexplored area, Pakistan. Ethnophytotherapeutic information was collected through well-planned questionnaire and interview methods by targeting 80 informants (70 males and 10 females), in the study area. Data was analyzed through quantitative tools like use value (UV) and credibility level (CL). A total of forty wound healing plant species, belonging to twenty-nine families, were being used in forty-six recipes. Herbs constitute (35%), shrubs (30%), trees (30%), and climbers (5%) in the treatment of multiple human injuries. For remedies preparations, leaves were most frequently utilized (52%) followed by whole plant, flowers, twigs, roots, bulb, bark, rhizome, resin, oil, leaf gel, latex, gum, and creeper. The most form of herbal preparation was powder (34.7%) and poultice (32.6%), followed by decoction, bandaged and crushed, in which 40% internally and 60 % externally applied. The drugs from these plants seem to be widely used to cure wounds: Acacia modesta, Aloe barbadensis, Azadirachta indica, Ficus benghalensis, Nerium oleander, and Olea ferruginea with higher use values (0.75). Local people are still connected with ethnophytotherapies practices for curing wounds for several reasons. This ethnomedicine and the wound healing plants are under severe threats; thus conservation must be considered. Further research should be directed towards implementing pharmacological activity on these invaluable botanical drugs.

Anticancer investigations on Carissa opaca and Toona ciliata extracts against human breast carcinoma cell line.

This study was aimed to determine the effectiveness of two ethnobotanically important plant species Carissa opaca and Toona ciliata against cancer cells. Antiproliferative activity of the plant extracts and their fractions was tested against MCF-7 breast cancer cell line using MTT assay. A concentration dependent inhibition was observed for both crude extracts. C. opaca crude extract showed 78.5% inhibition while T. ciliata showed 57% activity against cancer cells at 500 µg/mL. Fractions were tested at 200 µg/mL concentration and were more active than crude extracts. Chloroform fraction of C. opaca showed maximum inhibition 99% followed by ethyl acetate and methanol fraction of C. opaca exhibiting 96% and 94% inhibition, respectively. Ethyl acetate fraction of T. ciliata showed 78% inhibition of cancer cells at the same concentration. Preliminary phytochemical screening revealed the chemical composition of C. opaca extract containing alkaloids, flavonoids, tannins and saponins while T. ciliata had tannins and coumarins. Present investigation suggests that tested plant species possess potent anticancer compounds specially chloroform, ethyl acetate and methanol fractions of C. opaca and ethyl acetate fraction of T. ciliata can be an important source of anticancer drugs.

In vitro cytotoxic activity of Aesculus indica against breast adenocarcinoma cell line (MCF-7) and phytochemical analysis.

Aesculus indica (Linn.) (Sapindaceae) is an ethanobotanically important plant specie traditionally used against rheumatism, skin and vein complaints. Cytotoxic potential of Aesculus indica crude leaf extract and its fractions was investigated against MCF-7 cell line. Crude extract of Aesculus indica was prepared in methanol by maceration technique. Crude extract was fractionated into four organic and one aqueous fraction on polarity basis. MTT assay was used to evaluate the reduction of viability of MCF-7 breast cancer cell line. Cell viability was inhibited by Aesculus indica crude extract in a dose dependent manner ranging from 34.2% at 10 µg/ml to 94% at 500µg/ml. Activity was found in an ascending order from hexane showing 29.8% inhibition to aqueous fraction indicating maximum inhibition, 60%. Phytochemical analysis of crude and fractionated extracts revealed presence of flavonoids, saponins, coumarins and tannins upto varying degrees. Methanol and aqueous fraction of methanol extract of Aesculus indica can be good source of cytotoxic compounds.

Antibacterial activity of some selected medicinal plants of Pakistan.

BACKGROUND: Screening of the ethnobotenical plants is a pre-requisite to evaluate their therapeutic potential and it can lead to the isolation of new bioactive compounds. METHODS: The crude extracts and fractions of six medicinal important plants (Arisaema flavum, Debregeasia salicifolia, Carissa opaca, Pistacia integerrima, Aesculus indica, and Toona ciliata) were tested against three Gram positive and two Gram negative ATCC bacterial species using the agar well diffusion method. RESULTS: The crude extract of P. integerrima and A. indica were active against all tested bacterial strains (12-23 mm zone of inhibition). Other four plant's crude extracts (Arisaema flavum, Debregeasia salicifolia, Carissa opaca, and Toona ciliata) were active against different bacterial strains. The crude extracts showed varying level of bactericidal activity. The aqueous fractions of A. indica and P. integerrima crude extract showed maximum activity (19.66 and 16 mm, respectively) against B. subtilis, while the chloroform fractions of T. ciliata and D. salicifolia presented good antibacterial activities (13-17 mm zone of inhibition) against all the bacterial cultures tested. CONCLUSION: The methanol fraction of Pistacia integerrima, chloroform fractions of Debregeasia salicifolia &Toona ciliata and aqueous fraction of Aesculus indica are suitable candidates for the development of novel antibacterial compounds.