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BioMed Research International / 2014 / Article
AbstractIntroductionMaterials and MethodsResultsDiscussionConclusionReferencesCopyright
Volume 2014 |Article ID 783059 | 13 pages | https://doi.org/10.1155/2014/783059
Biological Activity of Blackcurrant Extracts (Ribes nigrum L.) in Relation to Erythrocyte Membranes
Dorota Bonarska-Kujawa ,1 Sylwia Cyboran,1 Romuald Żyłka,1 Jan Oszmiański,2 and Halina Kleszczyńska1
1Department of Physics and Biophysics, Wrocław University of Environmental and Life Sciences, Norwida 25, 50-375 Wrocław, Poland
2Department of Fruits, Vegetable and Cereals Technology, Wrocław University of Environmental and Life Sciences, Chełmońskiego 37/41, 51-630 Wrocław, Poland
Academic Editor: Pranee Winichagoon
Received29 Aug 2013
Accepted28 Nov 2013
Published16 Jan 2014
Compounds contained in fruits and leaves of blackcurrant (Ribes nigrum L.) are known as agents acting preventively and therapeutically on the organism. The HPLC analysis showed they are rich in polyphenol anthocyanins in fruits and flavonoids in leaves, that have antioxidant activity and are beneficial for health. The aim of the research was to determine the effect of blackcurrant fruit and leaf extracts on the physical properties of the erythrocyte membranes and assess their antioxidant properties. The effect of the extracts on osmotic resistance, shape of erythrocytes and hemolytic and antioxidant activity of the extracts were examined with spectrophotometric methods. The FTIR investigation showed that extracts modify the erythrocyte membrane and protect it against free radicals induced by UV radiation. The results show that the extracts do not induce hemolysis and even protect erythrocytes against the harmful action of UVC radiation, while slightly strengthening the membrane and inducing echinocytes. The compounds contained in the extracts do not penetrate into the hydrophobic region, but bind to the membrane surface inducing small changes in the packing arrangement of the polar head groups of membrane lipids. The extracts have a high antioxidant activity. Their presence on the surface of the erythrocyte membrane entails protection against free radicals.
Blackcurrant (Ribes nigrum L.) is a shrub commonly grown in various parts of the world of temperate climate. Its tasteful fruits are a rich source of vitamin C and other health beneficial substances such as: routine, organic acids, pectins, micro- and macronutrients and essential oils [1].
Blackcurrant fruits contain polyphenolic substances with antioxidant, antimicrobial, antiviral, and antibacterial properties [2–6]. Owing to these properties, polyphenols protect and support many functions of organs and systems and in particular the digestive [3, 7], nervous [8], and circulatory [9, 10] systems. In cells cultured in vitro, polyphenols exhibit anticancer activity, inhibiting the multiplication and growth of cancer cells by inducing apoptosis in them [11, 12]. Anthocyanins, in particular derivatives of cyanidin and delphinidin, which are the main polyphenols in fruit extract, are used in the treatment of eye defects and diseases of the eye [13, 14].
Contained in the leaves of blackcurrant, quercetin derivatives, as indicated in many studies, have a range of activities, including antimicrobial, anti-inflammatory, antiviral, antitoxic, antiseptic, and antioxidant effects, and are supposed to support the treatment of cancers [15–19]. Very good antioxidant properties of quercetin-3-O-glucoside in relation to biological membranes were showed in previous studies by the authors [20, 21].
For oxidation and destruction of biological systems are responsible high concentrations of free radicals that are formed either during metabolic processes or as a result of exposure to UV radiation. A conspicuous and important place of attack of free radicals is the cell membrane. Oxidation of its components and, in particular, the membrane lipids by free radicals causes disorder in the structure and function of the cell membrane, which leads to pathological changes in the organism. Development of many dangerous diseases linked directly with peroxidation of membrane lipids can be prevented by providing the organism with natural antioxidants, which are polyphenolic substances contained in different parts of the plant. Their protective effects, as scavengers of free radicals, depend on both the number of hydroxyl groups in the polyphenolic molecule as well as the number of molecules associated with the membrane. They protect the red blood cell membrane against oxidation and hemolysis induced by free radicals [22–26]. As indicated in our previous studies, the effectiveness of certain flavonoids, anthocyanins in particular, is much greater than the activity of vitamin E and its synthetic counterpart Trolox [20, 27].
High antioxidant activity of plant extracts and their ingredients was shown in numerous studies conducted around the world [4, 16, 17, 24]. The authors of many works consider the effective protection of biological membranes against oxidation dependent on the capabilities of the polyphenolic substances binding to membranes [22–26]. Due to the amphiphilic nature of polyphenols, it can be expected that they incorporate, mainly due to their structural similarity, at different depths into the lipid phase of biological membranes, changing their properties to varying degrees. However, there is little work on the impact of plant extracts and polyphenolic compounds on the properties of biological systems, and in particular of biological membranes. An important issue, therefore, seems to be the relationship between the high antioxidant activity of plant polyphenols in relation to biological membranes and the extent of physical changes that these substances induce in membranes. Such studies have been carried out to a minor extent for selected plant extracts in relation to membrane of erythrocytes and lipid membrane models [23, 24, 26, 28–31].
The present study aimed to determine the antioxidative activity of extracts from the fruit (BCF) and leaves (BCL) of blackcurrant, with respect to the biological membrane, and in particular in relation to membrane lipids. Given the rich polyphenolic composition of blackcurrant extracts, one could expect an effective protection of the membranes against oxidation-inducing agents. The research was conducted with respect to the membrane of erythrocytes, which was treated as an example and model of the biological membrane. Antioxidative activity of the extracts was determined on the basis of fluorimetric studies, with oxidation of erythrocyte membranes induced by UVC radiation and the AAPH compound. Also, the influence of extracts from fruits and leaves of blackcurrant on the properties of the erythrocyte membrane was examined, taking into account their possible negative impact. In particular, the hemolytic activity of the extracts and their impact on osmotic resistance of erythrocytes were assayed spectrophotometrically. Erythrocyte shapes were also examined in the presence of extracts, using the optical and electron microscopes. Using the Fourier transform infrared spectroscopy (FTIR) method, it was specified the location in the membrane of red blood cells of the phenolic substances contained in the extracts and their impact on its degree of hydration. Also, the protective effect of the extracts towards changes in the membrane of erythrocytes due to its exposure to UVC radiation was determined.
2.1. Materials
Plant extracts of fruits and leaves of blackcurrant (Ribes nigrum L.) were obtained from the Department of Fruit, Vegetable, and Cereal Technology, Wroclaw University of Environmental and Life Sciences. The percent content of polyphenols in the extracts was determined with liquid chromatography (HPLC).
The studies were conducted on pig erythrocytes and isolated erythrocyte membranes (ghosts), which were obtained from fresh blood using the Dodge method [32]. The content of erythrocyte membranes in the samples was determined on the basis of protein concentration, which was assayed using the Bradford method [33]. The choice of pig erythrocytes was dictated by the fact that this cell's percentage share of lipids is closest to that of the human erythrocyte, and the blood was easily available.
The fluorescent probe 3-(p-(6-phenyl)-1.3.5-hexatriene) propionic acid (DPH-PA) was purchased from Molecular Probes, Eugene, Oregon, USA. The oxidation inductor 2,2′-diazobis (2-amidinopropane) dihydrochloride (AAPH) and Trolox was purchased from Sigma-Aldrich, Steinheim, Germany.
Erythrocyte membranes were irradiated with UVC radiation from bactericidal lamp (3,5 mW/cm2).
2.2. Methods
2.2.1. HPLC-DAD and UPLC-ESI/MS
Polyphenols were isolated from leaves and fruits by extraction with water containing 200 ppm of SO2, the ratio of solvent to leaves or fruits being 3 : 1. The extract was absorbed on Purolite AP 400 (UK) for further purification. The polyphenols were then eluted out with 80% ethanol, concentrated, and freeze-dried. The percent content of polyphenols in individual preparations was determined by means of liquid chromatography HPLC and UPLC method. Phenolic compounds were identified with the HPLC/DAD method and the method of UPLC/ESI/MS analysis described in papers [34].
2.2.2. Antioxidant Activity of Extract
The DPH-PA probe was used in the fluorimetric experiments. Erythrocyte membranes with and without (control) addition of extracts were suspended in a phosphate buffer of pH 7.4 and UVC irradiated or treated with the chemical oxidation compound AAPH for 30 min. Free radicals, released in the process of membrane lipids irradiation, cause quenching of DPH-PA fluorescence, decreasing the fluorescence intensity. As a measure of the extent of lipid oxidation was assumed relative fluorescence, that is, the ratio of an UVC-irradiated or AAPH oxidized probe fluorescence to the initial fluorescence of the probe. Here, as a control was assumed the relative fluorescence of an erythrocyte membranes suspension that contained the DPH-PA probe, oxidized with UVC or AAPH compound, while the blank was the relative fluorescence of a suspension of the same concentration but not oxidized.
A spectrofluorimeter (Carry Eclipsce, Varian) was used to measure free radicals concentration in the samples. Excitation and emission wavelengths were nm and nm. The measure of lipid oxidation was the relative change of fluorescence intensity F/F0, where F0 is the initial fluorescence and F the one measured during an oxidation procedure described in a paper [35]. The percentage of lipid oxidation inhibition was calculated from the following formula: where is relative fluorescence of an UVC irradiated sample, or oxidized by AAPH, for 30 min in the presence of the compounds, is relative fluorescence of the control sample, oxidized by AAPH or UVC irradiated, measured after 30 min, and is relative fluorescence of the blank sample, not subjected to oxidation procedure, measured after 30 min.
The results of the assay were expressed in reference to Trolox, standard antioxidant.
2.2.3. Hemolytic Activity of Extracts and Osmotic Resistance of Erythrocytes
The hemolytic experiments were conducted on fresh, heparinized blood. For washing the erythrocytes, and in the experiments, an isotonic phosphate solution of pH 7.4 (131 mM NaCl, 1.79 mM KCl, 0.86 mM MgCl2, 11.79 mM Na2HPO4 × 2H2O, and 1.80 mM Na2H2PO4 × H2O) was used. Full blood was centrifuged for 3 min, 1251 g at 4°C to remove the plasma and leucocytes. Upon removing from plasma, the erythrocytes were washed four times in phosphate solution and then incubated in the same solution but containing proper amounts of the compounds studied. The modification was conducted at 37°C for 1 h, each sample containing 10 mL of erythrocyte suspension of 2% hematocrit was stirred continuously. After modification 1 mL samples were taken, centrifuged and the supernatant assayed for hemoglobin content using a UV-Vis spectrophotometer (Cary 300 Bio, Varian) at 540 nm wavelength. Hemoglobin concentration in the supernatant, expressed as percentage of hemoglobin concentration of totally hemolyzed cells, was assumed as the measure of the extent of hemolysis.
For osmotic resistance, upon removing from plasma the erythrocytes were washed three times with a cool (ca. 4°C), 310 mosm PBS isotonic solution. Next, a 2% red cells suspension containing plant extracts of 0.01 mg/mL concentration was prepared and left for 1 h at 37°C with continuous stirring. After this modification the suspension of erythrocytes was centrifuged for 15 min at room temperature in order to remove the cells from the extract solution. From the cell sediment were taken 100 μL samples of the extract-modified cells and suspended in test tubes containing NaCl solutions of 0.5–0.86% concentration and to an isotonic (0.9%) NaCl solution. In solutions of the same concentrations were also suspended unmodified red blood cells that constituted the control for osmotic resistance determinations. Then, the suspension was stirred and centrifuged under the above stated conditions. After that the percentage of hemolysis was measured with a spectrophotometer at nm. On the basis of the results obtained, the relation was determined between the percentage of hemolysis and NaCl concentration in the solution. Next, using the obtained plots, the NaCl percent concentrations (C50) that caused 50% hemolysis were found. The C50 values were taken as a measure of osmotic resistance. If a determined sodium chloride concentration is higher than that of control cells, the osmotic resistance of the erythrocytes is regarded to be lower, and vice versa.
2.2.4. Erythrocyte Shapes
For investigation with the optical microscope, the red cells separated from plasma were washed four times in saline solution and suspended in the same solution but containing 0.01 and 0.1 mg/mL of BCL and BCF extracts, respectively. Hematocrit of the erythrocytes in the modification solution was 2%, the modification lasting 1 h at 37°C. After modification the erythrocytes were fixed with a 0.2% solution of glutaraldehyde. After that the red cells were observed under a biological optical microscope (Nikon Eclipse E200) equipped with a digital camera. The photographs obtained made it possible to count erythrocytes of various shapes and then percent share of the two basic forms (echinocytes and stomatocytes) in a population of ca. 800 cells was determined. The individual forms of erythrocyte cells were assigned morphological indices according to the Bessis scale [36], which for stomatocytes assume negative values from −1 to −4 and for echinocytes-positive from 1 to 4.
For investigation with the electron microscope, the red cells separated from plasma were washed four times in saline solution and suspended in the same solution but containing 0.1 mg/mL of the blackcurrant leaf (BCL) and fruit (BCF) extracts. Hematocrit of the erythrocytes in the modification solution was 2%, the modification lasting 1 h at 37°C. After modification the erythrocytes were fixed for 48 h in 2.5% solution of glutaraldehyde. After that the preparations were washed in phosphate buffer for 20 min, and then the material was dehydrated in a rising series of acetone concentrations (30, 50, 60, 70, 80, 90 and 100%). Each sample was washed for 15 min in an appropriate concentration and the material remain in pure acetone for 30 min. Next, the erythrocytes were dried for 12 h at room temperature. Erythrocytes thus prepared were deposited on object stages and subjected to X-ray microanalysis by means of an X-ray analyzer, Brucker AXS Quantax, collaborating with the ESPRIT ver. 1.8.2. program. Next, the samples were coated with gold using the Scancoat 6 (Edwards, London) sprinkler. The material ultrastructure was analyzed using a scanning microscope (EVO LS15 ZEISS) with SE1 detector, under high vacuum and accelerating voltage EHT = 20 kV.
2.2.5. FTIR Investigation of Erythrocyte Membrane
Erythrocyte ghosts were washed three times in 0.9% NaCl solution. Next, the ghost suspension was incubated as 1 mL samples (150 μL ghosts + 850 μL physiological salt) for 24 h at 37°C. In that way the control sample with ghosts suspension was prepared, as well as samples with blackcurrant fruit and leaf extracts, obtaining 0.05 mg/mL concentration. After incubation, the samples were centrifuged for 15 min at 16000 g and IR spectra were recorded from a condensed membrane suspension. Tests were also conducted with samples containing erythrocyte membranes irradiated with UV, prepared in a similar way. After 24 h incubation with extracts, the ghost suspension was exposed to UV lamp for 1 h and then centrifuged and spectra taken in the IR range. The measurements were performed with a Thermo Nicolet 6700 MCT spectrometer, with ZnSe crystal, applied at room temperature. Each single spectrum was obtained from 128 records at 2 cm−1 resolution in the range 700–4000 cm−1. Preliminary elaboration of a spectrum was done using the EZ OMNIC v 8.0 program, also of the Thermo Nicolet firm. After filtering the noise out from the spectrum of the object studied, the spectrum of the NaCl solution was subtracted in order to remove the OH band of water and the base line was corrected.
3.1. HPLC-DAD and UPLC-ESI/MS
Detailed quantitative and qualitative contents of phenolic compounds in the extracts from leaves and fruits of blackcurrant are given in Table 1.
Compounds BCF BCL* BCF/BCL BCF/BCL BCF/BCL
Content (%) (min) λ max (nm) (MS) (m−)
Chlorogenic acid 0.12 1.15 10.71 320 353
p-Coumaroylquinic acid 0.11 0 7.18 311 341
Neochlorogenic acid 0 0.1 7.18 320 353
Cryptochlorogenic acid 0 0.11 10.87 320 353
Delphinidin-3-O-glucoside 4.58 0 15.29 526 465
Delphinidin-3-O-rutinoside 22.49 0 16.21 525 611
Cyanidin-3-O-glucoside 6.00 0 17.03 516 449
Cyanidin-3-O-rutinoside 21.18 0 18.03 516 595
Petunidin-3-O-rutinoside 0.24 0 19.17 527 625
Quercetin-3-O-rutinoside 0.11 0.33 23.17/19.04 355 611/609
Quercetin-3-O-galactoside 0.17 2.52 23.77/20.71 355 465/463
Quercetin-3-O-glucoside 0.15 0 24.46 355 465
Quercetin-3-(6′′-malonyl)-glucoside 0 1.91 21.29 355 549
Quercetin-3-O-glucosyl-6′′-acetate 0 5.53 23.17 354 505
Myricetin-3-O-rutinoside 0.04 0 19.04 355 627
Myricetin-3-O-galactoside 0.88 0 20.71 358 481
Myricetin-3-O-glucoside 0.08 0 21.29 356 481
Kaempferol-3-O-rutinoside 0 0.23 23.77 342 593
Kaempferol-3-O-galactoside 0 0.41 24.46 342 447
Kaempferol-3-O-glucosyl-6′′-acetate 0.96 26.96 346 489
Total 56.14 13.23
Published previously in Oszmiański et al., 2011 [34].
The percent content and characterization of phenolic compounds of the extract of blackcurrant fruits (BCF) and leaves (BCL) using their spectral characteristic in HPLC-DAD (retention time, λ max) and positive and negative ions in UPLC-ESI-MS [M–H]−.
The phenolic contents of blackcurrant leaves indicate that the dominant components in the leaves are quercetin derivatives, which constitute 80% of polyphenolic compounds in dry mass of extract. Very interesting though is the polyphenol composition of blackcurrant fruits, constituting over a half (56.14%) of total dry mass of extract.
The blackcurrant fruit extract was analyzed by UPLC-ESI-MS and HPLC-DAD systems and is summarized in Table 1 and Figures 1 and 2. A total of 13 kinds of polyphenolic compounds found in blackcurrant fruit extract were identified and quantified. Two hydroxycinnamates were detected: chlorogenic and p-coumaroylquinic acids. The compounds that had a [M − H]− at m/z 353 and 341 with 320 nm and 311 nm after a fragmentation yielded a caffeic and p-coumaric acid, respectively.
Direct injection ESI-MS of polyphenols in the blackcurrant fruit extract. Positively charged molecular ion are 449.24 cyanidin-3-O-glucoside, 465.28 delphinidin-3-O-glucoside, quercetin-3-O-glucoside and quercetin-3-O-galactoside, 595.30 cyanidin-3-O-rutinoside, 611.28 delphinidin-3-O-rutinoside and quercetin-3-O-rutinoside, and 625 petunidin-3-O-rutinoside.
HPLC profile (520 nm) of blackcurrant extracts tr. 15.29 min delphinidin-3-O-glucoside, 16.21 min delphinidin-3-O-rutinoside, 17.03 min cyanidin-3-O-glucoside, 18.03 min cyanidin-3-O-rutinoside, and 19.17 min petunidin-3-O-rutinoside.
Anthocyanins, delphinidin-3-O-glucoside (peak tr. 15.29 min), delphinidin-3-O-rutinoside (peak tr. 16.21 min), cyanidin-3-O-glucoside (peak tr. 17.03 min), cyanidin-3-O-rutinoside (peak tr. 18.03 min), petunidin-3-O-rutinoside (peak tr. 19.17 min) (Figure 2), were identified on the basis of reference compounds, UV-Vis spectra, mass spectra, and the literature. Delphinidin and cyanidin were the two major anthocyanidins, commonly known in blackcurrant fruit [37]. The minor peak was identified as petunidin aglycone.
Furthermore, a total of 6 flavonol glycosides were detected (Table 1). Three quercetin derivatives were detected: quercetin-3-O-rutinoside, quercetin-3-O-galactoside and quercetin-3-O-glucoside. Peaks with 23.17 min, 23.77 min and 24.46 min had of 355 nm, respectively. All compounds had a fragmentation yielded a quercetin ion at m/z 303. Galactosides and glucosides have the same molar masses and mass spectra, and they differ according to their retention times only. Analogously, myricetin galactoside 20.71 min was assumed to eluate before myricetin glucoside 21.29 min. Myricetin-3-O-rutinoside 19.04 min was also identified in our samples, as defined in earlier investigations [38–40]. The anthocyanidin derivatives were the most predominant phenolic group found in blackcurrant fruit extract and constituted 54.49% weight of powder. Among identified anthocyanidin derivatives the most important compounds were delphinidin and cyanidin rutinoside (43.67%) The other compounds such as hydroxycinnamic acids and flavonol derivatives accounted total for 1.66% of blackcurrant fruit extract.
3.2. Antioxidant Activity of Extract
The investigation of antioxidative activity of BCF and BCL extracts were conducted on erythrocyte membranes, based on the extent of membrane lipids oxidation. The oxidation was induced with UVC and the AAPH compound. The extent of lipid oxidation was assayed fluorimetrically, based on the kinetics of DPH-PA probe quenching caused by free radicals that arose during oxidation.
The relative fluorescence for the studied concentrations of BCF and BCL extracts decreases with time of oxidation with UVC radiation, which shows that the degree of the lipid oxidation inhibition increases. Quenching of DPH-PA fluorescence in the presence of BCF and BCL extracts at different concentrations was also observed when lipid oxidation was induced in erythrocyte membranes with the chemical compound AAPH at 60 μM.
Based on the kinetics of the oxidation curves obtained for various concentrations of both the extracts and compared with Trolox, the concentration responsible for 50% inhibition of membrane lipids oxidation (IC50) was found. The results from the fluorimetric method are given in Table 2 and Figure 3.
Extract/inducer Concentration IC50 (µg/mL) ± SD
UVC AAPH
Blackcurrant leaves (BCL)
Blackcurrant fruits (BCF)
Values of IC50 for BCL and BCF extracts, and Trolox that inhibit erythrocyte membrane lipids oxidation by 50%, determined with the fluorimetric method. The oxidation was induced with UVC radiation and the AAPH compound.
Relation between relative fluorescence intensity and time of UVC induced oxidation of erythrocyte membranes for control and test sample at different concentrations of blackcurrant extracts of leaves (a) and fruits (b).
The results indicate that the extracts protect erythrocyte membrane lipids against free radicals induced with UVC and the AAPH compound. Extract concentration responsible for 50% protection of the lipids is comparable with that of Trolox in the case of AAPH and smaller when oxidation was UVC induced.
As indicated by the IC50 values, BCL and BCF extracts protect membrane lipids against oxidation. The results obtained have shown that BCL extract protect the erythrocyte membrane against UVC-induced oxidation better than BCF extract but worse than Trolox. For protecting membranes against AAPH-induced oxidation, the BCL and BCF extracts were equal in antioxidant activity to Trolox. Such results suggest that the reaction with free radicals depends on the oxidation inducer. In the case of photo-oxidation the percentage of inhibition and the kinetics of the process are slower, whereas in the case of the AAPH compound the process is much faster and efficient in inhibiting oxidation. The results indicate that polyphenols, mainly of the flavonoid group, contained in blackcurrant extract have good antioxidative properties, being more effective towards free radicals that arise in the process of AAPH-induced oxidation of erythrocyte membranes than UVC-induced one.
3.3. Hemolysis and Osmotic Resistance of Erythrocytes
The results on the effect of blackcurrant extracts on hemolysis of erythrocytes compiled in Table 3 testify that in the presence of BCF and BCL extracts red blood cells do not undergo increased hemolysis compared with unmodified cells, hemolysis with extract at 0.1 mg/mL not exceeding 5% in respect to the control probe. The control probe contained suspension of unmodified erythrocyte cells in phosphate buffer (pH 7.4).
Extracts Percentage of hemolysis ± SD
Concentration (mg/mL) BCF BCL
Percent content of hemolyzed erythrocytes in the presence of BCL and BCF extracts of specific concentrations.
In the studies of the effect of the extracts on erythrocyte osmotic resistance no significant differences between the degree of hemolysis in the control cells and those modified with BCF extract was found, in aqueous solutions of various concentrations of sodium chloride. Concentrations (C50) of sodium chloride, expressed as a percentage, determined for control blood cells and those modified with BCL and BCF extracts of 0.01 mg/mL concentration are C50 control—0.725%, BCL—0.644%, BCF—0.714%. These results indicate an increase in osmotic resistance of blackcurrant leaf extract modified cells, suggesting that the red blood cell membrane treated with BCL extract is less sensitive to changes in osmotic pressure (Figure 4).
Percent of hemolysis of cells modified with blackcurrant leaf (BCL) and fruit (BCF) extract of 0.01 mg/mL concentration versus sodium chloride concentration.
3.4. Erythrocyte Shapes
Figure 5 shows the percent share of the various forms of cells in a population of erythrocytes modified with BCL and BCF extracts of 0.1 and 0.01 mg/mL concentration. As seen in Figures 5(a), 5(b), and 6, these extracts induce mostly various forms of echinocytes, mainly discoechinocytes. It can thus be assumed that the blackcurrant extracts concentrate mainly in the outer monolayer of the erythrocyte membrane when inducing echinocytes, and practically do not permeate into the inner monolayer of the membrane [41–44].
Percentage share of different shapes of erythrocytes induced by BCL and BCF extracts at 0.1 (black bar) and 0.01 (white bar) mg/mL concentration, control (grey bar). On the abscissa, there are morphological indices for the respective shapes of cells: spherostomatocytes (−4), stomatocytes II (−3), stomatocytes I (−2), discostomatocytes (−1), discocytes (0), discoechinocytes (1), echinocytes (2), spheroechinocytes (3), and spherocytes (4).
Shapes of unmodified erythrocytes (a) and modified with BCF (b) and BCL (c) at 0.1 mg/mL concentration, observed with electron microscope.
The work [43, 44] showed that the formation of echinocytes occurred when amphiphilic molecules were incorporated in the outer monolayer of the erythrocyte membrane. Compounds penetrating to the inner monolayer of the membrane induced formation of stomatocytes. We can therefore assume that BCF and BCL extract components concentrated mainly in the outer monolayer of the erythrocyte membrane.
3.5. FTIR Investigation of Erythrocyte Membrane
Spectroscopic studies in the near infrared allowed to take absorption spectra for unmodified and BCL and BCF modified erythrocytes. Spectra of unmodified and extract modified membranes irradiated with UVC were also taken. The results obtained allowed us to observe changes induced in the erythrocyte membrane by UVC radiation and phenolic components of the extracts. In the spectra, the characteristic frequency bands of the protein and lipid components were analysed. We identified the characteristic for erythrocytes frequency bands for individual membrane components: methyl groups and methylene hydrocarbon chains (2980–2830 cm−1), carbonyl groups of lipids (1770–1710 cm−1), the amide band I (1690–1590 cm−1) and amide band II (1580–1500 cm−1), and phosphate band (1280–1200 cm−1) and choline band (1000–940 cm−1) [45–48]. Analysis of the spectra of membranes treated with aqueous extracts of blackcurrant leaves and fruit showed no significant changes in the amide I and amide II bands (vibration bands of groups C=O and N–H proteins). There were also no changes in the vibrational bands of methyl and methylene groups, alkyl chains of lipids, and C=O vibrations of carbonyl groups of lipids. This result suggests that the polyphenolic compounds contained in the extracts do not significantly affect the structure of the proteins contained in the membrane of red blood cells, and they do not modify the hydrocarbon chains of lipid molecules. UVC radiation exposure of control unmodified membranes and membranes modifies with the extracts does not cause visible changes in the above mentioned bands. The presence of the extracts in erythrocytes membranes causes a slight shift of the phosphate band in the direction of smaller wave numbers, which points to a slight increase in the degree of hydration of PO2 group. A slightly larger displacement of this band is caused by the extract from BCL. Exposure of the membranes to UVC radiation results in a significant shift of the phosphate band towards higher values of wave numbers, suggesting a clear reduction in the number of OH groups associated with lipid phosphate groups via hydrogen bonds. The presence of the extracts in a suspension reduced the transfer effect of phosphate bands distinctly. Phosphate bands of irradiated membranes in the presence of extracts are shifted toward lower wave numbers relative to the irradiated ghosts, which is particularly visible with BCL extract. So we can assume that the presence of extracts in membranes exposed to UVC protects them against changes induced by the radiation, causing the membrane properties becoming closer to those of membrane not exposed to radiation. A particularly high level of membrane protection was observed in the presence of blackcurrant leaf extract (Figure 7).
Phosphate band of erythrocyte membrane modified with BCF and BCL extract (a), and of erythrocyte membrane modified with BCF and BCL extract, irradiated with UVC (b). Solid line, control membrane; dashed line, membrane with fruit extract; dotted line, membrane with leaf extract.
The presence of extract and exposure to UVC light caused visible changes also in the choline N+ (CH3) band of lipids. In the case of ghosts not exposed to UVC the presence of extracts caused distinct decrease in the half-width of the choline band, which suggests a change in the packing order in that part of membrane and can be linked with presence of sugar groups in the extract components. UV radiation caused shift of the band towards lower wave numbers. For ghosts irradiated in the presence of fruit extract the effect of light was markedly smaller, while in the presence of leaf extract no changes were observed compared with nonirradiated ghosts (Figure 8).
Choline band of erythrocyte membrane modified with BCF and BCL extract (a), and of erythrocyte membrane modified with BCF and BCL extract, irradiated with UVC (b). Solid line, control membrane; dashed line, membrane with fruit extract; dotted line, membrane with leaf extract.
Statistical analysis was carried out using Statistica 9.0 (StatSoft Inc.). All the experiments were performed at least in triplicate unless otherwise specified. Analysis of variance was carried out and significance between means was determined using Dunnett's post hoc test. Results are presented as mean ± SD. Significant levels were defined at .
The results of the presented research have shown that blackcurrant leaves (BCL) and blackcurrant fruits (BCF) extracts induce small changes in erythrocyte membranes. Their polyphenolic composition makes possible an interaction with the lipids of biological membranes. The main polyphenolic constituent of BCF extract are anthocyanins, which constitute over 98% of the total polyphenol content of the extract. It was shown that the beneficial properties of anthocyanins, cyanidins and their glycosides, are connected with their ability to scavenge free radicals, including the reactive forms of oxygen [49–51]. Cyanidins were found to bind with the biological membrane, protecting it against oxidation [21, 52–54]. It can thus be assumed that mainly anthocyanins are responsible for the protective action of blackcurrant fruit extracts with respect to the biological membrane.
BCL extract differs in polyphenolic composition from BCF extract, where quercetin derivatives dominate, which constitute ca. 80% of the total polyphenols content of the extract. An antioxidation activity of quercetin glycosides in relation to the erythrocyte membrane was shown in our previous studies [20]. However, it is not yet fully elucidated the molecular mechanism of the antioxidant action of these compounds in relation to the biological membrane.
The present study aimed to determine the antioxidative activity of blackcurrant extracts with respect to the membrane of red blood cells, treated as a model of the biological membrane. The impact of the extracts on erythrocyte membrane properties was also examined. Selected research methods helped to determine the location of the compounds contained in the extracts, based on erythrocyte shape changes and hemolytic and FTIR studies. The results of the investigations undertaken enabled us to determine the relationship between the antioxidative activity of the extracts and their affinity to the membrane.
Hemolytic tests showed that polyphenolic compounds contained in the extracts do not induce hemolysis, therefore do not cause lytic effect on red blood cells in the used range of concentrations. The lack of hemolysis and cytotoxicity for chosen flavonoids was also confirmed in the work [55]. One can therefore assume, on the basis of the studies [56–60], that the compounds do not embed deep into the hydrophobic membrane area. Hemolysis caused by various lytic compounds occurs when such substances penetrate deeply into the membrane, weakening the interaction between its components. Disturbed is the structure of the membrane and facilitated transport of water swells the cell, and the membrane is permanently damaged. Studies of osmotic resistance confirmed the hemolytic results, they showed that as a result of incorporation of the extract contained compounds the osmotic resistance does not change or is even greater in the case of BCL extract.
The extract from the fruit does not alter the osmotic resistance, that is, does not change the properties of the erythrocyte membrane, whereas the leaf extract causes an increase in the resistance, which means that substances present in the extract bind to the membrane and make it stronger, so that it becomes less sensitive to changes in osmotic pressure.
Both the extracts induced an alternation in the erythrocyte morphology, from normal discoid shape to an echinocytic form. BCF and BCL extracts are responsible for creation of varied forms of echinocytes, BCL mainly echinocytes and BCF echinocytes and spheroechinocytes. It can thus be assumed, according to the bilayer couple hypothesis [42], that the extracts concentrate mainly in the outer monolayer of the erythrocyte membrane when inducing various forms of echinocytes, and practically do not permeate into the inner monolayer of the membrane.
The changes observed in the IR spectrum suggest a superficial character of the interaction of BCF and BCL components with the hydrophilic part of the erythrocyte membrane. FTIR spectra for extract modified membranes showed no significant changes in the hydrophobic part of the hydrocarbon chains, but showed a change in the degree of hydration in the phosphate and choline band of membrane phospholipids. As the authors suggest [48, 61], as a result of the effect of polyphenols on lipid membrane, hydrogen bonds are formed between the hydroxyl groups of polyphenols and the polar groups of lipids. In addition, numerous studies showed that plant extracts, particularly those rich in polyphenol compounds with sugar residues, reduce the packing order of the hydrophilic area of membrane, causing increased hydration of this area.
As indicated by the results, after one hour of exposure to UVC radiation, changes occurred in the erythrocyte membrane, in particular in its hydrophilic region. This is shown by a clear shift of the phosphate band of polar heads of lipids, which may be related to oxidation of OH groups which are connected by hydrogen bonds with the phosphate groups or lipids. The presence of the extracts in ghosts suspension markedly reduced the effect of phosphate band transfer that was caused by exposure to UV radiation. In particular, such an effect was observed in the case of BCL extract, which caused a complete abolition of UVC induced changes in control ghosts, and even a visible widening of the band. This may suggest that during exposure to UVC radiation in the presence of the extracts (BCL and BCF) the membrane phosphate groups were not oxidized but had undergone hydration (BCL), which also indicates the presence of extract components in this part of the membrane. In addition, it is believed that the polyphenolic compounds, especially those occurring in the form of glycosides, cause hydration of the membrane, transporting numerous water molecules bound to them [31, 40, 48–51, 53, 54, 56–61]. Such protective action of the extracts can be attributed to their phenolic components that bind to the membrane surface, and containing numerous OH groups in their structure they, possibly, as the first undergo oxidation.
The location of the compounds in the hydrophilic part of membrane seems to constitute a protective shield of the cell against other substances, the reactive forms of oxygen in particular, which finds its reflection in their antioxidant properties. BCL and BCF extracts exhibited a high antioxidant activity towards reactive forms of oxygen, which developed as a result of membrane photo-oxidation induced by UVC radiation and by the AAPH compound.
The results obtained indicate that BCL and BCF extracts exert beneficial effect on the organism protecting the cell membrane against oxidation. The polyphenolic compounds contained in the extracts do not disturb the structure of the biological membrane, to which they bind only superficially, penetrating its hydrophilic region, and do not penetrate the membrane hydrophobic region, as is evident from the hemolytic, microscopic, and FTIR investigations. It can thus be expected that they do not disturb the function of the biological membrane.
In this context, a protection of the organisms against the harmful effects of free radicals and reactive oxygen species is made possible by providing the body with appropriate quantities of plant polyphenolic substances. Thus, the results of this research encourage the consumption of fruits and extracts from blackcurrant leaves to fight free radicals and thus increase our resistance to many diseases.
The authors confirm that the paper has been read and approved by all of the authors. They also confirm that there are no other persons who participated in the preparation of this paper other than the authors listed. In addition, they confirm that the order with which the authors are listed is agreed and approved by all authors. The authors confirm that there is no conflict of interests associated with this paper. Furthermore, this work has not been financially supported by other institutions, which could have influenced its outcome. The authors confirm that they have followed the regulations of Wroclaw University of Environmental and Life Sciences concerning intellectual property. They declare that there are no impediments to the publication of this work, because they have taken into account the protection of the intellectual property associated with this work. Furthermore, they declare that this paper is not under consideration for publication elsewhere.
This work was sponsored by the Ministry of Science and Education, scientific Project nos. N N312 422340 and N N304 173840.
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Bias alert: I'm a sucker for world instruments. They can add just the right amount of spice to otherwise ho-hum dance tracks. Elsewhere, hip-hop is folding in more ethnic elements (particularly Middle Eastern flavors). Bhangra is even working its way into the mainstream.
Ethno Instrument (EI) is a cross-platform plug-in/standalone instrument with a 4GB library of keyboard-mapped sounds and 4GB of loops (for the specs, visit www.motu.com). It's quality all the way: The samples are well-recorded, looped, and mapped.
EI searches for installed components during installation (but not afterward), so if you have big hard drives, the program may seem frozen. It isn't — be patient. As for copy protection, the program uses iLok.
EI is a one-stop instrument, with 64 parts and 17 stereo outs. If you need to instantiate multiple instances (which you can do), you probably need a course on arranging. As such, it's a "program within a program" when hosted in a DAW, and is also quite complete as a standalone device. Parts can stream from disk or load into RAM to strike the best balance of disk and RAM usage.
The selection of instruments covers a wide range of styles and geographical locations, and the browser makes it easy to select loops or instruments. Most of the instruments have limited ranges that match the instruments being sampled; I would have preferred to have the lower and upper limits stretched — I don't always want authenticity. In any event, transposing the samples is a workaround. Better yet, it would have been nice to fill up those empty keys with idiomatic riffs and articulations.
For those who do enjoy warping things, there's a synth engine with filter, amp, LFO, EQ, etc. Nice. And you can do MIDI automation, as well as "MIDI learn" for external control. Furthermore, you can apply velocity and keyswitching to layered instruments.
But the big deal to me is the way EI handles loops. The concept of slicing a loop into pieces and assigning each slice to a keyboard key is not new, but it's particularly applicable here as you can create expressive variations on loops. You can also drop and drag loops (as MIDI data or audio that tempo syncs — very cool) into your instrument tracks. After dragging MIDI data, you can edit it to create useful variations. Again, this isn't necessarily a new concept; many programs treat REX files this way. But the execution here is superb. Even with extreme stretching, sound quality remains intact. Granted, the loops are stretch-friendly to begin with, but that doesn't minimize the work that must have been put into the slicing process.
EI also includes a convolution reverb, and particularly in standalone mode, strengthens the "one-stop" vibe. It's bus-based, so each part can have different amounts of reverb, and adds value to the package.
Finally, it's worth adding that the 120-page printed documentation is excellent. It not only takes you through all aspects of the program, but almost half the manual is devoted to describing the instruments and listing the various sounds and patch mappings.
Just as cooking benefits from the occasional exotic spices, so does music. At this price, and with this kind of library, Ethno Instrument is a superb (and painless) way to get into world-class world sounds.
Product type: Plug-in/standalone instrument for MAS/VST/RTAS/DXi/AU; MacOS X 10.3.9 or later, Windows XP.
Target market: Studios needing easy-to-use, cost-effective ethnic sounds; stage musicians who use a computer.
Strengths: Handles loops exceptionally well. Lots of variety. Tempo-matching drag-and-drop loop integration with host for both audio and MIDI. Includes convolution reverb. Synth-based processors for editing.
Limitations: More samples of articulations would have been welcome.


Our state-of-the-art production facility is FDA certified and features environmentally controlled packaging suites for compliance with all FDA standards (21 CFR, parts 210 and 211) for the processing, packing or holding of bulk drugs and finished pharmaceuticals.
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Our experienced project team and trained production associates will work with you every step of the way to fully understand your custom packaging needs, marketing objectives, production timeframes and critical delivery deadlines to provide the greatest value and quality assurance. We are committed to providing you exceptional service and a high quality, cost-effective solution to all of your packaging needs.
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text:
/**
 * Module dependencies.
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var mongoose = require('mongoose');
var utils = require('../../lib/utils');

/**
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exports.load = function (req, res, next, id) {
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  utils.findByParam(photo.categorys, { id: id }, function (err, comment) {
    if (err) return next(err);
    req.category = comment;
    next();
  });
};

/**
 * Create category
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exports.create = function (req, res) {
  var photo = req.photo;
  var user = req.user;
  console.log(req.body);
  if (!req.body.category) return res.redirect('/photos/'+ photo.id);

  photo.addCategory(user, req.body, function (err) {
    if (err) return res.render('500');
    res.redirect('/photos/'+ photo.id);
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}

/**
 * Delete comment
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exports.destroy = function (req, res) {
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    if (err) {
      req.flash('error', 'Oops! The comment was not found');
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};

redpajama_set_name: RedPajamaGithub


It was pretty much the same quality as the rest only you get off Oahu just a bit, see a different angle of Chinaman's Hat and are then left on a small long strip of an island that you are free to explore on your own. Incredible views of dragon's tail and a nice, quiet guy there that was weaving headdresses out of palm tree stalks which you get to keep, LOL.
If it hadn't of been for my oldest son, I would have done horseback instead as the movie tour was just too generic for me and for whatever reason, he was a huge LOST fan and I guess figured he might see the actor's or something, LOL.


text:
Carriola saturnoides is een donsvlinder uit de familie van de spinneruilen (Erebidae). De wetenschappelijke naam van de soort is voor het eerst geldig gepubliceerd in 1879 door Snellen.

Donsvlinders

redpajama_set_name: RedPajamaWikipedia


The Engineering Design of Systems compiles a wealth of information from diverse sources, providing a unique, one-stop reference of current methods and models for systems engineering. This updated edition features important new information on Systems Modeling Language (SysML), more descriptive material on usage scenarios based on literature from use case development, updated homework assignments, and use of the software product CORE to generate the SysML figures. This book serves as an excellent introductory reference suitable for students and professionals alike.


… what is surprising is that OL seem to have been engrossed in counting the repetition of GNH instead of the substance of the address itself.
Dorji is right. We should pay attention to, and analyze, the substantive parts of the PM's address. So that's exactly what we'll do over the next few days. Here's the plan: I'll pick up some issues, one at a time, and we'll discuss them.
At the Conference of Parties Summit on Climate Change in Copenhagen (COP 15) last December, Bhutan declared that it will forever remain carbon neutral and serve as a net carbon sink.
Climate change is a reality. And, left unchecked, global warming could inflict dire consequences on Bhutan, a country that has become increasingly dependent on the Himalayan water resources.
So the government's promise sounds good. The "Declaration of the Kingdom of Bhutan – the Land of Gross National Happiness to Save our Planet", as the proclamation is called, sounds like a good idea.
Is it really a good idea? The government seems to think so. But we, the people, don't know. We don't know, because we were never consulted.
In its enthusiasm to make the carbon neutral promise at the Copenhagen Summit, the government seems to have done a hurried job – it signed the declaration on 11th December 2009, and, barely a week later, announced it in Copenhagen – without any consultations with the people. We don't know if experts were consulted, but we do know that our farmers were not consulted. Similarly, the private sector was not consulted. And, even though the Parliament was in session when the declaration was signed, even the parliament was not consulted.
Should the government have consulted the people and the Parliament? Yes. After all, the declaration is a momentous promise, one that will have far reaching consequences, one that has been made on our behalf, and our children's behalf, for all time to come.
And, as it turns out, one that we may find difficult to honour.
What I find ironic, is our government's climate change initiative is just lip service.
They say they want to charge higher taxes to largers vehicles, yet, they provide quota to people who can afford them, so that they won't have to pay taxes.
The latest news is that MPS are going to get to vehicle logos. Is that how you lead by example? Why don't all the MPs and their families get tattoos of MP logo on their forehead. I cannot believe the egos of those people.
While the rest of the world is trying to create equality and get rid of discrimination, Bhutan is moving backwards. In Bhutan, all they want is to create different classes of people. The red kabney's the MPs, everyone wants to be treated like they are class above other people.
Shame on them. No wonder Bhutan's human right records never improves.
As long as you don't go all out to sell the country to polluting industrialists with no consideration whatsoever for the environment, Bhutan can remain carbon neutral. It is people who are bent on supporting such industries inspite of the declaration and try to convince others that it is not possible that we should watch out for.
I think the government must have consulted the climate experts on agreeing with the carbon neutrality declaration. Same time, COP 15 has high expectations from the nations and our country also offered the most appropriate commitment in terms of fightnig with the global warming issue. This declaration to me gives enough oppurtunity for our country to further negotiate and garner support from developed countries in supporting sector development by aligning with the green growth concept.
Bhutans emission status at the moment is also negative becuase of our large forest coverage (constitution spells out 60 % at all times) but every one must be aware of the carbon neutral vision. We must explore alternatives for building eco friendly instead of timber fed houses to reduce deforestation. There are number of alternatives in each sector to maintain carbon neutral emissions and this is the right time to start working on it or we will miss these oppurtunities.
Above all, this declaration is a guide, a click to remind our policy makers to consider green growth development.(I hope they seriously adopt this).
However,even with our effort in remaining carbon neutral, the consequences from climate change will not spare us and we will be the most vulnerable rather becuase of our geographical location and adaptation capacity. (The change in climate is all becuase of the historical emissions made by the developed countries). Therefore, time has come for us to prepare ourselves to adapt to this change in climate and to initiate well planned response measures.
why didn't you raise the issue of public referendum or atleast the consensus of the parliament when the present government committed all our viable hydro-projects for next 10 years to India.Do they have the right to dicide on our future…esp..by 11 of them????


The Rye Garnets had a lot to play for when they met Yorktown last Friday night. First, the Class A semifinal would see the winner move on to play the defending champions from Somers High for all the Westchester/Rockland marbles. Second, it was a chance for Dino Garr's team to avenge the 66-42 thumping the Huskers administered in 2016 in this same semifinal game. Third (as if they needed a third piece of motivation), Yorktown defeated Rye back in September by a score of 34-30 in a game the Garnets led 21-7 at halftime.
Seemingly all that stood between the Garnets and a title shot was Brett Makar, Yorktown's Super 11 back. As it turned out, Makar was all the Huskers needed. He ran for 189 yards on the evening and scored a touchdown. He sacked Rye QB Declan Lavelle when the Garnets were mounting a comeback. And he intercepted a pass when it was do or die.
Here's how it all went down. With both teams determined not to engage in another shootout, the teams left Charlie Murphy Field at halftime with the Huskers up 9-0. Lavelle, Rye's phenomenal sophomore quarterback, hadn't completed a pass. And yet the Garnets were upbeat.
The change after the break was electric, as Rye scored twice to take the lead. Lavelle would throw for over 200 yards in the second half of play, and he scored on a 1-yard plunge. Then, 25 seconds into the final period, workhorse running back Billy Chabot gave Rye a 13-9 lead.
On the next series, the Huskers' Tommy Weaver scored on a QB keeper to put his team back on top. Then a crucial sack by Makar turned possession over and led to what proved to be the winning score. Facing a third down at the Rye 40, Weaver lofted a throw that just eluded a Garnet interception by inches. Instead, it nestled in the arms of Alex Ujkaj, the same guy who caught the winning pass with under a minute to go in the teams' earlier meeting.
Rye wasn't finished. Lavelle led the Garnet and Black in their hurry-up offense, connecting on an aerial to Dylan Concavage with 1:18 left, a score that trimmed the deficit to 23-19. Rye tried an onside kick that the Huskers recovered. But Makar, of all people, fumbled the ball away in that final minute. The Garnets made it across midfield, but Yorktown's all-everything player atoned for his boo-boo with a spectacular pick of Lavelle's Hail Mary.
So did the whole team.
(pic# 1517) The band of brothers comes together before the kickoff.
(pic# 1645) Gavin Kenney hauls in another pass from Declan Lavelle.


Pranic Healing® is a highly evolved and tested system of energy medicine developed by GrandMaster Choa Kok Sui that utilizes prana to balance, harmonize and transform the body's energy processes. Prana is a Sanskrit word that means life-force. This invisible vital energy keeps the body alive and maintains a state of good health. In acupuncture, the Chinese refer to this subtle energy as Chi.
Pranic Healing® is a simple yet powerful & effective system of no-touch energy healing. It is based on the fundamental principles that the body is a self-repairing living entity that possesses the ability to heal itself and that the healing process is accelerated by increasing this life force that is readily available from the sun, air and ground to address physical & emotional imbalances. Master Choa Kok Sui says "Life Energy or prana is all around us. It is pervasive; we are actually in an ocean of Life Energy. Based on this principle, a healer can draw in Pranic Energy or Life Energy from the surroundings."
Pranic Healing® requires no drugs, gadgets, not even physical contact with the subject. Physical contact is not required because the practitioner is working on the energy body and not directly on the physical body. This energy body, or aura, is the mold or blueprint that surrounds and interpenetrates the physical body. It is the energy body that absorbs life energy and distributes it throughout the physical body, to the muscles, organs, glands, etc. The reason Pranic Healing® works on the energy body is that physical ailments first appear as energetic disruptions in the aura before manifesting as problems in the physical body.


Electrical extension cords are a great way to extend circuits to areas that you are working, away from available outlets. Sometimes we only need a short extension cord to give us the ability to plug our tools in and complete the project. Then again, I've had many times that I had to tie many extension cords together to reach the area that I needed to reach. But how many extension cords can one tie together and still have the amount of power to power up those tools? Is it actually safe to keep adding extension cords to an existing circuit? Well, there are some other factors to consider as well. how many feet from the electrical circuit breaker panel is the outlet that you are connecting to? It could be 50 feet away on the other side of the house. What is size circuit breaker the circuit connected to? What size is wiring actually feeding the outlet that you are about to plug into? As you can see, there are many factors to consider beyond the reality of the extension cord.
The longer the run of wire, the more resistance, and thus, more heat. So how does this affect you? Well, power tools draw a certain amount of amperage to run both correctly and efficiently. This electrical load may be too great for the size of extension cord it is attached to. Because of wire size, the resistance of the wire, and the voltage drop due to the distance, these variables can damage both extension cords and the power tools that are connected to them.Be sure to choose wisely the extension cords by choosing a heavy gauge wire by following the chart that I have provided. You can plainly see the maximum amperage and wire gauge that can supply each over a number of feet of cord. As you add additional feet of cord, the amperage availability gets less.
Voltage drop in the cord often doesn't allow the power tools to run at full speed. This causes them to heat up and often inflicts damage on them. Wiring inside the tools can melt as well as inflicting damage to the contacts. As the power tools heat up, so does the extension cords that they are connected to. That, in turn, leads to circuit breakers tripping, often due to a heating up of the breaker and/or overloading of the circuit breaker.
To be safe, try not to exceed the extension cord chart below. Be safe, not sorry.


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West End Girls were a Canadian pop band and girl group formed in 1990 in Vancouver, British Columbia.

History
The band was formed in Vancouver by producers John Dexter and John Livingston, and consisted of three teenaged girls, Camille Henderson, Aimee MacKenzie and Silvana Petrozzi. After several months of performing lessons with Henderson's father Bill Henderson, the lead singer of rock group Chilliwack, the trio recorded and released their debut album, West End Girls, on Johnny Jet Records in 1991. That album produced the Canadian Top 10 hits "Not Like Kissing You" and "I Want U Back" (a remake of the Jackson 5 hit), as well as the Top 40 singles "Say You'll Be Mine" and "Show Me The Way".

Petrozzi left the band in 1992, and was replaced by Janele Woodley. In 1993, shortly before the release of the group's second album, Henderson also left and was replaced by Celia-Louise Martin. In 1993, the band's new lineup released the album We Belong Together, which included the singles  "R U Sexin' Me", "Pure", and "Sexy'". Their second album wasn't as successful as their first one and they broke up the following year. Shortly afterwards, the group's record label released a remix album of some of their earlier hits.

They received a Juno Award nomination for Most Promising New Group in 1992 and they also toured with Roxette on the Canadian leg of that group's 1992 Canadian tour.

MacKenzie later joined the Canadian R & B group D-Cru, and their single "Show Me" sampled the West End Girls single "Show Me The Way".  She is now a swimsuit model.

Petrozzi/Kane later formed Pacifika with guitarist Adam Popowitz, bassist Toby Peter, and percussionist Elliot Polsky.  Their first album, Asunción, was released in 2007 and their second, Super Magique, was released in 2010.  Kane released a solo album, La Jardinera, in November 2012.

Henderson later toured extensively as a backing vocalist for Sarah McLachlan and performed lead vocals on Delerium's track "Duende".  She is now a vocal coach.

Woodley later formed the pop rock duo A Perfect Day with Joseph Hrechka and they released the album All Over Everything in 2004.

Martin has done occasional acting gigs.

Discography

Albums
West End Girls, 1991 (Johnny Jet Records/A&M Records)
We Belong Together, 1993 (Johnny Jet Records/A&M Records)
Hits, Remixes and Other Cool Stuff, 1997 (Dexter Entertainment Group)
The Definitive Collection, 2000 (Dexter Entertainment Group)

Singles

Soundtrack use
 In 1999, "Sexy" was featured in the beginning of lesbian cult classic movie Better Than Chocolate, as was "Pure (You're Touching Me)".
Cover Version
In 1992, Hong Kong singer/actress Sammi Cheng Sau Man covered "Say You'll Be Mine". in her Cantonese album "Never Too Late".

Musical groups established in 1990
Musical groups disestablished in 1994
Musical groups from Vancouver
Canadian pop music groups
Canadian girl groups
1990 establishments in British Columbia
1994 disestablishments in British Columbia


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Make Christopher Plummer's Sexy Evil Santa Claus Your New Christmas Tradition
You know you've always wanted to see Ross's Dad and Baron von Trapp duke it out!
By Brittany KnupperDec 24th, 2022, 11:00 am
Picture it. Canada 1978. You have Elliott Gould at the peak of his popularity (and his sexiness.) You have Christopher Plummer as his nemesis. And you also have John Candy dating a hot blonde who wears novelty t-shirts. What follows is one of the greatest heist movies of all time.
And it's also a Christmas movie.
That's right I'm talking about the crime classic The Silent Partner, directed by Daryl Duke. In it, we watch Elliott Gould as Miles Cullen – a bored bank teller whose small bank is located inside a large shopping mall. When Miles figures out that his bank is about to be robbed by a man dressed as Santa Claus, he decides to seize upon an irresistible opportunity. Instead of contacting the police or warning his bosses, he starts emptying his cash drawer into a lunchbox.
When the Santa thief reaches Miles, he hands over the small remaining portion of bills… but tells the authorities later that the entire amount from the drawer was stolen. Miles gets to go home with the remaining cash and none are the wiser! It's the perfect crime! Until Arthur, the Santa thief, figures it out when the news announces the entire amount stolen.
And, it turns out, Arthur is a violent psychopath. And what follows is an increasingly tense and endlessly entertaining cat-and-mouse game in which both Miles and Arthur attempt to outwit and outmaneuver each other. Miles gets Arthur arrested only to have him set free, while Arthur sends in Elaine (Celine Lomez) to seduce Arthur into giving up the location of the hidden cash. The film is full of unexpected twists and turns, from the reveal that Arthur is a flamboyant and feminine, queer-coded man (and yes that means we get to see him in some fantastic fishnet tank tops and makeup and also drag! Yes, Christopher Plummer in drag!), to some shockingly brutal and violent deaths.
(Image: EMC/Toronto)
But it is definitely the two stars Plummer and Gould, that keep this in my rotation as a holiday must-watch. Their chemistry is fantastic and both of them excel at playing characters who know how to keep things close to the vest. It does dabble a bit in the extremely 70s and problematic "violent cross-dresser" trope, but I find Plummer's performance fascinating because he manages to mix the hyper-masculine (and misogynistic) elements of Arthur with the hyper-feminine (his clothing, nail polish, eyeliner, and his final "disguise") in such surprising ways that the performance feels incredibly subversive.
Meanwhile, Gould is such a slick charmer even as a bank-teller that you would think was in way over his head. But he plays Miles with so many layers, each one slowly pealing back until you realize that Miles might just be outplaying everyone. And did I mention that Christopher Plummer, even playing a violent psychopath, looks really hot in his lil fishnet tank! Not to mention, we love a movie that involves our predatory banking and loans system getting taken down a peg or two!
So this year, why not download the Kanopy app, or hit up the "to rent" sections of Apple TV and Amazon Prime, and give yourself a new Christmas (or Hanukkah!) tradition. Impress your parents by picking out a film from before you were born! It might be a 44-year-old film but it's new to you! So happy holidays from my holiday tradition to yours!
Image: EMC/Lionsgate
And don't forget, John Candy gets the hottie!
(Image: EMC/Lionsgate)
Christopher PlummerSanta Clausthe silent partner

redpajama_set_name: RedPajamaCommonCrawl


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R.I.P. 'Where The Wild Things Are' Creator Maurice Sendak (1928-2012)
Renowned children's book author and illustrator Maurice Sendak has died at the age of 83. The The Associated Press reports the creator of Where The Wild Things Are, In The Night Kitchen and Outside Over There passed away this morning due to complications from a stroke...
'Where The Wild Things Are' Creator Maurice Sendak Trolled by Stephen Colbert [Video]
The prolific and influential writer and illustrator Maurice Sendak is best known for Where The Wild Things Are, considered one of the all-time greats of children's literature. That classification as "children's literature" is just one of the many ideas to which the venerable and indeed hil…
'Where the Wild Things Are': When the Counterculture Makes Kid Flicks
I'm almost certainly not alone in expressing my guarded -- even conflicted -- anticipation at the forthcoming release of the "Where the Wild Things Are" film; it's a beloved children's classic filtered through a thrift store sheet-staining hipster's wet dream...
Brian Heater
Link Ink — Rejected Playboy Cover Girls, Google Maps Canvas, Maurice Sendak Invites Parent…
Art: The Etsy shop of "theGorgonist" has some great Alice in Wonderland illustrations, plus this totes adorbs Star Trek bro-down between Kirk and Spock performing an intergalactic fistbump. (Super Punch) Movies: "Where the Wild Things Are" author Maurice Sendak answers a question…
Toy Crossover: Where the Wild Things Are
The Spike Jonze movie adaptation of the classic children's book "Where the Wild Things Are" by Maurice Sendak is coming in October, and so are these two-inch tall Kubrick toys of Max and friends, available for $9.99. If that's not enough, there's also a second set of…


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Altered Carbon: A Carbon-air Battery as a Next-generation Energy Storage System
Imagine a stable power supply along with a high energy density! This is exactly what has been developed by scientists at Tokyo Institute of Technology (Tokyo Tech) in the form of a carbon-based energy storage system. This could be a potential alternative to large-scale hydrogen-based energy storage systems that suffer from low energy density, allowing us to efficiently harness the renewable energy sources to meet the demand for clean energy.
One of the barriers to generating electricity from wind and solar energy is their intermittent nature. A promising alternative to accommodate the fluctuations in power output during unfavorable environmental conditions are hydrogen storage systems, which use hydrogen produced from water splitting to generate clean electricity. However, these systems suffer from poor efficiency and often need to be large in size to compensate for it. This, in turn, makes for complex thermal management and a lowered energy and power density.
In a study published in Journal of Power Sources, researchers from Tokyo Tech have now proposed an alternative electric energy storage system that utilizes carbon (C) as an energy source instead of hydrogen. The new system, called a "carbon/air secondary battery (CASB)," consists of a solid-oxide fuel and electrolysis cell (SOFC/ECs) where carbon generated via electrolysis of carbon dioxide (CO2), is oxidized with air to produce energy. The SOFC/ECs can be supplied with compressed liquefied CO2 to make up the energy storage system.
"Similar to a battery, the CASB is charged using the energy generated by the renewable sources to reduce CO2 to C. During the subsequent discharge phase, the C is oxidized to generate energy," explains Prof. Manabu Ihara from Tokyo Tech.
As the carbon is stored in a confined space in the SOFCs/ECs, the energy density of the CASB is limited by the amount of carbon it can hold. Despite this limitation, the researchers found that the CASB had a higher volumetric energy density compared to hydrogen storage systems.
Another indicator of battery performance is the charge-discharge efficiency. To evaluate this metric, the researchers performed a charge-discharge experiment. They observed that the transformations between C and CO2 were due to "Boudouard reactions" characterized by a redox reaction of a mixture of carbon monoxide (CO), CO2 and C. Specifically, during the charging phase, C was deposited on the electrode via the electrochemical reduction of CO2 and the reduction of CO via the Boudouard decomposition. During the discharge phase, the C was oxidized to CO and CO2 via the Boudouard gasification reaction and electrochemical oxidation respectively. The researchers found that the C utilization for energy generation of the CASB depended on the equilibrium between the 3 different carbon species (C, CO2, CO), also known as the "Boudouard equilibrium."
The CASB system was able to utilize most of the carbon deposited on the electrode for energy generation, demonstrating a high Coulombic efficiency of 84%, indicating that most of the stored energy can be obtained during the discharge phase. Furthermore, it showed a superior power density of 80 mW/cm2 and a charge-discharge efficiency of 38% that was sustained over 10 charge-discharge cycles. This suggested that no degradation of the fuel electrode occurred.
"Compared to hydrogen storage systems, the CASB system is expected to have a smaller system size and higher system efficiency", says Prof. Ihara. Their new system could lay the foundation for compact and efficient carbon energy storage systems that could work alongside renewable energy sources for a fossil-fuel-free future.
Indeed, we might fight carbon with carbon!
Keisuke Kameda, Sergei Manzhos, and Manabu Ihara*
Carbon/air secondary battery system and demonstration of its charge-discharge
Department of Chemical Science and Engineering, School of Materials and Chemical Technology, Tokyo Institute of Technology
* Corresponding author's email: mihara@chemeng.titech.ac.jp
Press release "Altered Carbon: A Carbon-air Battery as a Next-generation Energy Storage System"
Monocrystalline silicon thin film for cost-cutting solar cells with 10-times faster growth rate fabricated | Tokyo Tech News
Tokyo Tech's Ookayama campus goes 'mega-solar' as part of the smart grid 'Ene-Swallow ver.3' project | Tokyo Tech News
Manabu Ihara - Creating an ambient energy society - Fighting global warming and growing the economy can go together! | Research Stories | Research
Environmental Energy Innovation Building | Research Stories | Research
Ihara Laboratory (Japanese)
Manabu Ihara | Researcher Finder - Tokyo Tech STAR Search
InfoSyEnergy Research and Education Consortium
Professor Manabu Ihara
School of Materials and Chemical Technology, Tokyo Institute of Technology
Email mihara@chemeng.titech.ac.jp


A medium amber gold in colour, Tullamore Dew Original is light bodied with sweetish spices, buttery and rounded with light citrus fruitiness. Warming on the palate this very drinkable whiskey exhibits pleasant maltiness with toasted wood and vanilla hints. The finish is smooth and gentle with complex flavours lingering.


Legacy Kickboxing returns to Houston for LK 3. The event takes place July 24 at the Bayou Music Center, and will feature some exciting matchups. Headlining bouts will feature Angel Huerta (5-1) versus Justin Houghton (6-2), and Valentina Shevchenko (56-1) versus Anke Van Gestel (22-3).
Also featured on the main card, the second leg of the light-heavyweight tournament. Andres Van Engelen (8-1) will take on UFC veteran, Danny Mitchell (4-1), and Glory veteran, Manny Mancha (11-2) will face Fernando Almeida (9-1).
UPDATE 6-10-15: Legacy has signed former UFC vet Sean Soriano to multi fight deal. Sean is 8-3, with all three losses in the UFC. Sean will be making his kickboxing debut at LK3. He will face Josh Glover will also be making his pro kickboxing debut.
Tickets are available here at MikeTheTruth.com.


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Canadian researchers to test if antiviral drug can stop COVID-19 outbreaks at nursing homes
Researchers in Toronto have been given the green light to launch Canada's first clinical trial into whether the antiviral drug favipiravir can help fight COVID-19 outbreaks in long-term care homes that have been devastated by the coronavirus.
Favipiravir, sold under the brand name Avigan, is typically used to treat viruses, like influenza, and works by stopping a cell's potential to become a factory for the virus and spread infection, according to the researcher leading the study.
"If we have a drug that you reliably use to stop outbreaks after they have started, that would be extraordinarily helpful," Dr. Allison McGeer, the director of infection control at Mount Sinai Hospital in Toronto, told Global News.
Nursing homes account for over 80 per cent of Canada's COVID-19 deaths, according to Canada's chief public health officer, Dr. Theresa Tam, with facilities in Quebec and Ontario hit particularly hard.
Japan approved the drug for influenza treatment in 2014 and added it to the country's stockpile of emergency medical supplies in case of an influenza outbreak, according to the Halifax-based biopharmaceutical company Appili Therapeutics Inc., which is sponsoring the drug trial.
McGeer said the drug is similar to the experimental drug remdesivir, which has been found to shorten the recovery time of COVID-19 patients from 15 to 11 days. But unlike remdesivir, which is administered intravenously, Favipiravir can be taken in tablet form, which makes it ideal for long-term care homes, according to McGeer.
2:03 Why non-profit nursing homes provide better care for residents
She is looking to enroll about 760 subjects, both residents and staff, at 16 long-term care homes in Ontario.
And with no vaccine currently available and a looming second wave of the virus approaching, she is hoping that favipiravir could work as effectively in stopping COVID-19 as oseltamivir, sold under the brand name Tamiflu, is in stopping influenza.
"It's obviously a lot to ask of residents and their social decisionmakers to be in a trial during a pandemic," she said. "But if it works, it will be brilliant. And the only way to find out if it works is to do the trials."
[ Sign up for our Health IQ newsletter for the latest coronavirus updates ]
Ontario long-term care facilities have been devastated by outbreaks of COVID-19, with more than 7,000 residents and staff becoming infected and over 1,700 residents deaths, according to the provincial government.
"Given the strikingly high fatality rate from COVID-19 in long-term care facilities, there is a particularly urgent need for randomized trials that explore ways of protecting the most vulnerable members of our society," Peter Jüni, a senior epidemiologist at St. Michael's Hospital and co-principal investigator, said in a statement.
The partially blinded, randomized trial — first approved by Health Canada last month — will evaluate the efficacy of favipiravir in nursing home residents compared with placebo to prevent COVID-19 outbreaks.
However, as the goal of the trial is to see whether the antiviral drug can control outbreaks of COVID-19 once they begin, McGeer said researchers are looking to enrol all 250 homes in Greater Toronto Area over the next six months so facilities can make quick decisions about whether they want to participate in the trial if they experience an outbreak.
1:39 Remdesivir: Drug shows promise as COVID-19 treatment
Remdesivir: Drug shows promise as COVID-19 treatment
Two small studies from researchers in Wuhan, China, published in February showed that favipiravir demonstrated some effectiveness against SARS-CoV-2, the virus that causes COVID-19.
And Russia announced in May that it had approved a version of favipiravir to treat COVID-19 after its trials found it could reduce the length of illness from 11 to four or five days.
McGeer said although there is limited research on the drug, there is only one way to find out if it's effective: conduct the trials.
"It's almost like it would be too good to be true. It makes me anxious just thinking about it," McGeer said. "But, you know, that's why we are doing it — because it might be true."
Derek Sloan has been ejected from the Conservative caucus
Coronavirus: Quebec to cover cost of recovering and recycling disposable masks from high schools
Community spread of new COVID-19 variants may be happening in Canada: Tam
1. Stock market
5. NBC
7. Earthquake
13. Ronald Reagan
14. Bill Clinton
15. Michelle Obama


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Sarah Michelle Gellar: Selma Blair facing multiple sclerosis 'with dignity, grace'
Geller expressed support for her longtime pal as her former "Cruel Intentions" co-star battles the debilitating disease.
Michael Kovac / Getty Images
Feb. 27, 2019, 5:29 PM UTC / Source: TODAY
By Eun Kyung Kim
Sarah Michelle Gellar showered her longtime friend Selma Blair with support, applauding the way her "Cruel Intentions" co-star has taken on her challenging battle with multiple sclerosis.
Geller took to Instagram to speak about Blair's very busy week, which started with her first public appearance since announcing her diagnosis and a pair of media interviews.
Selma Blair hit the red carpet Sunday for the Vanity Fair Oscar Party and her first public appearance since revealing she has multiple sclerosis. Getty Image
"To say I'm proud, would be a gross understatement," Geller wrote in the caption of a photo of Blair dressed up for the Vanity Fair Oscar party Sunday night. "When I have a cold, I want to hide from the world under my covers. But not Selma. She is facing this diagnosis, the way she faces everything, with dignity, grace and head on."
Geller, 41, has been friends with Blair, 46, since the pair starred together in the 1999 movie "Cruel Intentions."
Blair and Gellar at the 2000 MTV Movie Awards.Ron Galella, Ltd. / WireImage
"I know the support and encouragement that she has received on this platform has truly been a source of strength for her," Geller wrote in her most recent post Monday. "This is not an easy journey, but Selma will not let this define her."
https://www.instagram.com/p/BjhESM5liSQ
Over the weekend, Blair made her way down the red carpet at Sunday's Vanity Fair Oscar party with a cane inscribed with her monogram and featuring a pink diamond. It was her first public appearance since Blair revealed in October that she had been diagnosed with MS.
https://www.instagram.com/p/BuX1qOSn5VC
A television interview Blair conducted days before the Oscars then aired Tuesday, the same day Vanity Fair posted an interview in which she discussed why she wants to be open about the challenges of living with MS.
"There's no tragedy for me," she told the magazine for its March issue. "I'm happy, and if I can help anyone be more comfortable in their skin, it's more than I've ever done before."
Actress Selma Blair makes red carpet return after multiple sclerosis diagnosis
Feb. 25, 201901:04
Eun Kyung Kim