The histories were randomly selected, and comprised a broad crosssection

of patients, including those with moderate to severe cognitive and communication deficits who are often underrepresented in the literature (Macrae and Douglas 2008). Our findings may therefore be generalised to similar cohorts with due considerations to the study’s limitations. The study was a retrospective audit that relied on clinical documentation. However, compliance with documentation was found to be good, and the assessments were conducted in a standardised manner by trained therapists. It was likely that the broad approach taken to audit each history captured the majority of complaints of shoulder pain. For instance, the notes covered the 24-hour period Talazoparib and were written by staff who worked closely with each patient doing tasks requiring shoulder function. Nevertheless, the audit did not collate important aspects such as severity and nature of shoulder pain, nor did it attempt to evaluate management processes or treatment outcome. The observational study supports that post-stroke shoulder pain is common, and more likely to occur in CX-5461 in vivo patients

who have stiff and weak shoulders. Ethics: The study was approved by the Human Research and Ethics Committee at Austin Health (No H2008/03389). We are grateful to Associate Professor Leonid Churilov from the National Stroke Research Institute for statistical advice and guidance; to physiotherapists and occupational therapists from the neurology units at Austin Health-Royal Talbot Rehabilitation Centre, and to undergraduate physiotherapists undertaking a professional development elective from the University of Melbourne who assisted with data collection and management for the project; and the Health Information Management staff for supporting this project. “
“Summary of: Liu-Ambrose T, Nagamatsu LS, Graf P, Beattie BL, Ashe MC, Handy TC (2010) Resistance training and executive functions: a 12-month randomized nearly controlled trial.Arch Intern Med 170: 170–178. [Prepared by Nicholas Taylor, CAP

Co-ordinator.] Question: Does resistance training improve cognitive function in older women living in the community? Design: Randomised controlled trial with concealed allocation and blinded outcome assessment. Setting: A local fitness centre and research centre in Canada. Participants: Women aged 65 to 75 years living independently in the community and with a Mini-Mental state examination score of at least 24 were included. Having a medical condition for which exercise was contraindicated, participating in resistance training in the last 6 months, and having depression were exclusion criteria. Randomisation of 155 participants allocated 52 to once-weekly resistance training (1RT), 54 to twice-weekly resistance training (2RT), and 49 to twice-weekly balance and tone exercises (BAT).

It was centrifuged at low speed to clarify the extract. The supernatant corresponding to the concentration of 20 mg/20 μl was used for the assay. Zea mays leaves (1.0 g) were homogenized in approximately 1 ml of the solvents (methanol/chloroform). INCB024360 order The supernatant was collected and dried at 60 °C well protected from light. The residue obtained after drying the chloroform and methanol extracts were weighed and dissolved in a known amount of DMSO to yield a concentration of 20 mg/5 μl, DMSO was maintained at a minimum level to avoid DMSO-induced events, if any. Fibroblast cells were isolated from chick embryo and were cultured using Dulbeccos modified Eagles medium (DMEM). The cells were seeded into 25 cm2 tissue culture flasks

and were maintained in CO2 incubator with 5% CO2 and 95% humidity,

supplemented with DMEM and 10% FBS. Penicillin and streptomycin (PAA) was also added to the medium to 1× final concentration. Hydrogen peroxide at a concentration of 200 μM was used as oxidants. The concentration of plant extract used was 20 mg. The cells were treated with the oxidant, both in the presence and the absence of the leaf extracts. The exposure of hydrogen peroxide was given for 1 h at 37 °C. The time points were arrived at by conducting a time-related response analysis of each cell type. A total of 106/107 cells per Eppendorf were seeded into 96-well plates and exposed for 1 h to H2O2/plant extracts. Cytotoxicity of drugs was assessed by the MTT assay according to the procedure of Igarashi and Miyazawa (2001).3 SRB binds to basic amino Roxadustat molecular weight acid residues in TCA-fixed cells to provide a sensitive index of cellular protein content that is linear over a range of cell density.4 The cell survival was measured as the percent absorbance compared to the control (untreated) cells at 492 nm. The incubated cells were spread on the microscopic slides with a drop of diluted Giemsa stain. The slides were Rolziracetam mounted with cover slips and observed under the phase contrast microscope (Nikon, Japan) for morphological changes

as described by Chih et al (2001).5 The numbers of cells showing apoptotic morphological changes were counted in each experimental group per 100 cells in ten different fields and the experiment was repeated for 5 times. PI staining was employed to discriminate apoptotic from normal cells, which reflects the nuclear changes during apoptosis using the protocol developed by Sarker et al (2000).6 The apoptotic cells were detected using the green filter of a fluorescence microscope (Nikon, Japan). The treated cells were incubated for 5 min with 10 μl of ethidium bromide (50 μg/ml) and spread by placing a cover slip over it. The apoptotic cells were scored by counting the cells with condensed chromatin and fragmented nuclei under fluorescent microscope (Nikon, Japan) using UV 2A filter at 400× magnification. The ratios of apoptotic cells to normal cells were calculated in each staining method.

All statements were scored on a five-point ordinal scale (‘totally disagree’ to ‘totally agree’) and average domain scores were used for analyses.26 More information about the psychometric validity of the outcome measures, as well as detailed assessment procedures have been described elsewhere.13 and 18 The assessment procedure was as follows: at home, the parents and children completed the AQuAA, the Multidimensional Fatigue Scale, and the attitude questionnaires. At the hospital body height and weight were measured, and several family characteristics were determined (siblings, parental

marital status, parental educational level and sports frequency of the immediate family). Selective motor control was assessed with the INCB024360 cost modified Trost test, during which the ability of children to dorsiflex the ankle and extend the knee in an isolated movement was scored in four categories: N/A = not able to make the movement, 0 = completely synergistic, 1 = partly synergistic, 2 = no synergy.27 Scores for each joint and leg were added to obtain a total score for

selective motor control. Parents also indicated the sports frequency of immediate family members in five categories (from 1 = never to 5 = daily), from which a mean score was SB203580 nmr calculated. Children then completed mobility capacity assessments and fitness tests, after which the ca-librated accelerometer was provided to register walking activity for one week. Additionally, children and parents received a diary to record their daily activities and accelerometer registration time. Information on data processing and controlling data quality of the accelerometer has been described elsewhere.18 A priori sample size calculation indicated that 22 children were needed in each group to detect a clinically relevant difference of 1000 strides per day between groups.28 Power was set at 80%, significance level at 5% and the standard deviation of the difference was set at 1175 strides (unpublished pilot data of over Dutch children with cerebral palsy). To allow for 10% loss

to follow-up, 25 children were included in each group. To determine the intervention effect, intention-to-treat analyses were performed using linear regression, or logistic regression for dichotomous outcomes (p < 0.05). Outcomes at 4 months, 6 months, and 12 months were the dependent variables, and group allocation and the measured outcome at baseline were the independent variables in the analyses. To correct for performing statistical tests over multiple time points, the critical p-value was divided by the number of tests performed, resulting in an alpha < 0.025 for outcomes measured three times, and an alpha < 0.017 for outcomes measured four times. Variables with non-normally distributed residuals were logarithmically transformed prior to performing linear regression analyses, after which the results were transformed back, providing a between-group change ratio.

Both vaccines appeared to provide a significant effect in the i.p. challenge model that could not be detected when fish were challenged through the assumed natural challenge route, i.e. in the cohabitation model. The conflicting results observed for the two laboratory models are likely to result from the fact that the challenge virus is injected in the same spatial

area as the vaccine in the i.p. model. Thus the challenge virus is released into an area where there is a chronic and active inflammatory response [28]. These results highlight the importance of studying vaccines under various conditions to obtain a more complete understanding of their performance. The present vaccine situation in the European salmonid farming industry is suboptimal. Despite vaccination of the fish population in exposed areas, the SAV epizootics remain as a major loss-contributing factor to the industry [4]. Moreover, Screening Library mw the available SAV-vaccine must

be given as a separate injection from a multi-component vaccine, with at least 230 day degrees separating the injections. This is an additional stressor for the fish and costly to the farmer. The high level of protection combined with the possibility to include the ALV405 antigen in a multi-component vaccine could therefore represent a significant improvement for both fish health and farming economy. “
“Influenza pandemics buy Cobimetinib are caused by the introduction of new influenza A virus subtypes in the human population. The viruses either circulated in animal reservoirs and enter the human population by zoönotic infections or they emerged by genetic reassortment between human and animal influenza A viruses [1]. The virus causing the outbreak of pandemic influenza A (H1N1) 2009 was the result of a series of reassortments among

H1N1 swine influenza viruses, H1N1 avian influenza virus and H3N2 human influenza virus [2] and [3]. The reassorted virus crossed the species barrier from swine to humans and caused a severe disease outbreak partially due to a substantial antigenic drift of the swine H1 as compared to the H1 in the earlier circulating epidemic H1N1 virus. Generally, the Cediranib (AZD2171) human population is immunologically naïve to such zoönotic or reassorted strains. Accordingly, disease outbreaks usually affect large geographical areas involving many countries and can result in severe morbidity and mortality [4] and [5]. From both a public health and socio-economic point of view, vaccination stands as the primary strategy for the prevention and control of influenza virus infections [6]. Currently licensed influenza virus vaccines consist of whole inactivated virus or purified virus proteins derived from virus grown in embryonated chicken eggs. The manufacturing process is time-consuming and the production capacity is limited [7].

01% gelatin (opsonization buffer). The bacteria treated with hyperimune or control mice sera were harvested and incubated with 4 × 105 peritoneal cells at 37 °C for 45 min with shaking (220 rpm). Ten-fold dilutions of the samples were performed and 10 μL aliquots of each dilution were cultured on blood agar plates. The count live colonies were performed as previously described [33]. After 20 min, slides of the M1 strain opsonophagocitic assay were prepared by cytospin, stained with Instant-Prov (Newprov, Brazil), subsequently analyzed by light microscopy using an Axion Vision Zeiss Imager A1 and photographed by Axion Vision software (Zeiss, Germany).

Statistical analysis was performed using Kruskal–Wallis test. Heart tissue was obtained from the lysate of a postmortem normal human mitral valve, separated Selleck Volasertib by SDS–PAGE and blotted onto nitrocellulose membranes KPT-330 [31] and [32]. The blots were blocked with Tris-buffered

saline containing 5% skim milk. The membrane was sequentially treated with a pool (n = 6) of BALB/c or Swiss immunized mice sera and anti-mouse IgG alkaline phosphatase and revealed with NBT-BCIP solution (Invitrogen, USA). We observed that anti-StreptInCor antibodies from the BALB/c mice sera pool were able to cross-recognize both the M5 and M1 proteins in total protein extracts from each strain (Fig. 1). The anti-StreptInCor antibodies from Swiss mice were able to neutralize the M1, M5, M12, M22 and M87 strains by cross-recognizing the M protein on the bacterial surface with a Median Fluorescence Intensity (MFI) 2 or 3 times greater than the MFI of control sera (Fig. 2). Anti-StreptInCor antibodies from BALB/c and Swiss mice were able to promote opsonophagocytosis and death of the M1, M5, M12, M22 and M87 strains (Fig. 3a and b, respectively). The amino acid sequences alignment of the M protein C-terminal region of the strains used in this study had, on average, 72% identity with the StreptInCor amino acid sequence (Fig. 3c). The M1, M6 and M12 strains had an additional block of 7 amino acids, while the M87 strain contained two fewer amino

acids than the StreptInCor sequence. M1 strain was killed in peritoneal cells by phagocytosis 20 min after the opsonization assay as observed by optical microscopy (Fig. 4a–d). No autoreactive Electron transport chain antibodies against human heart mitral valve protein extracts were observed (Fig. 5). The development of a vaccine against multiple S. pyogenes strains without causing autoimmunity will bring numerous benefits to human health. A vaccine would prevent streptococcal infections and sequelae and could be more effective and longer-lasting than the currently used treatment. In addition to have broad coverage against strains, a vaccine should promote the production of neutralizing and opsonophagocytic antibodies, which are the body’s major defense lines against extracellular microorganisms. In the 70 and 80s several models of anti S.

The antioxidant potential of ABE and ABCNPs was investigated in the search for new bioactive compounds from natural resources. It has been used to evaluate the potential of various natural plants and vegetable extracts as antioxidants.15 The inhibition values were originate at 27.78%, 27.78% and 25.51% for

ABE, ABCNPs and ascorbic acid were observed at a concentration of 50, 100, 150, 200 and 250 μg/ml, respectively (Fig. 1(A)). For ABTS•+ radical cation was generated by the interaction of ABTS•+ (250 mM) CHIR-99021 nmr and K2S2O8 (40 mM) and observed different concentration of 50, 100, 150, 200 and 250 μg/ml, respectively (Fig. 1(B)). In ABE and ABCNPs the inhibitory concentration (IC50) was found to be 250 μg/ml. This suggests that antioxidant activity was retained even after the encapsulation of chitosan with ABE. Fig. 1(C) shows the reducing ability of the ABE and ABCNPs compared to that of ascorbic acid and increased dose dependently. At the concentration of 250 μg/ml, the AB mushroom extracts and its loaded chitosan nanoparticles were determined to have 81.97% and 78.13% reducing power relative to the ascorbic acid 73.52%, respectively. The extracts showed more scavenging

activity on hydroxyl radical and reducing power. Free radical scavenging is a generally accepted mechanism for phenolic antioxidants to inhibit lipids oxidation. The antioxidative activity of phenolics is generally directed by their chemical MRIP structures, the activity increases with increasing the number of hydroxyl groups and their location Vandetanib solubility dmso in the molecules involved.16 The amount of total phenolics was reported 1 g of sample contains 8.19 ± 1 mg of gallic acid by Folin–Ciocalteu method and total flavonoid analysis by the assay of aluminum chloride spectrophotometric reported 1 g of sample contains 10.3 ± 1 mg of quercetin in ABE and ABCNPs shown in Fig. 2(a) and (b). Pekkarien et al attributed the antioxidant activity of phenolic acids in a bulk lipid system to their DPPH radical scavenging activity.17A. bisporus

contained significant amounts of phenolic amino acids (tyrosine, L-glutaminyl-4-hydroxybenzene, 3, 4-dlhydroxyphenylalanine and L-glutaminyl-3,4-dihydroxybenzene), which may be responsible for the relatively high antioxidative activity. The acute lethal effect of ABE and ABCNPs on rats (Table 2 and Fig. 3(a) and (b)) shows that number of animal died within 72 h. After the major signs of toxicity noticed within 72 h included change in physical activity, difficulty in breathing, mortality, loss of appetite, general weakness, respiratory suffering and convulsions or coma. These signs were not seen in bellow 2747.25 mg/kg b.w. in ABE and 3178.86 mg/kg b.w. of ABCNPs, but progressed and became increasingly pronounced as the dose increased towards 4000 mg/kg b.w. of ABE and 5000 mg/kg b.w. of ABCNPs. The LD50, around 3000 mg/kg b.w. is thought to be safe as suggested by Lork.

2 Intrascrotal tuberculomas are very rare, with very few cases reported in the English literature. Tuberculosis of the spermatic cord is usually a disease that affects the sexually active man with a genitourinary contamination. But the few cases described in childhood imply the possibility of hematogenous spread of the bacillus. It can also affect patients with pulmonary infection

(<1%).3 Contamination by bacille Calmette-Guérin instillation for bladder cancer has also been described but is uncommon. Prostate is more usually involved4 Clinically, the patients show a painful unilateral Alpelisib research buy swelling of the scrotum. Voiding problems are usually absent when only the extraurinary organs are involved. As in our case, usual tuberculous signs such as fever, night sweats, and weight loss can be absent. Imaging findings including ultrasonography and computed tomographic scan are not specific. Search for bacillus in urine and semen should be performed in case of call signs

(hematuria, hemospermia, dysuria, and so forth). Polymerase chain reaction is very useful in this indication and gives quick detection. Differential diagnosis is represented by benign and malignant conditions. Scrotal tuberculoma is usually a peroperative discovery. BMS-354825 research buy Most patients are operated for genital suspect masses, and unfortunately, most of them undergo undue orchiectomy (75%).5 This kind of mistake can be avoided by a thorough preoperative checkup (clinical examination, tuberculosis skin test, ultrasonography, chest radiography, and urine and semen analyses) and a peroperative frozen section. Limited resection of the mass with preservation of the testes and epididymis must be performed once malignancy is excluded. The medical treatment consists in the combination Chlormezanone of powerful antituberculous drugs according to the regimen: 2 (rifampicin + isoniazid + pyrazinamide + ethambutol) + 4 (rifampicin

+ isoniazid) (R, rifampicin 10 mg/kg/J; H, isoniazid 5 mg/kg/J; Z, pyrazinamid 20-30 mg/kg/J). Though it is only 1 case, the tuberculosis of the spermatic cord is a rare condition that must be kept in mind, especially in developed countries where tuberculosis has known recrudescence in the last decades. A complete preoperative checkup with a peroperative frozen section (when available) must be performed to avoid an excessive surgery that can threaten the patient’s fertility. The authors declare that they have no relevant conflicts of interest. “
“A 45-year-old male patient presented to our institute with a history of left hip pain for 6 months and no past medical history of chronic diseases. No history of trauma is provided. The patient also went to a private clinic with the same complain and diagnosed to have osteoarthritis of the left hip joint and treated using nonsteroidal anti-inflammatory drugs.

His details have now been added. The authors apologize for any inconvenience caused. “
“Paratuberculosis is a highly prevalent chronic mycobacterial infection of the small intestine of ruminants. It causes substantial economic losses at farm level, particularly in cattle [1]. Transmission of the causative organism Mycobacterium avium subspecies paratuberculosis (MAP) amongst ruminants occurs by excretion via feces into the environment, where it may survive for prolonged periods of time [2]. When the disease progresses towards the clinical stage of infection, MAP can also be present in milk [3]. As a result of the latter it may represent a food safety issue given

the possible association between MAP and human Crohn’s disease [4]. Currently, a vaccine to control paratuberculosis

Selleck Navitoclax in cattle is not available, since the whole cell vaccine registered for use in sheep interferes with control programs against bovine tuberculosis. Individual MAP proteins as subunit vaccine candidates may overcome this interference. Dolutegravir datasheet In bovine paratuberculosis [5] and [6], similar to other mycobacterial diseases such as tuberculosis and leprosy, heat shock proteins (Hsp) elicit strong cell mediated and antibody responses. Our previous studies indicated that immune responsiveness to recombinant MAP Hsp70 proteins in naturally infected animals was predominantly cell mediated [6] and [7]. Since protective immunity to intracellular mycobacterial pathogens is thought to be cell mediated [8], recombinant MAP Hsp70 protein was used as a subunit vaccine in cattle concomitant with experimental infection with MAP. It induced protection as indicated by significantly reduced bacterial shedding [9]. In addition, Mannose-binding protein-associated serine protease MAP Hsp70 subunit vaccination did not interfere with current diagnostic methods to diagnose bovine TB [10]. Surprisingly, and in strong contrast with our previous observations in field cases of bovine paratuberculosis, this immunization-challenge study showed limited cell mediated responses against MAP Hsp70 and

pronounced MAP Hsp70 specific antibody production in the vaccinated animals [9]. The contribution of antibodies to protection against mycobacterial infections is disputed by some (reviewed in [11] and [12]), and supported by others (reviewed in [13]). Most of the recent studies on serum therapy of M. tuberculosis (MTb) infection report protective effects of antibodies specific for polysaccharide bacterial cell wall antigens such as the polysaccharide lipoarabinomannan (reviewed in [14]). In mice, a monoclonal antibody (Ig A) directed against a small surface-expressed mycobacterial heat shock protein (the 16 kD α-crystallin homologue) protected against early infection of murine lungs with MTb [15].

Furthermore the use of radiolabeled wood pulp NFC hydrogel as a potential biomedical device amongst other biomedical applications has not been demonstrated before. However, the biocompatibility and toxicity of bacterial and plant-derived cellulose materials have been documented both in vitro and in vivo use with of small animals ( Märtson et al., 1999, Vartiainen et al., 2011, BKM120 supplier Alexandrescu et al., 2013, Roman et al., 2010, Kovacs et al., 2010, Pértile et al., 2011, Helenius et al., 2006 and Moreira et al., 2009). In addition, we demonstrate a reliable and efficient method for NFC radiolabeling for the purpose of molecular imaging with a small animal SPECT/CT. To image NFC in animals by SPECT/CT,

NFC was labeled with 99mTc-NFC according to a previously described procedure for 99mTc-labeled carboxymethyl-cellulose (Schade et al., 1991) with slight modifications. 1.6% NFC stock hydrogel (GrowDex®, UPM-Kymmene Corporation, Finland) was used to prepare 1% NFC hydrogel with added stannous chloride stock (17.5 μg/ml in saline solution) and 99mTc-pertechnetate (99mTcO4−) stock (∼80 MBq/ml in saline solution) to a final volume of 1 ml. Briefly, 590 μl of http://www.selleckchem.com/products/NVP-AUY922.html the stock NFC was added to 285 μl of stannous chloride dehydrate solution (Angiocis®, IBA Molecular, Belgium) followed with 10 min incubation and mixing. Subsequently,

125 μl of 99mTcO4− was added to the reaction mixture to reach the NFC concentration of 1% and incubated while mixing for 30 min. To optimize the method for 99mTc-NFC labeling, various conditions were tested during the labeling

procedure, such as buffer pH ranging from 4.74 to 8.05, different incubation times for 99mTcO4−/NFC reaction mixture (5, 10, 15, 20, 25 and 30 min) and stannous chloride concentrations ranging from 50 to 0.05 μg/ml. The stability of the radiolabel was investigated in neutral isotonic pH by incubating the 1% 99mTc-NFC samples for 24 h. Samples were prepared in stock solutions as described above in saline or in fetal bovine serum found (FBS) (Sigma–Aldrich, Finland). Radiochemical purity and efficiency was tested at every time point (0, 15, 60, 120, 240 min and 24 h). TLC determined labeling efficiency and radiochemical purity of 99mTc-NFC with ITLC-SG chromatography plates (Agilent Technologies, Santa Clara, CA, USA) in methylethylketone (MEK) solvent system. Plates were cut in smaller equally sized pieces and placed in standard RIA tubes for radioactive measurement with a gamma counter (RiaCalc. WIZ, Wallac 1480 WIZARD® 3″, Finland). Animal studies were approved by the Finnish National Animal Experiment Board and performed in accordance with the Animal Welfare Act (247/1996) and Good Laboratory Practices for Animal Research. The release properties of plant-derived NFC implants were investigated with the use of radiolabeled small compounds. The use of 99mTc-NFC allows localization of the NFC in animals.

14 Butylated hydroxy anisole (BHA) (Himedia, India) was used as standard. The extract in methanol was tested at 20–250 μg/ml. DPPH solution was used at 20 μmol/l. DPPH dilution with methanol without extract was control. Percentage of scavenging was calculated as follows, DPPHscavengingactivity(%)=[(Acontrol−Asample)/Acontrol]×100 The data was presented as mean of triplicate. The concentration required for 50% reduction of DPPH radical (IC50) was determined graphically. Lipophilic antioxidants in the extract was measured selleck products using β-carotene–linoleic acid system.15

The extract and quercetin in DMSO were tested at 100 μg/ml, 500 μg/ml and 1000 μg/ml. Total reaction volume was 3 ml. The absorbance was recorded at 470 nm at regular time intervals from 0 to1500 min. The control contained 0.2 ml DMSO without extract. The reagent without β-carotene was served as blank. The data is presented as mean of triplicate readings. The antioxidant activity (AA) was expressed as percentage inhibition and calculated using the following equation: AA(%)=[(Degradationrateofcontrol−degradationrateofsample)/Degradationrateofcontrol]×100where

degradation rate = ln (a/b) × 1/t, where ln = natural log, a = initial absorbance (470 nm), b = absorbance (470 nm) after time ‘t’ (in min). A modified thiobarbituric acid Selinexor chemical structure reactive species (TBARS) assay was used.9 The extract and quercetin were tested at 60 μg/ml, 120 μg/ml, and 600 μg/ml in 250 μl aliquots. The absorbance was measured at 532 nm. The reaction without extract or quercetin served as the control. The test blank contained linoleic acid emulsion without peroxidation treatment. The assay was carried out as described previously with modifications.16 10 μl of extract or quercetin dilutions of 100 μg/ml, 200 μg/ml and 500 μg/ml concentrations incubated for 30 min with 5 μl of calf thymus CYTH4 DNA (Genei, India. 1 mg/ml) treated with Fenton reagent. Then, the reaction was terminated by adding 30 μl loading buffer (2.5 μg/ml bromophenol blue, 60% sucrose in 1 ml TBE buffer 10 mmol/l and pH 8.0) and 15 μl of which was electrophoresed at 60 eV potential for 30 min in submerged 1% agarose gel. The intact bands without shearing in

the electrophoretogram indicates the DNA protection. HPLC was performed using analytical HPLC system (Agilent Technologies assembled 1100 and 1200 series) equipped with quaternary pump and UV–visible detector. Reversed phase chromatographic analysis was carried out in isocratic conditions using RP-C18 column (4.6 mm × 250 mm) packed with 5 μm diameter particles. The separation was carried out in water-acetonitrile-acetic acid (80:20:3, v/v/v) as mobile phase at flow rate of 0.8 ml/min. Quercetin, gallic acid, 4-hydroxy benzoic acid, vanillic acid, epicatechin, ferulic acid, p-coumaric acid, phloroglucinol and chlorogenic acid (Sigma Aldrich, Germany) were used as reference standards at 300 ppm in methanol. The injection volume was 10 μl. Detection was done at 280 nm and 320 nm.