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Background: The minimum number of lymph nodes that should be evaluated in colon cancer to adequately categorize lymph node status is still controversial. The lymph node ratio (LNR) may be a better prognostic indicator. Materials & methods: We studied 1065 patients treated from 1 January 2000 to 31 August 2012. Results: Significant differences in survival were detected according to regional lymph nodes (pN) (p < 0.001) and LNR (p < 0.001). LRN and pN are independent prognostic factors. Spearman correlation analysis showed a significant correlation between the total number of dissected lymph nodes and pN (rs = 0.167; p < 0.001), but the total number of dissected lymph nodes is not significantly correlated with LNR (rs = -0.019; p = 0.550). Interpretation: In this study, LNR seems to demonstrate a superior prognostic value compared with the pN categories, in part due to its greater independence regarding the extent of lymphadenectomy.

Pork products are important sources of foodborne non-typhoidal Salmonella in Brazil where antibiotics are commonly used throughout the pork production process and this has the potential to selectively favor antibiotic-resistant strains. We characterized the genotypic and phenotypic diversity of S. enterica isolates (n = 41) that were isolated in Brazil. Isolates were collected from ten swine farms and one slaughterhouse. Whole-genome sequencing and in silico serotyping demonstrated that the S. enterica serovar Typhimurium was the most common serotype (n = 17), but eight additional servoars were identified. Isolates presented high similarity based on comparison of DNA sequences (minimum of 89.6%), and sequence variation grouped according to serotype. Eight multilocus sequence types were identified with ST19 being most common (n = 21). Several plasmids replicons were detected, with Col (RNAI) the most abundant (n = 30), followed by IncR (n = 22), IncI1 (n = 10) and IncA/C2 (n = 10). Minimum inhibitory concentration assays showed that the principle resistance phenotypes were for streptomycin (90.2%), tetracycline (87.8%), ampicillin (80.5%), chloramphenicol (70.7%) and ciprofloxacin (51.2%). Only two isolates were resistant to third-generation cephalosporins and no isolates were resistant to two tested carbapenems. Twenty-six unique antimicrobial-resistance genes were identified with bla TEM-1A and bla TEM-1B likely responsible for most beta-lactam resistance and floR responsible for most chloramphenicol resistance. Six strains were positive for mcr-1. At the time of collection, the sampled farms were adding ciprofloxacin to feed and this may have contributed to the high prevalence of resistance to this antibiotic. The high number of multidrug resistant Salmonella and the presence of multiple resistant genes and plasmids emphasize the diversity of Salmonella in the studied pork chain, specially from serotype Typhimurium., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

A low-cost polymer-based structure is proposed to improve the coupling between a fiber end section and photodetector active surface in optical links based on standard single-mode fiber (SSMF), which employs vertical cavity surface emitting lasers operating at 850 nm, i.e., below the SSMF cutoff wavelength. Considering receivers as small-area detectors, which are generally necessary to guarantee high-speed operation but at the same time are particularly subject to power fluctuations due to modal noise (whose impact is in turn enhanced in the presence of fiber-to-photodetector misalignment), significant achievements are demonstrated by employing the presented structure. Indeed, in the presence of a misalignment of $ \pm 4 $±4 to $ \pm 6\;{\unicode{x00B5}{\rm m}} $±6µm, which is nowadays typically achievable, the relative optical power fluctuations due to modal noise reduce in the presented case more than four times (2.5% from more than 10%) with respect to the case of butt-coupling, which implies an increase of the same factor in the output signal-to-noise ratio at the receiver end.

This study assessed an enzyme-linked immunosorbent assay (ELISA) based assay to detect Salmonella in swine as a potential tool to predict the presence of Salmonella in swine carcasses. The following samples were collected from 10 swine batches: blood ( n  = 100); environment (barn floor, n  = 10, and lairage floor, n  = 10); meat juice ( n  = 100, obtained after defrosting of diaphragm); tonsils ( n  = 100); mesenteric lymph nodes (MLNs) ( n  = 100); and carcasses after bleeding ( n  = 100), after singeing ( n  = 100), after evisceration ( n  = 100), and after final rinsing ( n  = 100). Blood and meat juice were subjected to ELISA to detect antibodies against Salmonella , and other samples were subjected to Salmonella detection by ISO 6579. Salmonella was detected in 3 samples from barn floors, 7 lairage floors, 45 tonsils, 43 MLNs and in 3 carcasses. Based on ELISA, Salmonella positive samples were: 86 and 46 blood serum (20% and 40% cut-offs) and 68 and 46 meat juice (20% and 40% cut-offs). Optical density readings from blood serum and meat juice presented a high and significant correlation ( r  = 0.93, p  < 0.001), and a substantial agreement for Salmonella detection ( K  = 0.69, ELISA 40% cut-off). The agreement between ELISA and microbiological analysis for Salmonella detection in pig carcasses were absent or poor, with the exception of results obtained by ELISA 40% cut-off from blood serum and meat juice with MLNs ( K  = 0.49 and 0.50, respectively) and tonsils ( K  = 0.29 and 0.30, respectively). Based on the obtained results, meat juice can be considered an alternative to blood serum as a matrix for ELISA for preliminary detection of Salmonella , allowing the identification of potential sources of contamination during slaughtering.

Listeria monocytogenes is a relevant pathogen usually associated with meat and ready-to-eat products. This study aimed to assess the distribution, adhesion, virulence and antibiotic resistance of L. monocytogenes in a pork production chain. Environment, carcass and food samples (n = 894) were obtained from different steps of a pork production chain over a 6-month period (10 samplings), including from farms and the slaughterhouse (reception, slaughtering, processing, storage and end products). L. monocytogenes was detected in samples from the reception (lairage floor, 1/10), slaughtering (drains, 2/20) and cutting room stages (conveyor belts in the final packing stage - 11/20, knife - 1/40, and cutting boards - 1/20). Positive results for conveyor belts were recorded in seven consecutive samplings. L. monocytogenes isolates (n = 87) were characterized as belonging to serogroup IVb and presented positive PCR results for inlA, inlB, inlC, inlJ, hlyA, plcA, actA and iap. Isolates were selected according to the original samples (n = 31) and subjected to Pulsed Field Gel Electrophoresis (PFGE), demonstrating their high clonal identity (98.4-100%). According to PFGE results and their original samples, isolates were selected (n = 16) and subjected to phenotypic assay to assess their adhesion potential and tested for resistance against 15 antibiotics; all tested isolates presented weak adhesion potential and were resistant to ampicillin. The present study demonstrated the persistence of L. monocytogenes in the pork processing facility, indicating the potential risk for cross-contamination with a potential virulent and resistant clone., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Pigs infected with Salmonella are an important source of contamination at slaughterhouses. We characterized the distribution, virulence genotypes and antimicrobial-resistance phenotypes for Salmonella isolates that were collected from different stages of a pork production chain. Each of ten pig lots were sampled for feed (n = 10), water (n = 10), barn floor (n = 10), lairage floor (n = 10), mesenteric lymph nodes (n = 100), tonsils (n = 100), processing environment (n = 120), pork cuts (n = 40) and carcasses after bleeding (n = 100), after singeing (n = 100), after evisceration (n = 100), and after final rinsing (n = 100). Salmonella was isolated according to ISO 6579, and after confirmation the isolates were subjected to serogrouping, macro-restriction digests and pulsed-field gel electrophoresis (PFGE), detection of virulence-related genes and antimicrobial-resistance phenotyping. Salmonella was recovered from barn floors from 3 pig farms (3/10), lairage floors (7/10), carcasses after bleeding (2/100) and final washing (1/100), palatine tonsils (45/100), mesenteric lymph nodes (43/100), utensils (3/120) and cuts (4/40). The most prevalent serogroup was O: 4 (82%) followed by O:3 (7.7%); O:9 (5.1%); O:8 (2.6%) and O:7 (2.6%). Recovered strains (n = 109) were classified into 24 different pulsotypes (XbaI restriction digest), which were arranged into five different clusters. Fourteen different virulence genotypes were observed based on 15 loci, and all isolates were positive for invA, sitC, pagC and tolC. There was a high prevalence of antimicrobial resistance against streptomycin (90.5%), tetracycline (88.1%), ampicillin (81.0%), chloramphenicol (71.4%), and ciprofloxacin (50.0%). No strain was resistant to ertapenem, meropenem or kanamycin. A majority (80.9%) of isolates were considered multidrug resistant (resistant to ≥3 antibiotic classes). This study provides valuable insight about the epidemiology of Salmonella in swine production, and despite the low presence of this pathogen in carcasses and meat cuts, the majority of isolates was multidrug resistant., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Inflammasomes are pivotal cytosolic molecular platforms involved in infection resistance. As multiprotein complexes, they consist of NOD-like receptors (NLRs), the adaptor proteins apoptosis-associated speck-like protein containing a CARD (ASC) and the effector molecules caspase-1 and caspase-11, whose assembly and activation depends on homotypic interactions. Here we describe WD repeat containing protein 90 (WDR90) as a new inflammasome component. We found that zebrafish wdr90 is highly induced by guanylate binding protein 4 (Gbp4) independently of inflammasome activation and caspase-1 activity. This gene encodes an evolutionarily conserved protein with unknown functions that contains several WD40 domains, which are involved in coordinating multiprotein complex assembly. Functional studies in zebrafish larvae showed that forced expression of wdr90 increased caspase-1 activity and inflammasome-dependent resistance to Salmonella enterica serovar Typhimurium infection. Wdr90 acted upstream of zebrafish caspase a (Caspa), the functional homolog of mammalian caspase-1, and Asc. Reconstitution experiments of the human inflammasome in HEK293 cells demonstrated that WDR90 was able to physically interact with and to alter the cellular distribution of NLRC4, but not of NLRP3 and AIM2. These results highlight the complexity of the inflammasome and the interest of studying fish immunity to understand not only the evolution of the immune system but also human immunity., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The extent of adulteration of dietary supplements has significantly increased in recent years. This situation worries health authorities and requires auxiliary analytical tools for the investigation of illegal substances purposely added. Ion exchange chromatography with conductivity detection is a consolidated analytical technique for the determination of inorganic compounds in various matrices. This technique has been applied to the pharmaceutical characterization of mainly impurities and degradation products. This work presents a new approach to ion exchange chromatography as a screening method to investigate the presence of amfepramone, femproporex, sibutramine, bisacodyl and amiloride in dietary supplements advertised for weight loss. The method was optimized and validated using a Metrosep C4 100/4.0 cation exchange column. The mobile phase consisted of 1.8 mm HNO 3 containing 2% acetonitrile (v/v), with a flow rate of 0.9 ml min -1 , and nonsuppressed conductivity detection was applied. The limits of detection and quantification varied from 1.01 to 3.62 mg L -1 and from 1.48 to 8.72 mg L -1 , respectively. The proposed method was successful applied to 78 solid dietary supplement samples, in two of which adulterations were found. Moreover, ion exchange chromatography with conductivity detection could be easily used for quality control without prior complex sample pre-treatment., (© 2019 John Wiley & Sons, Ltd.)

Control of Salmonella spp. in food production chains is very important to ensure safe foods and minimize the risks of foodborne disease occurrence. This study aimed to identify the prevalence and main contamination sources of Salmonella spp. in a pig production chain in southern Brazil. Six lots of piglets produced at different farms were tracked until their slaughter, and samples were subjected to Salmonella spp. detection. The obtained isolates were serotyped, subjected to antimicrobial resistance testing, and pulsed field gel electrophoresis (PFGE). Salmonella spp. was detected in 160 (10.2%) samples, and not detected in pig carcasses after final washing or chilling. Among the 210 Salmonella spp. isolates, S. Typhimurium was the most prevalent (n = 101) and resistant to at least one antimicrobial. High resistance rates were detected against tetracycline (83.8%), chloramphenicol (54.3%), and trimethoprim-sulfamethoxazole (33.3%). The isolates that were non-susceptible to three or more classes of antimicrobials (n = 60) were considered multidrug-resistant (MDR), and isolates resistant to up to six of the tested antimicrobials were found. PFGE allowed the identification of genetic diversity and demonstrated that farm environment and feed supply may be sources for the dissemination of Salmonella spp. along the production chain. The results revealed the sources of Salmonella contamination in the pig production chain and highlighted the risks of antimicrobial resistance spread.

Objective: The new regulatory framework for dietary supplements in Brazil prompted this analysis of the current outlook of these products and the challenges posed by the new guidelines., Methods: We conducted a qualitative, observational and descriptive study of dietary supplements commercialized in Brazilian online stores with the help of the Google® search tool. We considered the ingredients on the labels, the effects attributed to these products and the commercial claims used as a means of promoting them to assess the necessary changes for the legal framework in the new guidelines. Finally, with the help of a database, we compared the effects declared by the manufacturers and attributed to certain ingredients with the scientific evidence described in literature., Results: In total, we purchased 44 dietary supplements from Brazilian online stores (n = 7). Of the samples studied, 34.2% could not be classified in the category Dietary Supplements, as recommended by the new regulation of the Brazilian Health Regulatory Agency due to the presence of prohibited substances; 16% of products should be commercialized as medicines. Regarding the commercial appeals, 97.7% had banned expressions. Numerous claims of effects attributed to certain products were characterized as consumer fraud because they have no scientific evidence., Conclusions: The necessary changes represent a major regulatory and production challenge due to the wide range of dietary supplements and markets, an effort that aims to protect the consumers' health. Some previous gaps in the regulatory framework were not fully solved.

The consumption of dietary supplements is increasing every year all over the world and has been accompanied by an increased frequency of adulteration of these products with synthetic pharmaceuticals. Analytical methods that allow testing for the presence of synthetic drugs in dietary supplements are needed to detect such fraudulent practices. To investigate the adulteration of dietary supplements marketed for weight loss using different commercial appeals, we developed an analytical method using ultra-high-performance liquid chromatography-electrospray tandem mass spectrometry (UHPLC-ESI-MS/MS) for simultaneous determination of 32 drugs, including anorexics, anxiolytics, antidepressants, diuretics, laxatives and stimulants. Separation was accomplished in 19 minutes using a Zorbax SB-C18 column and a gradient elution program with 0.05% formic acid in water/acetonitrile as a mobile phase. Limits of quantification ranged from 0.14 to 3.92 μg L-1, and accuracy ranged from 80.00 to 119.48%. A simple extraction procedure was used in the pretreatment step by dissolving the samples in 100% methanol followed by a 1000 to 10,000-fold dilution in the mobile phase and filtration through a Teflon membrane (0.2 μm). The method was applied to the screening and quantification of the drugs in 108 formulations marketed as food supplements for slimming, weight loss, thermogenics, and supplements for meal replacement. Caffeine and p-synephrine were found as stimulants in 80 samples, listed or not on the label., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

New guidelines for the limits of elemental impurities in drug products were introduced by the International Council for Harmonization in 2014. While the guidelines define a limit for each element, the complete quantification of the 24 elements included is, in fact, unnecessary. An accurate "pass/fail" test to determine whether the threshold was exceeded or not could be valuable in this context. In this study, a screening procedure using the features of high-resolution continuum source graphite furnace atomic absorption spectrometry for the evaluation of 12 elements in three different drugs was developed. The three-dimensional absorbance spectrum including time and wavelength in the vicinity of the main line of the element allows for a pass/fail decision related to the presence or absence of the element in the sample. Additionally, the bi-dimensional absorbance-wavelength spectrum defines the elements captured in the window when additional peaks are seen in the spectrum. The analysis of the selected drugs included sample digestion, the definition of pyrolysis and atomization temperatures, determination of the limit of detection and other validation parameters for each element. The evaluation of the spectra, both three- and bi-dimensional, revealed that only three elements, Cr, Ni, and Cu, were present in the samples in amounts above the LOD and therefore "fail" in the test. Nevertheless, they were quantified, and the analysis revealed that their levels were below the permitted daily exposure, which are at least 6 times higher than the LOD of the selected elements. Operating in a routine mode, the proposed method is a good option for the evaluation of elemental impurities in drug active ingredients or drug final products., (Copyright © 2018. Published by Elsevier B.V.)

Persea americana, popularly known as avocado, has been empirically used as analgesic and anti-inflammatory including in the skin disorder treatment. Species of the genus Persea also show a photoprotective effect against UVB radiation. We investigated the antinociceptive and anti-inflammatory effects from a topical formulation containing the P. americana leaf extract in a UVB irradiation-induced burn model in mice and performed a gel-formulation stability study. The antinociceptive and anti-inflammatory effect was evaluated through mechanical allodynia, paw oedema, and inflammatory cell infiltration. Phenolic compounds were quantified by UHPLC-MS/MS. The gel-formulation stability study was performed analyzing organoleptic characteristics, pH, and viscosity. P. americana (3%) gel was able to prevent the UVB irradiation-induced mechanical allodynia on the 2nd and 3rd day after irradiation with maximum inhibition of 60 ± 12% at 2nd day. Such effect may be attributed, at least in part, due the presence of (+)-catechin (302.2 ± 4.9 μg/g) followed by chlorogenic acid (130 ± 5.1 μg/g) and rutin (102.4 ± 0.9 μg/g) found in the extract. The gel was not able to prevent the inflammatory parameters such as edema and leukocyte infiltration induced by UVB irradiation. No changes important were detected in the stability study, mainly in low temperature. Our results suggest that P. americana gel-formulation, which presented stability, ensuring its quality and the therapeutic effect, could be an interesting strategy for the treatment of the pain associated with sunburn; this effect could be attributed to its biological constituents, especially catechin, chlorogenic acid, and rutin.

A methodology to assay simultaneously iron and nickel present as contaminants in multimineral and multivitamin supplements was investigated. High-resolution continuum source graphite furnace atomic absorption spectrometry and direct solid sample analysis were used. Measurements were done with the secondary lines of Fe (352.604 nm) and Ni (352.454 nm) to avoid spectral interferences. The best temperatures for pyrolysis and atomization for Fe and Ni were 1000 and 2700 °C, respectively. Chemical modifiers were not necessary and no matrix effects were observed. Aqueous standard solutions were used for calibration. The limit of detection was 0.517 µg g -1 for Fe and 0.011 µg g -1 for Ni. The precision ranged from 4.3% to 17% and 4.4-20% for Fe and Ni, respectively. The method accuracy was confirmed by comparing statistically the results obtained by solid sampling with those of sample acid digestion. The proposed methodology was successfully applied to determine both metals in different multimineral and multivitamin supplements., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Flexible medical devices are primarily made of plasticized polyvinyl chloride (PVC). In recent times, to avoid undesired migration of the PVC plasticizers, ethyl vinyl acetate (EVA) and polypropylene (PP) has replaced PVC. Nevertheless, other additives are necessary to generate useful polymeric materials. Metallic species present in such additives can also leach out into the infusion solutions. The migration of barium (Ba), cadmium (Cd), lead (Pb), tin (Sn), and zinc (Zn) from devices made from PVC, EVA, and PP was evaluated. Bags and infusion sets were decomposed and their metallic contents analyzed. Glucose, NaCl, and Tween 80 were assessed as extraction media. These solutions were stored in PVC, EVA, and PP bags, heat-sterilized, and stored for 8 months at room temperature. Aliquots were taken before and after sterilization and then once per month to determine the contents of the metals. Commercial glucose and NaCl infusions were analyzed by taking aliquots of the solutions from the bags and from the administration set after their administration to patients. The three polymers contained the five metals. Ba was found in the highest concentration in all samples, with a mean of 8.0 mg/kg in PVC, 4.2 mg/kg in EVA, and 4.7 mg/kg in PP samples. Despite this, the only element that migrated into the glucose, NaCl, and Tween 80 solutions was Zn. The same result was found for the commercial glucose and NaCl infusions. Moreover, the Zn concentration in the administration sets was on average 52% higher than that found in the bags. LAY ABSTRACT: Flexible medical devices for infusions and artificial nutrition are made of plastics, such as polyvinyl chloride (PVC), ethyl vinyl acetate (EVA), and polypropylene (PP). These polymers contain additives necessary to generate useful materials. Metallic species present in these additives can leach out into the infusion solutions and come into contact with patients. To assess the risk of patient exposure to these metals, we evaluated the migration behavior of barium (Ba), cadmium (Cd), lead (Pb), tin (Sn), and zinc (Zn) from devices made from PVC, EVA, and PP. Bags and infusion sets were analyzed. Glucose, NaCl, and Tween 80 were investigated as extraction media. The three polymers contained the five metals. Ba was found in the highest concentration in all samples. Despite this, the only element that migrated into the glucose, NaCl, and Tween 80 solutions was Zn., (© PDA, Inc. 2019.)

Caffeine is present in products marketed for weight loss, with the purpose of increasing thermogenesis and lipid metabolism. The dosage declared by the product manufacturer, or even its presence, is not always correctly described on the label. This work aimed to investigate the undeclared synthetic caffeine in weight loss formulations by a high-performance liquid chromatography with diode array detection (HPLC-DAD) method. From one hundred products purchased through Brazilian e-commerce, seventeen contained caffeine, either naturally or synthetically added to formulation. The caffeine-containing samples were confirmed by an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method, and adulteration was clearly proven in five products. The content highest caffeine contained 448.8 mg per dose. Other irregularities were also found; nevertheless, the most serious was the addition of synthetic drugs without asking the consumers. Additional drugs expose the consumer to more possible side-effects as well as deleterious drug interactions. Intentional adulteration with any unlabeled substance is typically motivated by a desire to increase or alter the claimed effect of the marketed product to gain a commercial advantage.

Background and Objectives: Parenteral nutrition (PN) administered to newborns (NB) may be contaminated with polycyclic aromatic hydrocarbons (PAHs) and may therefore increase the contact with these toxicants in very early life stages. The aim of the study is to determine to what extent, if any, commercial products for PN are contaminated with PAHs and to determine whether these contaminants, when present in the bag content, are delivered to NB and whether 1-hydroxypyrene (1-HP), the pyrene metabolite, can be detected in the urine of exposed NB., Methods: Commercial products and the bags administered to 10 NB during their period in the NICU were analyzed for the 16 priority US Environmental Protection Agency PAHs. Urine samples were collected and analyzed for their 1-HP content. Urine samples of a control group composed of 8 breastfed NB were also analyzed for the determination of 1-HP., Results: From 9 different commercial products used to compound PN bags, 6 were contaminated with PAHs, with total concentrations varying from 0.02 to 10.56 mg/L. In the bags administered to the NB, this sum varied from 0.01 to 6.30 mg/L with a mean of 2.62 mg/L. Therefore, for each 100 mL PN, an average load of 0.26 mg PAHs was observed. The majority of the urine samples taken from NB in the study group (80%) contained 1-HP, but it was not detected in the urine of any baby in the control group., Conclusions: The contamination of PN with PAHs poses a critical toxicological risk. The elevated contaminant concentrations and the parenteral way of administration make this source of PAHs considerably worse than any other, including maternal exposure to environmental pollution or tobacco.

Introduction: The lung transplantation (LTx) program began in Ceará in 2011 and the first LTx was performed on June 11, 2011. The aim of this study was to present the initial results of the 6-year experience of our program., Methods: We retrospectively reviewed our experience on LTx from June 2011 to August 2017. Data on recipients and transoperative and postoperative outcomes were recorded in a database., Results: Twenty-two (56.4%) were single LTx, 15 (38.5%) were double, and 2 (5.1%) bilateral lobar. The mean age was 47.5 ± 15 years, and 26 (66.7%) were men. Twenty-eight (71.8%) had pulmonary fibrosis; 5 (12.8%) had pulmonary emphysema, 3 (7.7%) had bronchiectasis; 2 (5.1%) had pulmonary hypertension, and 1 (2.6%) had lymphangioleiomyomatosis. Complications occurred in 82% (32/39) and in-hospital mortality was 30.8% (single LTx = 27.8% and double LTx = 33.3%). The main complications were infection in 17 (43.5%) cases and primary graft dysfunction in 7 (17.9%). There was a significant improvement in pulmonary function in the first year of follow-up (forced expiratory volume pre-LTx = 37% ± 16% and 12 months post-LTx = 72% ± 22%, P = .001); and overall survival at 36 months was 59.0%, with no difference between single- and double-lung transplants., Conclusions: Idiopathic pulmonary fibrosis was the most common underlying disease and single LTx was the most commonly performed operation. There was a high incidence of postoperative complications and in-hospital mortality, but the 36-month follow-up showed a marked improvement in lung function and a global survival similar to the literature., (Copyright © 2018 Elsevier Inc. All rights reserved.)

An analytical method utilizing liquid chromatography coupled to mass spectrometry with electrospray ionization has been developed for the identification of prostaglandins (PGs) in cerebral tissues. The five compounds identified (thromboxane B2, prostaglandin E2, prostaglandin D2, 6-keto-prostaglandin F1 alpha and prostaglandin F2 alpha) are cellular mediators of inflammation and are involved in a variety of physiological and pathological processes by acting on membrane receptors on the surfaces of target cells. The parameters of the electrospray ionization interface were optimized to obtain the highest possible sensitivity for all compounds studied. The limits of detection ranged from 0.25 to 1.09 μg L -1 , and the limits of quantification ranged from 0.83 to 3.64 μg L -1 . The method was validated and applied to samples of brain tissue from five mice. The sample concentrations of the four prostaglandins quantified ranged from 375 ȵg L -1 for prostaglandin E2 to 6602 μg L -1 for prostaglandin D2. An advantage of this work that should be emphasized is the fast response of the method, which allows to obtaining the lipid profile after a 3 min chromatographic run., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: Rubber closures are the primary packaging material for sterile preparations intended for repeated use. Important features of rubber closures are achieved after additives are added to the elastomeric material that compounds the rubber. Among these additives is carbon black. Because of its origin, carbon black may contain polycyclic aromatic hydrocarbons (PAHs). The U.S. Environmental Protection Agency has identified 16 priority PAHs on the basis of concerns that they cause or might cause cancer in animals and humans. Regulatory agencies impose carbon black purity specifications based on limits for total PAHs (0.5 mg/kg) and benzo[a]pyrene (5 μg/kg) or benzo[a]pyrene only (250 μg/kg). PAHs in rubber packaging used for pharmaceutical formulations and in parenteral products stored in containers with rubber stoppers were investigated., Methods: To this end, the method proposed by the National Institute for Occupational Safety and Health-based on high-performance liquid chromatography with ultraviolet and fluorescence detection-was adapted to determine the levels of PAHs in rubber stoppers (gray and red) and in lipid emulsions and amino acid solutions stored in bottles with rubber stoppers., Results: The rubber materials were shown to contain 12 PAHs, in concentrations ranging from 0.25-3.31 µg/g. Only 1 of 18 samples (11 amino acid solutions and 7 lipid emulsions) was uncontaminated. The most prevalent contaminants were pyrene, benzo[a]pyrene, and fluoranthene. The total PAH concentrations in the samples ranged from 0.11-5.96 µg/mL., Conclusion: Components of parenteral nutrition may be contaminated with PAHs, and rubber stoppers represent a potential source of these contaminants.

HPLC coupled to UV diode array detection (DAD) is proposed for the determination of polycyclic aromatic hydrocarbons (PAHs) in pharmaceutical products for parenteral administration. Because rubber is a possible source of PAHs for these products, samples stored in containers with rubber parts were selected for the analysis. The basis for method optimization was EPA Method 8310, which determines 16 priority PAHs in ground water and wastewater by HPLC using both UV and fluorescence detection. Using DAD, two channels were selected for detection, with one operating at 254 nm for the detection of nine PAHs and the other at 225 nm for the detection of seven PAHs. This method allowed for the detection of PAHs using external calibration with LODs and LOQs ranging from 0.001 to 0.060 µg/mL and from 0.003 to 0.167 µg/mL, respectively. Within-day precision, expressed as RSD, varied from 1.24 to 7.76% for PAH concentrations from 0.05 to 0.50 µg/mL, and intraday precision varied from 3.10 to 9.40% for the same concentration range. Method accuracy was confirmed by recoveries of 75-120% of the spiked samples. This method was applied for the determination of PAHs in three commercial infusion solutions and in nine different medications stored in syringes prior to administration to patients. Twelve of 16 PAHs were found in these samples. Total PAH concentrations varied from 0.13 to 13.50 µg/mL. Pyrene was the most prevalent contaminant, being present in 11 of 12 samples in concentrations ranging from 0.17 to 4.80 µg/mL. This method presented good sensitivity for the measurement of PAH in the target samples, allowing for the determination of the 16 priority PAHs in one run and in 30 min.

A sensitive and accurate method, employing high-resolution continuum source atomic absorption spectrometry, for the determination of antimony (Sb) was developed. Conditions such as pyrolysis and atomization temperatures, the use of chemical modifiers, and sample pretreatment were optimized for the determination of Sb in pharmaceutical preparations and beverages stored in polyethylene terephthalate (PET) containers. In addition to container analysis, the influence of time of contact, temperature, and content composition on the migration of Sb was investigated. Twenty-six samples were periodically analyzed until finalize 1 year bottling. Eight mineral water samples were analyzed after storage at temperatures from 40 to 80°C for 48 h. Five different-colored PET samples were stored at 80°C for 20 days and periodically analyzed. Bottles, containing from 46.4 mg/kg Sb (amber PET) to 91.6 mg/kg Sb (colorless PET), leached Sb depending on the content ingredients, time of contact, and temperature. Although drinking water itself did not promote Sb migration at room temperature, juices and medicines did. After 1 year of storage, the Sb level in the samples ranged from 5 to 50 μg/L. The rate of leaching depended on the temperature, which was slow below 60°C, but rapidly increased at 60-80°C. The higher the Sb content in the bottle, the higher the amount of migration. Because Sb is a possible carcinogen to humans, containers for pharmaceutical use should be better controlled, and factors that increase Sb leaching in products that are widely consumed should be avoided.

Ethnopharmacological Relevance: Dolichandra unguis-cati L. is a native climbing plant of Brazil, popularly known as "unha de gato". It has been traditionally used mainly as an antipyretic, anti-inflammatory and anti-tumor agent, yet little toxicological information is found in the literature., Aim of the Study: To identify the chemical composition of the hydroethanolic extract obtained from the leaves of Dolichandra uniguis-cati and to evaluate the acute and subacute toxicity in male and female rats, in order to assess the safety profile of this plant., Materials and Methods: In the acute study, a single dose (2000mg/kg) of the extract was orally administered to male and female rats. In the subacute study, the extract was orally administered to male and female rats at doses 100, 200 and 400mg/kg for 28 days. Behavioral changes, catalase and tbars evaluations, biochemical, hematological and histopathological analysis were determined. The extract' chemical composition was accessed through UHPLC/MS., Results: Chlorogenic acid, caffeic acid, ferulic acid, vanillinic acid, p-coumaric acid, rosmarinic acid, trans-cinnamic acid, luteolin, apigenin, quercitrin and quercetin were identified in the extract. In the acute treatment, the extract was classified as safe (category 5), according to the OECD guide. In relation to the subacute study, females showed a reduction in AST (100, 200 and 400mg/kg), ALT (200mg/kg) and BUN (100 and 200mg/kg) levels, while male rats 400mg/kg presented an increase in AST levels. The Chol dosage significantly decreased in female rats in a dose-dependent manner, whereas for male rats this parameter showed no statistically significant reductions. No behavioral and histopathological changes were recorded., Conclusions: Our results indicate that the hydroethanolic extract of Dolichandra unguis-cati leaves did not present relevant toxic effects when administered orally to male and female rats. The extract also showed a potential hypocholesterolemic activity., (Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.)

Food and agricultural industries generate substantial quantities of phenolic-rich by-products that could be valuable natural sources of antioxidants. The aim of this study was to identify and quantify the phenolic compounds and radical scavenging activities of two by-products (pomace and lees) from Vitis vinifera L. cv Pinot noir. We found a different distribution of phenolic classes (flavanols, flavonols, phenolic acids and stilbenes) and singular scavenging activity against free radicals (hydroxyl, superoxide and peroxyl radicals). The major class of phenolics in pomace was flavanols and in lees was flavonols, with catechin (117.9 ± 2.5 μg g(-1)) and quercetin (42.4 ± 1.2 μg g(-1)) being the most abundant individual compounds. We also found high potential on scavenging activity against superoxide radicals in pomace (80% of scavenging activity) and radical peroxyl (67% scavenging activity). These results show the possibility of using Pinot noir by-products as promising additives or as a source for the development of new products in different segments of the food and cosmetic industries.

An analytical method using liquid chromatography-atmospheric pressure photoionization tandem mass spectrometry with toluene as a dopant was developed for the determination of triterpenes in medicinal plant extracts. The 12 compounds determined have been shown to exhibit biological activity, such as gastroprotective, hepatoprotective, anti-inflammatory, antiviral and anti-tumor effects. The parameters of the atmospheric pressure photoionization interface were optimized to obtain the highest possible sensitivity for all of the compounds. The limits of detection and quantification ranged from 0.4 to 157.9 µg l -1 and 1.3 to 526.4 µg l -1 , respectively. The method was validated and applied to extracts of five medicinal plants species (Mansoa alliacea (Lam.) A.H.Gentry, Bauhinia variegata var variegata, Bauhinia variegata var alboflava, Cecropia obtuse Trécul and Cecropia palmate Willd) from the Amazonian region. The concentrations of the six triterpenes quantified in the samples ranged from 0.424 mg kg -1 for ursolic acid to 371.96 mg kg -1 for β-amyrin, which were quantified by using the standard addition method (n = 3). Copyright © 2016 John Wiley & Sons, Ltd., (Copyright © 2016 John Wiley & Sons, Ltd.)

Nickel and cobalt were simultaneously assayed in vitamin B12 formulations by using atomic spectrometry. The proposed method is based on a compromise between the proximity of specific Ni and Co spectral lines and the relative abundances of the analytes in the samples. The analytes were found in concentrations ranging from 9.48 to 26.20µg L(-1) (Ni) and from 156.90 to 279.25mg L(-1) (Co) in the commercial samples of vitamin B12. The limits of detection and quantification were 1.21 and 3.64mg L(-1) for Co and 0.39 and 1.19µg L(-1) for Ni. Sample cleanup was not necessary for the determinations, and the interferences were discussed., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Methyl-xanthines and adrenergic stimulants, such as caffeine and synephrine, are commonly added to food supplements due to their stimulating and thermogenic effects. In addition, the abusive consumption of food supplements with ergogenic and aesthetic purposes has been observed worldwide. This work describes the study of caffeine, p-synephrine, hordenine, octopamine, tyramine, ephedrine and salicin as stimulants in dietary supplements marketed in Brazil for weight loss and physical fitness claims. A total of 94 different products were acquired from 30 Brazilian websites. Thus, the sampling of marketed supplements was performed in virtual commerce (e-commerce) with claims of weight loss, appetite reduction, fat burning and metabolism acceleration. The developed analytical method involved the separation of the stimulants by HPLC with diode array detection (HPLC-DAD) by using a gradient elution of flow rate (0.7-2.5 ml min(-1)) and mobile phase composition (0.1% H3PO4/methanol). The validated method was applied to the study of 46 dietary supplements. Caffeine, p-synephrine and ephedrine were found to be present as stimulants in 52% of the studied samples marketed as encapsulated or bulk forms. Caffeine was found to be present in concentrations that represent doses from 25.0 to 1476.7 mg day(-1). Synephrine was found in concentrations that represent doses from 59.1 to 127.0 mg day(-1). Ephedrine was found to be associated with caffeine in one formulation at a concentration representing a 26.1 mg day(-1) dosage.

The aim of this study was to investigate the occurrence of multidrug-resistant, extended spectrum beta-lactamase (ESBL) producing Salmonella spp. isolated from conveyor belts of broiler cutting rooms in Brazilian broiler processing plants. Ninety-eight strains of Salmonella spp. were analyzed. Multidrug resistance was determined by the disk diffusion test and the susceptibility of the isolated bacteria was evaluated against 18 antimicrobials from seven different classes. The double disk diffusion test was used to evaluate ESBL production. Of the 98 strains tested, 84 were multidrug resistant. The highest rates of resistance were against nalidixic acid (95%), tetracycline (91%), and the beta-lactams: ampicillin and cefachlor (45%), followed by streptomycin and gentamicin with 19% and 15% of strain resistance, respectively. By contrast, 97% of the strains were sensitive to chloramphenicol. 45% of the strains were positive for the presence of ESBL activity. In this study, high rates of multidrug resistance and ESBL production were observed in Salmonella spp., (Copyright © 2015 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)

Niemann-Pick disease (NPD) is a rare syndrome caused by abnormal intracellular sphingomyelin lipid storage in cells known as "Pick cells." NPD can start in childhood or develop insidiously, with a predilection for reticuloendothelial cells and the nervous system. NPD is a heterogeneous clinical, and biomolecular disorder which has 6 variants. There is no curable treatment for NPD. Generally, the treatment for all types of Niemann-Pick disease is to support. Type B NPD (NPD-B) is mostly characterized by hepatosplenomegaly, which can evolve to hepatic cirrhosis. In patients who progress to liver failure, liver transplantation may be improve liver function. The Transplant Service of Hospital Universitário Walter Cantídio performed its first liver transplants in patients with NPD-B with good results, demonstrating the efficacy of this procedure in selected cases., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Objectives: The aim was to evaluate the effect of high molecular weight protein fraction of Calotropis procera latex on edema formation and oxidative stress in carrageenan-induced paw inflammation., Methods: A sub-plantar injection of carrageenan was given to induce edema in the hind paw of the rat. The inhibitory effect of high molecular weight protein fraction of C. procera latex was evaluated following intravenous administration (5 and 25 mg/kg body weight) and was compared with that of diclofenac given orally (5 mg/kg). The levels of reduced glutathione (GSH), thiobarbituric acid reactive substances (TBARS) and myeloperoxidase (MPO) were measured in the inflamed paw tissue at the end of the study., Results: The high molecular weight protein fraction obtained from the latex of C. procera produced a dose-dependent inhibition of edema formation that was accompanied by normalization of levels of oxidative stress markers (GSH and TBARS) and MPO, a marker for neutrophils in the paw tissue., Conclusions: The high molecular weight protein fraction of C. procera latex ameliorates acute inflammation in the paw through its antioxidant effect.

In this work, the antioxidant and antimicrobial activities of Brassica oleracea var. capitata extracts obtained through ultrasound-assisted extraction are evaluated. The extracts obtained using the best extraction conditions were subjected to different hydrolysis conditions before their use in the biological tests. The crude and hydrolysed extracts were characterized using gas chromatography coupled with a mass detector. The use of ultrasound at 30 °C with 60% (by volume) solvent enabled obtaining a richer extract. All extracts had antioxidant activities against DPPH (13.0-80.0%), superoxide (35.2-63.2%) and peroxyl (89.3-99.5%) radicals, but the use of hydrolysed extracts considerably improved the antioxidant activities. Antimicrobial activities only of the hydrolysed extracts of Brassica oleracea var. capitata were detected. It was confirmed that antioxidant activity of vegetable extracts can be considerably increased when hydrolysis is applied as a pretreatment to their extraction.

Organochalcogens are promising pharmacological agents that possess significant biological activities. Nevertheless, because of the complexity of mammalian models, it has been difficult to determine the molecular pathways and specific proteins that are modulated in response to treatments with these compounds. The nematode worm Caenorhabditis elegans is an alternative experimental model that affords easy genetic manipulations, green fluorescent protein tagging and in vivo live analysis of toxicity. Abundant evidence points to oxidative stress in mediating manganese (Mn)-induced toxicity. In this study we challenged worms with Mn, and investigated the efficacy of inedited selenium- and tellurium-xylofuranosides in reversing and/or protecting the worms from Mn-induced toxicity. In addition, we investigated their putative mechanism of action. First, we determined the lethal dose 50% (LD50) and the effects of the xylofuranosides on various toxic parameters. This was followed by studies on the ability of xylofuranosides to afford protection against Mn-induced toxicity. Both Se- and Te-xylofuranosides increased the expression of superoxide dismutase (SOD-3). Furthermore, we observed that the xylofuranosides induced nuclear translocation of the transcription factor DAF-16/FOXO, which in the worm is known to regulate stress responsiveness, aging and metabolism. These findings suggest that xylofuranosides attenuate toxicity Mn-induced, by regulating the DAF-16/FOXO signaling pathway., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

In this work were extracted bioactive compounds from Brassica oleracea var capitata using supercritical CO2 and evaluated the antioxidant potential of the extracts. Five extractions were accomplished to investigate the influence of pressure (10-25 MPa) and temperature (20-60 °C) in the extraction yield, chemical composition and antioxidant potential towards peroxyl, superoxide and hydroxyl radicals. The highest extraction yield was obtained at 60 °C and 25 MPa, which was 0.47 wt% (run 2). In the characterisation of the extracts obtained was possible the identification of sulforaphane and iberin nitrile that present known biological properties. The extracts of all runs presented antioxidant activities towards the three radicals, but the highest activities for all radicals were using the extracts obtained in the run 2. The use of supercritical CO2 extraction to obtain bioactive compounds of B. oleracea var capitata showed to be a promising alternative to conventional extraction methods, since allowed the extraction of compounds with scientific and industrial interest., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The latex of Calotropis procera is a rich source of proteolytic activity. This latex is known to contain two distinct cysteine peptidases: procerain and procerain B. In this study, new cysteine peptidases were purified from C. procera latex. The enzymes were purified by two sequential ion-exchange chromatography steps (CM-Sepharose plus Resource S(®)) at pH 5.0 and 6.0. The purified enzymes had molecular mass spectra corresponding to CpCP-1=26,213, CpCP-2=26,133 and CpCP-3=25,086 Da. These enzymes exhibited discrete differences in terms of enzymatic activity at a broad range of pH and temperature conditions and contained identical N-terminal amino acid sequences. In these respects, these three new proteins are distinct from those previously studied (procerain and procerain B). Circular dichroism analysis revealed that the new peptidases contain extensive secondary structures, α(15-20%) and β(26-30%), that were stabilized by disulfide bonds. The purified enzymes exhibited plasma-clotting activity mediated by a thrombin-like mechanism. The set of results suggest the three isolated polypeptides correspond to different post-translationally processed forms of the same protein., (Copyright © 2013 Elsevier B.V. All rights reserved.)

A standard additions-dilution solid-state electrochemical method for the determination of psychoactive 1,4-benzodiazepine and antidepressants drugs used as adulterants in commercial slimming herbal formulations is described and compared with conventional standard addition method. The proposed method, based on the voltammetry of microparticles approach, permits quantify, via standard additions methodology, 1,4-benzodiazepine and antidepressants drugs in phytotherapeutic formulations with no need of sample dissolution using dilution with a reference electroactive compound. The method was used to measure 1,4-benzobenzodiazepines (clonazepam, flurazepam, alprazolam, midazolam, bromazepam, chlordiazepoxide, lorazepam and diazepam) and antidepressants (bupropion, sertraline, paroxetine and fluoxetine) in slimming formulations that have been commercialized in Brazil., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Science has a critical role to play in guiding more sustainable development trajectories. Here, we present the Sustainable Amazon Network (Rede Amazônia Sustentável, RAS): a multidisciplinary research initiative involving more than 30 partner organizations working to assess both social and ecological dimensions of land-use sustainability in eastern Brazilian Amazonia. The research approach adopted by RAS offers three advantages for addressing land-use sustainability problems: (i) the collection of synchronized and co-located ecological and socioeconomic data across broad gradients of past and present human use; (ii) a nested sampling design to aid comparison of ecological and socioeconomic conditions associated with different land uses across local, landscape and regional scales; and (iii) a strong engagement with a wide variety of actors and non-research institutions. Here, we elaborate on these key features, and identify the ways in which RAS can help in highlighting those problems in most urgent need of attention, and in guiding improvements in land-use sustainability in Amazonia and elsewhere in the tropics. We also discuss some of the practical lessons, limitations and realities faced during the development of the RAS initiative so far.

Inflammation, angiogenesis and cytokine production are common features of almost, if not all tumors. However, the extent of these processes induced by different types of tumors has not been evaluated. We investigated the growth pattern of the experimental metastatic tumors, B16F10 melanoma, CT26.WT colon and 4T1 mammary cells inoculated in the flank of syngeneic mice and determined the degree of inflammation, angiogenesis, and production level of pro-inflammatory and pro-angiogenic cytokines within the tumors. In addition, we have analyzed vascular changes in the interface between the tumors and the adjacent cutaneous tissue and levels of relevant pro-inflammatory and pro-angiogenic cytokines systemically. The weight of tumors 15 days post-inoculation of 10(6) cells was markedly different. Melanomas were 2 and 10-fold heavier than colon and mammary tumors, respectively. Locally, CT26.WT tumor cells induced more vessels in cutaneous tissue adjacent to the tumors but systemically, the plasma levels of VEGF were higher (approximately 2-fold) in 4T1 tumor-bearing mice compared with the other two tumors. Mammary tumors presented the most prominent inflammatory content as assessed by a range of markers (inflammatory enzymes and cytokines). The vascular index, as determined by the intra-tumor amount of hemoglobin and number of vessels in hot spot areas, was also higher (approximately 2-fold) in melanomas compared with the other two tumors. These findings showing that distinct tumor types determine differential grade of inflammation, angiogenesis and host interaction in mice may provide new insights to tailor differential therapeutic approach based on the status of tumor biomarkers., (Copyright © 2012 Elsevier Inc. All rights reserved.)

A solid state electrochemical method for screening different families of adulterant chemicals illegally added to commercial phytotherapuetic formulations is described. The proposed method, based on the voltammetry of microparticles approach, permits a fast and sensitive way to distinguish between anorexics (amfepramone, fenproporex, sibutramine), benzozodiazepinic anxiolytics (clonazepam, flurazepam, alprazolam, midazolam, medazepam, chlordiazepoxide, diazepam), antidepressants (bupropione, fluoxetine, sertraline, paroxetine), diuretics (hydrochlorothiazide, furosemide, chlortalidone, amiloride, spironolactone), and hypoglycemics (glimepiride, chlorpropamide, glibenclamide) based on characteristic voltammetric signals recorded on solid micro- or nanosamples attached to graphite electrodes immersed into aqueous electrolytes., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A reverse-phase (RP)-HPLC method is reported for determining L-tyrosine, p-octopamine, synephrine, tyramine and hordenine as chemical markers of the species Citrus aurantium in raw material, dry extracts and phytotherapeutic herbal formulations. Using RP-HPLC with diode array detection (DAD) and gradient elution, the amines were determined in 12 different products from different Brazilian states labelled as containing C. aurantium. The presence of the amines was confirmed by mass spectrometry using electrospray ionisation (ESI-MS/MS). This RP-HPLC method allowed the separation of the amines from complex mixtures containing caffeine, ephedrine, salicin and other raw materials (e.g. Garcinia camboja, Phaseolus vulgaris, Caralluma fimbriata, Cassia nomane, Ephedra sp. and Cordia ecalyculata). The method proved useful and selective for inspecting herbal medicines containing p-synephrine and structural analogues. The herbal products analysed had a p-synephrine content ranging from 0.005 to 4.0% (w/w).

A new method is described for the determination of the most common diuretic and laxative adulterants found in formulations of anorexics and antidepressants. The method is based on the separation of furosemide, hydrochlorothiazide, chlorthalidone and amiloride (diuretics), phenolphthalein (laxative), amfepramone (anorexic) and fluoxetine and paroxetine (antidepressants) by capillary zone electrophoresis with capacitively coupled contactless conductivity detection. The method showed a precision ranging from 1.9% to 6.9% for a concentration of 25 mg/L, 0.6% to 5.3% for a concentration of 50 mg/L and 1.6% to 6.0% for a concentration of 100 mg/L for all analytes. The accuracy was 99% for amiloride, 102% for chlorthalidone, 101% for hydrochlorothiazide, 101% for furosemide, 94% for phenolphthalein, 105% for fluoxetine, 114% for paroxetine and 117% for amfepramone. The method allowed the drugs to be determined in the formulations at concentrations higher than 5.1 mg/kg for amiloride, 7.7 mg/kg for chlorthalidone, 6.8 mg/kg for hydrochlorothiazide, 10.7 mg/kg for furosemide, 8.4 mg/kg for phenolphthalein, 11.0 mg/kg for fluoxetine, 9.4 mg/kg for paroxetine and 11.0 mg/kg for amfepramone. Three of the 26 analysed herbal formulations were found to be adulterated (not declared on the label) with the diuretic hydrochlorothiazide. Five other samples contained diuretics declared on the label on the formulation. Thus, a total of eight samples, which were marketed as natural products, contained diuretics (declared or not) on the formulation.

The aim of this study was to evaluate the efficiency of low oxygen tension (5% CO(2) , 5% O(2) and 90% N(2) ) on in vitro oocyte maturation using defined media (0.1% polyvinyl alcohol - PVA) or 10% porcine follicular fluid (PFF)-supplemented media. To achieve this goal, oocytes were evaluated regarding cortical granules (GCs) migration, nuclear maturation and sperm penetration. Oocytes were in vitro matured under different conditions: 5% or 20% O(2) atmosphere and 0.1% PVA- or 10% PFF-supplemented media and evaluated at 0 and 44 h of maturation. To evaluate the migration of CGs and nuclear maturation, by confocal microscopy, oocytes were incubated with 100 μg of FITC-PNA/ml and 10 μg/ml of propidium iodide. To address sperm penetration, after maturation, in vitro fertilization for 6 h and in vitro culture for 18 h, zygotes were incubated with 10 mg/ml Hoechst 33342. Pronuclei and polar bodies were quantified using an epifluorescence microscope. Atmosphere conditions did not affect the CGs migration, but media supplementation did. Oocytes matured in 10% PFF media had a higher percentage of CGs in the oocyte periphery than oocytes matured in PVA-supplemented media. However, this fact did not have effect on in vitro sperm penetration levels. No effect of atmosphere conditions and media supplementation was observed on the rates of metaphase II oocytes. Therefore, the use of low oxygen tension in association with PVA maturation media does not improve the in vitro maturation system of porcine oocytes, because its use did not improve nuclear maturation, CGs migration and zygotes monospermic rates., (© 2011 Blackwell Verlag GmbH.)

The objectives of the present study were to evaluate the spread of Salmonella Enteritidis to different cutting boards (wood, triclosan-treated plastic, glass, and stainless steel) from contaminated poultry skin (5 log CFU/g) and then to tomatoes and to analyze the effect of different protocols used to clean these surfaces to control contamination. The following procedures were simulated: (1) no cleaning after handling contaminated poultry skin; (2) rinsing in running water; (3) cleaning with dish soap and mechanical scrubbing; and (4) cleaning with dish soap and mechanical scrubbing, followed by disinfection with hypochlorite. The pathogen was recovered from all surfaces following procedure 1, with counts ranging from 1.90 to 2.80 log, as well as from the tomatoes handled on it. Reduced numbers of S. Enteritidis were recovered using the other procedures, both from the surfaces and from the tomatoes. Counts were undetectable after procedure 4. From all surfaces evaluated, wood was the most difficult to clean, and stainless steel was the easiest. The use of hypochlorite as a disinfecting agent helped to reduce cross-contamination., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Objective: To evaluate the frequency and clinical features of endemic and other opportunistic infections in liver or kidney transplant recipients in four transplant centres in different geographical areas of Brazil., Methods: Retrospective analysis of medical and laboratory records of four transplant centres on endemic and other opportunistic infections in liver or kidney transplant recipients. Analyses were performed with spss statistical software., Results: From 2001 to 2006, 1046 kidney and 708 liver transplants were registered in all centres. The average age was 42 years. Among 82 (4.7%) cases with infections, the most frequent was tuberculosis (2.0%), followed by systemic protozoal infections (0.7%), toxoplasmosis (0.4%) and visceral leishmaniasis (0.3%). Systemic fungal infections occurred in 0.6%, of which 0.4% were cryptococcosis and 0.2% were histoplasmosis. Dengue was the only systemic viral infection and was registered in two cases (0.1%), of which one was classified as the classic form and the other as dengue haemorrhagic fever. Nocardiosis was described in one case (0.05%). The infectious agents most frequently associated with diarrhoea were Blastocystis sp., Schistosoma mansoni and Strongyloides stercoralis., Conclusions: Opportunistic Infections in transplant patients have a wide spectrum and may vary from asymptomatic to severe infections with high mortality. A better understanding of the epidemiology of endemic pathogens and clinical manifestations can contribute to the establishment of an early diagnosis as well as correct treatment aimed at decreasing morbidity and mortality., (© 2011 Blackwell Publishing Ltd.)

Aim: To evaluate two NiTi wires with different carbon and oxygen contents in terms of mechanical resistance to rotary bending fatigue (RBF) under varied parameters of strain amplitude and rotational speed., Methodology: The wires produced from two vacuum induction melting (VIM) processed NiTi ingots were tested, Ti-49.81 at%Ni and Ti-50.33 at%Ni, named VIM 1 and VIM 2. A brief analysis related to wire fabrication is also presented, as well as chemical and microstructural analysis by energy dispersive spectroscopy (EDS) and optical microscope, respectively. A computer controlled RBF machine was specially constructed for the tests. Three radii of curvature were used: 50.0, 62.5 and 75.0 mm, respectively, R(1), R(2) and R(3), resulting in three strain amplitudes ε(a) : 1.00%, 0.80% and 0.67%. The selected rotational speeds were 250 and 455 rpm., Results: The VIM 1 wire had a high carbon content of 0.188 wt% and a low oxygen content of 0.036 wt%. The oxygen and carbon contents of wire VIM 2 did not exceed their maximum, of 0.070 and 0.050 wt%, according to ASTM standard (ASTM F-2063-00 2001). The wire with lower carbon content performed better when compared to the one with higher carbon content, withstanding 29,441 and 12,895 cycles, respectively, to fracture., Conclusions: The surface quality of the wire was associated with resistance to cyclic fatigue. Surface defects acted as stress concentrators points. Overall, the number of cycles to failure was higher for VIM 2 wires with lower carbon content., (© 2011 International Endodontic Journal.)

Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world and the third leading cause of cancer-related death. In this study, we sought to assess the outcome of patients with HCC who underwent orthotopic liver transplantation (OLT) in a center in the northeast of Brazil. Between May 2002 and July 2008, 294 OLTs were performed at our center. In 45 patients, HCC was confirmed by histological examination of the explant. Patients were predominantly men of ages ranging from 14-67 years. Hepatitis C virus was involved in 55.4% of the cases. Alpha fetoprotein (AFP) levels were normal in 65.2% of the patients and surpassed 100 ng/mL in only 10.4%. The median waiting time on the list was 10 months. Seventeen patients (37.7%) presented a solitary nodule, 19 (42.2%) had 2 or 3 nodules, and 9 patients (20%) had more than 3 nodules. The maximal diameter of the largest tumor was <3 cm in 26 patients (57.7%) and exceeded 5 cm in 6 patients (13.3%). Ten tumors were well differentiated, 32 were moderately differentiated, and 3 were poorly differentiated. Eleven tumors showed microvascular invasion. There have been 4 tumor recurrences. There was an association between microvascular invasion and tumor recurrence with a statistically significant relative risk. In conclusion, OLT is an excellent option for patients with HCC. The recurrence rate was low (<10%). However, we believe that more prospective studies are needed about OLT beyond the Milan criteria because our study suggested that microvascular invasion may be more important than tumor size or number.

We aimed to examine baseline predictors of treatment response in bulimic patients. 241 seeking-treatment females with bulimia nervosa completed an exhaustive assessment and were referred to a six-session psychoeducational group. Regression analyses of treatment response were performed. Childhood obesity, lower frequency of eating symptomatology, lower body mass index, older age, and lower family's and patient's concern about the disorder were predictors of poor abstinence. Suicidal ideation, alcohol abuse, higher maximum BMI, higher novelty seeking and lower baseline purging frequency predicted dropouts. Predictors of early symptom changes and dropouts were similar to those identified in longer CBT interventions.

Background: Hepatic artery thrombosis (HAT) is a serious complication after orthotopic liver transplantation (OLT) and remains a significant cause of graft loss. HAT following OLT has been reported in 3% to 9% of patients. Among the surgical factors considered to be associated with HAT, arterial reconstruction might be the most important. The goal of this study was to compare the incidence of HAT between interrupted suture (IS) and continuous suture (CS) techniques during hepatic artery reconstruction in liver transplantation., Methods: We performed a retrospective analysis of 200 consecutive liver transplantations occurring between May 2002 and December 2006, including medical records for: age, gender, cold ischemic time, warm ischemic time, type and number of arterial anastomosis. Hepatic artery anastomoses were performed using a 7-0 prolene with a running CS in the first 105 patients (CS group), and with an IS in the last 95 patients (IS group)., Results: Statistical analysis of age, gender, cold and warm ischemia time, and number of hepatic artery anastomoses was not different between the CS and IS groups. Eleven episodes of HAT were identified in the CS group (10%) and two episodes (2%) in the IS cohort, a significant difference (P = .0173)., Conclusions: Our results suggested that IS might be a better choice for hepatic artery anastomosis with a lower incidence of HAT.

Background: Agnathia is a rare malformation characterized by the absence of the mandible., Methods: A male rhesus monkey with malformations was found dead and studied by internal examination, radiographs and histopathology., Results: A case of a rare first branchial arch anomaly with agenesis of the mandible and tongue is presented. The animal also had visceral deformities. However, ears were normal in shape and only slightly low in position. The craniofacial malformations may reflect incomplete separation of the first branchial arch into its maxillary and mandibular processes., Conclusions: The association between the craniofacial and other corporal anomalies is unclear.

Arterial complications after liver transplantation are frequent. Hepatic artery thrombosis (HAT) is usually associated with biliary complications. Herein we have reported a case of a patient who was admitted for jaundice, itch, and elevated aspartate aminotransferase and alanine aminotransferase levels at 6 weeks after liver transplantation. HAT associated with a biloma was diagnosed and an urgent operation performed requiring a new biliodigestive anastomosis technique. Fourteen months after the first transplant, the patient was retransplanted. The operation performed may be an alternative to treat biliary complications due to late HAT.

Conclusion: Every patient with severe or profound hearing loss must have a temporal bone high-resolution computed tomography (CT) scan. Stapedotomy is a simple, safe and low-cost procedure compared with cochlear implantation and can provide very good results. This can justify our decision to propose stapedotomy at the initial treatment in patients with very far-advanced otosclerosis. In cases of hearing failure after stapes surgery, cochlear implantation is an option., Objective: This study aimed to find the best first intention treatment of very far-advanced otosclerosis., Materials and Methods: This was a retrospective study and included 14 patients with non-measurable preoperative bone and air conduction thresholds and otosclerosis on temporal bone high-resolution CT scan. Stapes surgery followed by a well fitted hearing aid was the initial treatment in 11 patients and cochlear implantation in 7 patients, including 4 patients who had poor results after stapedotomy. Objective and subjective audiometric results were studied and compared between stapedotomy and cochlear implantation groups., Results: Objective and subjective results were statistically better in the cochlear implant group than in the stapedotomy group. However, four patients in the stapedotomy group had comparable results to the patients with cochlear implants.