Additive effects of Artemisia capillaris extract and scopoletin on the relaxation of penile corpus cavernosum smooth muscle.

The objective was to investigate the cellular effect and action mechanism of Artemisia capillaris extract (ACE) and its component, scopoletin, on penile corpus cavernosum smooth muscle (PCCSM). In vitro study with PCCSM, the precontracted PCCSM with phenylephrine was treated with ACE or scopoletin. Cyclic nucleotides in the perfusate were measured by radioimmunoassay and expression of protein and mRNA of endothelial nitric oxide synthase (eNOS) and neuronal nitric oxide synthase in the perfused PCCSM were measured by western blot and real-time PCR, respectively. The interaction of ACE or scopoletin with udenafil was also evaluated. ACE and scopoletin exerted a significant and concentration-dependent relaxation in PCCSM. The perfusion with ACE or scopoletin significantly increased cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) and the perfusion with ACE or scopoletin increased the expression of eNOS mRNA and protein. Furthermore, ACE or scopoletin enhanced udenafil-inducing relaxation in PCCSM. ACE and scopoletin relaxed the PCCSM mainly by activating nitric oxide-cGMP system and cAMP pathway and they may be additive therapeutic candidates for ED patients who do not completely respond to udenafil.

Identification of anti-biofilm components in Withania somnifera and their effect on virulence of Streptococcus mutans biofilms.

AIMS: The aim of this study was to identify components of the Withania somnifera that could show anti-virulence activity against Streptococcus mutans biofilms. METHODS AND RESULTS: The anti-acidogenic activity of fractions separated from W. somnifera was compared, and then the most active anti-acidogenic fraction was chemically characterized using gas chromatography-mass spectroscopy. The effect of the identified components on the acidogenicity, aciduricity and extracellular polymeric substances (EPS) formation of S. mutans UA159 biofilms was evaluated. The change in accumulation and acidogenicity of S. mutans UA159 biofilms by periodic treatments (10 min per treatment) with the identified components was also investigated. Of the fractions, n-hexane fraction showed the strongest anti-acidogenic activity and was mainly composed of palmitic, linoleic and oleic acids. Of the identified components, linoleic and oleic acids strongly affected the acid production rate, F-ATPase activity and EPS formation of the biofilms. Periodic treatment with linoleic and oleic acids during biofilm formation also inhibited the biofilm accumulation and acid production rate of the biofilms without killing the biofilm bacteria. CONCLUSIONS: These results suggest that linoleic and oleic acids may be effective agents for restraining virulence of S. mutans biofilms. SIGNIFICANCE AND IMPACT OF THE STUDY: Linoleic and oleic acids may be promising agents for controlling virulence of cariogenic biofilms and subsequent dental caries formation.

The flavone eupatilin inhibits eotaxin expression in an NF-κB-dependent and STAT6-independent manner.

The CC chemokine eotaxin contributes to epithelium-induced inflammation in airway diseases such as asthma. Eupatilin (5,7-dihydroxy-3',4',6'-trimethoxyflavone), a bioactive component of Artemisia asiatica Nakai (Asteraceae), is reported to inhibit the adhesion of eosinophils to bronchial epithelial cells. However, little is known about the molecular mechanism of eupatilin-induced attenuation of bronchial epithelium-induced inflammation. In this study, we investigated the effect of eupatilin on expression of eotaxin-1 (CCL11), a potent chemoattractant for eosinophils. Eupatilin significantly inhibited eotaxin expression in bronchial epithelial cells stimulated with TNF-α, while NF-κB and IκBα kinase (IKK) activities declined concurrently. Eupatilin also inhibited mitogen-activated protein kinase (MAPK) activity; however, all of these anti-inflammatory activities were reversed by MAPK overexpression. In contrast, eupatilin did not affect the signal transducer and activator of transcription 6 (STAT6) signalling in bronchial epithelial cells stimulated with IL-4. Furthermore, eupatilin significantly attenuated TNF-α-induced eosinophil migration. These results suggest that the eupatilin inhibits the signalling of MAPK, IKK, NF-κB and eotaxin-1 in bronchial epithelial cells, leading to inhibition of eosinophil migration.

A laboratory study of the effects of processing blood through a cell salvage device and leucocyte depletion filter on levels of pro-inflammatory cytokines and bradykinin.

We investigated changes in concentrations of interleukin-1ß, interleukin-6, tumour necrosis factor-α and bradykinin in blood during passage through a cell salvage device and a leucocyte depletion filter, with or without application of subatmospheric pressure across the filter. Blood samples from 19 healthy women undergoing scheduled caesarean section showed concentrations of cytokines and bradykinin in blood filtered under gravity flow that were equal to or significantly lower than those of pre-operative venous blood samples. They were also significantly lower than that in postoperative orthopaedic shed blood, which is commonly reinfused after orthopaedic surgery. A minority of samples taken from blood that had been filtered using subatmospheric pressure showed raised interleukin-6 concentrations. We suggest that use of a leucocyte depletion filter for cell-salvaged blood with gravity flow is likely to be safe with regard to concentrations of cytokines and bradykinin. However, this may not hold true for the filter used with subatmospheric pressure. If transfusion of salvaged blood using a leucocyte depletion filter seems to induce hypotension, elevation of interleukin-6 should be suspected.

Diversity of morphology and function in arbuscular mycorrhizal symbioses in Brachypodium distachyon.

Brachypodium distachyon is a grass species that serves as a useful model for wheat and also for many of the grass species proposed as feedstocks for bioenergy production. Here, we monitored B. distachyon symbioses with five different arbuscular mycorrhizal (AM) fungi and identified symbioses that vary functionally with respect to plant performance. Three symbioses promoted significant increases in shoot phosphorus (P) content and shoot growth of Brachypodium, while two associations were neutral. The Brachypodium/Glomus candidum symbiosis showed a classic 'Paris-type' morphology. In the other four AM symbioses, hyphal growth was exclusively intracellular and linear; hyphal coils were not observed and arbuscules were abundant. Expression of the Brachypodium ortholog of the symbiosis-specific phosphate (Pi) transporter MtPT4 did not differ significantly in these five interactions indicating that the lack of apparent functionality did not result from a failure to express this gene or several other AM symbiosis-associated genes. Analysis of the expression patterns of the complete PHT1 Pi transporter gene family and AMT2 gene family in B. distachyon/G. intraradices mycorrhizal roots identified additional family members induced during symbiosis and again, transcript levels were similar in the different Brachypodium AM symbioses. This initial morphological, molecular and functional characterization provides a framework for future studies of functional diversity in AM symbiosis in B. distachyon.

Apoptotic effect of Polygonum Cuspidatum in oral cancer cells through the regulation of specificity protein 1.

OBJECTIVES: The aim of this study was to evaluate the growth inhibitory and apoptosis-inducing effects and mechanisms of Polygonum cuspidatum root in oral cancer cells. MATERIALS AND METHODS: The testing materials were separated by normal-phase silica gel liquid chromatography. The effect of P. cuspidatum root on apoptotsis and its mechanism were performed using 3-(4,5-dimethylthiazol-20yl)-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrazolium) (MTS) assay, western blot analysis, RT-PCR, promoter assay, and (4'-6-Diamidino-2-phenylindole) (DAPI) staining. RESULTS: The methanol extract of P. cuspidatum (MEPC) inhibited the proliferation of oral cancer cells by inducing caspase-dependent apoptosis. Protein and mRNA expression levels and the transactivation of Specificity protein 1 (Sp1) were markedly decreased in KB cells treated with MEPC. Ethyl acetate fraction (EA) from MEPC was more potent than aqueous fraction (AQ) from MEPC to induce apoptosis. F2, F3, and F4 from EA differentially inhibited the growth of KB cells, and it depends on the amount of Emodin in F2, F3, and F4. Moreover, Emodin inhibited oral cancer cell growth and induced caspase-dependent apoptosis by decreasing Sp1. MEPC also decreased an apoptosis-related downstream target of Sp1 protein, survivin. CONCLUSION: The results from this study strongly suggest that MEPC, its fraction, and Emodin may be potential bioactive materials to cause apoptosis mechanism via the down-regulation of Sp1 in oral cancer cells.

Bioactivity-guided separation of anti-acidogenic substances against Streptococcus mutans UA 159 from Polygonum cuspidatum.

OBJECTIVES: The aim of this study was to separate the anti-acidogenic substances against Streptococcus mutans UA 159 from Polygonum cuspidatum. MATERIAL AND METHODS: The anti-acidogenic substances were separated by a series of liquid-liquid fractionations followed by normal-phase silica gel liquid chromatography, based on high-performance liquid chromatography and glycolytic pH-drop assay. The effectiveness of the separated substances on the acidogenicity of Streptococcus mutans UA 159 was examined using sodium fluoride as a positive control. The chemical composition and quantities of the components of the substances was also assessed by qualitative-quantitative chromatographic analysis. RESULTS: Among the substances separated from P. cuspidatum, F3 showed the strongest inhibitory effect on the acidogenicity of S. mutans UA 159 in a dose-dependent manner without displaying any bactericidal activity. F3 decreased the acidogenicity of S. mutans even at 12.5 microg ml(-1) (P < 0.05). F3 consisted mainly of resveratrol and emodin (C(14)H(12)O(3) and C(14)H(4)O(2)(OH)(3)CH(3), respectively), which made up approximately 60% of the weight of F3. CONCLUSION: F3 can be considered as a promising agent for controlling the acidogenicity of S. mutans and subsequent dental caries formation.

Food protective effect of geraniol and its congeners against stored food mites.

The acaricidal activities of compounds derived from the oil of Pelargonium graveolens leaves against the storage food mite, Tyrophagus putrescentiae, were compared with the activity of a commercial acaricide, benzyl benzoate, in an impregnated fabric disk bioassay. Purification of the active constituent from P. graveolens was accomplished by silica gel chromatography and high-performance liquid chromatography. Structural analysis of the active constituent by 1H nuclear magnetic resonance (NMR), 13C-NMR, 1H-13C shift correlated spectroscopy NMR, and distortionless enhancement by polarization transfer NMR identified trans-3,7-dimethyl-2,6-octadien-1-ol (geraniol). Based on the 50% lethal dose values, the most toxic compounds against T. putrescentiae were geraniol (1.95 microg/cm3), which was followed by nerol (2.21 microg/cm3), citral (9.65 microg/cm3), benzyl benzoate (11.27 microg/cm3), and beta-citronellol (15.86 microg/cm3). Our results suggest that geraniol is more effective in controlling T. putrescentiae than benzyl benzoate is. Furthermore, geraniol, which is used as a flavoring for beverages, candies, ice creams, and baked goods and congeners (citral and nerol), may be useful for managing populations of T. putrescentiae.

HSPF-Paddy simulation of water flow and quality for the Saemangeum watershed in Korea.

The Hydrological Simulation Program-FORTRAN (HSPF) is a comprehensive model that was developed to simulate many processes related to water flow and quality in watersheds of almost any size and complexity. Paddy rice fields often dominate extensive portions of the landscape in the Asian monsoon region. The hydrological and environmental conditions of paddy fields differ somewhat from those of other land uses, and HSPF may not adequately simulate watersheds in paddy farming regions. HSPF was previously modified to HSPF-Paddy; here, we examined the applicability of the modified model. The model was applied to simulate the water flow and quality of the Saemangeum watershed (2523 km2) in Korea, where paddy rice fields comprise about one-third of the total watershed area. Long-term monitoring data (5 years for water flow, 10 years for water quality) were used in the calibration and verification processes. Model performance was in the range of "very good" and "good" based on model efficiency (R2) and percent difference. The accuracy of the daily simulation was lower than that of monthly simulation for water flow. The water-quality simulation results were encouraging for this sizable watershed with mixed land uses; HSPF-Paddy proved adequate, and its application is recommended to simulate watershed processes in paddy farming regions.

Water quality modeling to evaluate BMPs in rice paddies.

A water quality model applicable to rice paddies was developed using field data from 1999-2002. Use of the Dirac delta function efficiently explained the nutrient-concentration characteristics of ponded water. The model results agreed reasonably well with the observed data. The ponded-water quality was influenced primarily by fertilization; nutrient concentration was especially high during early cultivation periods. Reducing surface drainage during the fertilization period may substantially reduce nonpoint source loading from paddies. Increased weir heights and shallow irrigation methods were evaluated by the model as practical methods for reducing nutrient loading from paddies. These methods were effective in reducing surface drainage and are suggested as "best management practices" (BMPs) if applied based on site-specific paddy conditions.

Mass balance analysis and water quality model development for loading estimates from paddy fields.

Mass balance analysis and water quality model development for paddy field were performed using field experimental data during 2001-2002. About half (47-62%) of the total outflow was lost by surface drainage, with the remainder (490-530 mm) occurring by evapotranspiration. Most of nutrient inflow and outflow were mediated by fertilization and plant uptake, respectively. Nutrient outflow by surface drainage runoff was substantial about 15%-29% for T-N and 6%-13% for T-P. However, the responses of yield and drainage outflow to fertilization were not significant in this study. A water quality model applicable to paddy fields was developed and it demonstrates good agreement with observed data. The nutrient concentration of ponded water was high by fertilization at early culture periods, so reducing surface drainage during fertilization period can reduce nutrient loading from paddy fields. Shallow irrigation, raising the weir height in diked rice fields, and minimizing forced surface drainage are suggested to reduce surface drainage outflow.

Expression of the hepatitis B surface S and preS2 antigens in tubers of Solanum tuberosum.

In an attempt to develop an edible vaccine, we transformed a recombinant hepatitis B virus (HBV) gene encoding the middle protein of HBV that contains the surface S and preS2 antigen into potato by Agrobacterium-mediated transformation. The HBV gene was under control of either the CaMV 35S promoter, the double 35S promoter with the AlMV 5' non-translated leader sequence, or the tuber-specific patatin promoter. HBV mRNA levels were higher with the 35S promoter than with the double 35S and patatin promoters; however, the levels of the S and preS2 antigen in the transformed tubers were higher with the patatin promoter than with the CaMV 35S and double promoters. The levels of preS2 antigen produced are the highest reported to date. Transgenic potato tubers were fed to mice, and the mice showed an immune response against the HBV S antigen.

Improved glucose tolerance and insulin sensitivity after electrical stimulation-assisted cycling in people with spinal cord injury.

DESIGN: Longitudinal training. OBJECTIVES: The purpose was to determine the effect of electrical stimulation (ES)-assisted cycling (30 min/day, 3 days/week for 8 weeks) on glucose tolerance and insulin sensitivity in people with spinal cord injury (SCI). SETTING: The Steadward Centre, Alberta, Canada. METHODS: Seven participants with motor complete SCI (five males and two females aged 30 to 53 years, injured 3-40 years, C5-T10) underwent 2-h oral glucose tolerance tests (OGTT, n=7) and hyperglycaemic clamp tests (n=3) before and after 8 weeks of training with ES-assisted cycling. RESULTS: Results indicated that subjects' glucose level were significantly lower at 2 h OGTT following 8 weeks of training (122.4+/-10 vs 139.9+/-16, P=0.014). Two-hour hyperglycaemic clamps tests showed improvement in all three people for glucose utilisation and in two of three people for insulin sensitivity. CONCLUSIONS: These results suggested that exercise with ES-assisted cycling is beneficial for the prevention and treatment of Type 2 diabetes mellitus in people with SCI. SPONSORSHIP: Supported by Alberta Paraplegic Foundation, Therapeutic Alliance.

Quadriceps muscle deoxygenation during functional electrical stimulation in adults with spinal cord injury.

STUDY DESIGN: Cross-sectional study comparing healthy subjects with age and gender matched subjects with spinal cord injury (SCI, injury levels from C5 to T12). OBJECTIVES: To compare the acute cardiorespiratory responses and muscle oxygenation trends during functional electrical stimulation (FES) cycle exercise and recovery in the SCI and healthy subjects exercising on a mechanical cycle ergometer. SETTING: Seven volunteers in each group participated in one exercise test at the Rick Hansen Center, University of Alberta, Edmonton, Canada. METHODS: Both groups completed a stagewise incremental test to voluntary fatigue followed by 2 min each of active and passive recovery. Cardiorespiratory responses were continuously monitored using an automated metabolic cart and a wireless heart rate monitor. Tissue absorbency, an index of muscle oxygenation, was monitored non-invasively from the vastus lateralis using near infrared spectroscopy. RESULTS: The healthy subjects showed significant (P<0.05) increases in the oxygen uptake (VO2), heart rate (HR) and ventilation rate (VE) from rest to maximal exercise. The SCI subjects showed a twofold increase in VO2 (P>0.05), a threefold increase in VE (P<0.05) and a 5 beats/min increase in HR (P>0.05) from the resting value. The SCI subjects demonstrated a lesser degree (P<0.05) of muscle deoxygenation than the healthy subjects during the transition from rest to exercise. Regression analysis indicated that the rate of decline in muscle deoxygenation with respect to the VO2 was significantly (P<0.05) faster in the SCI subjects compared to healthy subjects. CONCLUSIONS: FES exercise in SCI subjects elicits: (a) modest increases in the cardiorespiratory responses when compared to resting levels; (b) lower degree of muscle deoxygenation during maximal exercise, and (c) faster changes in muscle deoxygenation with respect to the VO2 during exercise when compared to healthy subjects.

Functional electrical stimulation exercise increases GLUT-1 and GLUT-4 in paralyzed skeletal muscle.

The study purpose was to determine the effect of functional electrical stimulation (FES)-leg cycle ergometer training (30 minutes on 3 d/wk for 8 weeks) on the GLUT-1 and GLUT-4 content of paralyzed skeletal muscle. Biopsy samples of vastus lateralis muscle were obtained pre- and post-training from five individuals with motor-complete spinal cord injury ([SCI] four men and one woman aged 31 to 50 years, 3 to 25 years postinjury involving C5-T8). Western blot analysis indicated that GLUT-1 increased by 52% and GLUT-4 increased by 72% with training (P < .05). This coincided with an increase in the muscle oxidative capacity as indicated by a 56% increase in citrate synthase (CS) activity (P < .05) and an improvement in the insulin sensitivity index as determined from oral glucose tolerance tests (P < .05). It is concluded that FES endurance training is effective to increase glucose transporter protein levels in paralyzed skeletal muscle of individuals with SCI.