Establishment of methods to analyze the structural and functional integrity of the quail (Coturnix coturnix japonica) sperm plasma membrane.

1. The objectives of this study were to establish the use of the fluorophores Hoechst 33342 and propidium iodide for the evaluation of sperm plasma membrane integrity and to identify an adequate hypoosmotic solution for the evaluation of sperm membrane functionality in quails.2. Sperm samples were collected from the vas deferens of nine quails. After initial evaluation, the samples were subjected to a flash-frozen assay. Three treatments with the following proportions of fresh sperm and sperm subjected to flash freezing were prepared as follows: 100:0 (T100), 50:50 (T50), and 0:100 (T0). The hypoosmotic swelling test used distilled water (0 mOsm/l) and fructose solutions (50, 100, and 200 mOsm/l).3. Immediately after recovery, the samples showed 75.6 ± 5.0% motility with vigour of 3.7 ± 0.3 and 96.1 ± 0.5% of the sperm appeared normal. The membrane integrity test showed 62.2 ± 5.2% intact sperm at T100, 29.0 ± 4.1% at T50 and 0.1 ± 0.1% at T0. Moreover, a greater number of reactive sperm (74.7 ± 6.7%) were observed when incubated in distilled water (0 mOsm/l) in comparison to other solutions (P < 0.05).4. The association of fluorescent probes composed of Hoechst 33342 and propidium iodide provided an efficient assessment of the integrity of the plasmatic membrane of quail spermatozoa. However, the study identified that the hypoosmotic swelling test has little predictive value regarding sperm membrane functionality in this species.

Analysis of the association of clinical factors and IL4 gene polymorphisms with root resorption in avulsed teeth after 1 year of replantation.

AIM: To investigate the association of clinical variables and polymorphisms (tag SNPs) in the interleukin 4 (IL4) gene, with the prognosis of avulsed and replanted teeth. METHODOLOGY: Ninety-four patients who suffered avulsion and had their teeth replanted and endodontically treated were included. Periapical radiographs were obtained soon after tooth replantation and after 1 year. For genotypic IL4 gene analysis, the DNA from oral mucosa cells was collected, and polymorphisms were investigated by real-time PCR. Univariate and multivariate analyses were performed to verify the association of clinical and genetic variables and the outcome of the replanted teeth (P < 0.05). RESULTS: After multivariate analysis, extra-alveolar time longer than 1 h was significantly associated with external root resorption. No significant association was observed between IL4 gene polymorphisms and root resorption. CONCLUSION: No association between root resorption and IL4 gene polymorphisms was observed. An extra-alveolar time of more than 1 h was associated with a susceptibility for external root resorption. Replanting the tooth in its socket immediately is the most important factor to maintain a healthy root surface.

Freshwater shrimps (Macrobrachium depressimanum and Macrobrachium jelskii) as biomonitors of Hg availability in the Madeira River Basin, Western Amazon.

Total mercury (THg) concentrations measured in two freshwater shrimp species (Macrobrachium depressimanum and Macrobrachium jelskii) showed a relationship with the location of artisanal and small-scale gold mining (ASGM) from the Madeira River Basin, Western Amazon. Between August 2009 and May 2010, 212 shrimp samples were collected in the confluence of the Madeira River with three of its tributaries (Western Amazon). THg concentration was quantified in the exoskeleton, hepatopancreas and muscle tissue of the shrimps by cold vapor atomic absorption spectrophotometry. There were no significant differences between the two shrimp species when samples came from the Madeira River, but Hg concentrations were significantly lower in a tributary outside the influence of the gold mining area. Average THg concentrations were higher in the hepatopancreas (up to 160.0 ng g-1) and lower in the exoskeleton and muscle tissue (10.0-35.0 ng g-1 and < 0.9-42.0 ng g-1, respectively). Freshwater shrimps from the Madeira River respond to local environmental levels of Hg and can be considered as biomonitors for environmental Hg at this spatial scale. These organisms are important for moving Hg up food webs including those that harbor economic significant fish species and thus enhancing human exposure.

Complete physical mapping of IL6 reveals a new marker associated with chronic periodontitis.

BACKGROUND AND OBJECTIVE: Interleukin-6 (IL-6) is a powerful stimulator of osteoclast differentiation and bone resorption. Production of IL-6 is modulated by polymorphisms, and higher levels of this cytokine are found locally in patients with chronic periodontitis. In this study we performed a modern approach - Complete physical mapping of the IL6 gene - to identify the polymorphisms associated with chronic periodontitis in a southern Brazilian population sample. MATERIAL AND METHODS: One-hundred and nine individuals of both genders (mean age: 41.5 ± 8.5 years) were divided into a study group (56 participants with periodontitis) and a control group (53 individuals without periodontitis). After collection and purification of DNA, nine tag single nucleotide polymorphisms (SNPs; rs1524107, rs2069835, rs2069837, rs2069838, rs2069840, rs2069842, rs2069843, rs2069845 and rs2069849) covering the entire gene were selected according to the information available on the International HapMap Project website and evaluated using real-time PCR. RESULTS: Differences in the distribution of the following parameters were statistically significant between study and control groups: number of teeth (p = 0.030); probing depth (p < 0.001); clinical attachment level (p < 0.001); gingival index (p < 0.001); plaque index (p = 0.003); calculus index (p < 0.001); and dental mobility (p < 0.001). It was found that marker rs2069837 (located in intron 2 of IL6) under G dominant was associated with protection against chronic periodontitis in a Brazilian population in the presence of clinical variables, such as visible plaque, dentist visit frequency and dental floss use, and was suggested for the first time as a marker of susceptibility to chronic periodontitis. CONCLUSION: Complete physical mapping of IL6 (using tag SNPs) was carried out for the first time, unveiling allele G of polymorphism rs2069837 (located in the second intron of IL6) as a suggestive marker of protection against chronic periodontitis in a Brazilian population.

Egg Developmental Time and Survival of Chrysomya megacephala and Chrysomya putoria (Diptera: Calliphoridae) Under Different Temperatures.

Chrysomya megacephala (F.) and Chrysomya putoria (Wiedemann) (Diptera: Calliphoridae) are considered of forensic, medical, and veterinary importance in Brazil because of their necrophagous and synanthropic behaviour. The development of flies can be influenced by temperature, and species from the same genus usually have different responses to external variables. The egg development of blow fly can be a useful complementary technique to estimate the minimum postmortem interval. Thus, this study aimed to compare the egg developmental time and survival of C. megacephala and C. putoria at different temperatures to determine the optimal temperature for egg development and the linear regression for developmental time and temperature, thereby determining the minimum threshold (t) and thermal summation constant (K) for each species. Adults of both species were collected in the region of Campinas city, São Paulo state, Brazil. Eggs were incubated at eight constant temperatures between 05 ± 1°C and 35 ± 1°C and the egg developmental time and survival were evaluated. There was no egg survival at 5 and 10°C. The K for C. megacephala and C. putoria were 179.41 HD and 189.94 HD, respectively. The regression slopes and t (10°C) were similar for both species. The optimal temperature for egg survival was between 25 and 35°C, for C. megacephala and 20 and 30°C, for C. putoria. The present data were similar to most data available in the literature, but differences in the same species are a possibility.

Impact of drug formulation and free platinum/cisplatin ratio on hypersensitivity reactions to cisplatin: formulation matters.

WHAT IS KNOWN AND OBJECTIVE: Use of cisplatin can induce type I hypersensitivity reactions that may also be linked to the quality of the drug utilized. We observed cases of hypersensitivity that appeared to be associated with the brand of cisplatin used. The aim of this study was to compare two different brands of cisplatin in relation to type I hypersensitivity reactions. METHODS: Brand A was used in a tertiary care teaching hospital until 2012, and use of brand B started from January 2013, when the first hypersensitivity cases were observed. Patients were categorized based on symptom. Cisplatin of both brands was analysed by high-performance liquid chromatography (HPLC) and high-resolution electrospray ionization mass spectrometry (ESI-(+)-MS) and characterized according to US Pharmacopeia. RESULTS AND DISCUSSION: There were no cases of hypersensitivity associated with the use of cisplatin brand A, whereas four of 127 outpatients that used cisplatin brand B were affected. The two brands were in accordance with the US Pharmacopeia parameters, and there was no significant difference in the total platinum levels between the two brands when analysed by HPLC. However, high-resolution ESI-(+)-MS analyses show that brand B contains approximately 2.7 times more hydrolysed cisplatin than brand A. WHAT IS NEW AND CONCLUSION: The increase in the hydrolysed form of cisplatin found in brand B may be the cause of the hypersensitivity reaction observed in a subset of patients. We present the first study of the quality of drugs by high-resolution ESI-(+)-MS. Drug regulatory agencies and manufacturers should consider including measurement of hydrolysed cisplatin as a quality criterion for cisplatin formulations.

Concentration of mycotoxins and chemical composition of corn silage: a farm survey using infrared thermography.

This work evaluated the chemical composition and mycotoxin incidence in corn silage from 5 Brazilian dairy-producing regions: Castro, in central-eastern Paraná State (n=32); Toledo, in southwestern Paraná (n=20); southeastern Goiás (n=14); southern Minas Gerais (n=23); and western Santa Catarina (n=20). On each dairy farm, an infrared thermography camera was used to identify 3 sampling sites that exhibited the highest temperature, a moderate temperature, and the lowest temperature on the silo face, and 1 sample was collected from each site. The chemical composition and concentrations of mycotoxins were evaluated, including the levels of aflatoxins B1, B2, G1, and G2; zearalenone; ochratoxin A; deoxynivalenol; and fumonisins B1 and B2. The corn silage showed a highly variable chemical composition, containing, on average, 7.1±1.1%, 52.5±5.4%, and 65.2±3.6% crude protein, neutral detergent fiber, and total digestible nutrients, respectively. Mycotoxins were found in more than 91% of the samples, with zearalenone being the most prevalent (72.8%). All samples from the Castro region contained zearalenone at a high average concentration (334±374µg/kg), even in well-preserved silage. The incidence of aflatoxin B1 was low (0.92%). Silage temperature and the presence of mycotoxins were not correlated; similarly, differences were not observed in the concentration or incidence of mycotoxins across silage locations with different temperatures. Infrared thermography is an accurate tool for identifying heat sites, but temperature cannot be used to predict the chemical composition or the incidence of mycotoxins that have been analyzed, within the silage. The pre-harvest phase of the ensiling process is most likely the main source of mycotoxins in silage.

Evaluation of an ELISA for canine leishmaniasis immunodiagnostic using recombinant proteins.

The present work describes the isolation and purification of two Leishmania chagasi (= syn. Leishmania infantum) recombinant proteins, rLci2B and rLci1A, and their use in the development of an immunoassay for the diagnostic of canine leishmaniasis. After protein expression and cell disruption, rLci2B was purified by immobilized metal affinity chromatography followed by size exclusion chromatography, whereas rLci1A, expressed as an inclusion body, was treated with urea and purified by anion-exchange chromatography. Homogeneities were ascertained by denaturing gel electrophoresis (MW (rLci2B) = 46,370; MW(rLci1A) = 88,400), isoelectric focusing (pI (rLci2B) = 5·91; pI (rLci1A) = 6·01) and Western blot. An indirect ELISA was developed using the purified antigens rLci2B and rLci1A and a leishmaniasis canine serum panel (n = 256). The ELISA showed 100% sensitivity and 95% specificity for rLci2B and 96% sensitivity and 92% specificity for rLci1A. The purified proteins did not present cross-reactivity with sera from dogs infected with Trypanosoma caninum, Babesia canis and Ehrlichia canis. Cross-reaction was verified with sera from dogs infected with Leishmania brasiliensis (11·7% for rLci2B and 2·9% for rLci1A). Based on ELISA results, it is suggested the use of rLci2B and rLci1A as antigens in an alternative serological assay for diagnostic of canine leishmania.

Effect of nandrolone decanoate on the development of three species of Chrysomya (Diptera:Calliphoridae), flies of forensic importance in Brazil.

Necrophagous insects are valuable tools for postmortem interval (PMI) estimation or for determining the cause of death. Due to the increase in deaths related to drug abuse, it is crucial to know how these substances affect the development of flies that feed on corpses, to avoid errors in the PMI estimates. This study evaluated the effect of nandrolone decanoate, an anabolic androgenic steroid, on the development ofimmatures of Chrysomya megacephala (F.), Chrysomya putoria (Wiedemann), and Chrysomya albiceps (Wiedemann) (Diptera:Calliphoridae) when added to an artificial rearing diet. Four experimental groups were delineated: three of them were given diets containing 4.5, 22.5, or 45 mg/kg nandrolone decanoate; and a drug-free control group. Weights were recorded at 12-h intervals from larval eclosion to pupation. No statistically significant differences were observed in mean larval weights, emergence interval, or emergence rates for all groups. However, differences in the three species were observed during the larval development. Initially, C. putoria reared in the highest concentration of decanoate showed greater weight gain. However, at older ages, immatures reached lower mean weights than the control group. For C. albiceps, the highest concentration of decanoate contributed to an effective lack of weight gain during almost the entire course of larval development. Therefore, the influence of any drug on development should always be considered.

Resistance Sources and Antixenotic Factors in Brazilian Bean Genotypes Against Bemisia tabaci.

Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) Middle East Asia Minor 1 is one of the most important pests of the common bean, due to its potential of causing direct and indirect damage. This study aimed to evaluate 78 bean genotypes to verify the occurrence of resistance of antixenosis type against B. tabaci. Initially, multiple-choice trials were performed to evaluate the oviposition preference and nymphs' establishment at 3 and 15 days after infestation. Subsequently, 21 bean genotypes were selected, and a no-choice test was conducted. Colorimetric analyses were performed to establish correlations between leaf color and insect establishment. In multiple-choice trial, the genotypes BRS Ametista, BRS Estilo, BRS Esplendor, SCS 204 Predileto, BRS Notável, IPR Eldorado, CHIB 06, IPR Quero-Quero, Iapar 81, CHIP 338, IPR Garça, Arcelina 4, SCS 202 Guará, IAC Esperança, H96102-1-1-1-52, CHIP 348, Carioca Comum, CHIP 300, IAC Carioca Eté, IAC Ybaté, and Tybatã were the least used for oviposition and nymph establishment, demonstrating antixenosis or antibiosis. In the no-choice trial, most genotypes were less attractive to whitefly, and the genotypes CHIB 06, IPR Garça, CHIP 300, and IAC Esperança had less oviposition. The most attractive genotypes presented high luminosity and more intense green and yellow colors, indicating positive correlation. Therefore, the genotypes BRS Ametista, SCS 204 Predileto, BRS Estilo, IPR Eldorado, SCS-202 Guará, Carioca Comum, Arcelina 4, CHIP 348, and IAC Esperança showed the highest resistance stability in the no-choice trial, and they are promising sources of antixenotic factors for use in breeding programs to obtain whitefly-resistant common bean lines.

Adhesion of Candida tropicalis to polystyrene and epithelial cell lines: Insights of correlation of the extent of adherent yeast cells among distinct surfaces.

Candida tropicalis is an emerging fungal pathogen associated with high mortality. We aimed to compare adherence capability of C. tropicalis to polystyrene and epithelial cell lines (HeLa and Vero), and determine whether adherent blastoconidia is cell-type specific. Blastoconidia adhesion to epithelial cells and polystyrene were determined by crystal violet assay. The percentage of epithelial cells with adhered blastoconidia and the number of adhered blastoconidia per cell line were determined by light microscopy. The correlation between adhesion surfaces was assessed by Pearson's correlation coefficient. The adhesiveness of C. tropicalis to polystyrene was greater than that observed for ephitelial cells. High correlation values (r2 0.9999222, p 0.007941) were found for the adhesion capability between biotic and polystyrene surface for isolates 100.10 (obtained from blood) and 335.07 (obtained from tracheal secretion). The number of adherent blastoconidia per HeLa cell was greater in comparison to that observed for Vero cells (P<0.05). Further, high correlation (r2 1, p 0.0001) was found for the adhesion ability between HeLa cells and Vero cells. The results suggest a correlation of C. tropicalis adhesion capability among different surfaces, and that the adhesion to epithelial cells is specific to the cell type.

Maternal high fat diet consumption reduces liver alpha7 nicotinic cholinergic receptor expression and impairs insulin signalling in the offspring.

The activation of nicotinic acetylcholine receptor α7 subunit (α7nAChR) has been associated to anti-inflammatory response in macrophages. High-fat diet (HFD) consumption during pregnancy and lactation impairs the cholinergic anti-inflammatory pathway in liver and white adipose tissue of offspring. In order to evaluate the relationship between damage in the cholinergic anti-inflammatory pathway and insulin resistance (IR) development, the liver of offspring of obese dams was investigated. Additionally, the capacity of α7nAChR activation to reduce IR induced by saturated fatty acid was investigated in hepatoma cell line. Initially, female mice were subjected to either standard chow (SC) or HFD during pregnancy and lactation period. After weaning, only male offspring from HFD dams (HFD-O) and SC dams (SC-O) were fed with the SC diet. Hepatic α7nAChR expression was downregulated, and hepatic TNF-α, IL-1ß, and pIKK level, but not pJNK, were elevated in the HFD-O compared to SC-O mice. Besides, hepatic expression of TNF-α in response to lipopolysaccharide (LPS) was higher in HFD-O than SC-O mice. Insulin-stimulated phosphorylation of the AKT was lower in HFD-O compared to SC-O. Additionally, insulin-stimulated phosphorylation of the AKT in KOα7Alb-Cre mice fed HFD was lower than WT mice fed HFD. In hepatoma cell line, palmitate increased IL-6 and TNF-α expressions and pJNK level. These effects were accompanied by reduced capacity of insulin to stimulate AKT phosphorylation. PNU or nicotine reduced cytokine expression and JNK activation, but improved insulin resistance induced by palmitate. Our results suggest that maternal obesity impairs hepatic α7nAChR expression and AKT phosphorylation in the offspring. In vitro studies suggest that α7nAChR activation has potential to reduce deleterious effect of saturated fatty acids on insulin signalling.

Difficulty in detecting low levels of polymyxin resistance in clinical Klebsiella pneumoniae isolates: evaluation of Rapid Polymyxin NP test, Colispot Test and SuperPolymyxin medium.

Polymyxins are important therapeutic options for treating infections, mainly those caused by carbapenem-resistant Klebsiella pneumoniae. Specific chemical characteristics of polymyxins make it difficult to perform antimicrobial susceptibility testing, especially within the clinical laboratory. Here we aimed to evaluate the performance of three phenotypic methods: Rapid NP Polymyxin Test, ColiSpot test and the SuperPolymyxin medium. To accomplish this, 170 non-duplicate clinical K. pneumoniae isolates were analysed (123 colistin-resistant and 47 susceptible). The sensitivity and specificity obtained for Rapid Polymyxin NP Test, Colispot and SuperPolymyxin medium were, respectively, 90% and 94%, 74% and 100%, and 82% and 85%. Very major errors occurred more frequently in low-level colistin-resistant isolates (MICs 4 and 8 µg/mL). Rapid Polymyxin NP proved to be a method capable of identifying colistin-resistant strains in acceptable categorical agreement. However, major errors and very major errors of this method were considered unacceptable for colistin-resistance screening. Although the Colispot test is promising and easy to perform and interpret, the results did not reproduce well in the isolates tested. The colistin-containing selective medium (SuperPolymyxin) showed limitations, including quantification of mucoid colonies and poor stability. Nevertheless, Colispot and SuperPolymyxin medium methods did not present acceptable sensitivity, specificity and categorical agreement. It is essential to use analytical tools that faithfully reproduce bacterial resistance in vitro, especially in last-line drugs, such as polymyxins, when misinterpretation of a test can result in therapeutic ineffectiveness.

Contribution of lean, fat, muscle color and degree of doneness to pork and beef species flavor.

Flavor is an important contributor to consumer acceptability of meat, our objective was to characterize the impact of species-specific fat/lean sources, fat level, degree of doneness and muscle color are on pork and beef flavor. Three separate experiments were conducted. Patties were formulated differently for each experiment in order to evaluate the desired variables. Experiment. 1: Flavor from combination patties (same species lean/fat or combination of species lean/fat) was not impacted by degree of doneness (66°C vs. 71°C). Beef flavor was highest in samples made with beef lean, regardless of species fat type. Pork flavor was highest in samples made with pork lean and had higher flavor intensity scores. Experiment. 2: Beef flavor was not increased in all-beef patties formulated with higher fat levels. Pork patties formulated with higher fat content increased pork flavor. Experiment. 3: All-beef and all-pork patties formulated with light or dark lean did not impact flavor in either species.

Analysis of the association of IL4 polymorphisms with orthodontic mini-implant loss.

The aim of this study was to investigate the association of clinical characteristics and IL4 tag single nucleotide polymorphisms (SNPs; rs2227284 and rs2243268) with orthodontic mini-implant (MI) failure. The sample included 135 subjects of both sexes, mean age 48.7±10years (range 20-76years): 104 in the control group (patients without any MI loss) and 31 in the study group (patients presenting ≥1 MI loss). Genotypes were determined by real-time PCR. Bivariate and multivariate analyses were performed (P<0.05). No association was found between the selected tag SNPs and MI loss. The C allele of the IL4 rs2243268 polymorphism in the recessive model was more frequent in patients who had fewer MIs installed (≤2 vs. >2; P=0.043, odds ratio 0.65, 95% confidence interval 0.58-0.74). On multivariate analysis, smoking habit was significantly associated with the group with multiple MIs installed (P=0.036), however the significance of the association with rs2243268 was not maintained. No association was found between the socio-demographic, smoking, or genetic factors studied and MI loss. This study supports the interaction between host and environmental factors and its influence on susceptibility to orthodontic MI failure.

Analysis of the association of polymorphism in the osteoprotegerin gene with susceptibility to chronic kidney disease and periodontitis.

BACKGROUND AND OBJECTIVE: Chronic kidney disease (CKD) is a complex disorder, which results in several complications involving disturbance of mineral metabolism. Periodontal disease is an infectious disease that appears to be an important cause of systemic inflammation in CKD patients. Periodontal disease is characterized by clinical attachment loss (CAL) caused by alveolar bone resorption around teeth, which may lead to tooth loss. Osteoprotegerin (OPG) is a key regulator of osteoclastogenesis. Polymorphisms are the main source of genetic variation, and single nucleotide polymorphisms (SNPs) have been reported as major modulators of disease susceptibility. The aim of this study was to investigate the association of a polymorphism located at position -223 in the untranslated region of the OPG gene, previously known as -950, with susceptibility to CKD and periodontal disease. MATERIAL AND METHODS: A sample of 224 subjects without and with CKD (in hemodialysis) was divided into groups with and without periodontal disease. The OPG polymorphism was analyzed by polymerase chain reaction and restriction fragment length polymorphism. RESULTS: No association was found between the studied OPG polymorphism and susceptibility to CKD or periodontal disease. CONCLUSION: It was concluded that polymorphism OPG-223 (C/T) was not associated with CKD and periodontal disease in a Brazilian population. Studies on other polymorphisms in this and other genes of the host response could help to clarify the involvement of bone metabolism mediators in the susceptibility to CKD and periodontal disease.

Drought promotes increases in total mercury and methylmercury concentrations in fish from the lower Paraíba do Sul river, southeastern Brazil.

Bioaccumulation of mercury and methylmercury in fish represents a serious risk to human beings. Extreme climate events like droughts may increase the trophic transfer of contaminants and net methylation of mercury. The present study assessed the influence of the 2014 drought on total mercury and methylmercury levels in fish from the lower Paraiba do Sul river basin. Contaminant levels were compared for Pimelodus fur, Pachyurus adspersus, Pimelodella lateristriga, Oligosarcus hepsetus, and Crenicichla lacustris captured in five sites in 2013 (N = 212) and 2014 (N = 231). The results indicate that levels of contaminants were higher during the drought in most species. Rainfall was weakly and negatively correlated with total mercury levels in most of the species. The weak relationship between these two variables was due to the indirect influence of rainfall on mercury bioaccumulation. In summary, drought increased the levels of two contaminants in fish.

Antiangiogenic activity of a bevacizumab-loaded polyurethane device in animal neovascularization models.

PURPOSE: To evaluate the antiangiogenic activity of bevacizumab-loaded polyurethane using two animal models of neovascularization. METHODS: The percentage of blood vessels was evaluated in a chicken chorioallantoic membrane model (n=42) and in the rabbit cornea (n=24) with neovascularization induced by alkali injury. In each model, the animals were randomly divided into the groups treated with the bevacizumab-loaded polyurethane device, phosphate-buffered-saline (negative control) and bevacizumab commercial solution (positive control). Clinical examination, as well as histopathological and immunohistochemical evaluation, were performed in the rabbit eyes. Microvascular density in hot spot areas was determined in semi-thin sections of corneal tissue by hematoxylin-eosin staining and factor VIII immunohistochemistry. Immunohistochemical analysis was also performed to evaluate VEGF expression. RESULTS: In the evaluated models, the use of bevacizumab (Avastin®) and the bevacizumab-loaded polyurethane device led to similar results with regard to inhibition of neovascularization. In the chorioallantoic membrane model, the bevacizumab-loaded polyurethane device reduced angiogenesis by 50.27% when compared to the negative control group. In the rabbit model of corneal neovascularization, the mean density of vessels/field was reduced by 46.87% on analysis of factor VIII immunohistochemistry photos in the bevacizumab-loaded polyurethane device group as compared to the negative control (PBS) sections. In both models, no significant difference could be identified between the bevacizumab-loaded polyurethane device and the positive control group, leading to similar results with regard to inhibition of neovascularization. CONCLUSIONS: The present study shows that the bevacizumab-loaded polyurethane device may release bevacizumab and inhibit neovascularization similarly to commercial bevacizumab solution in the short-term.

Organotropism of methylmercury in fish of the southeastern of Brazil.

This is one of the first studies to evaluate the effect of biometric variables (total length and weight), diet, and abiotic matrices (sediment and water column) on the bioaccumulation of methylmercury in tissues (muscle, liver, and gills) of four fish (two carnivore-invertivores, Pimelodus fur and Pachyurus adspersus; one carnivore-piscivore, Oligosarcus hepsetus; and one omnivore, Pimelodella lateristriga) in the lower section of a river in southeastern Brazil. Samples of fish (n = 120), water (n = 5) and sediment (n = 5) were collected at five sites characterized by pollution with mercury due to the use of organomercury fungicides and stream bed gold mining, commonly carried out in that section of the river in the 1980s. The results show that biometric variables are strongly correlated with methylmercury levels in muscle (r = 0.61, p < 0.0005) of P. fur. As a rule, concentrations of total mercury and methylmercury did not vary considerably between the organs of the species of different food habits, because of the environmental conditions in the study area. Despite the low concentrations of mercury in sediments (<0.05 mg kg-1 wet. wt), this compartment is a representative source of this pollutant for the organisms investigated, due to the close contact these animals keep with it in view of the low water columns in that section of the river.

Risk of ingesting As, Cd, and Pb in animal products in north Rio de Janeiro state, Brazil.

This study evaluated the levels of As, Cd, and Pb in muscle and liver the cattle and chicken. The risk was estimated for the adult population of a midsized city in southeast Brazil, concerning the tolerable ingestion and cancer risk. Samples of muscle and liver (cattle and chicken) were collected (n = 250). Samples of mineral supplements for cattle (n = 4) and chicken feed samples (n = 4) were evaluated as one of many potential source of contamination. Muscle, liver, mineral supplement, and feed samples were dissolved in acid medium and analyzed by ICP-OES. Daily muscle and liver intake was estimated using a questionnaire (N = 427). Daily intake of trace elements by the population based on the consumption of cattle muscle, cattle liver, chicken muscle, and chicken liver was low, corresponding to 2.76%, 0.33%, 2.12%, and 0.22% of the Tolerable Intake defined by the WHO for As; 0.54%, 0.29% 0.55%, 0.01%, for Cd; and 0.80%, 0.07%, 0.62%, 0.02%, for Pb. The mean of total ingestion of As, Cd and Pb was 5.43%, 1.18% and 1.51%, respectively of Tolerable Intake defined by WHO. Cancer risk was lower than 5 × 10-5 year-1. The results indicate that the muscle and liver consumption is a source of As, Cd, and Pb. Consumers that ingest cattle and chicken muscle need attention in terms the risk of cancer related to intake of As and Cd. Feed and mineral supplementation remain as one of many sources of exposure of As, Cd, and Pb.