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The use of silver nanoparticles (AgNPs) in commercial products has increased significantly in recent years. However, findings on the toxic effects of the AgNPs are still limited. This paper reports an investigation on the cytotoxic and genotoxic potential of the AgNPs on root cells of Allium cepa. Germination (GI), root elongation (REI), mitotic (MI), nuclear abnormality (NAI), and micronucleus index (MNI) were determined for seeds exposed to various AgNPs diameters (10, 20, 51, and 73 nm) as well as to the silver bulk (AgBulk) (micrometer-size particles) at the concentration of 100 mg·L -1 . Transmission electron microscopy (TEM) provided the particle size distribution, while dynamic light scattering (DLS) was used to get the hydrodynamic size, polydispersity index, and zeta potential of the AgNPs. Laser-induced breakdown spectroscopy (LIBS) and inductively coupled plasma/optical emission spectrometry (ICP OES) were applied for quantifying the AgNPs content uptake by roots. Silver dissolution was determined by dialysis experiment. Results showed that the AgNPs penetrated the roots, affecting MI, GI, NAI, and MNI in meristematic cells. Changes in these indicators were AgNPs diameter-dependent so that cytotoxic and genotoxic effects in Allium cepa increased with the reduction of the particle diameter. The results also revealed that the AgNPs were the main responsible for the cytotoxicity and genotoxicity since negligible silver dissolution was observed., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The amphyphylic triazoanilines recently synthesized 1-(4-(3-aminophenyl)-1H-1,2,3- triazole-1-yl)-3-(3-pentadecylphenoxy)propan-2-ol (1) and 1-(4-(4-aminophenyl)-1H- 1,2,3-triazole-1-yl)-3-(3-pentadecylphenoxy)propan-2-ol (2), synthesized from cardanol and glycerol, have photophysical properties which allow their use in the development of fluorescent biomarkers with applicability in the biodiesel quality control. Based on this, the present research evaluated the toxic effects of both compounds in different biological models through the investigation of survival and mortality percentages as a measure of acute toxicity on Daphnia similis and Oreochromis niloticus, larvicidal assay against Aedes aegypti, and cytotoxic activity on mammary cells. Results demonstrate that these triazoanilines 1 and 2 have shown low acute toxicity to the biological models investigated in this study up to the following concentrations: 4.0 mg L-1 (D. similis), 4.0 mg L-1 (A. aegypti larvae), 1.0 mg L-1 (O. niloticus), and 1.0 mg mL-1 (mammary cells). This fact suggests the potential for safe use of compounds 1 and 2 as fluorescent markers for the monitoring of biodiesel quality, even in the case of environmental exposure. Besides all of that, the reuse of cardanol and glycerol, both industrial wastes, favors the maintenance of environmental health and is in agreement with the assumptions of green chemistry. Graphical abstract ᅟ.

Emerging contaminants (ECs) are synthetic or naturally occurring chemicals that are not commonly monitored despite having the potential of entering the environment and causing adverse ecological and/or human health effects. This study aimed to determine whether ECs are present in the surface waters of two rivers in Mato Grosso do Sul State, Brazil, and evaluate the effects of ECs mixtures at environmentally relevant concentrations on zebrafish (Danio rerio) gene expression. ECs concentrations were determined using solid-phase extraction and liquid chromatography-mass spectrometry. The ECs most frequently detected were caffeine, imidacloprid, 2-hydroxy atrazine, tebuthiuron, atrazine, and bisphenol A. We used these data to reconstruct ECs mixtures reflecting environmental concentrations, codenamed T1, T2, and T3. No effects were observed, so the concentrations were increased. After a preliminary evaluation of the No Observed Effect Concentration for each mixture, we analyzed changes in the expression of zebrafish target genes (cyp1a, hsp70, cat, sod1, tsh, cyp19a1a, cyp19a1b, cyp26b1, casp8, sox2, cyb561d2, and thrb). cat was overrepresented in T1 and underrepresented in the other treatments. All of the mixtures induced the expression of cyp19a1b, which is a marker for (xeno-)estrogen exposure, and two of them increased the expression of cyp1a, which is used to indicate the presence of dioxin-like compounds. The rivers studied had low EC concentrations, and there was no indication of any harmful effects on the zebrafish. However, intensive agricultural activity may result in unsuspected peaks of EC pollution, and subsequent negative effects on living organisms., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The streams and rivers of the Upper Paraná River Basin have been seriously affected by impacts of high population density around the basin area. Fishes are widely used as models to assess the health of aquatic ecosystems, being considered as bioindicators of environmental pollution. In this context, our objective was to assess the potential genotoxic and mutagenic effects of the polluted water in three native fish species (Astyanax lacustris, Hypostomus ancistroides, and Rhamdia quelen) from Tarumã Microbasin, Upper Paraná River, Brazil. We also investigated the concentration of metals in water and in fish muscle to verify bioavailability and bioaccumulation of metals. For both less impacted sites (LI) and impacted sites (IMP) of the microbasin, the concentrations of metals were above the maximum limit allowed by Brazilian legislation (Resolution CONAMA 357/2005), except for Pb, total Cr, and Cu at LI sites and total Cr at IMP sites. A. lacustris showed a higher frequency of micronuclei (MN) at IMP sites compared with LI sites (p < 0.0001). We found no significant differences in MN frequency between site classes for H. ancistroides and R. quelen (p > 0.05). There were no significant differences between site classes regarding to nuclear abnormalities in erythrocytes frequencies (p > 0.05). A. lacustris from IMP sites had higher concentrations of Pb, Cu, Fe, Zn, and Ni in muscle tissue (p < 0.05), whereas H. ancistroides from IMP sites had higher concentration of Cr, Cu, and Ni (p < 0.0001) and R. quelen showed higher concentration of Cd, Fe, and Ni at these sites (p < 0.0001). So, the chosen biomarkers are able to identify the environmental risk of the water pollution.

Zebu cattle (Bos taurus indicus) are highly adapted to tropical regions. However, females reach puberty after taurine heifers, which affects the economic efficiency of beef cattle breeding in the tropical regions. The aims of this study were to establish associations between the haplotype alleles of the bovine genome and age at first calving (AFC) in the Nelore cattle, and to identify the genes and quantitative trait loci (QTL) related to this phenotype. A total of 2,273 Nelore cattle (995 males and 1,278 females) genotyped using the Illumina BovineHD BeadChip were used in the current study. The association analysis included females with valid first calving records as well as open heifers. Linkage disequilibrium (LD) analysis among the markers was performed using blocks of 5, 10, and 15 markers, which were determined by sliding windows shifting one marker at a time. Then, the haplotype block size to be used in the association study was chosen based on the highest r2 average among the SNPs in the block. The five HapAlleles most strongly associated with the trait (top five) were considered as significant associations. The results of the analysis revealed four genomic regions related to AFC, which overlapped with 20 QTL of the reproductive traits reported previously. Furthermore, there were 19 genes related to reproduction in those regions. In conclusion, the use of haplotypes allowed the detection of chromosomal regions associated with AFC in Nelore cattle, and provided the basis for elucidating the mechanisms underlying this trait., Competing Interests: Haroldo H. R. Neves is an employee of GenSys Consultores Associados S/S Ltda. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

The objective of this research was to evaluate land use and occupation in the landscape surrounding two neotropical streams of the Upper Paraná River in Brazil and to investigate if the water from these streams induces erythrocyte nuclear abnormalities in Astyanax lacustris. We observed that the areas surrounding the streams are predominantly used for agricultural purposes (73.46% of the total area), followed by buildings (15.21%). Only 8.78% of the area consisted of forest fragments. We observed higher frequencies of micronuclei in erythrocytes of A. lacustris exposed to water from impacted sites (IMP) compared with less impacted sites and the negative control (NC) (p < 0.05). Analysis of nuclear alterations in A. lacrustis erythrocytes showed significant differences in the frequency of notched nuclei and nuclear pyknosis only between the NC and the IMP sites (p < 0.05). Our results indicate that water from the most impacted sites of the microbasin can induce mutagenic and genotoxic effects in A. lacustris.

Anthropic activities are directly related to the contamination of aquatic ecosystems owing to the release of numerous chemicals from agricultural and urban waste. These contaminants cause environmental degradation and a decrease in the availability of water quality. The objective of this search was to evaluate the efficiency of physicochemical, chemical, and microbiological tests; extraction of chlorophyll a; and genetic parameters to identify anthropic activities and weather condition effects on the stream water quality and the consequences of its use by the population. The physicochemical parameters were within the limits allowed by the Brazilian law. However, contamination by metals (Cd 0.510 mg L -1 , Co 0.405 mg L -1 , and Ni 0.316 mg L -1 ) has been found at various collection points to be more than the allowable values. The antibiotic oxytetracycline was detected in stream water in quantities of up to 89 μg L -1 . In relation to microbiological contamination, Escherichia coli and Pseudomonas spp. have been isolated. The averages of chlorophyll a were up to 0.15558 mg cm -2 . Genetic tools identified greater number of micronuclei and DNA damage in periods that showed lower rainfall rates and lower amounts of metals. The analysis used for monitoring was efficient to verify the interference that animal breeding and planting of different cultures have caused on that stream. Thus, the continued use of this water for drinking, irrigation of vegetables, and recreational activities makes the population susceptible to contamination by bacteria and creates conditions for the development of genetic alterations in the long run.

Purpose: Integra® dermal regeneration template is a bilayer membrane system that acts as a scaffold for regenerating dermal skin cells. It is used for wound reconstruction following burns, extensive injuries, and a large tumor excision in multiple parts of the body. The dermal layer is made of porous matrix of bovine tendon collagen and glycosaminoglycan. The epidermal layer is made of polysiloxane layer. In this study, the authors evaluated the use of Integra® dermal regeneration template for the immediate reconstruction of the orbital exenteration socket., Methods: Five patients who underwent exenteration and immediate reconstruction of the socket with Integra® dermal regeneration template were included in this study. Demographic and clinical features, healing time, complications, and follow-up time were recorded., Results: The study included 4 male patients and 1 female patient with a mean age of 74 years (range, 49-87 years). The primary diagnoses were orbital extension of conjunctival melanoma in 2 patients, squamous cell carcinoma in 1 patient, and uveal melanoma in 1 patient, and aggressive orbital Wegener granulomatosis in 1 patient. There was no postoperative infection, necrosis, hematoma, or fluid accumulation in any patients. The mean postoperative follow-up period was 20 months (range, 11-42 months). The sockets were completely granulated by 4 weeks, and epithelized, getting ready for the prosthesis in 8 weeks., Conclusions: Integra® dermal regeneration template can be used for the immediate reconstruction of the socket following exenteration. It is easy to use, and provides a short healing time without any need for any additional reconstructive procedures.

Background: Plants of the genus Campomanesia belong to the family Myrtaceae and are very abundant in Cerrado areas. Teas from leaves of species of this genus are used for intestinal infections, combating obesity, stomach pathology, fever and among others., Objective: The present study evaluated the chemical composition and antioxidant potential, cytotoxicity and genotoxicity of ethanolic extract from leaves of Campomanesia pubescens (Mart. ex DC.) O.Berg., Method: For the evaluation of antioxidant activity, the free radical DPPH and for determination of phenolic compounds Folin-Ciocalteau reagent were used. Identification of the substances was performed by HPLC-DAD by comparison of the retention times with standards analyzed under the same conditions and by evaluation of molecular absorption spectra in the ultraviolet and visible region. The cytotoxicity, genotoxicity were evaluated using Allium cepa bioassay., Results: In the ethanolic extract 7-hydroxy-6-methyl-5-methoxyflavanone, 5,7-dihydroxy-6-methylflavanone, 5,7-dihydroxy-8-methylflavanone, 2´,4´-dihydroxy-6´-methoxychalcone, 5,7-dihydroxy-6,8- dimethylflavanone, 2´,4´-dihydroxy-5´-methyl-6´-methoxychalcone and 2´,4´-dihydroxy-3´,5´-dimethyl-6´- methoxychalcone were identified. The extract showed antioxidant activity and cytotoxic effects on cell division and increased chromosomal alterations in Allium cepa test., Conclusion: These results showed antioxidant activity and suggest the cytotoxic and genotoxic effects in Allium cepa of ethanolic extract obtained from the leaves of Campomanesia pubescens., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.)

The present study evaluated mutagenic and genotoxic effects and metal accumulation in the liver and musculature of Pterygoplichthys ambrosetti and Prochilodus lineatus in the Amambai River, a tributary of the Upper Paraná River in Brazil. We also evaluated the potential for these fish species as environmental bioindicators and performed risk assessment for conservation purposes. We found that P. ambrosetti had a higher frequency of micronuclei compared to P. lineatus (p < 0.0001). There were no significant differences between species in other erythrocyte nuclear abnormalities (ENA) (p > 0.05). For both species, the liver contained a higher concentration of metals (Cd, Pb, Cr, Cu, Fe, Zn, and Ni) than the musculature did (p < 0.0001). Of the two species, P. ambrosetti was more suitable to evaluate mutagenic and genotoxic effects and metal accumulation in the liver and musculature, likely due to its resident behavior; hence, we highlight the potential of this species for use as an environmental bioindicator. The concentrations of non-essential metals observed in the fish confirm conditions of environmental stress in the Amambai River, possibly related to the discharge of pollutants and exacerbated by lack of native vegetation cover along the watercourse.

Nuclear abnormalities (micronuclei and meta-nuclear changes) have been used as biomarkers to identify cell damages. As children are more vulnerable to the adverse effects of pollution when compared to adults, assessing genetic damage caused by environmental influences is of great interest. As such, the objective was to determine metanuclear (karyolysis, pycnosis, karyorrhexis, binucleated cells, chromosome bridges and micronuclei) in cells from the oral mucosa of children associated with the school environment, gender, exposure to cigarette smoke and vehicular traffic. Analyses of nuclear abnormalities were performed in exfoliated buccal cells of children from two public schools located in Dourados - MS. The data were analyzed through Kruskal-Wallis test considering a significance level of 5% (p < .05). The results showed that children exposed to cigarette smoke presented higher levels of nuclear abnormalities than children who were not usually exposed to this type of mutagenic and genotoxic agent, suggesting that such contaminants are related to clastogenic and aneugenic effects on DNA. Moreover, female children had higher amounts of nuclear abnormalities when compared to male children. With regards to the school environment, the study results indicated statistical differences in of term chromosomal abnormalities for schools A and B. Thus, it was possible to determine that children exposed to cigarette smoke are susceptible to further genetic damage than unexposed children, and female children may be more susceptible to genotoxic and mutagenic agents. This study contributes to the current knowledge on the mutagenic characteristics of human cells, supporting the adoption of preventive Public Health measures.

In the present study, genotoxicity and mutagenicity were investigated in Tradescantia pallida exposed to vehicular traffic at different sites in a high-altitude tropical climate. During March, May, July, September, and November 2014, a comet assay and micronucleus bioassays were conducted on young inflorescences and leaves of T. pallida collected from twelve towns in the southern region of Mato Grosso do Sul with different amounts of vehicular traffic. Weather parameters (temperature, relative humidity and rainfall) were measured and vehicles were counted to determine traffic levels in each town. A higher frequency of genotoxic and mutagenic damage was observed in the municipality of Dourados. The highest frequency of genetic damage was observed in September and November according to both assays. Relative humidity and rainfall were inversely proportional to the frequency of genetic damage in T. pallida during the collection period. Based on these results, we conclude that the bioassays are efficient for assessing the effects of vehicular traffic in these towns with respect to weather conditions over time. These bioassays can be applied to identify risk areas, which are determined by climatic conditions and air pollutants released., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

This study aimed to evaluate DNA damage in animal and plant cells exposed to water from the Água Boa stream (Dourados, Mato Grosso do Sul, Brazil) by using bioassays, and to identify the chemical compounds in the water to determine the water quality in the area. Through the cytotoxicity bioassay with Allium cepa, using micronucleus test, and comet assay, using Astyanax altiparanae fish, the results indicated that biological samples were genetically altered. Micronuclei were observed in erythrocytes of A. altiparanae after exposure to water from locations close to industrial waste discharge. The highest DNA damage observed with the comet assay in fish occurred with the exposure to water from locations where the presence of metals (Cu, Pb, Cd, Ni) was high, indicating the possibility of genotoxic effects of these compounds. Thus, these results reinforce the importance of conducting genotoxicity tests for developing management plans to improve water quality, and indicate the need for waste management before domestic and industrial effluents are released into the rivers and streams.

Purpose: The purpose of this study is to compare the outcomes following femtosecond laser-assisted deep anterior lamellar keratoplasty (DALK) with 75% of stromal dissection (predescemetic group) and femtosecond laser-assisted DALK using big-bubble technique with total stromal resection (descemetic group) for the treatment of keratoconus., Subjects and Methods: Twenty eyes of 17 patients with keratoconus were studied. There were 10 eyes of 9 patients in predescemetic group and 10 eyes of 8 patients in descemetic group. The postoperative best-corrected visual acuity (BCVA), manifest refraction, keratometry, endothelial cell density (ECD), and central corneal thickness (CCT) were analyzed., Results: All surgeries were performed uneventfully. At 1 year after surgery, the BCVA, corneal astigmatism, keratometry, CCT, and ECD between two groups were not statistically significant (all P > 0.05). However, the mean manifest refraction was -9.43 ± 7.44 diopter (D) and -1.03 ± 1.13D in predescemetic and descemetic groups, respectively, which was statistically significant between two groups (P < 0.05)., Conclusions: The results of BCVA and corneal astigmatism, keratometry, ECD, and CCT were comparable between two groups. However, the mean postoperative manifest refraction was lower in descemetic group.

Myracrodruon urundeuva is a plant native to Brazil, which is used by the indigenous population for the treatment of candidiasis. The aims of this study were to evaluate the antifungal activity of extract against human vaginal Candida species and evaluate the possible toxicological activities of M. urundeuva. Initially, ethanol extracts, ethyl acetate fractions, and hydroalcoholic fractions of the bark and leaf of M. urundeuva were used to determine the minimum inhibitory concentration. The extracts that showed antifungal activity were characterized by liquid chromatography and subjected to toxicity assessment. Toxic, cytotoxic, genotoxic, and mutagenic testing were performed using Allium cepa and Ames assays with the ethanol extracts of the bark and leaves. Hemolytic activity was evaluated in erythrocytes and acute toxicity in rats. The ethanol bark extracts showed best activity against Candida albicans, C. krusei, and C. tropicalis ATCC (4-512 µg/mL). Chemical characterization indicated the presence of flavonoids and tannins in the extracts. Hemolytic activity, genotoxicity, and mutagenicity were not observed. The results of the Ames and A. cepa tests were also in agreement, ethanol bark extracts and ethanol leaf extracts of M. urundeuva showed absence of mutagenic activity. Similar results were observed in the A. cepa assay and acute toxicity test in rats. M. urundeuva bark extracts showed potential for the treatment of vaginal infections caused Candida species, as a topical.

Thyroid-associated ophthalmopathy (TAO) is a condition associated with a wide spectrum of ocular changes, usually in the context of the autoimmune syndrome, Graves' disease. In this topical review, we attempted to provide a roadmap of the recent advances in current understanding the pathogenesis of TAO, important aspects of its clinical presentation, its impact on the ocular surface, describe the tissue abnormalities frequently encountered, and describe how TAO is managed today. We also briefly review how increased understanding of the disease should culminate in improved therapies for patients with this vexing condition.

Use of groundwater from private wells in households has increased considerably, owing to a better cost/benefit ratio than that of water provided by local utilities for a fee. However, this water is usually untreated, which makes it a vehicle for diseases. Thus, monitoring this water is necessary to ensure its integrity and quality. We aimed to evaluate the physical, chemical, and microbiological parameters of untreated groundwater drawn from different types of wells, and the antimicrobial susceptibility profile of the bacteria isolated from this water. Wellwater samples were collected in two Brazilian cities. Although physical and chemical parameters of the water were suitable for drinking, Escherichia coli was detected in 33% of the samples. E. coli contaminated 65% of dug wells and 10.25% of drilled wells. Many bacteria isolated were resistant to multiple antibacterial agents, including β-lactams. Microbial contamination of this water was related to the well depth, and was more common in dug wells, making this water unfit for human consumption. Consumption of such contaminated and untreated water is a public health concern. Thus, individuals who regularly use such water must be alerted so they may either take preventive measures or connect to the water distribution system operated by local utilities., Competing Interests: The authors declare no conflict of interest. The funding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

The azoles are the class of medications most commonly used to fight infections caused by Candida sp. Typically, resistance can be attributed to mutations in ERG11 gene (CYP51) which encodes the cytochrome P450 14α-demethylase, the primary target for the activity of azoles. The objective of this study was to identify mutations in the coding region of theERG11 gene in clinical isolates of Candida species known to be resistant to azoles. We identified three new synonymous mutations in the ERG11 gene in the isolates of Candida glabrata (C108G, C423T and A1581G) and two new nonsynonymous mutations in the isolates of Candida krusei--A497C (Y166S) and G1570A (G524R). The functional consequence of these nonsynonymous mutations was predicted using evolutionary conservation scores. The G524R mutation did not have effect on 14α-demethylase functionality, while the Y166S mutation was found to affect the enzyme. This observation suggests a possible link between the mutation and dose-dependent sensitivity to voriconazole in the clinical isolate of C. krusei. Although the presence of the Y166S in phenotype of reduced azole sensitivity observed in isolate C. krusei demands investigation, it might contribute to the search of new therapeutic agents against resistant Candida isolates.

The aim of this study was to evaluate a genome wide association study (GWAS) approach to identify single nucleotide polymorphisms (SNPs) associated with fertility traits (early puberty) in Nellore cattle (Bos indicus). Fifty-five Nellore cows were selected from a herd monitored for early puberty onset (positive pregnancy at 18 months of age). Extremes of this phenotype were selected; 30 and 25 individuals were pregnant and non-pregnant, respectively, at that age. DNA samples were genotyped using a high-density SNP chip (>777.000 SNP). GWAS using a case-control strategy highlighted a number of significant markers based on their proximity with the Bonferroni correction line. Results indicated that chromosomes 5, 6, 9, 10, and 22 were associated with the traits of interest. The most significant SNPs on these chromosomes were rs133039577, rs110013280, rs134702839, rs109551605, and rs41639155. Candidate genes, as well as quantitative trait loci (QTL) previously reported in the Ensembl and Cattle QTLdb databases, were further investigated. Analysis of the regions close to the SNP on chromosomes 9 and 10 revealed that four QTL had been previously classified under the reproduction category. In conclusion, we have identified SNPs in close proximity to genes associated with reproductive traits. Moreover, U6 spliceosomal RNA was present on three different chromosomes, which is possibly associated with age at first calving, suggesting that it might be a strong candidate for future studies.

Schinus terebinthifolius Raddi is a perennial native from Atlantic forest. It is of high ecological plasticity and is used in traditional medicine. Based on promising reports concerning its bioactivity, it was included as a species of great interest for distribution through the National Health System. A number of agronomic studies to guide its crop production are therefore underway. This study examined diversity and phylogenetic relationships among native S. terebinthifolius populations from different Brazilian ecosystems: Cerrado; sandbanks; dense rainforest; and deciduous forest. The intergenic regions rpl20-5'rps12, trnH-psbA, and trnS-trnG were sequenced from cpDNA and aligned using BLASTn. There were few fragments for comparison in GenBank and so only region trnS-trnG was informative. There were variations among and within populations with intravarietal polymorphisms and three distinct haplotypes (HpSM, HpDDO, HpNE), once populations from NE (sandbanks and rainforest) clustered together. Sequences from HpSM, HpNE, and HpDDO returned greater similarity to haplotypes A (AY928398.1), B (AY928399.1), and C (AY928400.1), respectively. A network, built by median-joining among native haplotypes and 10 available on GenBank, revealed HpSM as the origin of all other haplogroups. HpDDO showed the most mutations and was closely related to haplogroups from Argentina. While this could indicate hybridization, we believe that the polymorphisms resulted from adaptation to events such as deforestation, fire, rising temperature, and seasonal drought during the transition from Atlantic forest to Cerrado. While more detailed phylogeographical studies are needed, these results indicate eligible groups for distinct climates as an important step for pre-breeding programs before field propagation.

In this study, the culture analysis of urine samples from patients hospitalized in the Central-West region of Brazil was performed, and the isolated microorganisms were phylogenetically identified as Trichosporon asahii. Maximum parsimony analysis of the IGS1 sequences revealed three novel genotypes that have not been described. The minimum inhibitory concentrations of the nine isolates identified were in the range of 0.06-1 μg/ml for amphotericin B, 0.25-4 μg/ml for fluconazole, and 0.03-0.06 μg/ml for itraconazole. Approximately 6/9 of the T. asahii isolates could form biofilms on the surface of polystyrene microplates. This study reports that the microorganisms isolated here as T. asahii are agents of nosocomial urinary tract infections. Furthermore, the IGS1 region can be considered a new epidemiological tool for genotyping T. asahii isolates. The least common genotypes reported in this study can be related to local epidemiological trends., (Copyright © 2015 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.)

To characterize the effect of vehicular traffic on air quality, the micronuclei of Tradescantia pallida tetrads were counted. Young inflorescences of T. pallida (Rose) D.R. Hunt var. purpurea were collected in 2010, 2011, and 2012, from three sites subjected to different intensities of vehicular traffic. The sites were located in the municipality of Dourados, in the State of Mato Grosso do Sul, Brazil. A standardized methodology was used to analyze the Tradescantia micronuclei, in order to evaluate the mutagenic potential of the local air pollutants. Statistical analyses using the Pearson's linear correlation were employed to determine the relationship between relative humidity and temperature, and the average number of micronuclei. In this study, an increase in the average number of passing vehicles was correlated with an increase in the frequency of micronucleus formation. Climatic factors also influenced micronucleus formation, although vehicular traffic remained the most important factor. Thus, the Tradescantia micronuclei assay may be a useful method of assessing air quality., (Copyright © 2015. Published by Elsevier Inc.)

Background: Candida tropicalis is an emerging major human pathogen in nosocomial infections, and it is considered the second or third species of Candida most isolated from urine cultures., Aims: The study aimed at characterizing genotypically C. tropicalis strains from patients with candiduria in a university hospital, and assessed the antifungal susceptibility profile., Methods: The study was conducted with hospitalized patients who developed urinary tract infection from C. tropicalis from June 2010 to June 2011 at the Grande Dourados University Hospital of the Federal University, Dourados, MS, Brazil. Susceptibility to the antifungal agents amphotericin B and fluconazole was determined by broth microdilution. The genotypic variability of isolates of C. tropicalis was analyzed by microsatellite markers and RAPD-PCR., Results: Only one isolate was resistant to amphotericin B (MIC→16μg/ml); the others were susceptible to fluconazole and amphotericin B. The genotypic variability by RAPD-PCR resulted in distinct profiles for RAPD markers. A total of 10 alleles were observed for the microsatellite loci, URA3 and CT14, which were grouped differently, and four associations were observed for locus URA3 and eight for locus CT14., Conclusions: C. tropicalis isolates from urine were susceptible to the antifungal agents tested. The genotyping techniques make possible proving the similarity and genetic diversity among isolates of C. tropicalis involved in nosocomial infections. This knowledge is important for the control and prevention of nosocomial infections caused by this yeast species., (Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.)

Water quality monitoring is used to determine the impact of human activities on the environment. We evaluated water quality in the Água Boa stream, located within the municipality of Dourados, State of Mato Grosso do Sul, Brazil, by analyzing physico-chemical, chemical, and microbiological parameters, as well as chlorophyll concentrations. Five sets of water samples were collected between December 2012 and November 2013 from three locations within the stream. The results showed the presence of Escherichia coli and antibiotic-resistant Pseudomonas spp. strains and high concentrations of organic matter (total dissolved solids), inorganic species (Mg, Ca, and Fe), and agrochemical residues (thiamethoxam). The main stream water contaminants are derived from urban, industrial, and agricultural activities within the watershed. Given the presence of contaminants, it is important that such findings are disseminated in order to highlight the risks that contact with this water may pose to human health. To preserve the environment and improve site conditions, people would need to participate by demanding that normative national and international standards be respected and that the situation be supervised by the competent governmental agencies; this would make it possible to reverse or minimize contamination problems within the Água Boa stream.

Introduction: The indiscriminate use of antimicrobials has selected for the emergence of resistant strains. Many mechanisms contribute to the spread of antimicrobial-resistant genes, and integrons play a key role in this process. The aim of this study was to describe the serotypes and resistance profiles, and to characterize the presence of integrons in Salmonella strains isolated from Dourados, Mato Grosso do Sul, Brazil., Methodology: Thirty-six isolates from different sources were used. To evaluate the resistance profiles, the determination of minimum inhibitory concentrations together with polymerase chain reaction were used to screen for the presence of class 1 and class 2 integrons., Results: The Infantis serotype of Salmonella was the most frequently isolated serotype. Minimum inhibitory concentrations showed that out of the 36 isolates, 11 (30.5%) were resistant to all the antimicrobials tested. These resistant isolates were separated into three groups: 4 clinical isolates (36.4%), 3 food isolates (36.4%), and 4 water isolates (27.2%). Class 1 integrons occurred in 31 (86.1%) isolates and were found in all 11 resistant isolates (35.5 %) and in 20 (64.5%) of the non-resistant isolates. Class 2 integrons were found in 3 (8.3%) isolates, which were all non-resistant., Conclusion: The presence of an integron did not necessarily confer resistance. Future studies will seek to identify the mechanism behind integron-mediated antimicrobial resistance.

Anatomical alterations in leaves and DNA damage in cells caused by the accumulation of atmospheric pollutants can be measured by epidermal leaf analyses and Tradescantia micronuclei assay with early pollen tetrad cells. The present study examined the feasibility of using somatic and germ cells of Tradescantia pallida for biomonitoring purposes in the city of Dourados, state of Mato Grosso do Sul (MS), Brazil. Stomatal, micronucleus and epidermal leaf analyses were performed, using standard methodologies, on plants growing at three locations during six different time periods. Tradescantia micronuclei data were analyzed using SAS 9.2 software package and stomatal data were analyzed using SANEST software. Analyses of stomatal characteristics and micronuclei examination in T. pallida were found to be an efficient tool for monitoring atmospheric pollution. The micronucleus assay suggested that the number of micronuclei in early pollen tetrad cells was related to the intensity of vehicular traffic. Increased number of epidermal cells and stomata and increased stomatal density observed at locations with greater vehicular traffic are likely physiological responses of those plants to the increased gas exchange in highly polluted environments.

The aim of the study was to detect polymorphisms in the leptin gene and to determine the association of these polymorphisms with growth and carcass traits in Nellore cattle. The single nucleotide polymorphisms (SNPs) -1457 (AJ571671:g.-1457A>G) and A59V (AF536174.1:g. 321C>T), as well as the microsatellite BM1500 (3.9 kb downstream), were genotyped. The measures of body weight and ultrasound examinations (rib eye area, back, and rump fat thickness) were performed in 3 different periods of animal management. During the first period, the animals were fed with grass and mineralized salt ad libitum. In the second period, they received grass and concentrate, and in the third, only concentrate. After the slaughter of animals, data were collected for classification and typification of carcasses. No significant association was found between the variables assessed and SNP -1457. Conversely, SNP A59V was associated with rump fat thickness and muscle color post-slaughter. BM1500 was associated with rump fat thickness in the first period (pre-slaughter), subcutaneous fat thickness in the second, weight of the animals in the third, and length of the carcass after slaughter. These results suggest that SNP A59V and the microsatellite BM1500 might be useful for marker-assisted selection in Nellore cattle.

Sheep of the Pantaneiro breed and seven other breeds, raised in the State of Mato Grosso do Sul, Brazil, were genotyped using eight microsatellite loci. The aim of the present study was to determine the genetic variability, phylogenetic relationship, and patterns of gene introgression and miscegenation among the animals surveyed, to obtain information about the genetic structure of locally adapted sheep in Mato Grosso do Sul. A total of 195 animals were used for genetic analysis. The Pantaneiro breed had the largest average number of alleles/locus (9.25), and higher allelic richness (6.95), while the Dorper population had the lowest values for these parameters (4.88 and 3.86, respectively). Analysis of genetic distance values and genetic structure between populations made it possible to characterize these animals with regard to distinct genetic groups. Average expected heterozygosity ranged from 0.72 (Pantaneiro) to 0.55 (Dorper), while average observed heterozygosity ranged from 0.63 (White Dorper) to 0.54 (Dorper). On the basis of the statistical parameters evaluated, it was possible to demonstrate that when compared to other populations, the Pantaneiro breed represented a reservoir of genetic diversity with rare and useful alleles for genetic improvement, emphasizing the importance of preserving the breed.

The present work sought to detect the presence of Pseudomonas spp. at different stages of an effluent treatment plant using the Australian system of stabilization ponds, and to determine the susceptibility of those isolates to different antimicrobials. Thirty-four isolates of Pseudomonas spp. derived from effluent treatment station water samples were collected near the transfer ducts between the ponds in November/2008 and december/2009. Among the Pseudomonas spp. isolates, 47.05 % showed susceptibility to all antimicrobials tested, 20.58 % were resistant to cefepime, and 24 % showed intermediate resistance to streptomycin. No Pseudomonas spp. isolates were found in the final pond, or in post-treatment effluents. The Pseudomonas spp. isolates did not exhibit multiresistance to the antimicrobials tested.

Advances in DNA technology have created biotechnological tools that can be used in animal selection and new strategies for increasing herd productivity and quality. The objective of the present work was to associate the genotypes of leptin gene exon 2 polymorphisms with productive traits in Nellore cattle. Blood was collected from Nellore males and PCR-RFLP reactions were performed with the restriction enzymes Cla I and Kpn 2I. The gene frequencies resulting from digestion by Cla I were 0.60 and 0.40 for allele A and T, respectively; the genotypic frequencies were AA = 0.20 and AT = 0.80. The gene frequencies from digestion by Kpn 2I were 0.81 for allele C and 0.194 for allele T; the genotypic frequencies were CC = 0.62 and CT = 0.38. The populations in both cases were not in Hardy-Weinberg equilibrium (p > 0.05), and the TT genotype was not found. Significant associations were noted between leptin gene exon 2 polymorphisms and five productive traits in Nellore cattle: carcass fat distribution, the intensity of red muscle coloration, pH, marbling, and post-slaughter fat thickness.

The present study describes a simple and efficient method utilizing high performance liquid chromatography (HPLC) coupled to fluorescence detection for the determination of kinetic parameters of glutamate uptake in nervous tissue. Retinal tissue obtained from 7-day-old chicks was incubated with known concentrations of glutamate (50-2000 μM) for 10 min, and the levels of the o-phtaldehyde (OPA)-derivatized neurotransmitter in the incubation medium were measured. By assessing the difference between initial and final concentrations of glutamate in the medium, a saturable uptake mechanism was characterized (K(m)=8.2 and V(max)=9.8 nmol/mg protein/min). This measure was largely sodium- and temperature-dependent, strongly supporting that the mechanism for concentration decrements is indeed uptake by high-affinity transporters. Added to this, our results also demonstrated that zinc chloride (an inhibitor of glutamate/aspartate transporters) evoked a concentration-dependent decrease in glutamate uptake, demonstrating the specificity of our methodology. Overall, the present work characterizes an alternative methodology to evaluate glutamate uptake in nervous tissue using HPLC. This approach could be an important tool for studies associated to the characterization of minute alterations in glutamate transport related with central nervous system injury., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Purpose: This study aimed to associate the intensity of vehicular traffic in the city of Dourados (Mato Grosso do Sul State, Brazil) with mutagenic effects and alterations in leaf physiology as measured by the quantity of micronuclei and the leaf surface parameters of Tradescantia pallida., Methods: Five collections of inflorescences were undertaken for 24 weeks to determine the quantities of micronuclei using the Tradescantia Micronuclei (Trad-MCN) bioassay. Leaf surface parameters, including stomatal index (SI), stomatal density, and the size of the stomatal ostiole opening size (SO), were evaluated in addition to Trad-MCN. Collections were made at four sampling points with different vehicular traffic intensities. Statistical analyses were performed with SAS software using the Tukey's and Kruskal-Wallis test. Additionally, associations of the characteristics were verified using Pearson's simple correlation analysis., Results: Significant effects were observed with the Trad-MCN bioassay (p < 0.01) that were related to the collection period and location, as well as significant differences (p < 0.05) for the effects of the collection points using the Kruskal-Wallis test. In general, the locations with greatest vehicular traffic had plants with the greatest stomatal density values. The characteristics SI and SO did not demonstrate significant differences (p > 0.05) in relation to the collection sites. The simple correlation analysis demonstrated a negative association (-0.65) between SI and Trad-MCN (p < 0.05)., Conclusion: Plants growing in localities with more intense vehicular traffic had greater quantities of micronuclei as well as higher frequencies and average numbers of stomata than localities with less traffic, indicating the presence of atmospheric contaminants that damaged their DNA.

Adult zebrafish were treated acutely with methylmercury (1.0 or 5.0 μg g(-1), i.p.) and, 24h after treatment, were tested in two behavioral models of anxiety, the novel tank and the light/dark preference tests. At the smaller dose, methylmercury produced a marked anxiogenic profile in both tests, while the greater dose produced hyperlocomotion in the novel tank test. These effects were accompanied by a decrease in extracellular levels of serotonin, and an increase in extracellular levels of tryptamine-4,5-dione, a partially oxidized metabolite of serotonin. A marked increase in the formation of malondialdehyde, a marker of oxidative stress, accompanied these parameters. It is suggested that methylmercury-induced oxidative stress produced mitochondrial dysfunction and originated tryptamine-4,5-dione, which could have further inhibited tryptophan hydroxylase. These results underscore the importance of assessing acute, low-level neurobehavioral effects of methylmercury., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Purpose: To report the results of Descemet stripping endothelial keratoplasty (DSEK) using the TAN EndoGlideTM device to facilitate the insertion of the endothelial membrane., Methods: Prospective clinical study that included nine patients presenting corneal edema secondary to endothelial dysfunction. Best corrected visual acuity, refraction, central corneal thickness, endothelial cell density and complications were analyzed after a six-month follow-up., Results: There was a significant improvement in the corneal edema and visual acuity in 7 patients (77.78%). The best corrected visual acuity ranged between 20/40 and 20/200. The average density of endothelial cells in six months varied between 1,305 cells/mm² and 2,346 cells/mm² with an average loss of 33.14% cells. Detachment of part of the graft was observed in one eye (11.11%) and primary failure of the endothelial transplantation occurred in 2 eyes (22.22%)., Conclusion: The device TAN EndoGlideTM facilitates the introduction of the graft in Descemet stripping endothelial keratoplasty.

The myostatin gene, also known as GDF8 (growth differentiation factor 8), is located on bovine chromosome 2 (BTA2); it has three exons and two introns. Myostatin is specifically expressed during embryonic development and in adult skeletal muscle, functioning as a negative regulatory protein. Several cattle breeds (Piedmontese, Belgian Blue and Blond'Aquitaine, and others) show polymorphisms in this gene; these polymorphisms are directly related to the double muscling phenotype. We looked for polymorphisms in the Nellore cattle myostatin gene and compared them with those known for taurine breeds. Seven regions, covering the three exons of this gene, were amplified by polymerase chain reaction and sequenced, including the untranslated region. DNA from 30 adult Nellore animals was collected; DNA sequencing revealed three, seven and four polymorphisms in exons 1, 2 and 3, respectively. We found previously reported polymorphisms, as well as several new ones; for instance, 37 polymorphisms were found in the untranslated region segment, and in introns 1 and 2 there were one and three polymorphisms, respectively. The high degree of allelic heterogeneity in the myostatin gene could be related to its high mutation rate; it also could be the result of a long history of artificial selection for meat production, which has probably favored such modifications and maintained them in cattle populations. These polymorphisms identified in Nellore cattle could be useful for breeding programs.

The myostatin gene, known as Growth Differentiation Factor 8 (GDF8), located at chromosome 2 (BTA2) in cattle, is specifically expressed during embryo development and in the adult skeletal muscle. Molecular analysis of this gene reveals the presence of three exons and two introns. Several cattle breeds, such as Piedmontese, Belgian Blue, Blond'Aquitaine, among others, show polymorphisms in this gene, which are directly related to double muscling phenotype. Piedmontese cattle shows a nucleotide transition G --> A (G938A) at exon 3, resulting in the substitution of cysteine to tyrosine, leading to a protein structure change, which determines myostatin inactivation and consequent muscular hypertrophy. The objective of this work was to implant the polymorphism G938A, naturally existent in Piedmontese breed, into in vitro propagated foetal myoblasts, from Nellore cattle. Single strand DNA (ssDNA) oligonucleotides were used to direct the same nucleotidic transition (G938A) to exon 3. Two transfection protocols (cationic lipid solution and electroporation) were tested and, 48 hours after transfection, RNA and DNA were extracted from myoblasts. Reverse transcription and polymerase chain reaction (PCR) were performed, using primers flanking the mutation region. The PCR products were cloned and analyzed by DNA sequencing, and it was possible to detect the nucleotidic CT transition at position 938, in the electroporated myoblasts. The existence of a positive signal in the transfection indicates that ssDNA oligonucleotides can be used to introduce this point mutation in specific functional gene sites.

The microsatellite loci FCA045, FCA077, FCA008, and FCA096 are highly variable molecular markers which were used to determine the genetic diversity in 148 captive Leopardus sp. The PCR-amplified products of microsatellite loci were characterized in ABI Prism 310 Genetic Analyzer. Allele numbers, heterozygosity, polymorphism information content, exclusive allele number, and shared alleles were calculated. Sixty-five alleles were found and their sizes ranged from 116 to 216 bp in four microsatellite loci. The heterozygosity ranged from 0.36 to 0.81 in Leopardus pardalis, 0.57 to 0.67 in L. tigrinus and 0.80 to 0.92 in L. wiedii. The polymorphism information content was from 0.80 to 0.88 in L. pardalis, 0.76 to 0.88 in L. tigrinus and 0.77 to 0.90 in L. wiedii. The margay (L. wiedii) showed the highest index of polymorphism among the three species in this study. These results imply that microsatellite DNA markers can help in the study of the genetic diversity of Leopardus specimens.