Antioxidant activities of different parts of Gnetum gnemon L.

Analyses on biological activities of Gnetum gnemon were done to determine the total phenolic and antioxidants of the plant. Four parts of G. gnemon were used in this study, which were leaf, bark, twig, and seeds of the plant. All parts were extracted in methanol, ethanol, hexane, chloroform and hot water using reflux. The total phenolic content of the plant extracts were determined by using Folin-Ciocalteu method. The results demonstrated that the bark from hot water extract showed the highest total phenolic at 10.71?±?0.01 mg GAE/ FDW, while the lowest was chloroform extract of seed at 2.15?±?0.01 mg GAE/ FDW. The antioxidant activity of the plant extracts were determined by using DPPH and FRAP assays, respectively. The DPPH results showed that all plant extracts demonstrated weak free radical scavenging activity tested at the final concentration of 300 μg/ml. In contrast, the methanolic twig extract showed strong reducing power activity (FRAP) at 83.55?±?1.05%, while the hot water seed extract showed the least activity at 41.86?±?4.22% tested at the final concentration of 300 μg/ml. However, there were no correlation between total phenolics and both antioxidant assays tested.     

Total phenolic and flavonoid contents and antioxidant activity of ginger (Zingiber officinale Rosc.) rhizome,callus and callus treated with some elicitors

The present study was aimed at determining total phenolic and flavonoid contents and studying the antioxidant activity of ginger (Zingiber officinale Rosc.) rhizome and callus, 6-gingerol and 6-shogaol and callus treated with elicitors. Petroleum ether (PE) and chloroform: methanol (1:1, v/v) (CM) extracts were prepared by maceration. Highest total phenolic content was obtained from the CM extract (60.34?±?0.43?mg gallic acid/g) of rhizome while callus showed lower content detected in the CM extract (33.6?±?0.07?mg gallic acid/g). Flavonoids were only detected in rhizome (CM extract 40.25?±?0.21?mg quercetin/g). Both rhizome extracts exhibited good antioxidant activity with higher activity recorded in PE extract (IC₅₀ value 8.29?±?1.73?μg/mL). Callus extracts revealed lower antioxidant activity (IC₅₀ value 1265.49?±?59.9?μg/mL obtained from CM extract). 6-gingerol and 6-shogaol displayed high antioxidant activity in both assays with IC₅₀ 4.85?+?0.58_DPPH and 5.35?±?0.33_ABTS μg/mL for the former and IC₅₀ 7.61?±?0.81_DPPH and IC₅₀ 7.05?±?0.23_ABTS μg/mL for the latter. Treatment of callus with elicitors showed significant (p?<?0.05) effects in enhancing phenolic content and related antioxidant activity. The highest significant increase in phenolic content (37% and 34%) and antioxidant activity in DPPH assay (34% and 30%) was observed in callus treated with 100?mg/L yeast extract and 50?mg/L salicylic acid respectively. Therefore, studying the effect of the elicitation of ginger cultured tissues in phenolic accumulation would be of immense importance for pharmacological, cosmetic and agronomic industries.     

Propolis protects against high glucose-induced vascular endothelial dysfunction in isolated rat aorta

While propolis is known to have abundant bioactive constituents and a variety of biological activities, it is not clear whether propolis has beneficial effects on high glucose-mediated vascular endothelial impairment. The aim of the present study was to investigate the potential protective effect of propolis extract against the acute vascular endothelial dysfunction resulting from exposure to high glucose load and to elucidate its underlying mechanism. Rat aortic rings were incubated with normal glucose (11 mM), high glucose (44 mM), or mannitol (44 mM) for 3 h with or without propolis extract (400 μg/ml). Contraction to phenylephrine (Phe, 10^?9–10^?5 M) and relaxation to acetylcholine (ACh, 10^?9–10^?5 M) and sodium nitroprusside (SNP, 10^?9–10^?5 M) were measured before and after incubation. Changes in malondialdehyde (MDA), reduced glutathione (GSH), and superoxide dismutase (SOD) were also measured. Phe-induced contraction was impaired by high glucose as the E _max decreased from 138.87?±?11.43 to 103.65?±?11.5 %. In addition, ACh-induced relaxation was impaired as the E _max decreased from 99.80?±?7.25 to 39.20?±?6.5 %. SNP-induced relaxation was not affected. Furthermore, high glucose decreased the levels of both SOD (by 6 U/ml) and GSH (by 68 %) and increased levels of MDA (by 85 %). Propolis extract prevented high glucose-induced impairment of Phe and ACh responses and increased both SOD and GSH, leading to decreased MDA levels. In conclusion, propolis can protect against high glucose-induced vascular dysfunction by reducing oxidative stress.     

Mannosylated liposomes bearing Amphotericin B for effective management of visceral Leishmaniasis

The cationic and mannosylated liposomes were prepared using the cast film method and compared for their antileishmaniasis activity. The surface of the Amphotericin B (Amp B)-bearing cationic multilamellar liposomes was covalently coupled with p-aminophenyl-α-D-mannoside using glutaraldehyde as a coupling agent, which was confirmed by agglutination of the vesicles with concanavalin A. The prepared liposomes were characterized for shape, size, percent drug entrapment, vesicle count, zeta potential, and in vitro drug release. Vesicle sizes of cationic and mannosylated liposomes were found to be 2.32?±?0.23 and 2.69?±?0.13?µm, respectively. Zeta potential of cationic liposomes was higher (30.38?±?0.3 mV), as compared to mannosylated liposomes (17.7?±?0.8 mV). Percentage drug release from cationic and mannose-coupled liposomes was found to be 45.7%?±?3.1 and 41.9%?±?2.8, respectively, after 24 hours. The in vivo antileishmanial activity was performed on Leishmania donovani–infected golden hamster, and results revealed that Amp B solution was reduced by 42.5?±?1.8% in the parasite load, whereas the placebo cationic liposomes and drug-containing cationic liposomes showed a reduced parasite load (i.e., 28.1?±?1.5 and 61.2?±?3.2%, respectively). The mannose-coupled liposomes showed a maximum reduction in parasite load (i.e., 78.8?±?3.9%). The biodistribution study clearly showed the higher uptake of mannosylated liposomes in the liver and spleen and hence the active targeting to the reticular endothelial system, which, in turn, would provide a direct attack of the drug to the site where the pathogen resides, rendering the other organs free and safe from the toxic manifestations of the drug.     

Composition, antimicrobial and antioxidant activity of the extracts of Eryngium palmatum Pančić and Vis. (Apiaceae)

The chemical composition, antimicrobial and antioxidant activity of Eryngium palmatum, an endemic plant species from the Balkan Peninsula, were investigated. The flavonoids apigenin (9.5±0.3 mg g^?1) and apigenin 7-O-glucoside (2.4±0.1 mg g^?1) were determined in a methanol extract of aerial parts using HPLC analysis. The methanol extract of roots contained catechin (5.0±0.1 mg g^?1), epicatechin (2.9±0.1 mg g^?1), chlorogenic acid (1.6±0.0 mg g^?1), gallic acid (0.9±0.0 mg g^?1) and rosmarinic acid (0.9±0.2 mg g^?1). GC-FID and GCMS analysis of a chloroform extract of aerial parts showed that the main volatile constituents were falcarinol, linoleic acid, hexadecanoic acid and methyl linoleate (comprising 32.6%; 24.4%; 19.9; 13.2% of the volatile fraction, respectively), while octanoic acid, tetradecanol and dodecanol dominated in the chloroform extract of the roots (34.9%; 25.8%; 22.2% of the volatile fraction, respectively). Investigation of antimicrobial activity by broth microdilution showed that the methanol and chloroform extracts of aerial parts and roots exerted a significant effect (MIC 3.5–15.6 μg mL^?1) against tested Gram-positive and Gram-negative bacteria. The methanol extracts of aerial parts or roots exerted moderate ferric reducing antioxidant power, DPPH radical scavenging activity and hydroxyl radical scavenging activity.     

Characterization of the interaction between liposomal formulations and Pseudomonas aeruginosa

The interactions between three liposomal formulations and Pseudomonas aeruginosa cells were evaluated by a lipid mixing assay and electron paramagnetic resonance (EPR) spectroscopy. The effect of the bacteria on the liposomal phase characteristics, the release of the liposomes’ content, and the uptake rate of gentamicin by bacteria were monitored as a function of time, using EPR spectroscopy. The [16-DSA uptake]_Total from DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) liposomes reached 93?±?12% over a 3-hour assay period, of which 9% crossed the bacterial inner membrane. A small amount of 16-DSA uptake from DPPC/Chol (cholesterol) vesicles was found throughout the 3-hour period of time. Although DPPC/DMPG (dimyristoylphosphatidylglycerol) vesicles showed a smaller value of [16-DSA uptake]_Total with respect to that of DPPC vesicles, they appeared to be effective in disrupting the bacterial membrane, resulting in a greater accumulation of 16-DSA inside the inner membrane. Exposure to bacteria caused the DPPC/Chol, DPPC, and DPPC/DMPG formulations to release 4.6?±?1.5, 17.6?±?1.2, and 34?±?3.7% of their content, respectively. Time-dependent fluid regions were developed within the vesicles when mixed with bacteria, and their growth over time depended on liposomal formulations. Incubation of gentamicin with bacteria for 3 hours resulted in 87?±?3% of the drug crossing the bacterial inner membrane. In conclusion, interaction between the liposome drug carriers and the bacterial cells result in vesicle fusion, disruption of the bacterial membrane, release of the liposomal content in the close vicinity of the bacteria cells, and the subsequent intracellular uptake of the released liposomal content.     

The membraneless bioelectrochemical reactor stimulates hydrogen fermentation by inhibiting methanogenic archaea

The membraneless bioelectrochemical reactor (Ml-BER) is useful for dark hydrogen fermentation. The effect of the electrochemical reaction on microorganisms in the Ml-BER was investigated using glucose as the substrate and compared with organisms in a membraneless non-bioelectrochemical reactor (Ml-NBER) and bioelectrochemical reactor (BER) with a proton exchange membrane. The potentials on the working electrode of the Ml-BER and BER with membrane were regulated to ?0.9 V (versus Ag/AgCl) to avoid water electrolysis with a carbon electrode. The Ml-BER showed suppressed methane production (19.8?±?9.1 mg-C·L^?1·day^?1) and increased hydrogen production (12.6?±?3.1 mg-H·L^?1·day^?1) at pH_out 6.2?±?0.1, and the major intermediate was butyrate (24.9?±?2.4 mM), suggesting efficient hydrogen fermentation. In contrast, the Ml-NBER showed high methane production (239.3?±?17.9 mg-C·L^?1·day^?1) and low hydrogen production (0.2?±?0.0 mg-H·L^?1·day^?1) at pH_out 6.3?±?0.1. In the cathodic chamber of the BER with membrane, methane production was high (276.3?±?20.4 mg-C·L^?1·day^?1) (pH_out, 7.2?±?0.1). In the anodic chamber of the BER with membrane (anode-BER), gas production was low because of high lactate production (43.6?±?1.7 mM) at pH_out 5.0?±?0.1. Methanogenic archaea were not detected in the Ml-BER and anode-BER. However, Methanosarcina sp. and Methanobacterium sp. were found in Ml-NBER. Prokaryotic copy numbers in the Ml-BER and Ml-NBER were similar, as were the bacterial community structures. Thus, the electrochemical reaction in the Ml-BER affected hydrogenotrophic and acetoclastic methanogens, but not the bacterial community.     

Phytochemical composition and antimicrobial,antioxidant and cytotoxic activities of <Emphasis Type="Italic">Anchusa officinalis</Emphasis> L. extracts

The aim of this study is to examine phytochemical composition and evaluation of antimicrobial, antioxidant and cytotoxic activity of Anchusa officinalis plant extracts in different solutions: ethanol, chloroform, petroleum, acetone and ethyl acetate. A comparative analysis has shown that ethanol extract had the highest concentration of phenols (104.03?±?0.63 mgGA/g), and the highest concentration of flavonoids (30.26?±?0.40 mgRU/g). The highest concentration of the condensed tannins recorded in chloroform extract (74.65?±?0.57 mg GA/g). The ethanol extract showed the strongest antioxidant and the best antimicrobial activity as compared to all other tested extracts, while the chloroform and the acetone extracts showed the best cytotoxic activity on cell line of mouse fibroblast carcinoma (L2OB). This is the first report of citotoxic activity of extract A. officinalis plant from Balkan region on tumor cell lines. HPLC analysis of A. officinalis plant extracts confirmed that the predominant polyphenol components were: rosmarinic acid, chlorogenic acid, naringenin, lutein-glycoside and rutin.     

Characterization,inhibitory activity and mechanism of polyphenols from faba bean (gallic-acid and catechin) on α-glucosidase: insights from molecular docking and simulation study

Abstract

The chemo-profiling of ethanolic extract of faba beans seeds was performed and explored as an α-glucosidase inhibitor. The inhibition of α-glucosidase is one of the alternatives approach to control postprandial hyperglycemia by, resulting in the delay of the carbohydrate digestion of absorbable monosaccharides. Ethanolic seed extract showed phenolic compounds, flavonoid such as gallic acid (m/z [M–?H]?=?169.0124,C₇H₆O₅) ellagic acid derivatives epigallocatechin (m/z [M–?H?=?305.0644,C₁₅H₁₄O₇),catechin (m/z [M–?H]?=?289.0656,C₁₅H₁₄O₆), epigallocatechin gallate (m/z [M–?H]?=?457.0578,C₂₂H₁₈O₁₁) and epicatechin monogallate (m/z [M–?H]?=?441.081, C₂₂H₁₈O₁₀). The extract was found to exert inhibitory activity (88.28?±?2.67%) (IC₅₀ value of 2.30?±?0.032?mg/mL) with a mixed mode of inhibition (Km, apparent = 0.54?±?0.020?mM and Vmax, apparent 0.136?±?0.04?mM/min). Molecular docking studies of gallic acid and catechin on α-glucosidase proposed productive binding modes having binding energy (?6.58?kcal/mol and ?7.25?kcal/mol) with an effective number of hydrogen bonds and binding energy. Tyr63, Arg197, Asp198, Glu 233, Asn324, Asp 326 of α-glucosidase participated in binding events with gallic acid and catechin. Molecular dynamics simulation studies were performed for both complexes i.e. gal:α-glucosidase and cat:α-glucosidase along with apo state of α-glucosidase, which revealed stable systems during the simulation. These findings of the present study may give an insight into the further development of the novel antidiabetic drug from the seeds of faba beans.     

Life table of Gaeolaelaps aculeifer (Acari: Laelapidae) feeding on larvae of Lycoriella auripila (Diptera: Sciaridae) with stage-specific estimates of consumption

Gaeolaelaps aculeifer (Canestrini, 1883) is a soil-dwelling predatory mite with potential for use as a biological control agent of fungus gnats (Diptera: Sciaridae) in mushroom production. The life table, predation rate and population growth rate of G. aculeifer on a diet of larvae of the sciarid fly, Lycoriella auripila, at 23?±?1°C, 60?±?5% RH and a photoperiod of 0:24 (L:D)?h was investigated. The results revealed that the duration of egg, larva, protonymph, deutonymph, females and males of G. aculeifer were 3.8?±?0.1, 1.4?±?0.1, 3.9?±?0.1, 4.1?±?0.1, 67.7?±?2.8 and 60.3?±?3.1 days, respectively. Net reproductive rate (R₀) was 54.8?±?7.1 offspring, intrinsic rate of increase (r) was 0.12?±?0.01 offspring day^?1, finite rate of increase (λ) was 1.13?±?0.01 day^?1and mean generation time (T) was 32.3?±?0.6 days. The predator consumed a mean of 0.08?±?0.05, 1.73?±?0.18, 3.16?±?0.28 and 75.9?±?7.1 third instar L. auripila larvae during the larval (1.3?±?0.1 days), protonymph (3.9?±?0.1 days), deutonymph (4.1?±?0.1 days) and adult (52.6?±?2.2 days) stages. Population parameters and consumption rates suggest that G. aculeifer has good potential as a biological control agent of L. auripila in mushroom production.     

Attractive Effects of American Serpentine Leafminer, Liriomyza trifolii (Burgess), to Light-Emitting Diodes

Phototactic responses of Liriomyza trifolii adults to six different light-emitting diodes (LEDs) were investigated, and their responses were compared to that using a luring lamp (BLB) under laboratory conditions. Based on the attraction rate under optimal light conditions, the green LED (520?±?5 nm) showed the highest attraction rate (99.7 %), followed by the yellow LED (590?±?5 nm, 96.1 %), the red LED (625?±?10 nm, 91.4 %), the blue LED (470?±?10 nm, 91.2 %), the UV LED (365 nm, 71.0 %), and the IR LED (730 nm, 5.6 %). Moreover, the green LED was approximately 1.4 times more attractive than BLB (71.1 %) to L. trifolii adults. These results suggest that the green LED was the most useful for monitoring of L. trifolii adults under optimal conditions.     

Decreased paraoxonase 2 enzymatic activity in monocyte/macrophages cells. A comparative in vivo and in vitro study for diabetes

Aim: To investigate peripheral blood monocytes/macrophages (Mo/M?) paraoxonase 2 (PON2) in diabetes and the factors modulating its activity.

Methods: One hundred and eighteen patients with newly diagnosed uncomplicated type 2 diabetes mellitus were compared regarding clinical, biochemical and oxidative stress parameters with 80 healthy subjects. The capacity of the peripheral blood mononuclear cells (PBMNC) to release pro-oxidants and to neutralise them was determined by measuring the respiratory burst (RB) and the intracellular antioxidant enzyme PON2. In vitro experiments were conducted on a differentiated monocytes cell line (dU937) that was exposed to serum deprivation followed by addition of isolated lipoproteins (VLDL or LDL).

Results: Paraoxonase 2 activity in Mo/M? was significantly lower in type 2 diabetes patients (0.042?±?0.044 vs 0.165?±?0.133U lactonase activity/mg protein in controls, p?1c) and insulin resistance (HOMA-IR). In multivariate regression models, 15–34% of the PON2 variance was explained by diabetes. The in vitro addition of VLDL normalised the RB of serum deprived dU937 cells, S? (to 82?±?18% of the cells incubated with serum, S+) and PON2 activity (from 0.524?±?0.061 in S???to 0.298?±?0.048?U/mg protein). In contrast, when LDL was added, the RB remained lower (61?±?12% of S+, p?=?.03) and PON2 higher (0.580?±?0.030?U/mg protein, p?=?.003).

Conclusions: The decrease in monocyte/macrophage PON2 enzymatic activity observed in type 2 diabetes cannot be totally explained by abdominal obesity and insulin resistance. The underlying molecular mechanisms need to be identified.     

Sulphate transport in the cyanobacterium Synechococcus R-2 (Anacystis nidulans, S. leopoliensis) PCC 7942

Synechococcus R-2 (PCC 1942) actively accumulates sulphate in the light and dark. Intracellular sulphate was 1.35 ± 0.23 mol m^?3 (light) and 0.894 ± 0.152 mol m^?3 (dark) under control conditions (BG-11 media: pH_o, 7.5; [SO₄^2?]_o, 0.304 mol m^?3). The sulphate transporter is different from that found in higher plants: it appears to be an ATP-driven pump transporting one SO₄^2?/ATP [ΔμSO₄^2?_i,o=+ 27.7 ± 0.24 kJ mol^?1 (light) and + 24 ± 0.34 kj mol^?1 (dark)]. The rate of metabolism of SO₄^2?at pH_o, 7.5 was 150 ± 28 pmol m^?2 s^?1 (n = 185) in the light but only 12.8 ± 3.6 pmol m^?2 s^?1 (n = 61) in the dark. Light-driven sulphate uptake is partially inhibited by DCMU and chloramphenicol. Sulphate uptake is not linked to potassium, proton, sodium or chloride transport. The alga has a constitutive over-capacity for sulphate uptake [light (n= 105): K_m= 0.3 ± 0.1 mmol m^?3, V_max, = 1.8 ± 0.6 nmol m^?2 s^?1; dark (n= 56): K_m= 1.4 ± 0.4 mmol m^?3, V_max= 41 ± 22 pmol m^?2 s^?1]. Sulphite (SO₃^2?) was a competitive inhibitor of sulphate uptake. Selenate (SeO₄^2?) was an uncompetitive inhibitor.     

Progesterone-loaded nanosized transethosomes for vaginal permeation enhancement: formulation,statistical optimization,and clinical evaluation in anovulatory polycystic ovary syndrome

Bio-identical progesterone (PRG) is an exogenous female steroidal hormone which is used for treatment of polycystic ovary syndrome (PCOS). However, it suffers from poor bioavailability due to hepatic metabolism and poor solubility. The target of this work was to evaluate and statistically optimize PRG-loaded nanovesicle transethosomes (NVTEs) based in mucoadhesive gel for transvaginal delivery of PRG as potential luteal-phase support. A 2⁴ full factorial design was used to explore the effect of phosphatidylcholine (PC), Tween 80, cetyltrimethyl ammonium bromide and ethanol concentration on particle size, entrapment efficiency (EE%), % in vitro PRG release after 24?h and transvaginal flux. PRG-loaded NVTEs were prepared by injection sonication method. The results revealed that the mean particle sizes ranged from 133.3?±?3.42 to 349.5?±?1.24?nm, zeta potential ranged from –23.5?±?3.84 to +74.6?±?4.97?mV, EE% ranged from 87.93?±?3.58 to 97.05?±?2.61%, % PRG release ranged from 50.9?±?2.75 to 90.69?±?2.07 and transvaginal flux ranged from 0.274?±?0.03 to 0.531?±?0.04?mg/cm²/h. The optimized formulation was subjected to transmission electron microscope for morphological examination and then incorporated in the mucoadhesive vaginal gel using Carbopol 974, hydroxyl propyl methylcellulose and sodium alginate. The optimized formulation was clinically studied in anovulatory PCOS and showed a significant increase in the serum PRG, endometrial thickness, echogenicity degree and the pregnancy rate. Briefly, PRG-loaded NVTEs vaginal gel might be a promising formulation for luteal phase support and increase pregnancy rate in anovulatory PCOS.     

Analysis of metabolic pathways and fluxes in a newly discovered thermophilic and ethanol‐tolerant Geobacillus strain

A recently discovered thermophilic bacterium, Geobacillus thermoglucosidasius M10EXG, ferments a range of C5 (e.g., xylose) and C6 sugars (e.g., glucose) and is tolerant to high ethanol concentrations (10%, v/v). We have investigated the central metabolism of this bacterium using both in vitro enzyme assays and ¹³C‐based flux analysis to provide insights into the physiological properties of this extremophile and explore its metabolism for bio‐ethanol or other bioprocess applications. Our findings show that glucose metabolism in G. thermoglucosidasius M10EXG proceeds via glycolysis, the pentose phosphate pathway, and the TCA cycle; the Entner–Doudoroff pathway and transhydrogenase activity were not detected. Anaplerotic reactions (including the glyoxylate shunt, pyruvate carboxylase, and phosphoenolpyruvate carboxykinase) were active, but fluxes through those pathways could not be accurately determined using amino acid labeling. When growth conditions were switched from aerobic to micro‐aerobic conditions, fluxes (based on a normalized glucose uptake rate of 100 units (g DCW)^?1 h^?1) through the TCA cycle and oxidative pentose phosphate pathway were reduced from 64 ± 3 to 25 ± 2 and from 30 ± 2 to 19 ± 2, respectively. The carbon flux under micro‐aerobic growth was directed to ethanol, L ‐lactate (>99% optical purity), acetate, and formate. Under fully anerobic conditions, G. thermoglucosidasius M10EXG used a mixed acid fermentation process and exhibited a maximum ethanol yield of 0.38 ± 0.07 mol mol^?1 glucose. In silico flux balance modeling demonstrates that lactate and acetate production from G. thermoglucosidasius M10EXG reduces the maximum ethanol yield by approximately threefold, thus indicating that both pathways should be modified to maximize ethanol production. Biotechnol. Bioeng. 2009;102: 1377–1386. © 2008 Wiley Periodicals, Inc.     

Callus mediated shoot organogenesis and regeneration of cytologically stable plants of Ledebouria revoluta: An ethnomedicinal plant with promising antimicrobial potency

Ledebouria revoluta are important ethnomedicinal plant found in India and South Africa. Micropropagation via indirect shoot organogenesis had been established from three types of explant (i.e. scale leaf, leaf lamina and root) of L. revoluta. Scale leaf was found superior as compared to leaf lamina and root explant with respect to their organogenic callus induction potentiality. Murashige and Skoog (1962) [MS] media supplemented with 3.0?mg?L^?1 2,4-dichlorophenoxyacetic acid, 0.75?mg?L^?1 β-naphthoxyacetic acid were best effective for inducing organogenic callus. Maximum 17.0?±?0.52 bulblets were induced from about 500?mg of callus within 42–46?days sub-culturing on a medium containing 0.75?mg?L^?1 kinetin. The bulblets were matured (86.7% success) after one month culture on the same medium composition. The best result of in vitro root induction with 100% response and 8.4?±?0.31 roots per bulb was achieved after 18?days of implantation on MS medium containing 2.0?mg?L^?1 indole-3-butyric acid. Plantlets were acclimatized with a 96.0% survival rate. Chromosomal studies revealed cytological stability of callus cells and all regenerants containing 2n?=?30 chromosomes, same as parental plants. Antimicrobial activity of L. revoluta was tested against two Gram-positive bacteria, three Gram-negative bacteria and two fungi. The methanol and ethanol extract proved more effective against bacteria, whereas acetone and chloroform extract shows potential anti-fungal activities. Present protocol can be applied reliably to produce uniform planting materials in large scale. In addition, this efficient indirect regeneration pathway via callus culture opens a way for improvement through genetic transformation.     

Chemical composition and ethanol production potential of Thai seaweed species

This study evaluated the potential use of several Thai seaweed species for ethanol production. The high biomass of the green algae Ulva intestinalis and Rhizoclonium riparium and the red algae Gracilaria salicornia and Gracilaria tenuistipitata in an earthen pond culture led us to select these species for our study. The seaweed species were analyzed for chemical composition, resulting in ash contents of 37.62?±?0.15 % and fiber of 11.93?±?0.16 %, with the highest values in R. riparium. Low lipid values were found in all species, with the highest value (p?<?0.05) in G. salicornia (1.69?±?0.07 %) and the lowest in R. riparium (0.28?±?0.01 %) and G. tenuistipitata (0.26?±?0.01 %). The highest carbohydrate contents were found in G. tenuistipitata (54.89 %), and the lowest were in R. riparium (29.53 %). G. tenuistipitata (8.58?±?0.36 %) and U. intestinalis (8.24?±?0.28 %) had higher sulfate contents compared with G. salicornia (4.69?±?0.04 %) and R. riparium (1.97?±?0.20 %). The monosugar algal tissue components were analyzed by HPLC; rhamnose, xylose, fucose, arabinose, mannose, glucose, and galactose were used as reference sugars. Total sugar was found to be highest in G. tenuistipitata (98.21 %). Arabinose, glucose, and galactose were the main sugar components in all species. Glucose obtained from G. tenuistipitata (6.55 %) and R. riparium (6.52 %) was higher than in G. salicornia (0.27 %) and U. intestinalis (2.78 %). G. tenuistipitata fermentation gave a higher yield of ethanol (4.17?×?10^?3 g ethanol g^?1 sugars; 139.12 μg ethanol g^?1 glucose) than R. riparium (0.086?×?10^?3 g ethanol g^?1 sugars; 33.84 μg ethanol g^?1 glucose), U. intestinalis (0.074?×?10^?3 g ethanol g^?1 sugars; 9.98 μg ethanol g^?1 glucose), and G. salicornia (0.031?×?10^?3 g ethanol g^?1 sugars; 1.43 μg ethanol g^?1 glucose).     

Prostaglandin Endoperoxide H Synthase-2 (PGHS-2) Variants and Risk of Obesity and Microvascular Dysfunction Among Tunisians: Relevance of rs5277 (306G/C) and rs5275 (8473T/C) Genetic Markers

The purpose of this study was to determine the impact of six PGHS-2 genetic variants on obesity development and microvascular dysfunction. The study included 305 Tunisian subjects (186 normal weights, 35 overweights and 84 obeses). PCR analyses were used for allelic discrimination between polymorphisms. Prostaglandin (PGE2, PGI2), leptin, and matrix metalloproteinase (MMP1, 2, 3, 9) levels were evaluated by ELISA. Fatty acid composition was performed by gas chromatography–mass spectrometry. Our results revealed that subjects carrying the PGHS-2 306CC (rs5277) and 8473CC (rs5275) genotypes present higher anthropometric values compared to wild-type genotypes (306GG, BMI (Kg/m²): 27.11?±?0.58; WC (cm): 93.09?±?1.58; 306CC, BMI: 33.83?±?2.46; WC: 109.93?±?5.41; 8473TT, BMI: 27.75?±?0.68; WC: 93.96?±?1.75; 8473CC, BMI: 33.72?±?2.2; WC: 117.89?±?2.94). A reduced microvascular reactivity and a higher PGE2 level were also found in individuals with the 306CC and 8473CC genotypes in comparison to 306GG and 8473TT carriers (306GG, Peak Ach-CVC (PU/mmHg): 0.46?±?0.03; PGE2 (pg/ml): 7933.1?±?702; 306CC, Peak Ach-CVC: 0.24?±?0.01; PGE2: 13,380.3?±?966.2; 8473TT, Peak Ach-CVC: 0.48?±?0.05; PGE2: 7086.41?±?700.31; 8473CC, Peak Ach-CVC: 0.23?±?0.01; PGE2: 13,175.7?±?1165.8). Fatty acid analysis showed a significant increase of palmitic acid (PA) (34.2?±?2.09 vs. 16.82%?±?1.76, P?<?0.001), stearic acid (SA) (25.76?±?3.29 vs. 9.05%?±?2.53, P?<?0.001), and linoleic acid (LA) (5.25?±?1.18 vs. 0.5%?±?0.09, P?<?0.001) levels in individuals carrying the PGHS-2 306CC genotype when compared to GG genotype individuals. Subjects with the 8473CC genotype showed also a significant increase of PA, SA ,and LA levels when compared to TT genotype carriers (PA: 38.02?±?1.51 vs. 12.65%?±?1.54, P?<?0.001; SA: 32.96?±?1.87 vs. 1.38%?±?0.56, P?<?0.001; LA: 26.84?±?2.09 vs. 3.7%?±?1.54, P?<?0.001). Logistic regression analysis revealed that PGHS-2 306CC and 8473CC variants are significantly associated with obesity status (OR 6.25, CI (1.8–21.6), P?=?0.004; OR 3.01, CI (1.13–8.52), P?=?0.03, respectively). Haplotypes containing the C₃₀₆:T₈₄₇₃ (OR 2.91; P?=?0.01) and G_306:C₈₄₇₃ (OR 5.25; P?=?0.002) combinations were associated with an enhanced risk for obesity development in the studied population. In conclusion, our results highlight that PGHS-2 306G/C and 8473T/C variants could be useful indicators of obesity development, inflammation, and microvascular dysfunction among Tunisians.

     

Plumieride from Allamanda cathartica as an inhibitory compound to plant pathogenic fungi

Allamanda leaf extract (Allamanda cathertica) was made in water at room temperature (25?± 2?°C) as well as in a number of less polar to highly polar solvents like methylene chloride, benzene, chloroform and ethyl acetate at their boiling point, that means, at refluxing temperature (40?± 2?°C). Methylene chloride, benzene, chloroform, ethyl acetate and water extracts were applied to determine their growth inhibition against Phomopsis vexans, Phytophthora capsici, Fusarium oxysporum, Rhizoctonia solani and Sclerotium rolfsii. Results of these extracts showed that refluxing methanol, ethanol and ethyl acetate extracts of Allamanda were statistically similar for inhibition of mycelial growth of all fungi tested. But effect of 50% ethanol extract is different; it inhibited 100% mycelial growth of P. vexans, P. capsici and F. oxysporum; 83.33% of R. solani and 88.63% of S. rolfsii. Effort was also made to find out the compound in Allamanda to be responsible for such antifungal activity. Thin layer chromatography (TLC) of Allamanda extracts showed the presence of a number of compounds having polarity very high to low. The R_f values of compounds in 37–42 fractions were calculated and from these six fractions, crystals were separated. These crystals were more or less white. Melting point of these crystals was determined by ordinary and digital melting point apparatus that ranged from 145.5–162 C. Structural determination of the compound was done by Infra-red (IR) spectral study. The finger print region was 700–1400?cm^?1. The strong band at 1612.4, 1633.6, 1693.4, 1655 and 2850.6?cm^?1 indicated the presence of conjugated double bond (–C=C–C=C–), non-conjugated double-bond (–C=C–C–C–C=C–), carbonyl group attached to carbon–carbon double (–CO–C=C), ester (–COOR) and C–H stretching, respectively. Mass spectra of separated compounds gave molecular weight 470. All these characters are typical to pumieride as described previously. Again, In vitro screening of plumieride against P. vexans, P. capsici, F. oxysporum, R. solani and S. rolfsii were found effective in inhibiting radial mycelial growth of these fungi at 1:2 w/v concentration.     

Protection of cowpea,Vigna unguiculata L. (Walp.) with Newbouldia laevis (Seem.) extracts against infestation by Callosobruchus maculatus (Fabricius)

Evaluation of protectant ability of Newbouldia laevis (Seem.) extracts against infestation by Callosobruchus maculatus in cowpea, Vigna unguiculata L. (Walp.) was carried out in the laboratory at ambient temperature of 28?±?2?°C and 70?±?5% relative humidity. Extracts from wood ash, leaf, stem and root bark were tested at different concentrations of 0, 1, 2, 3, 4 and 5%. One hundred per cent mortality of adult beetles was achieved at all concentrations within 72?h of treatment with extracts except in wood ash at 1% concentration, but they were significantly different (p?<?0.05) from the controls. All the extracts were still able to cause high beetle mortality after one, two and three months of cowpea storage at high concentrations (4 and 5%) except wood ash, although there was a slight decrease in mortality during the period of storage. All the extracts significantly (p?<?0.05) reduced oviposition and adult emergence of C. maculatus when compared with the controls although the reduction was higher at 5% concentration than others. Adult beetle emergence was completely prevented at higher concentrations (4 and 5%) except in wood ash. Both oviposition and adult emergence increased during the months of storage probably because of the slight reduction in the effectiveness of the extracts. The root bark extract was much more effective in reducing oviposition and adult emergence than others throughout the period of storage. The plant extracts of N. laevis was able to protect the cowpea seeds from damage and prevent weight loss. Cowpea seed damage and weight loss was significantly more (p?<?0.05) in the controls than other for the three-month duration probably because of the more adult emergence. The extracts from N. laevis did not adversely affect the germination of the protected seeds and seed germination ranged from 86.7 to 100%. It has been shown in this study that the extracts of N. laevis were effective against C. maculatus in cowpea although the root bark extract seemed to be the most effective while the wood ash extract was least effective. Their effectiveness, however, slightly decreased during the period of storage. N. laevis could be incorporated into pest management of stored cowpeas since the products are ecologically safe.