A VALIDATED HPLC ANALYTICAL METHOD FOR THE ANALYSIS OF SOLASONINE AND SOLAMARGINE IN IN VITRO SKIN PENETRATION STUDIES

To assess topical delivery studies of glycoalkaloids, an analytical method by HPLC-UV was developed and validated for the determination of solasonine (SN) and solamargine (SM) in different skin layers, as well as in a topical formulation. The method was linear within the ranges 0.86 to 990.00 mu g/mL for SN and 1.74 to 1000.00 mu g/mL for SM (r = 0.9996). Moreover, the recoveries for both glycoalkaloids were higher than 88.94 and 93.23% from skin samples and topical formulation, respectively. The method developed is reliable and suitable for topical delivery skin studies and for determining the content of SN and SM in topical formulations.

Exergy efficiency analysis of chemical and biochemical stages involved in liquid biofuels production processes

Liquid biofuels can be produced from a variety of feedstocks and processes. Ethanol and biodiesel production processes based on conventional raw materials are already commercial, but subject to further improvement and optimization. Biofuels production processes using lignocellulosic feedstocks are still in the demonstration phase and require further R&D to increase efficiency. A primary tool to analyze the efficiency of biofuels production processes from an integrated point of view is offered by exergy analysis. To gain further insight into the performance of biofuels production processes, a simulation tool, which allows analyzing the effect of process variables on the exergy efficiency of stages in which chemical or biochemical reactions take place, were implemented. Feedstocks selected for analysis were parts or products of tropical plants such as the fruit and flower stalk of banana tree, palm oil, and glucose syrups. Results of process simulation, taking into account actual process conditions, showed that the exergy efficiencies of the acid hydrolysis of banana fruit and banana pulp were in the same order (between 50% and 60%), lower than the figure for palm oil transesterification (90%), and higher that the exergy efficiency of the enzymatic hydrolysis of flower stalk (20.3%). (C) 2011 Elsevier Ltd. All rights reserved.

Qualitative SEM/EDS analysis of microleakage and apical gap formation of adhesive root-filling materials

Objective: The aim of this study was to compare the correspondence between gap formation and apical microleakage in root canals filled with epoxy resin-based (AH Plus) combined or not with resinous primer or with a dimethacrylate-based root canal sealer (Epiphany). Material and Methods: Thirty-nine lower single-rooted human premolars were filled by the lateral condensation technique (LC) and immersed in a 50-wt% aqueous silver nitrate solution at 37 degrees C (24 h). After longitudinal sectioning, epoxy resin replicas were made from the tooth specimens. Both the replicas and the specimens were prepared for scanning electron microscopy (SEM). The gaps were observed in the replicas. Apical microleakage was detected in the specimens by SEM/energy dispersive spectroscopy (SEM/EDS). The data were analyzed statistically using an Ordinal Logistic Regression model and Analysis of Correspondence (alpha=0.05). Results: Epiphany presented more regions containing gaps between dentin and sealer (p<0.05). There was correspondence between the presence of gaps and microleakage (p<0.05). Microleakage was similar among the root-filling materials (p>0.05). Conclusions: The resinous primer did not improve the sealing ability of AH Plus sealer and the presence of gaps had an effect on apical microleakage for all materials.

Effect of pulse repetition rate and number of pulses in the analysis of polypropylene and high density polyethylene by nanosecond infrared laser induced breakdown spectroscopy

Pulse repetition rates and the number of laser pulses are among the most important parameters that do affect the analysis of solid materials by laser induced breakdown spectroscopy, and the knowledge of their effects is of fundamental importance for suggesting analytical strategies when dealing with laser ablation processes of polymers. In this contribution, the influence of these parameters in the ablated mass and in the features of craters was evaluated in polypropylene and high density polyethylene plates containing pigment-based PbCrO4. Surface characterization and craters profile were carried out by perfilometry and scanning electron microscopy. Area, volume and profile of craters were obtained using Taylor Map software. A laser induced breakdown spectroscopy system consisted of a Q-Switched Nd:YAG laser (1064 nm, 5 ns) and an Echelle spectrometer equipped with ICCD detector were used. The evaluated operating conditions consisted of 10, 25 and 50 laser pulses at 1, 5 and 10 Hz, 250 mJ/pulse (85 J cm(-2)), 2 mu s delay time and 6 mu s integration time gate. Differences in the topographical features among craters of both polymers were observed. The decrease in the repetition rate resulted in irregular craters and formation of edges, especially in polypropylene sample. The differences in the topographical features and ablated masses were attributed to the influence of the degree of crystallinity, crystalline melting temperature and glass transition temperature in the ablation process of the high density polyethylene and polypropylene. It was also observed that the intensities of chromium and lead emission signals obtained at 10 Hz were two times higher than at 5 Hz by keeping the number of laser pulses constant. (C) 2011 Elsevier B. V. All rights reserved.

Analysis of slow- and fast-alpha band asymmetry during performance of a saccadic eye movement task: Dissociation between memory- and attention-driven systems

This study aimed at analyzing the relationship between slow- and fast-alpha asymmetry within frontal cortex and the planning, execution and voluntary control of saccadic eye movements (SEM), and quantitative electroencephalography (qEEG) was recorded using a 20-channel EEG system in 12 healthy participants performing a fixed (i.e., memory-driven) and a random SEM (i.e., stimulus-driven) condition. We find main effects for SEM condition in slow- and fast-alpha asymmetry at electrodes F3-F4, which are located over premotor cortex, specifically a negative asymmetry between conditions. When analyzing electrodes F7-F8, which are located over prefrontal cortex, we found a main effect for condition in slow-alpha asymmetry, particularly a positive asymmetry between conditions. In conclusion, the present approach supports the association of slow- and fast-alpha bands with the planning and preparation of SEM, and the specific role of these sub-bands for both, the attention network and the coordination and integration of sensory information with a (oculo)-motor response. (C) 2011 Elsevier B.V. All rights reserved.

Hypotheses, rationale, design, and methods for prognostic evaluation of cardiac biomarker elevation after percutaneous and surgical revascularization in the absence of manifest myocardial infarction. A comparative analysis of biomarkers and cardiac magnetic resonance. The MASS-V Trial

Background: Although the release of cardiac biomarkers after percutaneous (PCI) or surgical revascularization (CABG) is common, its prognostic significance is not known. Questions remain about the mechanisms and degree of correlation between the release, the volume of myocardial tissue loss, and the long-term significance. Delayed-enhancement of cardiac magnetic resonance (CMR) consistently quantifies areas of irreversible myocardial injury. To investigate the quantitative relationship between irreversible injury and cardiac biomarkers, we will evaluate the extent of irreversible injury in patients undergoing PCI and CABG and relate it to postprocedural modifications in cardiac biomarkers and long-term prognosis. Methods/Design: The study will include 150 patients with multivessel coronary artery disease (CAD) with left ventricle ejection fraction (LVEF) and a formal indication for CABG; 50 patients will undergo CABG with cardiopulmonary bypass (CPB); 50 patients with the same arterial and ventricular condition indicated for myocardial revascularization will undergo CABG without CPB; and another 50 patients with CAD and preserved ventricular function will undergo PCI using stents. All patients will undergo CMR before and after surgery or PCI. We will also evaluate the release of cardiac markers of necrosis immediately before and after each procedure. Primary outcome considered is overall death in a 5-year follow-up. Secondary outcomes are levels of CK-MB isoenzyme and I-Troponin in association with presence of myocardial fibrosis and systolic left ventricle dysfunction assessed by CMR. Discussion: The MASS-V Trial aims to establish reliable values for parameters of enzyme markers of myocardial necrosis in the absence of manifest myocardial infarction after mechanical interventions. The establishments of these indices have diagnostic value and clinical prognosis and therefore require relevant and different therapeutic measures. In daily practice, the inappropriate use of these necrosis markers has led to misdiagnosis and therefore wrong treatment. The appearance of a more sensitive tool such as CMR provides an unprecedented diagnostic accuracy of myocardial damage when correlated with necrosis enzyme markers. We aim to correlate laboratory data with imaging, thereby establishing more refined data on the presence or absence of irreversible myocardial injury after the procedure, either percutaneous or surgical, and this, with or without the use of cardiopulmonary bypass.

Mechanical analysis of conventional and small diameter conical implant abutments

PURPOSE. The aim of the present study was to evaluate if a smaller morse taper abutment has a negative effect on the fracture resistance of implant-abutment connections under oblique compressive loads compared to a conventional abutment MATERIALS AND METHODS. Twenty morse taper conventional abutments (4.8 mm diameter) and smaller abutments (3.8 mm diameter) were tightened (20 Ncm) to their respective implants (3.5 x 11 mm) and after a 10 minute interval, implant/abutment assemblies were subjected to static compressive test, performed in a universal test machine with 1 mm/min displacement, at 45 degrees inclination. The maximum deformation force was determined. Data were statistically analyzed by student t test. RESULTS. Maximum deformation force of 4.8 mm and 3.8 mm abutments was approximately 95.33 kgf and 95.25 kgf, respectively, but no fractures were noted after mechanical test. Statistical analysis demonstrated that the evaluated abutments were statistically similar (P=.230). CONCLUSION. Abutment measuring 3.8 mm in diameter (reduced) presented mechanical properties similar to 4.8 mm (conventional) abutments, enabling its clinical use as indicated. [J Adv Prosthodont 2012;4:158-61]

Automated analysis of lidocaine and its metabolite in plasma by in-tube solid-phase microextraction coupled with LC-UV for pharmacokinetic study

A sensitive, selective, and reproducible in-tube solid-phase microextraction and liquid chromatographic (in-tube SPME/LC-UV) method for determination of lidocaine and its metabolite monoethylglycinexylidide (MEGX) in human plasma has been developed, validated, and further applied to pharmacokinetic study in pregnant women with gestational diabetes mellitus (GDM) subjected to epidural anesthesia. Important factors in the optimization of in-tube SPME performance are discussed, including the draw/eject sample volume, draw/eject cycle number, draw/eject flow rate, sample pH, and influence of plasma proteins. The limits of quantification of the in-tube SPME/LC method were 50 ng/mL for both metabolite and lidocaine. The interday and intraday precision had coefficients of variation lower than 8%, and accuracy ranged from 95 to 117%. The response of the in-tube SPME/LC method for analytes was linear over a dynamic range from 50 to 5000 ng/mL, with correlation coefficients higher than 0.9976. The developed in-tube SPME/LC method was successfully used to analyze lidocaine and its metabolite in plasma samples from pregnant women with GDM subjected to epidural anesthesia for pharmacokinetic study.

Accelerated Solvent Extraction for Gas Chromatographic Analysis of Nicotine and Cotinine in Meconium Samples

Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)

Analysis of dispersive and dissipative media with optical resonances

In this paper we analyze the problem of light-matter interaction when absorptive resonances are imbedded in the material dispersion. We apply an improved approach to aluminum (Al) in the optical frequency range to investigate the impact of these resonances on the operating characteristics of Al-based nanoscale devices. Quantities such as group velocity, stored energy density, and energy velocity, normally obtained using a single resonance model [Wave Propagation and Group Velocity (Academic Press, 1960), Nat. Mater. 11, 208 (2012)], are now accurately calculated regardless of the medium adopted. We adapt the Loudon approach [Nat. Mater. 11, 208 (2012)] to media with several optical resonances and present the details of the extended model. We also show pertinent results for Al-based metal-dielectric-metal (MDM) waveguides, around spectral resonances. The model delineated here can be applied readily to any metal accurately characterized by Drude-Lorentz spectral resonance features. (C) 2012 Optical Society of America

Single-cell expression analysis of BMP15 and GDF9 in mature oocytes and BMPR2 in cumulus cells of women with polycystic ovary syndrome undergoing controlled ovarian hyperstimulation

To detect expression of bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) in oocytes, and their receptor type 2 receptor for BMPs (BMPR2) in cumulus cells in women with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization (IVF), and determine if BMPR2, BMP15, and GDF9 expression correlate with hyperandrogenism in FF of PCOS patients. Prospective case-control study. Eighteen MII-oocytes and their respective cumulus cells were obtained from 18 patients with PCOS, and 48 MII-oocytes and cumulus cells (CCs) from 35 controls, both subjected to controlled ovarian hyperstimulation (COH), and follicular fluid (FF) was collected from small (10-14 mm) and large (> 18 mm) follicles. RNeasy Micro Kit (Qiagen(A (R))) was used for RNA extraction and gene expression was quantified in each oocyte individually and in microdissected cumulus cells from cumulus-oocyte complexes retrieved from preovulatory follicles using qRT-PCR. Chemiluminescence and RIA assays were used for hormone assays. BMP15 and GDF9 expression per oocyte was higher among women with PCOS than the control group. A positive correlation was found between BMPR2 transcripts and hyperandrogenism in FF of PCOS patients. Progesterone values in FF were lower in the PCOS group. We inferred that BMP15 and GDF9 transcript levels increase in mature PCOS oocytes after COH, and might inhibit the progesterone secretion by follicular cells in PCOS follicles, preventing premature luteinization in cumulus cells. BMPR2 expression in PCOS cumulus cells might be regulated by androgens.

DNA repair gene excision repair cross complementing-group 1 (ERCC1) in head and neck squamous cell carcinoma: analysis of methylation and polymorphism (G19007A), protein expression and association with epidemiological and clinicopathological factors

Aims: To evaluate the associations of excision repair cross complementing-group 1 (ERCC1) (DNA repair protein) (G19007A) polymorphism, methylation and immunohistochemical expression with epidemiological and clinicopathological factors and with overall survival in head and neck squamous cell carcinoma (HNSCC) patients. Methods and results: The study group comprised 84 patients with HNSCC who underwent surgery and adjuvant radiotherapy without chemotherapy. Bivariate and multivariate analyses were used. The allele A genotype variant was observed in 79.8% of the samples, GG in 20.2%, GA in 28.6% and AA in 51.2%. Individuals aged more than 45 years had a higher prevalence of the allelic A variant and a high (83.3%) immunohistochemical expression of ERCC1 protein [odds ratio (OR) = 4.86, 95% confidence interval (CI): 1.2-19.7, P = 0.027], which was also high in patients with advanced stage (OR= 5.04, 95% CI: 1.07-23.7, P = 0.041). Methylated status was found in 51.2% of the samples, and was higher in patients who did not present distant metastasis (OR = 6.67, 95% CI: 1.40-33.33, P = 0.019) and in patients with advanced stage (OR = 5.04, 95% CI: 1.07-23.7, P = 0.041). At 2 and 5 years, overall survival was 55% and 36%, respectively (median = 30 months). Conclusion: Our findings may reflect a high rate of DNA repair due to frequent tissue injury during the lifetime of these individuals, and also more advanced disease presentation in this population with worse prognosis.

Identification and expression analysis of microRNAs and targets in the biofuel crop sugarcane

Abstract Background MicroRNAs (miRNAs) are small regulatory RNAs, some of which are conserved in diverse plant genomes. Therefore, computational identification and further experimental validation of miRNAs from non-model organisms is both feasible and instrumental for addressing miRNA-based gene regulation and evolution. Sugarcane (Saccharum spp.) is an important biofuel crop with publicly available expressed sequence tag and genomic survey sequence databases, but little is known about miRNAs and their targets in this highly polyploid species. Results In this study, we have computationally identified 19 distinct sugarcane miRNA precursors, of which several are highly similar with their sorghum homologs at both nucleotide and secondary structure levels. The accumulation pattern of mature miRNAs varies in organs/tissues from the commercial sugarcane hybrid as well as in its corresponding founder species S. officinarum and S. spontaneum. Using sugarcane MIR827 as a query, we found a novel MIR827 precursor in the sorghum genome. Based on our computational tool, a total of 46 potential targets were identified for the 19 sugarcane miRNAs. Several targets for highly conserved miRNAs are transcription factors that play important roles in plant development. Conversely, target genes of lineage-specific miRNAs seem to play roles in diverse physiological processes, such as SsCBP1. SsCBP1 was experimentally confirmed to be a target for the monocot-specific miR528. Our findings support the notion that the regulation of SsCBP1 by miR528 is shared at least within graminaceous monocots, and this miRNA-based post-transcriptional regulation evolved exclusively within the monocots lineage after the divergence from eudicots. Conclusions Using publicly available nucleotide databases, 19 sugarcane miRNA precursors and one new sorghum miRNA precursor were identified and classified into 14 families. Comparative analyses between sugarcane and sorghum suggest that these two species retain homologous miRNAs and targets in their genomes. Such conservation may help to clarify specific aspects of miRNA regulation and evolution in the polyploid sugarcane. Finally, our dataset provides a framework for future studies on sugarcane RNAi-dependent regulatory mechanisms.

Physicochemical analysis of blood and urine in the course of acute kidney injury in critically ill patients: a prospective, observational study

Abstract Background Sequential physicochemical alterations in blood and urine in the course of acute kidney injury (AKI) development have not been previously described. We aimed to describe these alterations in parallel to traditional renal and acid–base parameters. Methods One hundred and sixty eight consecutive critically ill patients with no previous kidney disease, who had an indwelling urinary catheter at ICU admission and who remained with the catheter for at least two days without dialysis were included. A sample of blood and spot urine were collected simultaneously, once daily, until catheter removal or dialysis requirement. Traditional acid–base and renal parameters were sequentially evaluated in parallel to blood and urinary physicochemical parameters. Patients were classified during this period as having or not AKI and, for patients with AKI, duration (transient or persistent) and severity (creatinine-based AKIN stage) were evaluated. Results One hundred and thirteen patients (67.3%) had AKI: 92 at ICU admission and 21 during the observation period. AKI development was characterized in blood by increased values of phosphate and unmeasured anions (SIG), decreased albumin, and in urine by decreased values of sodium (NaU), chloride (ClU) as well as high urinary strong ion difference (SIDu). These alterations began to occur before AKI diagnosis, and they reverted in transient AKI but remained in persistent AKI. NaU, ClU and albumin decreased, and phosphate, SIG and SIDu increased with AKI severity progression. NaU and ClU values increased again when AKIN stage 3 was reached. Conclusions Simultaneous physicochemical analysis of blood and urine revealed standardized alterations that characterize AKI development in critically ill patients. These alterations paralleled AKI duration and severity. Future studies should consider including sequential evaluation of urine biochemistry as part of the armamentarium for AKI diagnosis and management.

A validated HPLC analytical method for the analysis of solasonine and solamargine in in vitro skin penetration studies

To assess topical delivery studies of glycoalkaloids, an analytical method by HPLC-UV was developed and validated for the determination of solasonine (SN) and solamargine (SM) in different skin layers, as well as in a topical formulation. The method was linear within the ranges 0.86 to 990.00 µg/mL for SN and 1.74 to 1000.00 µg/mL for SM (r = 0.9996). Moreover, the recoveries for both glycoalkaloids were higher than 88.94 and 93.23% from skin samples and topical formulation, respectively. The method developed is reliable and suitable for topical delivery skin studies and for determining the content of SN and SM in topical formulations.