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Evidence-Based Complementary and Alternative Medicine
Volume 2012 (2012), Article ID 420715, 15 pages
Phytochemical Analysis and Antimicrobial, Antinociceptive, and Anti-Inflammatory Activities of Two Chemotypes of Pimenta pseudocaryophyllus (Myrtaceae)
1Unit of Exact and Technologic Sciences, Goiás State University, Anápolis, Brazil
2Institute of Tropical Pathology and Public Health, Federal University of Goiás, Goiânia, Brazil
3Faculty of Pharmacy, Federal University of Goiás, Goiânia, Brazil
4Department of Physiological Sciences, Institute of Biological Sciences, Federal University of Goiás, Goiânia, Brazil
Received 9 June 2012; Accepted 4 August 2012
Academic Editor: Vincenzo De Feo
Copyright © 2012 Joelma Abadia Marciano de Paula et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preparations from Pimenta pseudocaryophyllus (Gomes) L.R. Landrum (Myrtaceae) have been widely used in Brazilian folk medicine. This study aims to evaluate the antimicrobial activity of the crude ethanol extracts, fractions, semipurified substances, and essential oils obtained from leaves of two chemotypes of P. pseudocaryophyllus and to perform the antinociceptive and anti-inflammatory screening. The ethanol extracts were purified by column chromatography and main compounds were spectrally characterised (1D and 2D 1H and 13C NMR). The essential oils constituents were identified by GC/MS. The broth microdilution method was used for testing the antimicrobial activity. The abdominal contortions induced by acetic acid and the ear oedema induced by croton oil were used for screening of antinociceptive and anti-inflammatory activities, respectively. The phytochemical analysis resulted in the isolation of pentacyclic triterpenes, flavonoids, and phenol acids. The oleanolic acid showed the best profile of antibacterial activity for Gram-positive bacteria (), followed by the essential oil of the citral chemotype (). Among the semipurified substances, Ppm5, which contained gallic acid, was the most active for Candida spp. () and Cryptococcus spp. (). The crude ethanol extract and fractions from citral chemotype showed antinociceptive and anti-inflammatory effects.
Pimenta pseudocaryophyllus (Gomes) L.R. Landrum (Myrtaceae) is a plant popularly known in Brazil as pau-cravo, louro-cravo, louro, craveiro, craveiro-do-mato, chá-de-bugre, and cataia [1–6]. In folk medicine, the leaves have been used to produce a refreshing drink with calming, diuretic, and aphrodisiac properties, as well as to treat colds and their complications and digestive and menstrual problems [2, 4–6]. It is the only Pimenta species native to Brazil [1, 3], and recent studies have shown the occurrence of different chemotypes for this species; these are characterised, for example, by the predominance of citral or ()-methyl isoeugenol in the essential oils .
Invasive infections caused by Candida spp. and Cryptococcus neoformans have increased significantly in recent years [8–11]. The cause of this rise is often related to immunodeficiency associated with transplantation  and acquired immunodeficiency syndrome (AIDS) , as well as the use of intravascular catheters , dialysis, and abusive use of glucocorticoids and broad-spectrum antibiotics . The drugs available to treat these infections are often not selective, are toxic, or have narrow action spectra ; moreover, some species are resistant to antifungal agents .
In the pharmacotherapy of bacterial diseases, the use of antibiotics in recent decades has significantly reduced the incidence of many infectious diseases. On the other hand, the severe side effects from many of these substances and the emergence of multiresistant microorganisms have stimulated research on the development of new antibacterial agents that are more specific, effective, and safe [24, 25].
There is thus a consensus on the need for further research on new alternative treatments for bacterial and fungal infections. Efforts have been focused on investigating the antimicrobial properties of products from plants [12, 25]. In addition to extensive use in folk medicine in diseases related to the common cold, which often involve microbial and/or inflammatory processes, experimental data show that plants of the genus Pimenta (Myrtaceae) have antimicrobial potential [26, 27]. The essential oil of P. pseudocaryophyllus leaves collected in two geographical areas in the state of São Paulo was active against strains of C. albicans, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus . In a previous study, we described the antimicrobial activity of the crude ethanol extract of leaves of this species collected in the Brazilian Cerrado against Gram-positive bacteria and Candida albicans , but we have not performed phytochemical studies for the isolation and identification of substances accountable for this activity.
Moreover, scientific studies have shown that Pimenta species, widely used in folk medicine, have analgesic and anti-inflammatory activities and are nontoxic in typical dosages [29–32]. Thus, the aims of this work were to carry out the phytochemical study, evaluate the antimicrobial activity of the crude ethanol extracts, fractions, semipurified substances, and essential oils obtained from leaves of two chemotypes of P. pseudocaryophyllus, and perform the screening of the antinociceptive and anti-inflammatory activities.
2. Material and Methods
2.1. General Experimental Procedures
The 1H and 13C one-dimensional and two-dimensional NMR spectra (Heteronuclear Single Quantum Coherence (HSQC) and Heteronuclear Multiple Bond Correlation (HMBC)) were obtained using a Bruker Avance III-500 spectrometer running at 500 MHz (1H) and 125 MHz (13C), using deuterated chloroform (CDCl3) and deuterated dimethyl sulfoxide (DMSO-d6) as solvents for nonpolar and polar samples, respectively. Tetramethylsilane (TMS) was used as an internal reference standard for chemical shifts (, ppm), and in some cases, the peak from the solvent was used for reference.
Essential oil samples were analysed by GC/MS using a QP5050A instrument (Shimadzu, Kyoto, Japan) with a capillary column of CBP-5 melted silica (30 m × 0.25 mm × 0.25 m; 5% phenyl-methylpolysiloxane film) (Shimadzu, Kyoto, Japan). Additionally, helium was used with a flow rate of 1 mL min−1 as a carrier gas, and a thermal profile of 60°C to 240°C with a gradient of 3°C min−1 followed by a gradient of 10°C min−1 up to 280°C, keeping a 5 min isotherm, was used. The ionisation energy of the detector was kept at 70 eV, and the sample injection volume following dilution in hexane (10%) was 0.5 L. The analysis was carried out in the scanning mode, with a 40–400 m/z mass interval and 1 : 5 injection ratio. The quantitative analysis was obtained by integrating the total ion chromatogram (TIC). The identification of the components was performed by comparing the mass and retention indices (RI) calculated using values for the mass and retention indices available in the literature . The retention indices were calculated by coinjection with a mixture of hydrocarbons, C8–C32 (Sigma, MO, USA), applying the Van Den Dool and Kratz equation .
The mass spectra of the flavonoids were collected using a coupled LC/EM/EM: Varian 1200L (Walnut Creek, CA, USA) system with a quadrupole ion analyser and ionisation through electron impact, 70 eV in positive mode m/z [M + H]+. The m/z scanning spectrum was 100–900, and the ionisation chamber was kept at room temperature.
The HPLC was held on Waters equipment (MA, USA) equipped with quaternary pump, e2695 separation module, 2998 diode array detector (PDA), and Empower 2.0 data processing system. The Varian C-18 (250 × 4.6 mm) column was used at room temperature. The detection system used was monitored at 360 nm for detection of flavonoids. The injection volume was of 30 L and the run in isocratic mode was used as mobile phase 75% of A [acetonitrile/water (0.1% acetic acid)—90 : 10] and 25% of B [MeOH (0.1% acetic acid)] at a constant flow of 1 mL min−1. The maximal running time was 20 min. Quercitrin (Sigma) was used as reference standard. The samples were previously filtered through Millex (Millipore, MA, USA) membrane and the mobile phase in 0.45 m PVDF membrane (Millipore).
For analytical and preparative thin-layer chromatography (ATLC, PTLC), chromatography plates prepared with G60 F254 Vetec silica gel, or F254 silica plates manufactured by Merck, RJ, Brazil, were used. Mixtures of organic solvents with the proper polarities for each analysed fraction were used as mobile phases. For detection of the components, the plates were observed under UV light at 254 nm and 365 nm and developed by the spraying of sulphuric vanillin followed by heating or exposure to a solution of 1% ethylamine diphenyl borate in methanol (NP) .
The column chromatography used silica gel G60 0.05–0.2 mm (Vetec, Brazil) and Sephadex LH-20 (GE Healthcare, Uppsala, Sweden) as stationary phases. As mobile phases, mixtures of organic solvents in increasing order of polarity were used, according to the polarity profiles of the fractions undergoing the process.
2.2. Plant Material
The leaves of P. pseudocaryophyllus of the (E)-methyl isoeugenol and citral chemotypes were collected in São Gonçalo do Abaeté, MG, Brazil, in February 2008, at 18°20′58.4′′S, 45°55′23.4′′W, and 864 m altitude. These two chemotypes are the most common found in the region of São Gonçalo do Abaeté and therefore were selected for this study.
The plant material was identified by Professor Carolyn Elinore Barnes Proença, Ph.D., of the Universidade de Brasília, and a voucher specimen (UFG-27159) was deposited at the herbarium of the Universidade Federal de Goiás.
2.3. Preparation of Extracts and Fractions
Air-dried leaves were ground in a knife mill. The crude ethanol extract was obtained by maceration of the powdered material (1 : 5 w/v) of each chemotype in 95% EtOH (v/v) at room temperature, followed by filtration, and concentration on a rotary evaporator at a temperature below 40°C. The extracts were concentrated to a constant weight and named crude ethanol extract of (E)-methyl isoeugenol chemotype (EEM) and crude ethanol extract of citral chemotype (EEC).
Fifty grams of each extract (EEM and EEC) were dissolved in 250 mL MeOH/H2O (7 : 3) and subjected to liquid/liquid partitioning with solvents of increasing polarity (hexane, dichloromethane, and ethyl acetate). The final MeOH/H2O residual was concentrated in a rotary evaporator for elimination of the MeOH, and the resulting aqueous fraction was lyophilised. Therefore, four fractions were obtained from the crude extract of each chemotype: the hexane fraction (HFM and HFC), the dichloromethane fraction (DFM and DFC), the ethyl acetate fraction (EAFM and EAFC), and the aqueous fraction (AFM and AFC).
For obtaining of essential oils, leaves from each chemotype were subjected to hydrodistillation for 3 h in a Clevenger apparatus. The essential oils were dried with anhydrous Na2SO4, packaged in amber glass vials, and stored at −20°C until use. They were named essential oil of the citral chemotype (EOC) and essential oil of the (E)-methyl isoeugenol chemotype (EOM).
2.4. Isolation of the Chemical Constituents
The HFC (2.5 g) was fractionated on a silica gel column (1 : 40) eluted with a gradient of hexane-EtOAc (5–100%), EtOAc-MeOH (1 : 1), and MeOH (100%). Seventy-five 20 mL fractions were collected and evaluated by TLC [hexane-EtOAc mixtures (10–30%)], observed under 254/365 nm UV light, and detected with sulphuric vanillin reagent. Similar fractions were pooled, resulting in 14 new fractions (CH-1 to CH-14). Fractions CH-8 through CH-11 were rechromatographed on a silica gel column eluted isocratically with CH2Cl2-petroleum ether (7 : 3), resulting in Ppc-1 (133.5 mg).
The DFC (1.0 g) was fractionated on a silica gel column (1 : 40) and eluted with a gradient of CH2Cl2-petroleum ether (7 : 3 and 9 : 1), CH2Cl2-EtOAc (5–100%), EtOAc-MeOH (1 : 1), and MeOH (100%). One hundred and one 5 mL fractions were collected. Using TLC [mobile phases composed of mixtures of CH2Cl2-EtOAc (75 : 25, 80 : 20, and 95 : 5)] with 254/365 nm UV light, and sulphuric vanillin reagent for detection, the fractions were combined into 20 new fractions (CD-1 to CD-20). Fractions CD-9, CD-10, and CD-11 were fractionated on a silica gel column eluted isocratically with petroleum ether-EtOAc (90 : 10). The semipurified fractions were rechromatographed on a silica gel column eluted isocratically with petroleum ether-acetone (40 : 10), resulting in Ppc-2 (22.6 mg).
In a column packed with Sephadex LH-20, 1.5 g of EAFC was subjected to chromatography, using MeOH (100%) as the mobile phase. Two hundred and sixty-one 10–20 mL fractions were collected. The fractions were evaluated by TLC [EtOAc-formic acid-acetic acid-H2O (100 : 11 : 11 : 26)], observed under 254/365 nm UV light and developed with sulphuric vanillin and NP reagents. Similar fractions were pooled, resulting in 12 new fractions (CEA-1 to CEA-12).
CEA-12 showed a single spot at close to 1, resulting in Ppc-3 (59.7 mg). Fractions CEA-7, CEA-8, and CEA-9 showed typical spots of flavonoids with similar values, so they were gathered and rechromatographed on a Sephadex LH-20 column eluted isocratically with MeOH-H2O (1 : 1). From this column, 175 fractions were collected and lyophilised after MeOH evaporation, evaluated by TLC, and grouped into 17 fractions (CEA-A to CEA-Q). The CEA-N fraction was subjected to preparative TLC [EtOAc-formic acid-acetic acid-H2O (100 : 11 : 11 : 26)], resulting in Ppc-4 (18.7 mg). CEA-K, CEA-L, and CEA-M were gathered and subjected to preparative TLC, and the semipurified fraction was rechromatographed on a Sephadex LH-20 column eluted with a gradient of acetone-H2O (1 : 1, 6 : 4, and 7 : 3). From this column, 93 1 mL fractions were collected and subjected to solvent evaporation, lyophilisation, and monitoring by TLC, which led to Ppc-5 (73.2 mg).
The AFC (1.2 g) was chromatographed on packed column with Sephadex LH-20 eluted with MeOH (100%). Approximately 180 fractions of 1–10 mL were collected. The fractions were evaluated by TLC [isobutanol-acetic acid-water (14 : 1 : 5)] and exposed with UV light and 1% FeCl3 in 0.5 M hydrochloric acid. Fractions with the same chromatographic profiles were pooled, resulting in 11 new fractions (AC-1 to AC-11). AC-9 was reactive to FeCl3, resulting in Ppc-6 (83.1 mg).
The HFM (2.5 g) was fractionated on a silica gel column (1 : 40) and eluted with 100% hexane-EtOAc (5–100%), EtOAc-MeOH (1 : 1), and MeOH (100%). One hundred 10 mL fractions were collected and evaluated by TLC [hexane- EtOAc (10–30%)], observed under UV light, and detected with sulphuric vanillin reagent, resulting in 14 new fractions (MH-1 to MH-14). Fractions MH-4 and MH-5 were rechromatographed on a silica gel column eluted isocratically with CH2Cl2-petroleum ether (7 : 3), resulting in Ppm-1 (106.2 mg) and Ppm-2 (75.8 mg). Fractions MH-9 and MH-10 were gathered and subjected to isocratic silica gel column chromatography [hexane-CH2Cl2-MeOH (10 : 10 : 1)], which resulted in Ppm-3 (36.9 mg).
The low output from DFM hindered the use of chromatography columns in isolating possible compounds. Thus, TLC of the DFM was carried out with some of the semipurified fractions for comparison to verify whether some of the previously isolated compounds could also be found in this fraction. Runs were performed with DFM, Ppc-1, Ppm-1, and Ppm-2 in CH2Cl2-petroleum ether (7 : 3), DFM and Ppc-2 in petroleum ether-acetone (40 : 10), and DFM and Ppm-3 in hexane-CH2Cl2-MeOH (10 : 10 : 1).
The EAFM (1.2 g) was subjected to chromatography on a packed column with Sephadex LH-20 [MeOH (100%)]. Approximately 159 1-mL fractions were collected and analysed by TLC [EtOAc-formic acid-acetic acid-H2O (100 : 11 : 11 : 26)]. Based on observation in UV light and detection with NP reagent, the fractions were pooled into four new fractions (MEA-1 through MEA-4). Fractions MEA-1 and MEA-2 showed typical spots of flavonoids with yellow-orange and greenish-yellow fluorescence after spraying with NP and observation under 365 nm UV light, with similar values; thus, they were pooled and rechromatographed on a Sephadex LH-20 column eluted with a sequence of acetone-H2O mixtures (1 : 1, 6 : 4, and 7 : 3). From this column, 99 fractions were collected and, after evaluation by TLC, resulted in Ppm-4 (273.4 mg).
The AFM (1.2 g) was subjected to chromatography in a packed column with Sephadex LH-20 [MeOH (100%)]. Approximately 117 1-mL fractions were collected. The fractions were monitored by TLC [isobutanol-acid acetic-water (14 : 1 : 5)] with use of the UV light and 1% FeCl3 in 0.5 M hydrochloric acid. Fractions with the same chromatographic profiles were pooled, resulting in nine new fractions, and one of the most reactive to the 1% FeCl3 was named Ppm-5 (203.6 mg).
2.5. Antimicrobial Activity
The extracts, fractions, essential oils, citral (Sigma), and semipurified substances were subjected to the microdilution test in broth for determining the minimum inhibitory concentration (MIC) in sterile 96-well microplates with “U-”shaped wells, as recommended by the Clinical and Laboratory Standards Institute (CLSI) [36, 37]. The experiments were performed in duplicate and repeated twice, independently, except for the semipurified substances.
American Type Culture Collection (ATCC) standard strains and clinical isolates were used (Table 1); these are kept in the Laboratories for Bacteriology and Mycology at the Institute of Tropical Pathology and Public Health, Federal University of Goiás, Goiânia, GO, Brazil.
Prior to testing, to reactivate the microbial cultures, the bacteria were transferred to Casoy broth and incubated at 37°C for 24 h, then transferred to inclined Casoy agar, and incubated at 37°C for an additional 24 h. The fungi were transferred to Sabouraud dextrose agar and incubated at room temperature for 24 to 48 h (Candida spp.) or 48 to 72 h (Cryptococcus spp.).
The culture medium used in the antibacterial activity test was 2x Müeller Hinton broth (MH) and that used in the antifungal activity test was RPMI 1640. The samples were solubilised in 10% dimethyl sulfoxide (DMSO) and diluted in MH broth (antibacterial activity) to obtain a concentration of 2000 g mL−1 or in RPMI (antifungal activity) to obtain a concentration of 1000 g mL−1. The semipurified substances were used at concentrations according to the available amount of the substance. For the preparation of samples with essential oils, 0.02% Tween 80 was added. Vancomycin (Sigma; 32 g mL−1) and gentamicin (Sigma; 128 g mL−1) were used as controls for Gram-positive and Gram-negative bacteria, respectively, and itraconazole (Sigma) at an initial concentration of 16 g mL−1 was used as a control for fungi.
The microplates inoculated with bacteria were incubated at 35°C 2°C for 16–20 h and for 24 hours at this temperature for Staphylococcus. One hour before the end of the incubation period, each well received 20 L of 0.5% triphenyl tetrazolium chloride (TTC), and the microplates were reincubated for approximately 30 min. The appearance of reddish colour was considered as proof of bacterial growth. The microplates inoculated with fungi were incubated at room temperature for 48 h (Candida spp.) and 72 h (Cryptococcus spp.). Fungal growth was checked visually, and the MIC was defined as the lowest concentration (g mL−1) of the sample fully capable of inhibiting bacterial and fungal growth.
2.6. Antinociceptive and Anti-Inflammatory Activity
These experiments were approved by the Animal Ethics Committee of the Federal University of Goiás, Protocol number 146/2008. Male, young adult mice, weighing between 25 and 30 g, were transferred to the experimental room two days before the tests, kept in a light/dark cycle of 12 h at 22 2°C in a noise-free facility, with water and food ad libitum. The food was taken away 12 h before the test, keeping the water available ad libitum at all times.
2.6.1. Evaluation of Antinociceptive Activity by Testing Abdominal Contortions Induced by Acetic Acid
The evaluation of abdominal contortions induced by acetic acid was carried out according to Koster et al.  and Lapa et al. . Test groups consisting of 10 mice per dose of extract, fraction, standard drug or vehicle were used. The mice from the different experimental groups received intraperitoneal injections of 1.2% acetic acid solution (v/v, 10 mL kg−1) 1 h after oral intake (gavage) of the extracts or fractions (in different doses) or indomethacin (standard drug) or vehicle (control). They were then placed under glass funnels and the contortions, contractions, and rotation of the abdomen followed by the extension of one or both back paws were counted over the subsequent 30 min.
The extracts, fractions, and essential oils were tested in the following doses: EEM and EEC—2000, 1000, and 500 mg kg−1; FHC—400 mg kg−1; DFC—240 mg kg−1; EAFC—1160 mg kg−1; AFC—480 mg kg−1; EOC—60, 200, 600 mg kg−1. Indomethacin was used at a dose of 10 mg kg−1. The control groups received the vehicle used in the solubilisation of each extract, fraction, or essential oil (10 mL kg−1), so there were three control groups that received either 10% propylene glycol in CMC gel, 10% DMSO in water, or only water.
2.6.2. Anti-Inflammatory Activity by Testing Ear Oedema Induced by Croton Oil
Tests of ear oedema induced by croton oil were carried out according to Zanini et al. . Groups of mice () were treated (v.o.) with dexamethasone (2.0 mg kg−1), vehicle (10% propylene glycol in CMC gel, 10 mL kg−1), or EEC (2000, 1000, and 500 mg kg−1). One hour after treatment, each animal received 20 L of a freshly prepared solution of croton oil (2.5% v/v) in acetone on the surface of the right ear. The left ear received the same volume of acetone. After 4 h, the animals were sacrificed and identical segments were taken from both ears. The formations and intensities of the oedema were expressed as the mean of differences in weight between the segments of the animals’ ears: the smaller the weight difference, the greater the potential for inhibition.
2.7. Statistical Analysis
The antinociceptive effects of the different extracts, fractions, and essential oils were expressed as the means (SEM) and are shown as percentages relative to the control group (vehicle). The significant differences between treated and control groups (vehicle) were assessed by an ANOVA and a multiple comparison by Tukey-Kramer. values 0.05 were considered statistically significant. The data analysis used the application GraphPad Prism 3.0.
3. Results and Discussion
3.1. Phytochemical Analysis
The phytochemical analysis of the fractions obtained from the crude ethanol extracts of P. pseudocaryophyllus leaves of the citral and (E)-methyl isoeugenol chemotypes enabled the identification of pentacyclic triterpenes (lupeol, -amyrin, -amyrin, oleanolic acid, betulinic acid, and ursolic acid), flavonoids (quercetin 3-O--L-rhamnopyranoside, quercetin 3-O--glucopyranoside, kaempferol 3-O--L-rhamnopyranoside, quercetin 3-O--arabinofuranoside, quercetin 3-O--arabinopyranoside, quercetin 3-O--arabinopyranoside, and catechin), gallic acid, and ellagic acid (Table 2). These constituents, except ursolic acid, were found for the first time in this species. They were identified based on spectra of 1D and 2D 1H and 13C NMR (HSQC and HMBC) and with comparison with data in the literature (copies of the original spectra can be obtained from the corresponding author). The flavonoid quercitrin was also identified based on the mass spectrum and analysis by HPLC in comparison with an authentic sample.
The results from the qualitative and quantitative analysis of volatile oils of P. pseudocaryophyllus, with the volatile constituents listed in order of elution, are found in Table 3. A total of 31 compounds were identified, accounting for 94–100% of the volatile components. There was a predominance of phenylpropanoid derivatives (97.5%) among the volatile components of the (E)-methyl isoeugenol chemotype, and almost all of the oil consisted of (E)-methyl isoeugenol (93.9%). The essential oil of the citral chemotype consisted mainly of oxygenated mono- and sesquiterpenes (69.65% and 13.7%, resp.), and the monoterpene aldehydes neral and geranial, which are referred to as citral when their isomers are mixed, were the major components (27.59% and 36.49%, resp.).
3.2. Antimicrobial Activity
The MIC values of extracts, fractions, and essential oils, as well as citral (Sigma), are described in Table 4. Table 5 shows the MIC of the semipurified substances and of vancomycin, gentamicin, and itraconazole, which were used as controls. The results were discussed taking into account the classification by Holetz et al. , which has also been adopted by other authors [42–44], where a MIC lower than 100 g mL−1 represents good antimicrobial activity; a MIC from 100 to 500 g mL−1 represents moderate antimicrobial activity; a MIC from 500 to 1000 g mL−1 represents weak activity; a MIC above 1000 g mL−1 suggests that the substance is inactive. EEC showed the lower MIC compared to EEM for Gram-positive bacteria, and both were inactive for Gram-negative bacteria (Table 4). EEC was the most active for fungal strains, with a MIC between 7.8 and 62.5 g mL−1. The lowest MIC of this extract was found for the strains of Cryptococcus (MIC = 7.8–15.6 g mL−1), while EEM showed good-to-moderate activity (MIC = 15.6–125 g mL−1) (Table 4) for the yeasts studied.
Regarding the fractions and semipurified substances, the dichloromethanic fractions of both chemotypes (DFM and DFC) were the most active for Gram-positive bacteria (MIC = 250–500 g mL−1) and were less active for the fungal strains (MIC 500 g mL−1) (Table 4). The antibacterial activity of DFC is due to substance Ppc-2, which had the greatest inhibitory effect on Gram-positive bacteria of the compounds studied (MIC = 31.2 g mL−1 for S. aureus ATCC 6538, S. epidermidis ATCC 12229, and B. cereus ATCC 14579; MIC = 62.5 g mL−1 for S. aureus ATCC 25923, M. roseus ATCC 1740, and B. atrophaeus ATCC 6633; MIC = 125 g mL−1 for M. luteus ATCC 9341) (Table 5). The phytochemical analysis showed that Ppc-2 is oleanolic acid, a pentacyclic triterpene with antibacterial activity [45–47], which was especially effective against Gram-positive bacteria. The TLC analysis of the DFM showed the presence of oleanolic, betulinic, and ursolic acids, which probably contributed to the observed antibacterial activity of this fraction [45–47]. Ppm-4, isolated from EAFM and composed of quercitrin, afzelin, isoquercitrin, avicularin, guaijaverin, catechin, and gallic acid, showed activity against C. neoformans L2 and L1 (CIM = 62.5 g mL−1) but was not active against Candida (Table 5). Studies have shown that catechin and quercitrin, the main components of Ppm-4, inhibit the growth of some pathogenic fungi [48–50]. Ppm-5, obtained from the AFM, showed an activity profile similar to this fraction, except for C. parapsilosis 11A (Table 5). The phytochemical analysis carried out on Ppm-5 verified the presence of gallic acid and its derivatives, which are known for their antimicrobial activities [51–53], as well as the presence of sugars. The Ppc-4 isolated from EAFC, which was composed of quercitrin, showed moderate inhibition of Candida strains (MIC = 250 g mL−1) and good-to-moderate inhibition of the strains of Cryptococcus (MIC = 62.5 to 250 g mL−1) (Table 5), partly contributing to the antifungal activity of this fraction. Ppc-5, which was also isolated from fraction EAFC, consisted of a mixture of flavonoids of which the major component is quercitrin and did not show activity under the conditions tested. Ppc-6 contributed in part to the antifungal activity of AFC, with MICs ranging from 62.5 to 125 g mL−1 for Candida strains (Table 5). The phytochemical analysis of Ppc-6 showed ellagic acid to be the major component. Similar results for Candida spp., due to ellagic acid, were reported by Silva et al. . These findings are promising in the search for new options against infections caused by Candida spp. and Cryptococcus spp., particularly with the continuous increase of opportunistic fungal resistance to available treatments, the emergence of rare species of Candida , and the increasing number of infections by Cryptococcus neoformans and C. gattii .
The essential oil of the citral chemotype (EOC) showed good activity (MIC = 62.5 μg mL−1) against B. cereus ATCC 14579 and moderate activity (MIC = 125–250 mL g g mL−1) against the remaining Gram-positive bacteria; in some cases, this essential oil showed better inhibition than citral (MIC = 125–250 g mL−1) (Table 4). Citral, the major component of the EOC, is a monoterpene aldehyde mixture of the isomers neral and geranial. Aldehydes, such as formaldehyde and glutaraldehyde, are known to have strong antimicrobial activity, and several researchers have demonstrated the antimicrobial effect of citral [57–62]. It is suggested that the aldehyde group conjugated to the carbon-carbon double bond, which is present in neral and geranial, provides a highly electronegative chemical structure that may explain its activity . The EOM was inactive against bacteria under the conditions tested. Its major component, the phenylpropanoid (E)-methyl isoeugenol, comprises almost all of the oil (93.9%). Structurally, (E)-methyl isoeugenol differs from eugenol because it has a methoxyl group at the C1 position of the ring instead of a hydroxyl group, and the double bond in its propenyl group is in a different position. The absence of the hydroxyl group in the phenolic structure of (E)-methyl isoeugenol may have contributed to the lack of activity of this compound. Griffin et al.  observed that the methylation of the hydroxyl group in eugenol, producing methyleugenol, resulted in a loss of activity for Gram-negative bacteria.
The EOC showed good activity against Cryptococcus (MIC = 15.6 g mL−1 to 62.5 g mL−1), but the EOM showed moderate activity against strains of Cryptococcus (MIC = 125 g mL−1 to 250 g mL−1) and showed no activity for species of Candida obtained from clinical isolates (MIC 500 g mL−1) (Table 4). The structure-activity relationship of the components of certain essential oils has been researched in phytopathogenic fungi, and there is little information on fungal pathogens in humans. For example, carbonyl ,-unsaturated compounds, such as the monoterpenic aldehydes neral and geranial, have strong antifungal activity [27, 64, 65], similar to their antibacterial activity. Little information regarding the antifungal mechanism of action of these aldehydes is available. The evaluation of the effects of citral on the membranes, organelles, and intracellular macromolecules of Aspergillus flavus spores has shown that citral damages the cell walls and membranes of spores, reducing their elasticity .
3.3. Antinociceptive and Anti-Inflammatory Screening
The EEM (v.o. 2000, 1000, and 500 mg kg−1) significantly reduced the number of abdominal contortions caused by the intraperitoneal injection of acetic acid in mice compared to animals that received only vehicle (control group) (Figure 1). There was no significant difference observed between the analgesic effects of the three doses used (data not shown).
The EEC (v.o. 2000 and 1000 mg kg−1) was able to inhibit, significantly and in a dose-dependent manner, the abdominal contortions in mice induced by the intraperitoneal acetic acid compared with the control group and showed no significant difference compared to the group of animals treated with indomethacin (10 mg kg−1) (Figure 2).
Given the dose-dependent antinociceptive effect shown by EEC (Figure 2), this extract was selected to perform the screening of the antinociceptive activity of its fractions and essential oils, as well as to evaluate its anti-inflammatory activity by testing ear oedema induced by croton oil. HFC (400 mg kg−1), DFC (240 mg kg−1), and EAFC (1160 mg kg−1) significantly decreased the number of abdominal contortions induced by the intraperitoneal injection acetic acid compared with the control group. However, there was no significant difference in the antinociceptive effect shown by the three fractions. The group of animals that received AFC (480 mg kg−1) showed a number of contortions similar to that of the group that received vehicle (Figure 3).
The EOC at doses of 60, 200, and 600 mg kg−1 (v.o.) showed significant dose-dependent inhibitory effects on the abdominal contortions induced by intraperitoneal acetic acid in mice compared to the control group (Figure 4).
The ear oedema induction test showed that the EEC has antiedematogenic activity that is significantly greater than that of the control at all doses tested (Figure 5). Pretreatment with EEC (2000, 1000, and 500 mg kg−1) reduced the oedema from 14.3 0.4 mg (control) to 10.9 0.5, 11.3 0.4, and 10.5 0.5 mg, respectively.
The results obtained from this study showed that both EEM and EEC have antinociceptive activity. It was not possible to infer whether this action involves peripheral and/or central mechanisms, as the model of abdominal contortions induced by the intraperitoneal injection of acetic acid in mice is sensitive to analgesic substances that act centrally and/or peripherally and that show a wide range of mechanisms of action . Similar results were observed in other species of the genus Pimenta [29, 30, 32].
The mixture of triterpenes lupeol and - and -amyrin isolated from HFC could be accountable for the observed activity. Several in vivo experiments have shown that, among the pharmacological activities attributed to these compounds, analgesic and anti-inflammatory effects are predominant [31, 67–72].
Oleanolic acid, which is widely distributed in the vegetable kingdom, has strong anti-inflammatory effects  that may have contributed to reducing the number of abdominal contortions induced by chemical stimulation in mice, as observed for the DFC.
Data from the literature report the antinociceptive activity of citral [76, 77], a major component of the EOC. Moreover, sedative, anxiolytic, and anticonvulsant effects were observed from the essential oils of Cymbopogon citratus in mice through the use of motor activity tests (“rota-rod” and “open-field”), a hypnosedative activity test (sleep induced by barbiturate), an anxiolytic action test (“plus-maze” and “light-dark box”), and an anticonvulsant action test (seizures chemically induced by pentylenetetrazole) . Therefore, the inhibition by the EOC of the contortions induced by chemical stimulation in mice in this study may be due to both peripheral and central mechanisms. However, the sedative and relaxing effects of citral, as well as the increase in sleep time induced by barbital, especially at high doses (200 mg kg−1) , may have exerted a greater influence on reducing the number of contortions than the analgesic effects themselves.
Until now, no systematic phytochemical and biological study on citral and the (E)-methyl isoeugenol chemotypes of P. pseudocaryophyllus had been reported. Among the isolated substances, oleanolic acid obtained from the dichloromethane fraction of the citral chemotype showed the best profile of antibacterial activity against the Gram-positive microorganisms used in this research (MIC = 31.2 to 125 g mL−1), followed by the essential oil of the citral chemotype, which showed good activity (MIC = 62.5 g g mL−1) against B. cereus ATCC 14579 and moderate activity (MIC = 125–250 g mL−1) against the other Gram-positive microbes.
The extracts, fractions, and essential oils from P. pseudocaryophyllus leaves showed several levels of antifungal activity against Candida spp. and Cryptococcus spp. The antifungal activity shown by the ethyl acetate and aqueous fractions from both chemotypes, especially against Candida albicans (MIC = 31.2 to 62.5 g mL−1) and C. parapsilosis (MIC = 15.6 to 62.5 g mL−1), showed the potential of this species as a source of new antifungal alternatives.
In the models of abdominal contortions induced by acetic acid and ear oedema induced by croton oil in mice, the crude extract of the citral chemotype showed antinociceptive and anti-inflammatory effects, respectively. These effects may be related to the presence of the pentacyclic triterpenes lupeol, -amyrin, and -amyrin and the flavonoids quercetin, quercitrin, and afzelin.
Conflict of Interests
The authors have no conflict of interests to declare.
The authors acknowledge the financial support of the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and Fundação de Amparo à Pesquisa do Estado de Goiás (FAPEG). Thanks are due to MCT/FINEP/CT-INFRA for financial support for Laboratório de Ressonância Nuclear Magnética of the Instituto de Química-UFG.
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domain: biology


Every March 8 International Womens Day is a bit of a blur. There are so many great events celebrating that day and, although I try, I cant get to all of them. I must say that I do wish two somewhat contradictory things that these events were stretched out over a longer period of time and that we wouldnt need to think about a womens day or month because the importance of women is so ingrained in all the work that we do.
Having said that (phew), I did go to an amazing array of events in less than a week: the US State Departments Women of Courage Awards, the International Republican Institutes Womens Democracy Network lunch, the Daily Beast/Newsweek Women of the World Summit, and an event I moderated at the Sewall-Belmont House on women and the Arab Spring.
And, so while overwhelmed by so many courageous women, and two Nobel Prize winners, I was struck by how many of these women were courageous for a deceptively simple act. Women like Zin Mar Aung from Burma, who was imprisoned for 11 years for delivering documents; like Hana El Hebskhi from Libya for documenting the violence and tumult of the Libyan revolution; like Noorjahan Akbar, from Afghanistan, who is a co-founder of a group that started the first womens internet caf in Kabul.
We often see people as courageous in hindsight. One lesson I learned from listening to all of the stories is that we all have the capacity for taking that first courageous step and moving forward to challenge barriers that restrict women and men from fully participating in their families, communities and countries. I hope that we all learn that lesson ourselves and pass it along to everyone we can.


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Hear more about where else you can find fresh oysters in California on an episode of the California Now podcast.
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domain: geography


text:
Want to maximize your gardens full potential? Then you should definitely try your hand at composting. This natural process can be very beneficial to your garden soil and plants, as compost made from organic materials can be made into a rich organic-based fertilizer. So if you happen to be generating a heap load of household wastes in a matter of days, then you should consider doing this process by means of a handy tumbleweed composter. This garden equipment can help you recycle and compost organic wastes in next to no time.
The Tumbleweed Composter
You can enjoy recycling by making good use of the tumbleweed composter. It’s very earth-friendly as it doesn’t require using electricity to make it work. This plastic-made compost bin is easy to look after and very cheap to maintain, as you’ll only be needing a supply of organic wastes from your home. You can easily fill it up with scraps from your kitchen, and of other organic waste materials from your garden.
The process of setting-up the composter
Every newly purchased tumbleweed composting bin comes with an instruction manual. Nonetheless, this compost tumbler is typically assembled in more or less 15 minutes. All you’ll need to start with this set-up is your screwdriver. Aside from the bin and its center axle, it usually comes with a rod and a few tube-shaped legs that are made out of stainless steel material.
Just like any other outdoor composting bin, you can load this composter with organic materials. Amongst the other things that you can throw into the bin are garden wastes and kitchen scraps. These organic wastes can be in the form of dried leaves (brown or green leaves can also be composted separately), grass clippings, presoaked newspaper strips or shreds, coffee grounds, crushed egg shells, fruit peels, and uncooked vegetable parts.
The spinning composter
This composting bin works through spinning. By pushing it down, the contents of the tumbler gets mixed and turned consistently (gets aerated). So rather then doing the mixing and turning manually by hand, the tumbleweed does the majority of the work for you. The breaking down of the organic materials can be expected to pass in results a lot faster than expected. By giving it a regular spin and maintenance, you can possibly produce a supply of rich compost in less than a month.
Composting indoors with the Tumbleweed Composter
If you happen to have no garden space to place a composter such as the Tumbleweed, then consider placing it inside your house. You can probably set it up by your balcony, inside your garage, or inside your basement. It can definitely be made into an indoor composting bin despite the space constraint. Other than that, the tumbleweed composter is also equipped to be pest proof. So securing your compost, and keeping it unspoiled is something that you can count on.
Uncle Jim’s recommends the Tumble Weed Composter
Reap the benefits of composting by using our Tumble Weed Composter. It’s made out of a UV-protected polypropylene plastic material that comes with a galvanized steel tube, stainless steel central breaker bars, and zinc plated steel fasteners. Order yours today to experience this equipments full potential.

domain: agronomy


Poll: Majority of Americans Say Religion Is Answer to Today's Problems
A new Gallup poll revealed that a small percentage of Americans consider religion old fashioned while the majority of Americans think it can actually solve today's world problems.
The recent poll asked over 1,000 adults about their views on the topic and found that 57 percent of individuals surveyed are optimistic about religion and only 30 percent find it to be out of date.
"The majority of Americans continue to believe that religion can answer today's problems, another indicator that the nation, by far, remains a religious country," said Frank Newport, Gallup's editor-in-chief, in a statement. "And, with the trend leveling off in recent years, it appears this aspect of the secularization of U.S. society may have slowed, if not halted, for the foreseeable future."
Despite the high percentage, the 57 percent figure reflects a decline over the last 40 years in the number of Americans who believe religion is the answer.
In 1957, 82 percent of Americans believed in religion's ability to solve today's problems but that number dropped to 62 percent when Gallup asked the same question in the 1970s. There was an increase in 2000 to 68 percent but that number continued to decline over the last 14 years.
"The 82 percent choosing the 'can answer today's problems' options in 1957 is in line with a number of other measures from that decade showing a high level of religiosity, including religious service attendance, importance of religion, and the percentage of Americans with a formal religious identity," said Newport.
Meanwhile, the percentage of Americans who believe religion is old fashioned and out of date has continued to increase since 2000 when it was 19 percent.
People over age 65, women, political conservatives, and those living in the South are found to be among the most religious groups, based on measures of church attendance and importance of religion, and are the most likely to consider religion as an answer to today's questions. However, younger, politically liberal individuals living in the Eastern part of the country are more likely to consider religion outdated.
The Gallup poll surveyed 1,028 adults in 50 states and the District of Columbia, May 8-11, 2014. The margin of error is plus or minus 5 percentage points.


After having restricted Zimbabwe to 151-9 in their 20 overs, the Aussies chased down the total with one ball to spare to complete their third win in four matches in the tournament.
Glenn Maxwell was the hero with the bat for Australia with the right-handed batsman slamming a 36-ball 58 in his side’s successful run chase. Maxwell’s crucial innings included five sixes and a boundary.
Earlier, Solomon Mire was in good touch with the bat once again for the hosts after his career-best 94 in Zimbabwe’s previous loss to Pakistan. The opener struck a 52-ball 63 on Friday to lead his side’s charge.
However, three wickets for seamer Andrew Tye and two-a-piece for Billy Stanlake and Jhy Richardson ensured that Zimbabwe could only muster 151-9 after electing to bat first.
Australia’s chase got off to the worst possible start with skipper Aaron Finch and Alex Carey dismissed inside the first five overs. Maxwell then took control of the chase with his counter-attacking innings but his dismissal at the hands of Blessing Muzarabani triggered a late collapse for the Aussies who lost three further wickets in as many overs following his departure.
It took an unbeaten cameo of 12 (7) from all-rounder Marcus Stoinis to get the Aussies over the finish line in the penultimate delivery of the match.
Australia and Pakistan will now lock horns in the final of the tri-series to be held on Sunday.
Know more about Sport360 Application


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Food nerds get a bit wiggly in the knees when you mention the Fancy Food Show — two gut-busting events held each year (in NY and SF) that have a major impact on what turns up on your local store shelves. Call it a crystal ball to your food future.
Hundreds of purveyors — small and large — from around the globe cram into the Moscone Center hawking the latest and greatest specialty products. At the West Coast show, there’s a dense concentration of artisan purveyors from Northern California and specifically Sonoma County.
It’s easy to see the obvious…lots of chocolate, tea and BBQ sauce. What’s not so obvious are subtle changes in what’s emerging–the little guys who are latching onto the next cultural zeitgeist.
Stay tuned for my interviews with some new faces on the SoCo scene, but meantime, here’s a peek at what’s on the rise, on the wane…and just plain weird.
(Keep in mind that some of these products aren’t yet on the market
and may not be released until later this year, but whenever possible
I’ve added local retailers who carry the brands. If you see them, keep
us informed in the Retail Spy Forum).
Biggest Star: Glammed up water and soda
If (like me) you’re addicted to the fizzy stuff and hate plain old
water, 2009 is your year to be better hydrated. The hottest new food
isn’t food at all, but gourmet-flavored beverages. No. Not Vitamin Water, but really swanky small-production flavors that are less about sweet and more about flavor. Take a gander at
these and throw your 20-ouncer out the window.
- Honeydew Hibiscus, Pear, Cucumber water/Hint Water
Chocolate Mint Water/MetroMint (Olivers)
Wine grape soda/Vignette Wine Country Soda (Whole Foods)
- Juniper Berry Soda/Dry Soda (Whole Foods, Bevmo, Rosso, Sea Thai)
Red Hill Pomegranate, Yakima Valley Grape and Costa Rican
Pineapple sparkling water/Fizzy Lizzy(Whole Foods,
Oliver’s, Girl and the Fig)
Victorian-inspired flavors of Dandelion & Burdock, Ginger Beer
and Shandy/Fentiman’s (Canada, not yet available in California)
Also Rising: Yogurt/Probiotics
Also hot are foods that do something good for your body. Yogurt is on
the rise as one of nature’s true miracle foods, knocking out the
short-lived goji insanity.
Lebanese yogurt gelato/Ciao Bella Gelato (Whole Foods)
Straus Creamery Organic Yogurt (Whole Foods, Pacific Market)
- Bellwether Farms Sheep milk Yogurt (Whole Foods, Pacific Market)
- Yacult, a fermented pro-biotic dairy drink from Japan (FoodMaxx, Lucky)
Alternative sweets & regional honey
Refined sugars are out and alternative sweetners like Stevia, honey and
agave are very in. Double points if you add something healthy like acai.
Lemon, lime and raspberry flavored cane sugar/Essential Cane
Q Tonic water sweetened with Agave (Bevmo)
Tupelo Honey/Savannah Bee
Pomegranate and Blueberry Acai Gummy Pandas/Bissinger
Ethnic foods are hot, but nothing is hotter than curry-mania. From
microwavable tikka massala to curry-spiked ice cream, stay tuned for
the Bollywoodization of your palate.
- Naga Sweet Indian curry,coconut and custard ice cream/Vosges Haut Chocolat (Dean and Deluca)
Ancient Organics Ghee
Sukhi’s Gourmet Indian Foods(Whole Foods, Oliver’s)
Also rising: Flavored nuts, foods with aloe as an ingredient and anything gluten-free.
Can you ever get too much of a good thing? Uh, yes. The following have hit the point where fascinating contenders may emerge, but copy-cats are starting to ruin the experience.
The New Coffee: Tea
From the hordes of tea-sellers on the floor, the gourmet tea leaf
market seems to be hitting its saturation point. One highlight from
Novato’s Republic of Tea are the Little Citizens’ Herb Teas, a line of
caffeine free organic teas flavored with strawberry, tangerine and
cherry. Get ’em young.
Everyone and their sister has started a line of fair trade, organic,
bittersweet, artisan chocolates. Enough already. Chocolate is delish, but just because you CAN make it doesn’t mean you should. Scariest contenders:
Enchanted Mushroom chocolate bar from Vosges and Dark
Chocolate/Jalapeno gelato from Ciao Bella Gelato. Hey, at least they
Oddly entertaining (or just odd)
Organic Gin from Juniper Green
Fartless Factory Bean Soups
- Sophia Loren pasta sauce in the same booth as NASCAR popcorn
John Wayne Stock & Supply Beef Jerky
A 9-year-old Australian Rock Lobster the size of a small car

domain: food


text:
Ph.D., 1975, Pennsylvania State University, The Milton S. Hershey College of Medicine
Mechanisms of Gene Induction by Peroxisome Proliferators
Dr. Geoghegan's lab is studying the effects of DHEA on gene expression. DHEA is a precursor for the gonadal steroids, but has independent effects as a modulator of gene expression. Dr. Geoghegan collaborates with Dr. Russ Prough who has demonstrated that DHEA is a peroxisome proliferator in rodents. Peroxisome proliferation involves induction of a number of genes associated with peroxisomal fatty acid oxidation as well as microsomal P450s involved in omega hydroxylation of fatty acids. These effects are mediated by activation of PPAR-alpha, a member of the nuclear hormone superfamily of transcription factors. We have also demonstrated that DHEA induces expression of cytochrome P450 3A, a major inducible drug metabolizing enzyme in the liver. This occurs by mechanisms that appear to be independent of PPAR. Using gene array analysis we have recently identified a number of other novel genes whose expression is affected in the liver by treatment with DHEA. Ultimately, identification of these genes and an understanding of how they are regulated , will help to elucidate the myriad physiological and biochemical affects associated with DHEA.
Webb SJ, Geoghegan TE, Prough RA, Michael Miller KK. (2006)The biological actions of dehydroepiandrosterone involves multiple receptors. Drug Metab Rev. 2006;38(1-2):89-116. Review.
S Gu, SL Ripp, RA Prough, and TE Geoghegan. Dehydroepiandrosterone Affects theExpression of Multiple Genes in Rat Liver Including 11b–Hydroxysteroid Dehydrogenase Type I: A cDNA Array Analysis. Mol. Pharm. 63, 722-731, (2003).
Parimal Bhattacherjee, P Mukhopadhyay, SL Tilley, BH Koller, T.Geoghgan and CA Paterson. Blood-Aqueous Barrier in Prostaglandin EP2 Receptor Knockout Mice. J. Ocular Immunology and Inflamation.10; 187-96 (2002)
Regulation of the rat glutathione S-transferase A2 gene by glucocorticoids: involvement of both the glucocorticoid and pregnane X receptors. Cameron K. Falkner, J.A. Pinaire, G-H. Xiao, T.E. Geoghegan and R.A. Prough.(2001) Mol. Pharm. 60, 611-619.
Cytochrome P-450 mRNAs are modulated by dehydroepiandrosterone, nafenopin, and triiodothyronine. Singleton DW, Lei XD, Webb SJ, Prough RA, Geoghegan TE. (1999) Drug Metab Dispos. 27:193-200.
Expression of prostaglandin receptors EP4 and FP in human lens epithelial cells. (1999). Mukhopadhyay P, Bhattacherjee P, Andom T, Geoghegan TE, Andley UP, Paterson CA. Invest Ophthalmol Vis Sci 1999 Jan;40(1):105-12.
Detection of EP2, EP4, and FP Receptors in Human Ciliary Epithelial and Ciliary Muscle Cells. (1997). Mukhopadhyay, P., T. E. Geoghegan, R. V. Patil, P. Bhattacherjee and C.A. Paterson. Biochemical Pharmacology 53: 1249-1255.
Regulation of CTP4A Expression in Rat by Dehydroepiandrosterone and Thyroid Hormone. (1996). S.J. Webb, G.-H. Xiao, T. E. Geoghegan, and R.A. Prough. Molecular Pharmacology 49, 276-287.
Regulation of Cytochromes P450 by DHEA and Its Anticarcinogenic Action. Prough, R.A., X.-D. Lei, G.-H. Xiao, H.-Q. Wu, T. E. Geoghegan, and S.J. Webb (1995) Ann New York Acad Sci 774, 187-199.


Baht slumps as economy in recession
- Published: 20/08/2013 at 11:22 AM
- Online news:
Thailand’s baht slumped to a one- year low after Southeast Asia’s second-largest economy entered a recession for the first time since 2009. Government bonds were little changed.
Gross domestic product unexpectedly decreased 0.3% in the three months through June from the previous quarter, when it contracted a revised 1.7%, the National Economic and Social Development Board said Monday.
The agency cut its 2013 expansion forecast to 3.8% to 4.3% from 4.2% to 5.2%. The MSCI Asia-Pacific Index of shares fell for a fourth day amid speculation the US Federal Reserve will cut stimulus that inflated emerging-market asset prices.
“Risk sentiment is hurt due to declines in stocks,” said Hideki Hayashi, a researcher at the Japan Center for Economic Research in Tokyo. “Amid weak risk appetite, growth concern is further weighing on the currency.”
The baht slumped 0.9%, the most since June 20, to 31.66 per dollar as of 9.07am in Bangkok, according to data compiled by Bloomberg. It reached 31.76 earlier, the weakest level since July 25, 2012. One-month implied volatility, a measure of expected moves in the exchange rate used to price options, climbed 19 basis points, or 0.19 percentage point, to 6.37%.
The Bank of Thailand will keep its benchmark interest rate at 2.5% on Wednesday, according to all 20 economists surveyed by Bloomberg.
BNP Paribas SA cut its 2013 growth forecast for Thailand to 3% from 4.5%, Philip McNicholas, a senior economist in Hong Kong, wrote in a research note dated Tuesday.
The yield on the 3.625% sovereign notes due June 2023 was steady at 3.98%, data compiled by Bloomberg show.
About the author
Writer: Bloomberg News


|Publication number||US8096077 B1|
|Application number||US 12/386,069|
|Publication date||Jan 17, 2012|
|Filing date||Apr 13, 2009|
|Priority date||Apr 13, 2009|
|Publication number||12386069, 386069, US 8096077 B1, US 8096077B1, US-B1-8096077, US8096077 B1, US8096077B1|
|Inventors||Clayton R. Caywood|
|Original Assignee||Caywood Clayton R|
|Export Citation||BiBTeX, EndNote, RefMan|
|Patent Citations (20), Referenced by (9), Classifications (10), Legal Events (3)|
|External Links: USPTO, USPTO Assignment, Espacenet|
(a) Field of the Invention
This invention relates to a rifle recoil absorbing strap assembly for holding a rifle, a pistol and other types of firearms thereon during long range, target shooting, distance sighting, hunting and other firing activities and more particularly, but not by way of limitation, to a rifle recoil absorbing strap assembly that is lightweight, portable and versatile. The strap assembly is adapted to be quickly attached to a target shooting bench, a tree trunk and branch, a hunting blind, a vehicle tailgate, a off-highway vehicle cargo rack, a fence post, on top of a ground surface, a monopod, a bipod, a tripod and other similar permanent and non-permanent rifle support objects when firing a high powered rifle, pistol or any firing mechanism.
(b) Discussion of Prior Art
Heretofore, there have been a variety of rifle recoil absorbing devices, recoil suppressing supports, and methods of attenuating firearm recoil. In U.S. Pat. Nos. 2,729,975 to Hawthorne et al. and 7,313,884 to Eddins, two different types of recoil suppressing rifle supports are disclosed and shown permanently mounted on support frames. Also, U.S. Pat. Nos. 4,799,329 to Nodo and 5,811,720 to Quinnell et al. illustrate a firearm recoil attenuator and a shooting rest with recoil reduction system mounted permanently on a support member. In U.S. Pat. No. 5,758,447 to Venetz, a recoil absorbing device is described having a rifle rest member permanently attached to a horizontal surface and a first strap member with weighted bag and a second strap member used to hold a rifle butt in place during the firing of the rifle.
None of the above mentioned patented recoil absorbing devices and systems disclose the unique features and structure of the subject flexible strap assembly that can be quickly attached to various objects for absorbing rifle recoil when firing a high powered rifle.
In view of the foregoing, it is a primary objective of the subject invention to provide a rifle recoil absorbing strap assembly for absorbing rifle recoil, preventing bruising of the shoulder and without having to permanently mount the strap assembly on a target shooting bench, a table or a horizontal support member.
Another object of the strap assembly is the invention is rugged in construction, simplistic in design, adapted for different sizes and shapes of rifles, pistols, and other firearms, and can be easily and quickly mounted on different types of horizontal and vertical support objects such as a target shooting bench, a hunting blind, a tree trunk and branch, a vehicle tailgate, an off-highway vehicle cargo rack, a fence post, on a ground surface, a monopod, bipod, tripod and other support objects.
Yet another object of the invention is the strap assembly includes a rifle butt cradle, made of strap material, for holding a rifle butt securely therein. The rifle butt cradle includes a horizontal, reinforcing member in the bottom of the butt cradle to prevent the strap assembly from slipping and folding over an edge of a target shooting bench and other horizontal surfaces when attached thereto.
The subject strap assembly is adapted for engaging and holding a rifle in place during target shooting and hunting. The strap assembly includes an elongated, flexible, horizontal first strap member with adjustable and releasable fastener. The adjustable and releasable fastener is used for securing together a first end and a second end of the first strap member. Also, the strap assembly includes a flexible, vertical second strap member with a first end and an opposite second end. The first and second end of the second strap member are adapted for receipt around a front portion of the rifle and secured thereto for preventing upward movement of the rifle when firing. Further, the strap assembly includes a rifle butt cradle adapted for receiving and securing a rifle butt of the rifle therein. Whereby, when the strap assembly is secured to a rifle securing object and the rifle is held thereon, the strap assembly greatly reduces rifle recoil during the firing of the rifle.
These and other objects of the present invention will become apparent to those familiar with rifle recoil absorbing devices and methods when reviewing the following detailed description, showing novel construction, combination, and elements as herein described, and more particularly defined by the claims, it being understood that changes in the embodiments to the herein disclosed invention are meant to be included as coming within the scope of the claims, except insofar as they may be precluded by the prior art.
The accompanying drawings illustrate complete preferred embodiments of the present invention according to the best modes presently devised and in which:
The strap assembly 10 includes an elongated, flexible, horizontal first strap member 24 with adjustable and releasable fastener 26. The adjustable and releasable fastener 26 is used for securing together a first end 28 and a second end 30 of the first strap member 24. In this drawing, the first strap member 24 is shown looped around the top and bottom of the bench 12 and tightened thereon using the adjustable and releasable fastener 26. The fastener 26 can also be used as a grommet for securing the strap assembly 10 to a support object.
Also, the strap assembly 10 includes a flexible, vertical second strap member 32 with a first end 34 having loop fasteners 36 and a second end 38 having hook fasteners 40. The first end 34 with loop fasteners 36 and the second end 38 with hook fasteners 40 are shown clearly in
Further, the strap assembly 10 includes a rifle butt cradle 44, make of strap material, adapted for receiving and securing the rifle butt 22 of the rifle 16 therein. As mentioned above, the strap assembly 10 can be secured to various types of permanent or moveable rifle securing objects with the rifle 16 held thereon. The rifle butt cradle 44 can include a foam shoulder pad 46 to further reduce any chance of shoulder bruising, when using the strap assembly 10.
Also in this drawing, the rifle butt cradle 44 is shown in greater detail and including a horizontal reinforcing member 48, shown in dashed lines, received in a horizontal sleeve 50 in a horizontal bottom portion 52 of the cradle 44. The cradle 44 also includes a “U” shaped vertical portion 54 and angled support straps 56. The “U” shaped vertical portion 54 of the cradle 44 is sized to receive different sizes of rifle butts. Further, a top of the vertical portion 54 includes a flap 58 with loop fasteners 36 for securing the flap to hook fasteners 40 on one side of the vertical portion 54 and holding the rifle butt 22 securely when firing the rifle 16. The cradle 44 along with the flap 58 can encompass the rifle butt 22 when using the strap assembly 10 with or without using the bench 12, as shown in
In this drawing, the first strap member 24 includes the grommet 68 positioned for being below the front portion 42 of the rifle 16 as shown in
While the invention has been particularly shown, described and illustrated in detail with reference to the preferred embodiments and modifications thereof, it should be understood by those skilled in the art that equivalent changes in form and detail may be made therein without departing from the true spirit and scope of the invention as claimed except as precluded by the prior art.
|Cited Patent||Filing date||Publication date||Applicant||Title|
|US3501074 *||May 31, 1968||Mar 17, 1970||Robert David Emerick||Gun carrier|
|US4012860 *||May 28, 1975||Mar 22, 1977||Auger Arnold J||Adjustable rifle rest|
|US5070636 *||Jul 13, 1990||Dec 10, 1991||Evan Mueller||Gun sighting device|
|US5669170 *||Jul 26, 1996||Sep 23, 1997||Norris; Terry Bruce||Hands-free sling for carrying a long gun or other elongated article|
|US5682700 *||Jan 24, 1997||Nov 4, 1997||Sandberg; Steven Robert||Rifle cleaning gradle|
|US5685103 *||Apr 19, 1996||Nov 11, 1997||Charles Eric Rabon||Gun support|
|US5758447 *||Apr 1, 1997||Jun 2, 1998||Venetz; Louis M.||Recoil absorbing device and method|
|US5811720 *||Jun 16, 1997||Sep 22, 1998||Quinnell; Glenn D.||Shooting rest with recoil reduction system|
|US5881487 *||Aug 3, 1998||Mar 16, 1999||Chalker; Dennis||Multi-purpose, multi-weapon tactical sling/harness|
|US5913668 *||Jan 2, 1998||Jun 22, 1999||Messer; Jerry Wayne||Weapon rest|
|US6266910 *||Sep 17, 1999||Jul 31, 2001||Ray L. Horine||Lap weapon holder|
|US7823318 *||Nov 12, 2008||Nov 2, 2010||Hall Thomas W||Rifle recoil absorption system|
|US20060137233 *||Dec 29, 2004||Jun 29, 2006||Paul Meeks||Stabilizing apparatus and method|
|US20060230664 *||Apr 18, 2005||Oct 19, 2006||Wallace Eddins||Recoil suppressing gun support|
|US20070278261 *||May 31, 2006||Dec 6, 2007||Gallagher Richard N||Rifle sling with stabilizing loop for elbow|
|US20070278262 *||May 31, 2006||Dec 6, 2007||Gallagher Richard N||Sling for automatic rifle to prevent injury by accidental discharge|
|US20080086928 *||Oct 12, 2006||Apr 17, 2008||Pereksta Michael E||Gun support for hunters|
|US20090025267 *||Jul 26, 2007||Jan 29, 2009||Alliant Techsystems Inc.||Firearm Rest|
|US20090049731 *||Aug 23, 2007||Feb 26, 2009||Jo Won Seuk||Firearm support device|
|US20100018102 *||Jun 5, 2009||Jan 28, 2010||Duffy's Hunting Products||Tree mounted apparatus|
|Citing Patent||Filing date||Publication date||Applicant||Title|
|US9163893 *||May 19, 2014||Oct 20, 2015||Tommy Joe Gutierrez||Adjustable rifle support|
|US9250034 *||Mar 15, 2013||Feb 2, 2016||Evergreen Asset Management, Llc||Multipurpose firearm securing systems, kits and methodologies|
|US9546839 *||May 31, 2016||Jan 17, 2017||Precision Accuracy Solutions, Inc.||Shooting rest adapted for mimicking hand-held shooting|
|US9618292 *||Dec 7, 2016||Apr 11, 2017||Precision Accuracy Solutions, Inc.||Shooting rest adapted for mimicking hand-held shooting|
|US9702653 *||Oct 9, 2015||Jul 11, 2017||Battenfeld Technologies, Inc.||Firearm shooting rest|
|US9733036||Dec 10, 2013||Aug 15, 2017||Evergreen Asset Management, Llc||Multipurpose securing systems, kits and methodologies|
|US9797673||Sep 30, 2016||Oct 24, 2017||Tommy J Gutierrez||Window-mount firearm rest|
|US20120285062 *||May 14, 2012||Nov 15, 2012||Cama Mark R||Recoil reduction and sighting-in system for a firearm|
|US20140157643 *||Mar 15, 2013||Jun 12, 2014||Evergreen Asset Management, Llc||Multipurpose firearm securing systems, kits and methodologies|
|U.S. Classification||42/94, 224/150, 73/167|
|Cooperative Classification||F41C33/005, F41A23/005, F41A23/16|
|European Classification||F41A23/16, F41A23/00B, F41C33/00F2|
|Aug 28, 2015||REMI||Maintenance fee reminder mailed|
|Jan 17, 2016||LAPS||Lapse for failure to pay maintenance fees|
|Mar 8, 2016||FP||Expired due to failure to pay maintenance fee|
Effective date: 20160117


text:
Hot pear cider, apple and cranberry juice plus sloe gin with cinnamon - you can't get a better winter cocktail than this
Put all the ingredients into your biggest saucepan or casserole dish. When you’re ready to serve, heat to just below simmering point, then ladle into glasses.
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text:
There is much lore and legendary tradition to hiking the Appalachian Trail, including the so-called "Maryland Challenge:" hiking the Maryland section of the Appalachian Trail in ONE DAY! You may miss some things hurrying to meet the Maryland Challenge, so this virtual hike can be done in less than one day and still see plenty of the special sites available in Maryland.
Join me hiking from the Mason-Dixon Line to Harper’s Ferry, or the other way if you prefer, and see all the sites in a much more relaxed and comfortable way.
Visit the Appalachian Trail, starting from any point in the listing below, and virtually hike from place to place — click on the up arrow () to hike toward Maine, or click on the down arrow () to hike toward Georgia; the up and down arrows are located at the left side of each Appalachian Trail page.
Below is a listing of places you can visit that correspond with places on the map marked by large red dots.
The Catoctin Mountain Range in Maryland, which is one of the most wonderful recreational treasures within driving distance of Washington, DC. People in the area of course use it for hiking, but also in certain places for camping, for trail-biking, for fishing, and even for boating and swimming. The southern end of this mountain range begins in the south just west of Frederick, MD, and runs about 30 miles north, roughly parallel to South Mountain, which is more to the west, and roughly parallel to MD-15, which is just east of it. On both sides it is surrounded by productive farm land. Over its length, it comprises a ridge that dips a few times into passes. It runs through four parks, as follows from north to south:
Gambrill State Park, which includes several well blazed looping trails in addition to the non-looping Catoctin Trail. The looping feature of these trails is nice … you can park at the trailhead, and follow the loop trails right back to the trailhead. Some of the trails are limited to foot traffic, while others also offer the excitement of mountain biking. [Click here to see the sites in Gambrill State Park].
Strictly speaking, this mountain range does continue south into Virginia, but at much lower elevations and with fewer notable hiking opportunities to explore.
The Catoctin Trail … Each of the four separate parks include many hiking trails. Additionally, there is one long trail, which is a wonderful resource, the Catoctin Trail, maintained by the Potomac Appalachian Trail Club (PATC). This trail is 26.6 miles long, and the northern end is just three miles from the Appalachian Trail on South Mountain. In addition to hiking and mountain biking, the trail is also the site for the annual 50K Catoctin Trail Run, which runs out-and-back from the Gambrill Tea Room to the Manor Area Visitors Center.
This photo journey provides you with an opportunity to virtually “hike” this trail in either direction. [Click here to see the sites and virtually hike the Catoctin Trail].
Maryland is sometimes called “America in Miniature,” because nearly every sort of geography found in America, except deserts, can also be found in Maryland, although not always on the same scale. There are many historic sites, and some very pleasant hiking opportunities. Come join me on some Maryland photo journeys not far from our Nation’s Capital, Washington, DC.


Saturday, 1 November 2008
Fall colours: the Carolina in the Fall stage at night
Thanks to Lilly Pavlak of Switzerland and the Czech Republic for these photos (and many others) and a full report on the Carolina in the Fall Festival, held two weeks ago at Shepherds Farm, North Wilkesboro, North Carolina, USA, at which the main guests were the Kruger Brothers, together with many local and national musicians including stars of the calibre of Bobby Hicks, Terry Baucom, and Ron Block.
9-year-old Samantha Snyder plays fiddle with Bobby Hicks and Jens Kruger; her 12-year-old brother Zeb is on guitar
The 'Uwe Kruger dream band': Bobby Hicks, Ron Block, Joel Landsberg, Adam Steffey, Uwe Kruger
The full text of Lilly's report can be read here.


Very-high-resolution tandem Fabry-Perot etalon cylindrical beam volume hologram spectrometer for diffuse source spectroscopy.
We demonstrate a compact and slitless spectrometer with high resolution formed by cascading a Fabry-Perot etalon (FPE) and a cylindrical beam volume hologram (CBVH). The most significant advantage of this combined spectrometer is that we can independently encode spectral information of a diffuse beam in a 2D plane. Also, we show that in this slitless configuration we can simultaneously benefit from the advantages of both elements: the high resolution of the FPE and the large spectral range of the CBVH. Here, we report on the experimental demonstration of a spectrometer with better than 0.2 nm resolution.
Badieirostami, M; Momtahan, O; Hsieh, C; Adibi, A; Brady, DJ
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Post-print of: Johnston, SG, Slavich, PG & Hirst, P 2005, 'Opening floodgates in coastal floodplain drains: effects on tidal forcing and lateral transport of solutes in adjacent groundwater', Agricultural Water Management, vol. 74, no. 1, pp. 23-46.
Agricultural Water Management home page available at www.elsevier.com/locate/agwat
Publisher's version of article available at http://dx.doi.org/10.1016/j.agwat.2004.10.010
The effects of opening tidal barriers (floodgates) upon water table levels and lateral transport of solutes adjacent drains was investigated at two sites on a coastal floodplain. The sites had contrasting geomorphology, soil texture and sediment hydraulic properties. The site with lower hydraulic conductivity (0.3–0.9 m day−1) soils (Romiaka) also had a higher elevation and hydraulic gradients towards the drain. While floodgate opening at Romiaka enhanced the amplitude of pre-existing tidal interaction with adjacent shallow groundwater, altered hydraulic gradients and caused some salt seepage, lateral solute movement from the drain was highly attenuated (<10>m). The site with very high hydraulic conductivity soils (Shark Creek; >125 m day−1) had a lower elevation and seasonally fluctuating hydraulic gradients. The introduction of a tidal pressure signal into the drain by opening the floodgate at Shark Creek caused tidal forcing of groundwater over 300 m from the drain. Floodgate opening at this site also caused changes in groundwater hydraulic gradients, leading to incursion of saline drain water into shallow groundwater over 80 m from the drain. Lateral movement of solutes was relatively rapid, due to macropore flow in oxidised acid sulfate soil horizons, and caused substantial changes to shallow groundwater chemical composition. Conversely, when groundwater hydraulic gradients were towards the drain at this site there was substantial lateral outflow of acid groundwater into drains. This study highlights the importance of assessing the hydraulic properties of soils next to drains on coastal floodplains prior to opening floodgates, particularly in acid sulfate soil backswamps, in order to prevent unintended saline intrusion into shallow groundwater.


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Analysis of Mercury adsorption onto activated Carbon originating from cow bone.
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This project was aimed at providing a solution to significant amounts of mercury released during laboratory work through laboratory wastewater drainages to the environment. This project was however limited to studying the effectiveness of activated carbon obtained from cow bone in removing a known concentration of mercury from the waste water. The carbon was obtained from burning the bones in limited oxygen and later activated using orthophosphoric acid. The activated carbon was then soaked in this wastewater overnight to adsorb the mercury onto the activated carbon surface. The analysis was carried out using an Inductively Coupled Plasma Mass Spectrometer (ICP-MS). This implied that the cow bone-activated carbon was able to remove up to 99.9969% of the mercury concentration within this water. It was therefore concluded that activated carbon from cow bone is an alternative and environmentally friendly method of reducing pollution due to mercury.


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Microdermabrasion is an effective skin care treatment that helps you to get rid of acne, discoloration, and clogged pores. It is a safe treatment that can be used by people of all ages, even those with sensitive skin.
Removing Dead Skin Cells
Microdermabrasion is a safe, non-surgical procedure that can remove dead skin cells and resurface your skin. It can also stimulate the production of collagen and elastin, which keep your skin healthy and youthful.
Removing dead skin will allow your skincare products to penetrate the surface of your skin more deeply. This will also increase your body’s cellular turnover and help remove toxins. Ultimately, your new, healthier skin will be more vibrant and smooth. Several microdermabrasion benefits include a brighter complexion, improved elasticity, and increased absorption of skincare products. Those who are interested in a microdermabrasion Virginia Beach treatment should consult with their doctor. They may recommend a series … Read the rest


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Lindough Premieres Nomalizo Ft. DJ Catzico
Over a week ago, Lindough announced the imminence of his song “Nomalizo” with DJ Catzico as the sole guest. He had urged fans to look forward to the song on 23 October. That day is here, and the song is here as well.
Well, we can assure you the wait for “Nomalizo” hasn’t been a waste after all. The song is a lyrical charmer and sure to get you vibing immediately. The song should resonate with just about any music lover out there.
Lindough has been on Mzansi’smusic horizon for a while during which time he has limned several fine numbers. He continues his fine offerings with the song in review. “Nomalizo” is the ideal number for a fine jig. Alone or in the company of a lover, you’ll surely love this song.
A fine artiste himself, Catzico is one of the Rainbow Nation’s fine DJ hands. The disc jockey and producer is particular famous as a serial collaborator with DJ Vista. The pair released “Ama Snax” with uBiza Wethu and Mr Thela in August.
Well, how would you rate the song? You might want to stream it below and join the conversation by dropping your thoughts in the comment section.
Nomalizo (feat. Dj Catzico) – Single
- Genre: Afro House
- Music Official Release Date: 2020-10-23
- Explicitness: notExplicit
- Number Of Tracks: 1

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