Development of miconazole nitrate containing chitosan microcapsules and their anti-Aspergillus niger activity

In this article, we report the development of chitosan/miconazole nitrate microcapsules. Four miconazole nitrate ratios including 12.5, 25, 50 and 100?mg were performed in the chitosan-based microencapsulation system. Chitosan microcapsules with the drug input of 25?mg showed the highest encapsulation efficiency (52.47%) and acceptable mean particle size (5.65?μm) when compared with those of 12.5, 50 and 100?mg. Fourier transform infrared spectroscopic spectrum proved the entrapment of miconazole nitrate into chitosan microcapsules. The antifungal result demonstrated that microcapsules containing 75?μg miconazole nitrate possessed comparable anti-Aspergillus niger activity as the commercial ointment. The growth inhibition of miconazole nitrate containing chitosan microcapsules towards human skin keratinocytes was found to be dose dependent. A total of 75?μg of miconazole nitrate containing microcapsules revealed about 25% of growth inhibition while that of 150?μg showed approximately 70% of growth inhibition. Special monitoring should be taken if a higher dose of miconazole nitrate was used to develop the microcapsules.     

Inactive Vibrio cholerae whole-cell vaccine-loaded biodegradable microparticles: in vitro release and oral vaccination

An approach is proposed using Vibrio cholerae (VC)-loaded microparticles as oral vaccine delivery systems for improved vaccine bioavailability and increased therapeutic efficacy. The VC-loaded microparticles were prepared with 50:50 poly(DL-lactide-co-glycolide) (PLG), 75:25 poly(DL-lactide-co-glycolide) and poly(lactide acid) (PLA)/PEG blend copolymers by the solvent evaporation method. VC was successfully entrapped in three types of microparticles with loading efficiencies and loading levels as follows: 50:50 PLG systems: 97.8% and 55.4 ± 6.9 µg/mg; 75:25 PLG systems: 89.2% and 46.5 ± 4.4?µg/mg; PLA/PEG-blended systems: 82.6% and 53.7 ± 5.8?µg/mg. The different distributions of VC in the core region and on the surface were as follows: 50:50 PLG systems 25.7 ± 1.9 and 6.2 ± 0.9?µg/mg; 75:25 PLG systems: 25.8 ± 2.2 and 3.6 ± 0.4?µg/mg; PLA/PEG-blended systems: 32.4 ± 2.1 and 5.2 ± 1.0?µg/mg, respectively. In vitro active release of VC was affected mainly by matrix type and VC-loaded location in microparticles. The therapeutic immunogenic potential of VC loaded with 50:50 PLG, 75:25 PLG and PLA/PEG-blended microparticles was evaluated in adult mice by oral immunization. Significantly higher antibody responses and serum immunoglobin Ig G, IgA and IgM responses were obtained when sera from both VC-loaded 75:25 PLG and PLA/PEG-blended microparticles immunized mice were titrated against VC. The most immunogenicity in evoking serum IgG, IgA and IgM responses was immunized by VC-loaded PLA/PEG-blended microparticles, and with VC challenge in mice, the survival rate (91.7%).     

Phase-separated chitosan–fibrin microbeads for cell delivery

Matrix-enhanced delivery of cells is a promising approach to improving current cell therapies. Our objective was to create cell-laden composite microbeads that combine the attractive features of the natural polymers chitosan and fibrin. Liquid polydimethylsiloxane was used to emulsify a chitosan–fibrinogen solution containing suspended human fibroblast cells, followed by initiation of thrombin-mediated polymerization of fibrin and thermal/pH-mediated gelation of chitosan. Chitosan/fibrin weight percent (wt%) ratios of 100/0, 75/25, 50/50 and 25/75 were investigated. Microbead diameters ranged from 275?±?99?µm to 38?±?10?µm using impeller speeds from 600 to 1400?rpm. Fibroblasts remained viable on day 1 post-fabrication in all matrices, but cell viability was markedly higher in high-fibrin microbeads by day 8 post-fabrication. Cell spreading and interaction with the extracellular matrix was also markedly increased in high-fibrin matrices. Such composite microbeads containing viable entrapped cells have potential for minimally invasive delivery of cells for a variety of tissue repair applications.     

Comparative study of the anti-inflammatory and antinociceptive effects of two varieties of Punica granatum

Context: Studies have shown that pomegranate, Punica granatum Linn. (Lythraceae), has remarkable biological and medicinal properties.

Objective: This work aimed to explore and compare the analgesic and anti-inflammatory activities of the methanol extract (MoE) obtained from fruit peels of two varieties of pomegranate: Amrouz (MoEA) and Sefri (MoES).

Materials and methods: Antinociceptive activity of MoEA and MoES was examined using four models of pain. The extracts were administered by the intraperitoneal route (i.p.) in writhing (50, 100 and 150?mg/kg) and formalin tests (25, 50 and 100?mg/kg) and by intra-cerebroventricular injection (i.c.v.) in hotplate and tail-immersion tests (10, 25 and 50 µg/3 µl/rat). anti-inflammatory activity was studied using the hind paw egg albumin test (50, 100 and 150?mg/kg, i.p.).

Results: In the writhing test, the index of pain inhibition (IPI) was 52% for MoEA (150?mg/kg, i.p.) and 29% for MoES (150?mg/kg, i.p.). In the formalin test, the IPI of early and late phase were, respectively, 75% and 82% for MoEA (100?mg/kg, i.p.) and 8% and 63% for MoES (100?mg/kg, i.p.). In the hotplate and tail-immersion test, MoEA and MoES increased in a dosedependent manner the reaction latency to the thermal stimuli. MoEA seems to be more potent than MoES. Only the analgesic effect of MoEA was partially inhibited by pretreatment with naloxone. Both extracts exerted a significant anti-inflammatory effect.

Discussion and conclusions: The results demonstrated that P. granatum contains active constituents, which possess antinociceptive and anti-inflammatory activity, justifying its popular uses.     

Magnolia dealbata seeds extract exert cytotoxic and chemopreventive effects on MDA-MB231 breast cancer cells

Context: Cancer prevention remains a high priority for the scientific world. Magnolia dealbata Zucc (Magnoliaceae), a Mexican endemic species, is used for the empirical treatment of cancer.

Objective: To evaluate the cytotoxic and cancer chemopreventive effects of an ethanol extract of Magnolia dealbata seeds (MDE).

Materials and methods: The cytotoxic effect of MDE, at concentrations ranging from 1 to 200?µg/ml, on human cancer cells and human nontumorigenic cells was evaluated using the MTT assay for 48?h. The apoptotic activities of MDE 25?μg/ml on MDA-MB231 breast cancer cells were evaluated using the TUNEL assay and the detection of caspase 3 using immunofluorescence analysis for 48?h, each. The chemopreventive effect was evaluated by administrating different doses of MDE, between 1 and 50?mg/kg, injected intraperitoneally daily into athymic mice which were implanted with MDA-MB231 cells during 28 days. The growth and weight of tumors were measured.

Results: MDE showed cytotoxic effects on MDA-MB231 cells (IC₅₀?=?25?µg/ml) and exerted pro-apoptotic activities as determined by DNA fragmentation in MDA-MB231 cells. MDE 25?µg/ml also induces the activation of caspase 3 in MDA-MB231 cells. These results suggest that Magnolia dealbata may be an optimal source of the bioactive compounds: honokiol (HK) and magnolol (MG). MDE 50?mg/kg i.p. exerted chemopreventive effects by inhibiting the growth of MDA-MB231 tumor by 75% in athymic mice, compared to the control group.

Conclusions: MDE exerts cytotoxic, apoptotic and chemopreventive activities on MDA-MB231 human cancer cells.     

Adjuvant anti-angiogenic therapy enhances chemotherapeutic uptake in a murine model of head and neck cancer*

Abstract

Intratumoural metabolic demands result in excessive angiogenic cytokine release leading to unorganised vasculature. Resultant fluid dynamics oppose blood flow and drug penetration due to a marked increase in interstitial fluid hydrostatic pressure. It is hypothesised that anti-angiogenic therapy may function to ‘prune’ vasculature and lead to improved chemotherapeutic penetration. Subcutaneous, OSC19 tumour bearing mice (n?=?5/dose/agent) were administered varying doses of an anti-mouse VEGFR2 (DC101) or an anti-mouse VEGFR3 (31C1) –3 d, –1 d, 0 d, +1 d and +3 d prior to 200?µg of cetuximab fluorescently labelled with IRDye800CW. Fluorescence imaging of tumours was performed 10 d post cetuximab-IRDye800CW dose to monitor therapeutic uptake. Co-administration of dual anti-angiogenic agents at 50–50%, 75–25% and 25–75% using optimal dose and time (–1 d 10?mg/kg anti-VEGFR2 and –1 d 40?mg/kg anti-VEGFR3) was also evaluated. In order to establish vessel normalisation, NG2 (pericyte marker) and CD31 (endothelial cells) ratios were assessed during immunohistochemical staining of tumour sections. Twenty-mg/kg anti-VEGFR3?+?5?mg/kg anti-VEGFR2 significantly (p?<?.0005) reduced tumour size (–73%) compared to control (59%). The 20?mg/kg anti-VEGFR3?+?5?mg/kg anti-VEGFR2 and 30?mg/kg anti-VEGFR3?+?2.5?mg/kg anti-VEGFR2 significantly (p?<?.0004) improved percent-injected cetuximab-IRDye800CW dose/gram tumour tissue compared to other groups. Adjuvant, dual anti-angiogenic therapy targeting VEGFR2 and VEGFR3 significantly enhances tumour chemotherapeutic uptake compared to control.     

Double-effective chitosan scaffold-PLGA nanoparticle system for brain tumour therapy: in vitro study

Abstract

The chitosan scaffold, which has both of anticancer and antivascularization effects, was developed for using in local therapy of brain tumours. This is why, poly-lactic-co-glycolic acid (50:50) nanoparticles (～200?nm) including an anticancer drug, 5-fluorouracil (5-FU), were prepared by emulsion-solvent evaporation method. Then, these nanoparticles and antivascularization agent, bevacizumab, were loaded into the scaffold during manufacturing by freeze-drying and embedding after freeze-drying, respectively. The idea behind this system is to destroy tumour tissue by releasing 5-FU and to prevent the proliferation of tumour cells by releasing bevacizumab. In addition, 3D scaffold can support healthy tissue formation in the tumourigenic region. In vitro effectiveness of this system was investigated on T98G human glioblastoma cell line and human umbilical vein endothelial cells. The results show that the chitosan scaffold containing 100?µg 5-FU and 100?µg bevacizumab has a potential to prevent the tumour formation in vitro conditions.     

Determination of patulin in fruit juices and compote of apple and pear

Abstract

Patulin is a mycotoxin that is produced by Penicillium, Aspergillus and Byssochlamys. This toxin is found in fruits juice. In this study, 32 samples of fruit juice and apple and pear compotes were randomly collected at supermarkets in Mazandaran, in spring 2012. High performance liquid chromatography (HPLC) method was applied to measure patulin. Of the 8 apple juice samples 37.5% were contaminated with >50?µg/L and 62.5% were contaminated with <50?µg/L. 12.5% of pear juices samples was contaminated with <50?µg/L. In compote of apple samples 12.5% was contaminated with <50?µg/L and none of the compote of pear samples were contaminated with this toxin.     

Radical scavenging,prolyl endopeptidase inhibitory,and antimicrobial potential of a cultured Himalayan lichen Cetrelia olivetorum

Context: Lichens are source of natural bioactive compounds which are traditionally used to cure a variety of ailments.

Objective: The objective of this study is to assess free radical scavenging, prolyl endopeptidase inhibitory (PEPI), and antimicrobial potential of a high altitude lichen species Cetrelia olivetorum (Nyl.) W. L. Culb. & C. F. Culb (Parmeliaceae).

Materials and methods: Lichen C. olivetorum has been cultured in vitro, and optimized culture conditions were implemented in bioreactor to obtain high quantity of biomass for the study of radical scavenging, PEPI, and antimicrobial activities. Radical scavenging activity of methanol extract of Cetrelia olivetorum (MECO) was tested at 100?µg/mL, PEPI activity at 25 and 50?µg/mL, and antimicrobial activity at 5, 25, 50, and 100?µg/mL conc. All the biological activities of natural thallus extract and its derived culture extract were evaluated spectrophotometrically.

Results: Murashige and Skoog medium supplemented with 3% glucose and 100?ppb indole-3-butyric acid (IBA) supported biomass growth at flask level and yielded 5.095?g biomass in bioreactor. MECO of both the cultured and the natural lichen exhibited half inhibiting concentration (IC₅₀) for radical scavenging activities in the range of 50–60?µg/mL, whereas the IC₅₀ value of standard antioxidants was found to be in the range of 12–29?µg/mL. The IC₅₀ value of lichen extract for PEPI activity was 144–288?µg/mL, whereas the IC₅₀ value of standard prolyl endopeptidase inhibitor, Z-pro-prolinal, was 57.73?µg/mL. As far as the antimicrobial activity of MECO is concerned, minimum inhibitory concentration (MIC) value of lichen extracts against tested microorganisms was obtained in the range of 50–104?µg/mL and found to be more effective than commercially available standard erythromycin.

Discussion: Murashige and Skoog medium containing IBA was found to be suitable for maximum biomass production of C. olivetorum under bioreactor conditions. The cultured lichen biomass extract also showed antioxidant, PEPI, and antimicrobial potential.

Conclusion: The present study indicates therapeutic potential of Himalayan lichen C. olivetorum against neurodegenerative diseases owing to its radical scavenging, PEPI, and antimicrobial activities. Further, the result encourages its commercial exploitation through mass culture for production of its bioactive components and their use in pharmaceutical and nutraceutical industries.     

Pharmacological evaluation for anti-asthmatic and anti-inflammatory potential of Woodfordia fruticosa flower extracts

Context: Woodfordia fruticosa Kurz. (Lythraceae) flowers are ethnopharmacologically acclaimed in the Indian medicinal system to treat asthma.

Objective: To evaluate W. fruticosa flower extracts for anti-asthmatic effect.

Materials and methods: Ethyl acetate, acetone, methanol, and hydro-alcohol extracts of W. fruticosa flowers were obtained successively and standardized. Ability of extracts to stabilize free radicals and compound-48/80-induced mast cell degranulation was evaluated. In vitro anti-inflammatory potential of extracts at 100 and 200?µg/ml by membrane stabilization and in vivo inhibition of rat paw edema up to 5?h (100 and 200?mg/ml; p.o.) was evaluated. In vitro bronchorelaxant effect was examined against histamine- and acetylcholine (1?µg/ml; independently)-induced guinea pig tracheal contraction. Extracts were evaluated for bronchoprotection (in vivo) ability against 0.1% histamine- and 2% acetylcholine-induced bronchospasm in guinea pigs at 100 and 200?mg/ml; p.o.

Results: Standardization studies revealed that the methanol extract exhibited highest polyphenolic (62.66 GAE), and flavonoid (6.32 RE) content and HPLC fingerprinting confirmed the presence of gallic acid (R_t 1.383). IC₅₀ values for DPPH scavenging and metal chelation by the methanol extract were 40.42 and 31.50?µg/ml. Methanol and ethyl acetate extracts at 100?µg/ml exhibited 06.52 and 07.12% of histamine release. Methanol, ethyl acetate, and hydro alcohol extracts at 200?mg/kg demonstrated 32.73, 29.83, 26.75% and 32.46, 9.38, 26.75% inhibition of egg albumin and carrageenan-induced inflammation, respectively. Methanol extract exhibited 100% bronchorelaxation and 48.83% bronchoprotection.

Conclusion: Woodfordia fruticosa flower (WFF) extracts exhibited anti-asthmatic effect by demonstrating bronchoprotection, bronchorelaxation, anti-inflammatory, antioxidant, and mast cell stabilization ability.     

Nano titanium exposure induces dose- and size-dependent cytotoxicity on human epithelial lung and colon cells

The productions as well as use of Titanium dioxide nanoparticles (TNPs) were rapidly increasing in the present nano-world. The TNP becomes an inevitable part our daily life in the form of cosmeceutical, bio-medical, and nano-pharmaceutical applications. The TNPs are either inhaled or ingested into the human body through common routes of exposure like the lungs and the oral-gastrointestinal tract (GIT). Human lung and colon were exposed to test particles, TNP 18?nm (TNP 18), TNP 30?nm (TNP 30), and TNP 87?nm (TNP 87) with a dose range 0.1–100?µg/ml. The effect of exposure was determined using MTT, LDH, and DCFH-DA methods. The TNP 18, TNP 30, and TNP 87 significantly (p?<?0.001) reduced cell viability in a dose- and a size-dependent manner in 60 and 100?µg/ml. The lowest IC₅₀ values 21.80 and 24.83?µg/ml were observed in A549 and Caco-2 for the smallest size, TNP 18. Further, for TNP 30, IC₅₀ values were 23.30 and 28.59?µg/ml compared to Nano QTZ 43.82 and 45.86?µg/ml. The EC₂₅ values of LDH leakage were 5.83 and 9.50?µg/ml for TNP 18 in lung and colon cells. Besides, ROS levels increased significantly at doses 60 (p?<?0.01) and 100 (p?<?0.001) µg/ml in two cells. The smaller size particle, TNP 18 has produced a significant (p?<?0.05) toxic effect at the lowest dose i.e., 10?µg/ml. Therefore, we conclude that TNP 18, TNP 30, and TNP 87 induced a dose- and size-dependent cytotoxicity via decreased cell viability, increased LDH and ROS levels by in vitro methods.     

Effects of Amburoside A and Isokaempferide,Polyphenols from Amburana cearensis,on Rodent Inflammatory Processes and Myeloperoxidase Activity in Human Neutrophils

Abstract: The present study evaluated the anti‐inflammatory activity of amburoside A (a phenol glucoside) and isokaempferide (a flavonol) isolated from the trunk bark of Amburana cearensis, a medicinal plant used in northeast Brazil for the treatment of asthma. Animals (male Wistar rats or Swiss mice) pre‐treated with amburoside A (25 and 50 mg/kg) or isokaempferide (12.5, 25 and 50 mg/kg), orally or intraperitoneally, showed a significant inhibition of the paw oedema induced by carrageenan (1%), prostaglandin E₂ (30 nmol/paw), histamine (200 µg/paw) or serotonin (200 µg/paw). Histological and morphometric evaluations of the rat paw oedema induced by carrageenan showed that amburoside A and isokaempferide also inhibited the accumulation of inflammatory cells. Amburoside A reduced significantly the paw oedema and the increase in vascular permeability induced by dextran, as related to the control group. Similar results were observed with the isokaempferide pre‐treatment. Furthermore, amburoside A or isokaempferide inhibited both leucocyte and neutrophil migrations, in mouse peritoneal cavity, after the carrageenan injection. The polyphenols were not cytotoxic and blocked N‐formyl‐methyl‐leucyl‐phenylalanine‐induced myeloperoxidase release and activity in human neutrophils. In addition, amburoside A and isokaempferide at 50 and 100 µg/ml concentrations reduced significantly the lipopolysaccharide‐mediated increase in tumour necrosis factor‐α (TNF‐α) levels. These results provide, for the first time, evidence to support the anti‐inflammatory activity of amburoside A and isokaempferide that seems to be related to an inhibition of inflammatory mediators, such as TNF‐α, as well as histamine, serotonin and prostaglandin E₂, besides leucocyte infiltration in a dose‐ or concentration‐dependent manner. These anti‐inflammatory effects can be explained, at least in part, by the ability of these compounds to reduce neutrophil degranulation, myeloperoxidase activity, mediators as well as TNF‐α secretion.     

Toxicity evaluation of magnetic iron oxide nanoparticles reveals neuronal loss in chicken embryo

Magnetic iron oxide nanoparticles (IONs) display the ability to cross blood???brain barrier and are envisioned as diagnostic and therapeutic applications, but there are few studies on their potential embryonic toxicity in higher vertebrates. This study investigates interaction of IONs with egg albumen and its subsequent toxicity on chicken embryo. Physicochemical interactions of IONs with egg albumen revealed alterations in friccohesity and secondary structural changes due to weak Vander Waals forces. Toxicity assessment of IONs (10, 25, 50, 100, and 200?µg/ml doses) on chicken embryo accounted for 100% mortality at 200?µg/ml dose due to Fe²⁺ ions overload. However, lower doses (50 and 100?µg/ml) recorded decrement in whole weights and crown-rump lengths of chicken embryo possibly due to ION–albumen interactions. Histology of brain tissue revealed degeneration of neurons (50–60%) at 10–100?µg/ml dose range of IONs. Toxicity studies of IONs with diverse animal models are needed to set a toxicity benchmark for preventing embryonic toxicity prior to its use in biomedical applications. This is the first study on toxicity assessment of IONs in chicken embryo.     

The plant extract of Pao pereira potentiates carboplatin effects against ovarian cancer

Context: Herbal preparation of Pao pereira [Geissospermum vellosii Allem (Apocynaceae)] has long been used by oncologic patients and Integrative Medicine practitioners in South America. However, its anticancer activities have not been systematically studied.

Objective: To investigate the anticancer effects of β-carboline alkaloids-enriched extract from Pao pereira (Pao), either alone or in combination with carboplatin, in preclinical ovarian cancer models.

Materials and methods: Cytotoxicity of Pao (0–800?µg/ml) against different ovarian cancer cell lines and an immortalized epithelial cell line was detected by flow cytometry, MTT assay and colony formation in soft agar. Combination of Pao and carboplatin, a primary chemotherapeutic drug for ovarian cancer, was evaluated using Chou-Talalay’s methods. Mice bearing intraperitoneally spread ovarian cancer were treated with 20 or 50?mg/kg/day Pao by i.p. injection. Carboplatin at 15?mg/kg/week i.p. was compared and combined to Pao treatments.

Results: Pao selectively inhibited ovarian cancer cell growth with IC₅₀ values of 180–235?µg/ml, compared to 537?µg/ml in normal cells. Pao induced apoptosis dose- and time-dependently and completely inhibited colony formation of tumor cells in soft agar at 400?µg/ml. Pao greatly enhanced carboplatin cytotoxicity, with dose reduction (DRIs) for carboplatin at 1.2–10 fold. In vivo, Pao alone suppressed tumor growth by 79% and decreased volume of ascites by 55%. When Pao was combined with carboplatin, tumor inhibition reached 97% and ascites was completely eradicated.

Discussion and conclusion: Pao possess potent antitumor activity and could enhance carboplatin effect, and therefore holds therapeutic potential in the treatment of ovarian cancer.     

An approach to enhanced stability: Formulation and characterization of Solanum lycopersicum derived lycopene based topical emulgel

Focus of the study was to design a novel and cost effective extraction technique for the lycopene from Lycopersicum esculentum L. fruit and to develop and characterize a stable emulgel formulation containing lycopene as an active ingredient as well as to design an analytical method to determine lycopene concentration in emulgel. Emulgel formulation was prepared and evaluated for its stability at different storage conditions, 8?°C, 25?°C, 40?°C, 40?°C?+?75% relative humidity (RH) and 50?°C, for 6?months. Results were statistically analyzed using two way ANOVA, Post-Hoc test and paired sample t-test at 5% significance level. Designed extraction technique presented comparable yield, 154.83?mg/Kg of tomato fruit, with all recoveries in the range of 145–156?mg/Kg of tomato. “P-values” calculated for different levels of stability parameters were <0.05, except at 50?°C and time points of 60th day and later. Analytical method designed was having linear range of lycopene 1–10?µg/mL with limit of detection 0.11?µg/mL and limit of quantification 0.34?µg/mL. All inter-day and intra-day recoveries were in the range of 94–105% while in all measurements RSD % was ≤5.36. It can be concluded that the extraction technique was cost effective with comparable results and analytical method was simple, robust, specific and sensitive enough to be used for lycopene concentration determination in emulgel formulation. Furthermore, designed formulation was stable even at high temperature of 40?°C and RH 75%.     

Evaluation of cytotoxic and genotoxic effects of Benodanil by using Allium and Micronucleus assays

The aim of this study was to evaluate the potential cytotoxic effects of Benodanil fungicide by employing both mitotic index (MI) and mitotic phases on the root meristem cells of Allium cepa and genotoxic effects by using in vitro micronucleus assay (MN) in human peripheral blood lymphocyte. In the Allium root growth inhibition test, the EC₅₀ value was first determined as 25?ppm. Then, 2?×?EC₅₀ value (50?ppm), EC₅₀ value (25?ppm), and 1/2?×?EC₅₀ value (12.5?ppm) were tested with different treatment periods (24, 48, and 72?h). Both negative and positive controls were also used in parallel experiments. We obtained that mitotic index and prophase index decreased when compared with the control in all concentrations. In the micronucleus assay, lymphocytes were treated with various concentrations (250, 500, 750, and 1000?µg/ml) of Benodanil for 24 and 48?h. The results showed that Benodanil did not induce MN frequency in all concentrations of both treatment periods. Additionally, it was determined that this pesticide decreased nuclear division index (NDI) significantly. It was concluded that Benodanil has a cytotoxic effects depending on decreasing of MI and NDI.     

Genoprotective and neuroprotective effects of Daphne gnidium leaf methanol extract,tested on male mice

Methanol extract of Daphne gnidium leaves was assessed for its antigenotoxic and neuroprotective effects through antioxidant and antibutyrylcholinesterase activities. Antigenotoxic activity was evaluated against methyl methanesulfonate injected intraperitoneally to mice, using the comet assay. The protective effect of D. gnidium reached 99.12%, at the lowest tested dose (44?mg/kg b.w.) in kidney cells, and 92.16% at the dose of 88?mg/kg b.w. in blood cells. The extract was dissolved in water and administrated to mice by intraperitoneal injection. Antioxidant activity was tested against DPPH radicals. It reached a maximum of 74.52% with an IC₅₀ value of 45?µg/ml. Anticholinesterase activity was determined against butyrylcholinesterase, an enzyme linked to Alzheimer disease. The extract exhibited antibutyrylcholinestrase effect with an inhibition percentage of 35.82% at the lowest tested dose (44?mg/kg b.w.).     

Antidiabetic effects of Mukia maderaspatana and its phenolics: An in vitro study on gluconeogenesis and glucose uptake in rat tissues

Context: Traditional medicine is used by over 60% of the world’s population for health care. Mukia maderaspatana (L.) M. Roem. (Cucurbitaceae) (Mukia) is extensively used in folklore medicine as an antidiabetic plant. It is rich in phenolics that contribute to its medicinal properties.

Objective: Mukia extract and phenolics such as quercetin and phloroglucinol are investigated for their in vitro antidiabetic activity.

Materials and methods: Quercetin, phloroglucinol, and methanol extract of the dried whole plant (0.25 and 0.5?mg/ml) were studied for the inhibition of gluconeogenesis in rat liver slices and glucose uptake in isolated rat hemi-diaphragm (50 and 100?µg/ml). Phenolics of Mukia were analyzed by HPLC.

Results and discussion: Glucose (1.2?mg/g/h) was synthesized from pyruvate and the synthesis was completely inhibited by insulin (1?U/ml). Quercetin at 0.25 and 0.5?mg/ml caused 65% and 89% inhibition (0.42?mg/g/h and 0.13?mg/g/h glucose). Addition of insulin did not increase inhibition. Phloroglucinol inhibited 100% glucose production with or without insulin. Mukia (0.25?mg/ml) inhibited gluconeogenesis (0.65?mg/g/h) by 45%, and with insulin, inhibition increased to 50% (0.59?mg/g/h). At 0.5?mg/ml, glucose production was stimulated by1.2-fold, but with insulin it was inhibited by 89% (0.13?mg/g/h glucose). Mukia had no effect on glucose uptake, but potentiated the action of insulin mediated glucose uptake (152.82?±?13.30?mg/dl/g/30?min) compared with insulin control (112.41?±?9.14?mg/dl/g/30?min) (p?0.05). HPLC analysis revealed the presence of phenolics.

Conclusion: Results provide scientific rationale for the use of Mukia in folk medicine as an antidiabetic nutraceutical.     

Effects of fibrates on human organic anion-transporting polypeptide 1B1-, multidrug resistance protein 2- and P-glycoprotein-mediated transport

The effects of different fibric acid derivatives (bezafibrate, clofibrate, clofibric acid, fenofibrate, fenofibric acid and gemfibrozil) on human organic anion transporting-polypeptide 1B1 (OATP2, OATP-C, SLC21A6), multidrug resistance protein 2 (MRP2/ABCC2) and MDR1-type P-glycoprotein (P-gp/ABCB1) were examined in vitro. Cyclosporin A (a known inhibitor of OATP1B1 and P-gp), MK-571 (a known inhibitor of MRP2) and cimetidine (an organic cation) were also tested. Bezafibrate, fenofibrate, fenofibric acid and gemfibrozil showed concentration-dependent inhibition of estradiol 17-β-D-glucuronide uptake by OATP1B1-stably transfected HEK cells, whereas clofibrate and clofibric acid did not show any significant effects up to 100?µM. Inhibition kinetics of gemfibrozil, which exhibited the most significant inhibition on OATP1B1, was shown to be competitive with a K_i?=?12.5?µM. None of the fibrates showed any significant inhibition of MRP2-mediated transport, which was evaluated by measuring the uptake of ethacrynic acid glutathione into MRP2-expressing Sf9 membrane vesicles. Only fenofibrate showed moderate P-gp inhibition as assessed by measuring cellular accumulation of vinblastine in a P-gp overexpressing cell-line. Cyclosporin A significantly inhibited OATP1B1 and P-gp, whereas only moderate inhibition was observed on MRP2. The rank order of inhibitory potency of MK-571 was determined as OATP1B1 (IC₅₀: 0.3?µM)?>?MRP2 (4?µM)?>?P-gp (25?µM). Cimetidine did not show any effects on these transporters. In conclusion, neither MRP2- nor P-gp-mediated transport is inhibited significantly by the fibrates tested. Considering the plasma protein binding and IC₅₀ values for OATP1B1, only gemfibrozil appeared to have a potential to cause drug–drug interactions by inhibiting OATP1B1 at clinically relevant concentrations.     

Effect of hydroalcoholic extract of Aegle marmelos fruit on radical scavenging activity and exercise-endurance capacity in mice

Context: Aegle marmelos L. Corr (Rutaceae) is an important Indian Ayurvedic medicinal plant used for the treatment of various ailments. However, little information is available on the anti-fatigue properties of its fruit.

Objective: Evaluation of the physical endurance and exercise-induced oxidative stress modulating properties of A. marmelos fruit in mice.

Material and methods: Radical scavenging activity of the fruit hydroalcoholic extract was evaluated using in vitro systems. The extract was further evaluated for its endurance-enhancing properties at three oral doses (100, 200 and 400?mg/kg?b.wt) in BALB/c mice for 21?d using a swimming test.

Results and discussion: The extract exhibited significant scavenging activity against DPPH (IC₅₀, 351?±?37?µg/ml) and ABTS radicals (IC₅₀, 228?±?25?µg/ml), respectively, with the polyphenol content of 95?µg/mg extract. It also inhibited AAPH radical-induced oxidation of biomolecules such as BSA protein (63%), plasmid DNA (81%) and lipids (80.5%). Administration of extract resulted in an increase in the duration of swimming time to exhaustion by 23.4 and 47.5% for medium and higher doses, respectively. The extract significantly normalized the fatigue-related biochemical parameters and also down-regulated the swim stress-induced over-expression of heat shock protein-70 and up-regulated the skeletal muscle metabolic regulators (GLUT-4 and AMPK1-α) by 2- and 3-fold, respectively, at the higher dose in muscle tissues.

Conclusion: Our study demonstrates the anti-fatigue properties of A. marmelos fruit, most probably manifested by delaying the accumulation of serum lactic acid, increasing the fat utilization and up-regulating the skeletal muscle metabolic regulators.