Anti-inflammatory effect of Spiranthera odoratissima A. St.-Hil. leaves involves reduction of TNF-alpha

Spiranthera odoratissima A. St.-Hil., 'manaca', is a medicinal species used in Brazil, especially in central region, for the treatment of several diseases such as pain and inflammation. In this study, the methanol/aqueous phase of the ethanol extract of the leaves of 'manaca' (MAP), at the doses of 50, 150 and 500 mg/kg was used to evaluate the anti-inflammatory and/or antinociceptive effects and the possible anti-inflammatory mechanism. The antinociceptive and anti-inflammatory activities of MAP were assessed using formalin test, carrageenan-induced paw oedema. The myeloperoxidase activity, capillary permeability, leukocyte migration and tumour necrosis factor alpha (TNF-alpha) levels were evaluated in pleural exudate. The MAP reduced the licking time only in the later phase of formalin test, and showed anti-inflammatory activity by reducing the paw oedema, migration cell, myeloperoxidase activity, capillary permeability and TNF-alpha levels. In conclusion, we confirmed the inflammatory activity of MAP and affirm that this effect involves the reduction of TNF-alpha level.

Intestinal permeability parameters in obese patients are correlated with metabolic syndrome risk factors

Background & aims: Altered intestinal permeability has been shown to be associated with metabolic alterations in animal models of obesity, but not in humans. The aim of this study was to assess intestinal permeability in obese women and verify if there is any association with anthropometric measurements, body composition or biochemical variables. Methods: Twenty lean and twenty obese females participated in the study. Anthropometric measurements, body composition and blood pressure were assessed and biochemical analyses were performed. Administration of lactulose and mannitol followed by their quantification in urine was used to assess the intestinal permeability of volunteers. Results: The obese group showed lower HDL (p < 0.05), higher fasting glucose, insulin, HOMA index and lactulose excretion than the lean group (p < 0.05), suggesting increased paracellular permeability. Lactulose excretion showed positive correlation (p < 0.05) with waist and abdominal circumference. Blood insulin and the HOMA index also increased with the increase in mannitol and lactulose excretion and in the L/M ratio (p < 0.05). L/M ratio presented a negative correlation with HDL concentration (p < 0.05). Conclusions: We demonstrated that intestinal permeability parameters in obese women are positively correlated with anthropometric measurements and metabolic variables. Therapeutic interventions focused on intestine health and the modulation of intestinal permeability should be explored in the context of obesity. (C) 2012 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

Oxidative Stress and Modification of Renal Vascular Permeability Are Associated with Acute Kidney Injury during P. berghei ANKA Infection

Malaria associated-acute kidney injury (AKI) is associated with 45% of mortality in adult patients hospitalized with severe form of the disease. However, the causes that lead to a framework of malaria-associated AKI are still poorly characterized. Some clinical studies speculate that oxidative stress products, a characteristic of Plasmodium infection, as well as proinflammatory response induced by the parasite are involved in its pathophysiology. Therefore, we aimed to investigate the development of malaria-associated AKI during infection by P. berghei ANKA, with special attention to the role played by the inflammatory response and the involvement of oxidative stress. For that, we took advantage of an experimental model of severe malaria that showed significant changes in the renal pathophysiology to investigate the role of malaria infection in the renal microvascular permeability and tissue injury. Therefore, BALB/c mice were infected with P. berghei ANKA. To assess renal function, creatinine, blood urea nitrogen, and ratio of proteinuria and creatininuria were evaluated. The products of oxidative stress, as well as cytokine profile were quantified in plasma and renal tissue. The change of renal microvascular permeability, tissue hypoxia and cellular apoptosis were also evaluated. Parasite infection resulted in renal dysfunction. Furthermore, we observed increased expression of adhesion molecule, proinflammatory cytokines and products of oxidative stress, associated with a decrease mRNA expression of HO-1 in kidney tissue of infected mice. The measurement of lipoprotein oxidizability also showed a significant increase in plasma of infected animals. Together, our findings support the idea that products of oxidative stress, as well as the immune response against the parasite are crucial to changes in kidney architecture and microvascular endothelial permeability of BALB/c mice infected with P. berghei ANKA.

Chronic VEGF Blockade Worsens Glomerular Injury in the Remnant Kidney Model

VEGF inhibition can promote renal vascular and parenchymal injury, causing proteinuria, hypertension and thrombotic microangiopathy. The mechanisms underlying these side effects are unclear. We investigated the renal effects of the administration, during 45 days, of sunitinib (Su), a VEGF receptor inhibitor, to rats with 5/6 renal ablation (Nx). Adult male Munich-Wistar rats were distributed among groups S+V, sham-operated rats receiving vehicle only; S+Su, S rats given Su, 4 mg/kg/day; Nx+V, Nx rats receiving V; and Nx+Su, Nx rats receiving Su. Su caused no change in Group S. Seven and 45 days after renal ablation, renal cortical interstitium was expanded, in association with rarefaction of peritubular capillaries. Su did not worsen hypertension, proteinuria or interstitial expansion, nor did it affect capillary rarefaction, suggesting little angiogenic activity in this model. Nx animals exhibited glomerulosclerosis (GS), which was aggravated by Su. This effect could not be explained by podocyte damage, nor could it be ascribed to tuft hypertrophy or hyperplasia. GS may have derived from organization of capillary microthrombi, frequently observed in Group Nx+Su. Treatment with Su did not reduce the fractional glomerular endothelial area, suggesting functional rather than structural cell injury. Chronic VEGF inhibition has little effect on normal rats, but can affect glomerular endothelium when renal damage is already present.

From Sample Processing to Quantification: A Full Electrochemical Approach for Neutral Analyte Derivatization, Capillary Electrophoresis Separation, and Contactless Conductivity Detection

A thin-layer electrochemical flow cell coupled to capillary electrophoresis with contactless conductivity detection (EC-CE-(CD)-D-4) was applied for the first time to the derivatization and quantification of neutral species using aliphatic alcohols as model compounds. The simultaneous electrooxidation of four alcohols (ethanol, 1-propanol, 1-butanol, and 1-pentanol) to the corresponding carboxylates was carried out on a platinum working electrode in acid medium. The derivatization step required 1 min at 1.6 V vs. Ag/AgCl under stopped flow conditions, which was preceded by a 10 s activation at 0 V. The solution close to the electrode surface was then hydrodynamically injected into the capillary, and a 2.5 min electrophoretic separation was carried out. The fully automated flow system operated at a frequency of 12 analyses per hour. Simultaneous determination of the four alcohols presented detection limits of about 5 x 10(-5) mol As a practical application with a complex matrix, ethanol concentrations were determined in diluted pale lager beer and in nonalcoholic beer. No statistically significant difference was observed between the EC-CE-(CD)-D-4 and gas chromatography with flame ionization detection (GC-FID) results for these samples. The derivatization efficiency remained constant over several hours of continuous operation with lager beer samples (n = 40).

Quantification of terbinafine in pharmaceutical tablets using capillary electrophoresis with contactless conductivity detection and batch injection analysis with amperometric detection

Terbinafine hydrochloride (TerbHCl) is an allylamine derivative with fungicidal action, especially against dermatophytes. Different analytical methods have been reported for quantifying TerbHCl in different samples. These procedures require time-consuming sample preparation or expensive instrumentation. In this paper, electrochemical methods involving capillary electrophoresis with contactless conductivity detection, and amperometry associated with batch injection analysis, are described for the determination of TerbHCl in pharmaceutical products. In the capillary electrophoresis experiments, terbinafine was protonated and analyzed in the cationic form in less than 1 min. A linear range from 1.46 to 36.4 mu g mL(-1) in acetate buffer solution and a detection limit of 0.11 mu g mL(-1) were achieved. In the amperometric studies, terbinafine was oxidized at +0.85 V with high throughput (225 injection h(-1)) and good linear range (10-100 mu mol L-1). It was also possible to determine the antifungal agent using simultaneous conductometric and potentiometric titrations in the presence of 5% ethanol. The electrochemical methods were applied to the quantification of TerbHCl in different tablet samples; the results were comparable with values indicated by the manufacturer and those found using titrimetry according to the Pharmacopoeia. The electrochemical methods are simple, rapid and an appropriate alternative for quantifying this drug in real samples. (C) 2012 Elsevier B.V. All rights reserved.

Capillary electrophoretic enantioselective determination of zopiclone and its impurities

A capillary electrophoretic enantioselective method with UV detection was developed and validated for the simultaneous quantification of zopiclone enantiomers and its impurities, zopiclone-N-oxide enantiomers, and 2-amino-5-chloropyridine, in tablets. The analytes were extracted from the tablets using ACN and were separated in an uncoated fused-silica capillary (50 mu m, 42 cm effective length, 50 cm total length) using 80 mM sodium phosphate buffer pH 2.5 and 5 mM carboxymethyl-beta-cyclodextrin as running buffer. The analytes and the internal standard (trimethoprim) were detected at 305 and 200 nm, respectively. A voltage of 27 kV was applied and the capillary temperature was maintained at 25 degrees C. All enantiomers were analyzed within 8 min and linear calibration curves over the concentration range of 0.40.8 mg mL-1 for each zopiclone enantiomer, 0.81.6 mu g mL-1 for 2-amino-5-chloropyridine and 0.40.8 mu g mL-1 for each zopiclone-N-oxide enantiomer were obtained. The coefficients of correlation obtained for the linear curves were greater than 0.99. The intra-day and inter-day accuracy and precision were lower than 2% for all analytes. This validated method was employed to study the degradation and racemization of zopiclone under stress conditions. This application demonstrated the importance of a stability-indicating assay method for this drug.

Permeability of riparian forest strips in agricultural, small subtropical watersheds in south-eastern Brazil

If riparian buffer zones are ineffective in preventing C-4 plant carbon from upland areas reaching the stream sediment, the composition of stream fauna can be significantly altered. The permeability of riparian forest strips in agricultural, small subtropical watersheds in south-eastern Brazil was measured in nine watersheds categorised according to the predominant land cover of the legally required 30-m buffer riparian zone. Four watersheds with well preserved riparian forest along the 30-m buffer zone were designated as FOREST watersheds; three watersheds, with a predominance of C-4 grasses from sugarcane to pasture, mixed with preserved riparian forests, were designated MIXED watersheds; and two watersheds were termed PASTURE-SUGAR because their entire 30-m buffer zone was covered by C-4 plants. Stable carbon (delta C-13) isotopes were used as tracers of upland C-4 carbon in sediments, suspended particulate organic carbon, terrestrial and aquatic invertebrates and two species of neotropical fish. Although the intact 30-m buffer zone of riparian forests did not entirely prevent the input of C-4 to the river environment and food web, there was a significant increase in C-4 carbon in those watersheds where the buffer zone was not covered by riparian forests. These findings emphasise the importance of riparian forests in mitigating disturbance in streams and support efforts to preserve such riparian corridors.

Capillary bioreactors based on human purine nucleoside phosphorylase: A new approach for ligands identification and characterization

The enzyme purine nucleoside phosphorylase (PNP) is a target for the discovery of new lead compounds employed on the treatment severe T-cell mediated disorders. Within this context, the development of new, direct, and reliable methods for ligands screening is an important task. This paper describes the preparation of fused silica capillaries human PNP (HsPNP) immobilized enzyme reactor (IMER). The activity of the obtained IMER is monitored on line in a multidimensional liquid chromatography system, by the quantification of the product formed throughout the enzymatic reaction. The Km value for the immobilized enzyme was about twofold higher than that measured for the enzyme in solution (255 +/- 29.2 mu M and 133 +/- 114.9 mu M, respectively). A new fourth-generation immucillin derivative (DI4G: IC50 = 40.6 +/- 0.36 nM), previously identified and characterized in HsPNP free enzyme assays, was used to validate the IMER as a screening method for HsPNP ligands. The validated method was also used for mechanistic studies with this inhibitor. This new approach is a valuable tool to PNP ligand screening, since it directly measures the hypoxanthine released by inosine phosphorolysis, thus furnishing more reliable results than those one used in a coupled enzymatic spectrophotometric assay. (C) 2011 Elsevier B.V. All rights reserved.

In vitro effects of Er,Cr:YSGG laser on dentine hypersensitivity. Dentine permeability and scanning electron microscopy analysis

The aim of the present study was to determine clinical parameters for the use of Er,Cr:YSGG laser in the treatment of dentine hypersensitivity. Two antagonist areas were determined as control and experimental areas for irradiation in 90 premolar roots. Each surface was conditioned with 24% EDTA (sub-group 1) and 35% phosphoric acid (sub-group 2) and irradiated with the following settings: 1) Er:YAG, 60 mJ, 2 Hz, defocused; groups 2 to 9: irradiation with Er,Cr:YSGG laser, 20 Hz, Z6 tip, 0% of air and water: 2) Er,Cr:YSGG 0.25 W; 3) 0.5 W; 4) 0.75 W; 5) 1.0 W; 6) 1.25 W, 7) 1.50 W, 8) 2 W; 9) 2 W. After irradiation, samples were immersed in methylene blue solution and included in epoxy resin to obtain longitudinal cuts. The images were digitalized and analyzed by computer software. Although the samples irradiated with Er:YAG laser showed less microleakage, sub-group 1 showed differences between the groups, differing statistically from groups 3, 6, and 9. The results of sub-group 2 showed that the mean values of Er:YAG samples showed a negative trend, however, no differences were detected between the groups. For scanning electron microscopy analysis, dentine squares were obtained and prepared to evaluate the superficial morphology. Partial closure of dentinal tubules was observed after irradiation with Er:YAG and Er,Cr:YSGG laser in the 0.25 and 0.50 W protocols. As the energy densities rose, open dentinal tubules, carbonization and cracks were observed. It can be concluded that none of the parameters were capable of eliminating microleakage, however, clinical studies with Er:YAG and Er,Cr:YSGG lasers should be conducted with the lowest protocols in order to determine the most satisfactory setting for dentine hypersensitivity.

In Situ Effect of Dentifrices Associated to CO2 Laser in the Permeability of Eroded Root Dentin

Objective: The aim of this study was to evaluate, through a crossover 2 x 2 in situ trial, the effect of a desensitizing dentifrice associated with CO2 laser irradiation to control the permeability of eroded root dentin. Background data: Facing the increased prevalence of erosive lesion and the need for preventive means to control painful symptoms related to them. Methods: Eighty slabs of bovine root dentin were subjected to initial erosive challenge (citric acid 0.3%, 2 h), followed by a remineralizing period in artificial saliva (24 h). Specimens were then divided according to dentin treatment: desensitizing dentifrice, desensitizing dentifrice + CO2 laser, fluoride anticavity dentifrice. and fluoride anticavity dentifrice + CO2 laser. After a 2-day lead-in period, 10 volunteers wore an intraoral palatal appliance containing four root dentin slabs, in two phases of 5 days each. During the intraoral phase, one side of the appliance was immersed in 0.3% citric acid, and the opposite side was immersed in deionized water, four times a day. One hour after the immersions, all specimens were brushed with dentifrice slurry provided by the researcher. After a 7-day washout period, volunteers were crossed over on the different dentifrice group. Each phase having been completed, the specimens were evaluated for permeability through an optical microscope. Results: Data were analyzed using ANOVA and no significant difference (p = 0.272) was found between the surface treatments performed on bovine root dentin. Conclusions: It can be concluded that fluoride anticavity or desensitizing dentifrice, regardless of the association with the CO2 laser irradiation, was able to control the permeability of eroded root dentin.

Reducing Water and Chloride Penetration Through Silicate Treatments for Concrete as a Mean to Control Corrosion Kinetics

There are currently many types of protective materials for reinforced concrete structures and the influence of these materials in the chloride diffusion coefficient and water penetration still needs more research. The aim of this work is to analyze the contributions regarding the typical three surface concrete protection systems (coatings, linings and pore blockers) and discusses the results of three pore blockers (sodium silicate) tested in this work. To this end, certain tests were used: one involving permeability mechanism (low pressure-immersion absorption), one involving capillary water absorption, and the last, a migration test used to estimate the effective chloride diffusion coefficient in saturated condition. Results indicated reduction in chloride diffusion coefficients and capillary water absorption, therefore, restrictions to water penetration from external environmental. As a consequence, a reduction of the corrosion kinetics and a control of the chloride ingress are expected.

AEROBIC EXERCISE TRAINING CORRECTS CAPILLARY RAREFACTION AND ALTERATIONS IN PROPORTIONS OF THE MUSCLE FIBERS TYPES IN SPONTANEOUSLY HYPERTENSIVE RATS

Aerobic exercise training (ET) has been established as an important non-pharmacological treatment of hypertension, since it decreases blood pressure. Studies show that the skeletal muscle abnormalities in hypertension are directly associated with capillary rarefaction, higher percentage of fast-twitch fibers (type II) with glycolytic metabolism predominance and increased muscular fatigue. However, little is known about these parameters in hypertension induced by ET. We hypothesized that ET corrects capillary rarefaction, potentially contributing to the restoration of the proportion of muscle fiber types and metabolic proprieties. Twelve-week old Spontaneously Hypertensive Rats (SHR, n=14) and Wistar Kyoto rats (WKY, n=14) were randomly assigned into 4 groups: SHR, trained SHR (SHR-T), WKY and trained WKY (WKY-T). As expected, ten weeks of ET was effective in reducing blood pressure in SHR-T group. In addition, we analyzed the main markers of ET. Resting bradycardia, increase of exercise tolerance, peak oxygen uptake and citrate synthase enzyme activity in trained groups (WKY-T and SHR-T) showed that the aerobic condition was achieved. ET also corrected the skeletal muscle capillary rarefaction in SHR-T. In parallel, we observed reduction in percentage of type IIA and IIX fibers and simultaneous augmented percentage of type I fibers induced by ET in hypertension. These data suggest that ET prevented changes in soleus fiber type composition in SHR, since angiogenesis and oxidative enzyme activity increased are important adaptations of ET, acting in the maintenance of muscle oxidative metabolism and fiber profile.

Stability-indicating methods for the enantioselective determination of dihydropyridines by high performance liquid chromatography and capillary electrophoresis

This paper presents simple, rapid, precise and accurate stability-indicating HPLC and CE methods, which were developed and validated for the determination of nitrendipine, nimodipine and nisoldipine. These drugs are calcium channel antagonists of the 1,4-dihydropyridine type which are used in the treatment of cardiovascular diseases. Experimental results showed a good linear correlation between the area and the concentration of drugs covering a relatively large domain of concentration in all cases. The linearity of the analytical procedures was in the range of 2.0-120.0 mu g mL-1 for nitrendipine, 1.0-100.0 mu g mL(-1) for nimodipine and 100.0-600.0 mu g mL(-1) for nisoldipine, the regression determination coefficient being higher than 0.99 in all cases. The proposed methods were found to have good precision and accuracy. The chemical stability of these drugs was determined under various conditions and the methods have shown adequate separation for their enantiomers and degradation products. In addition, degradation products produced as a result of stress studies did not interfere with the detection of the drugs' enantiomers and the assays can thus be considered stability-indicating.

Effect of pH, mucin and bovine serum on rifampicin permeability through Caco-2 cells

Rifampicin, a poorly soluble drug, has great importance in therapeutics as it is the main drug used to treat tuberculosis. The characterization of its permeability and the factors that influence it represent an important tool for predicting its bioavailability. Caco-2 cell monolayers were used as models of the intestinal mucosa to assess the uptake and transport of rifampicin and the effects of various experimental conditions were investigated, in order to establish the influence of these variables on rifampicin permeability. Different pHs (5.8, 6.8 and 7.4) in the apical medium, the presence or absence of mucin (3.0% w/v) in the donor site and the presence or absence of bovine serum albumin (4.0% v/v) in the receptor chamber were the evaluated conditions. The quantification of rifampicin in the apical or basolateral chambers was performed by a validated HPLC-UV method. The change in the donor chamber pH showed that permeability values were greater at pH 6.8, although this increase does not result in an alteration of the qualitative classification of rifampicin, which has high permeability. Mucin and bovine serum showed no effects on the permeability of rifampicin at the concentration tested. Overall, the current study suggests that pH, artificial mucin and bovine serum proteins have no influence on rifampicin permeability. Copyright (c) 2012 John Wiley & Sons, Ltd.