Under flow impedimetric measurements using magnetic particles for label-free detection affinity target

The necessity to adapt sensors based on electrochemical techniques for high throughput analysis control increases the interest to develop new analytical systems able to perform measurements under buffer now. In this report we explored the possibility of employing a new system to make impedimetric measurements to detect the interaction between proteins and small molecules. The well-known biotin-streptavidin interaction was adopted to evaluate the proposed assembly. This system allows us to perform experiments under flow. Magnetic beads functionalized with streptavidin were used and first characterized using AFM and FTIR. Non-faradic impedance spectroscopy allowed the detection of the biotin-streptavidin interaction. Using our new system and under a flow of PBS buffer, 5 10-5 M of biotin was detected with a stable signal. (c) 2007 Elsevier B.V. All rights reserved.

Identification of cause of impairment in spiral drawings, using non-stationary feature extraction approach

Parkinson’s disease is a clinical syndrome manifesting with slowness and instability. As it is a progressive disease with varying symptoms, repeated assessments are necessary to determine the outcome of treatment changes in the patient. In the recent past, a computer-based method was developed to rate impairment in spiral drawings. The downside of this method is that it cannot separate the bradykinetic and dyskinetic spiral drawings. This work intends to construct the computer method which can overcome this weakness by using the Hilbert-Huang Transform (HHT) of tangential velocity. The work is done under supervised learning, so a target class is used which is acquired from a neurologist using a web interface. After reducing the dimension of HHT features by using PCA, classification is performed. C4.5 classifier is used to perform the classification. Results of the classification are close to random guessing which shows that the computer method is unsuccessful in assessing the cause of drawing impairment in spirals when evaluated against human ratings. One promising reason is that there is no difference between the two classes of spiral drawings. Displaying patients self ratings along with the spirals in the web application is another possible reason for this, as the neurologist may have relied too much on this in his own ratings.

Non-invasive strategy in assessing asthma through biofluids metabolomics exploration: exhaled breath and urine potentialities

Asthma is a significant health issue in the pediatric population with a noteworthy growth over the years. The proposed challenge for this PhD thesis was the development of advanced methodologies to establish metabolomic patterns in urine and exhaled breath associated with asthma whose applicability was subsequently exploited to evaluate the disease state, the therapy adhesion and effect and for diagnostic purposes. The volatile composition of exhaled breath was studied combining headspace solid phase microextraction (HS-SPME) with gas chromatography coupled to mass spectrometry or with comprehensive two-dimensional gas chromatography coupled to mass spectrometry with a high resolution time of flight analyzer (GC×GC–ToFMS). These methodologies allowed the identification of several hundred compounds from different chemical families. Multivariate analysis (MVA) led to the conclusion that the metabolomic profile of asthma individuals is characterized by higher levels of compounds associated with lipid peroxidation, possibly linked to oxidative stress and inflammation (alkanes and aldehydes) known to play an important role in asthma. For future applications in clinical settings a set of nine compounds was defined and the clinical applicability was proven in monitoring the disease status and in the evaluation of the effect and / or adherence to therapy. The global volatile metabolome of urine was also explored using an HSSPME/GC×GC–ToFMS method and c.a. 200 compounds were identified. A targeted analysis was performed, with 78 compounds related with lipid peroxidation and consequently to oxidative stress levels and inflammation. The urinary non-volatile metabolomic pattern of asthma was established using proton nuclear magnetic resonance (1H NMR). This analysis allowed identifying central metabolic pathways such as oxidative stress, amino acid and lipid metabolism, gut microflora alterations, alterations in the tricarboxylic acid (TCA) cycle, histidine metabolism, lactic acidosis, and modification of free tyrosine residues after eosinophil stimulation. The obtained results allowed exploring and demonstrating the potential of analyzing the metabolomic profile of exhaled air and urine in asthma. Besides the successful development of analysis methodologies, it was possible to explore through exhaled air and urine biochemical pathways affected by asthma, observing complementarity between matrices, as well as, verify the clinical applicability.

A non-invasive method for silencing gene transcription in honeybees maintained under natural conditions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A Novel Diagnostic Target in the Hepatitis C Virus Genome

BackgroundDetection and quantification of hepatitis C virus (HCV) RNA is integral to diagnostic and therapeutic regimens. All molecular assays target the viral 5'-noncoding region (59-NCR), and all show genotype-dependent variation of sensitivities and viral load results. Non-western HCV genotypes have been under-represented in evaluation studies. An alternative diagnostic target region within the HCV genome could facilitate a new generation of assays.Methods and FindingsIn this study we determined by de novo sequencing that the 3'-X-tail element, characterized significantly later than the rest of the genome, is highly conserved across genotypes. To prove its clinical utility as a molecular diagnostic target, a prototype qualitative and quantitative test was developed and evaluated multicentrically on a large and complete panel of 725 clinical plasma samples, covering HCV genotypes 1-6, from four continents (Germany, UK, Brazil, South Africa, Singapore). To our knowledge, this is the most diversified and comprehensive panel of clinical and genotype specimens used in HCV nucleic acid testing (NAT) validation to date. The lower limit of detection (LOD) was 18.4 IU/ml (95% confidence interval, 15.3-24.1 IU/ml), suggesting applicability in donor blood screening. The upper LOD exceeded 10(-9) IU/ml, facilitating viral load monitoring within a wide dynamic range. In 598 genotyped samples, quantified by Bayer VERSANT 3.0 branched DNA (bDNA), X-tail-based viral loads were highly concordant with bDNA for all genotypes. Correlation coefficients between bDNA and X-tail NAT, for genotypes 1-6, were: 0.92, 0.85, 0.95, 0.91, 0.95, and 0.96, respectively; X-tail-based viral loads deviated by more than 0.5 log10 from 5'-NCR-based viral loads in only 12% of samples (maximum deviation, 0.85 log10). The successful introduction of X-tail NAT in a Brazilian laboratory confirmed the practical stability and robustness of the X-tail-based protocol. The assay was implemented at low reaction costs (US$8.70 per sample), short turnover times (2.5 h for up to 96 samples), and without technical difficulties.ConclusionThis study indicates a way to fundamentally improve HCV viral load monitoring and infection screening. Our prototype assay can serve as a template for a new generation of viral load assays. Additionally, to our knowledge this study provides the first open protocol to permit industry-grade HCV detection and quantification in resource-limited settings.

Evidences of non-additive effects of multiple parasitoids on Diatraea saccharalis Fabr. (Lep., Crambidae) populations in sugarcane fields in Brazil

Biological control is a relatively benign method of pest control. However, considerable debate exists over whether multiple natural enemies often interact to produce additive or non-additive effects on their prey or host populations. Based on the large data set stored in the Sao Joao and Barra sugarcane mills (state of São Paulo, Brazil) regarding the programme of biological control of Diatraea saccharalis using the parasitoids Cotesia flavipes and tachinid flies, in the present study the author investigated whether the parasitoids released into sugarcane fields interfered significantly with the rate of parasitized D. saccharalis hosts. The author also observed whether there was an additive effect of releasing C. flavipes and tachinids on the rate of parasitized hosts, and looked for evidence of possible negative effects of the use of multiple parasitoid species in this biological control programme. Results showed that C. flavipes and the tachinids were concomitantly released in the Barra Mill, but not in the Sao Jao Mill. Furthermore, in the Barra Mill there was evidence that the parasitoids interacted because the percentage of parasitism did not increase after the release of either C. flavipes or tachinids. In the Sao Joao Mill, when both parasitoid species were released out of synchrony, both the percentage of parasitism by C. flavipes as well as that of the tachinids increased. When large numbers of tachinids were released in the Barra Mill, they caused a significant lower percentage of parasitism imposed by C. flavipes. The implications of the results as evidence of non-additive effects of C. flavipes plus tachinids on D. saccharalis populations are discussed.

Shikimate Kinase (EC 2.7.1.71) from Mycobacterium tuberculosis: Kinetics and Structural Dynamics of a Potential Molecular Target for Drug Development

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Host specificity of Metriona elatior, a potential biological control agent of tropical soda apple, Solanum viarum, in the USA

The leaf beetle Metriona elatior from Brazil-Argentina was screened in the Florida (USA) State quarantine facility as a potential biological control agent of tropical soda apple, Solanum viarum, a recently arrived weed species. Multiple-choice host-specificity tests were conducted in small cages (60 cm x 60 cm x 60 cm) using 95 plant species in 29 families. Adults fed heavily on the main target weed (S. viarum), and on turkeyberry, Solanum torvum (noxious weed of Asiatic origin); fed moderately on red soda apple, Solanum capsicoides (weed of South American origin), and eggplant, Solanum melongena (economic crop); and fed lightly on aquatic soda apple, Solanum tampicense (weed of Mexican-Caribbean-Central American origin), and on silverleaf nightshade, Solanum elaeagnifolium (native weed widely distributed). M. elatior adults laid 84 to 97% of their egg masses on S. viarum, and 3 to 16% on S. melongena. Non-choice host-specificity tests were also conducted in quarantine in which M. elatior adults and neonate larvae were exposed to 17 and 19 plant species, respectively. Tests with the neonates indicate that this insect was able to complete its development on S. viarum, S. torvum, S. melongena, and S. capsicoides. Although some adult feeding and oviposition occurred on S. melongena in quarantine on potted plants in small cages, no feeding or oviposition by M. elatior was observed in field experiments conducted in Brazil. Surveys in unsprayed S. melongena fields in Argentina and Brazil indicated that M. elatior is not a pest of S. melongena in South America. The evidence obtained from the South-American field surveys, Brazil open-field experiments, and Florida quarantine host specificity tests indicate that M. elatior causes significant feeding damage to S. viarum, and does not represent a threat to S. melongena crops in the USA. Therefore an application for permission to release M. elatior against S. viarum in the USA was submitted in October 1998.

miRNApath: A database of miRNAs, target genes and metabolic pathways

MicroRNAs (miRNAs) are small non-coding RNAs that regulate target gene expression and hence play important roles in metabolic pathways. Recent studies have evidenced the interrelation of miRNAs with cell proliferation, differentiation, development, and diseases. Since they are involved in gene regulation, they are intrinsically related to metabolic pathways. This leads to questions that are particularly interesting for investigating medical and laboratorial applications. We developed an miRNApath online database that uses miRNA target genes to link miRNAs to metabolic pathways. Currently, databases about miRNA target genes (DIANA miRGen), genomic maps (miRNAMap) and sequences (miRBase) do not provide such correlations. Additionally, miRNApath offers five search services and a download area. For each search, there is a specific type of input, which can be a list of target genes, miRNAs, or metabolic pathways, which results in different views, depending upon the input data, concerning relationships between the target genes, miRNAs and metabolic pathways. There are also internal links that lead to a deeper analysis and cross-links to other databases with more detailed information. miRNApath is being continually updated and is available at http://lgmb.fmrp.usp.br/mirnapath. ©FUNPEC-RP.

Connective tissue graft plus resin-modified glass ionomer restoration for the treatment of gingival recession associated with non-carious cervical lesions: Microbiological and immunological results

Objectives: It was previously reported the clinical results of placing subgingival resin-modified glass ionomer restoration for treatment of gingival recession associated with non-carious cervical lesions. The aim of this study was to evaluate the influence of this treatment on the subgingival biofilm and gingival crevicular fluid (GCF) inflammatory markers. Materials and methods: Thirty-four patients presenting the combined defect were selected. The defects were treated with either connective tissue graft plus modified glass ionomer restoration (CTG+R) or with connective tissue graft only (CTG). Evaluation included bleeding on probing and probing depth, 5 different bacteria targets in the subgingival plaque assessed at baseline, 45, and 180 days post treatments, and 9 inflammatory mediators were also assessed in the GCF. Results: The levels of each target bacterium were similar during the entire period of evaluation (p > 0. 05), both within and between groups. The highest levels among the studied species were observed for the bacterium associated with periodontal health. Additionally, the levels of all cyto/chemokines analyzed were not statistically different between groups (p > 0. 05). Conclusion: Within the limits of the present study, it can be concluded that the presence of subgingival restoration may not interfere with the subgingival microflora and with GCF inflammatory markers analyzed. Clinical relevance: This approach usually leads to the placement of a subgingival restoration. There is a lack of information about the microbiological and immunological effects of this procedure. The results suggest that this combined approach may be considered as a treatment option for the lesion included in this study. © 2012 Springer-Verlag.

Dispersion and evaporation of droplets amended with adjuvants on soybeans

Quantification of the effects of adjuvants on droplet behaviour on plant surfaces is needed to improve pesticide spray application efficiency for soybeans. Dispersion and evaporation of single 300-μm diameter droplets amended with each of four spray adjuvants at five concentrations were investigated for four soybean plant surfaces (abaxial and adaxial leaflet surfaces, petiole, basal stem). The four adjuvants were a crop oil concentrate (COC), a modified seed oil (MSO), a non-ionic surfactant (NIS) and an oil surfactant blend (OSB). A single-droplet generator was used to produce and deposit 300-μm diameter droplets on target surfaces under controlled environmental conditions. Adjuvants significantly increased the dispersion (or wetted area) of droplets on plant surfaces. Droplet-wetted areas increased with increased adjuvant concentrations but not in direct proportion. The average increases of wetted areas across the four soybean plant surfaces were 443, 462, 416, or 343% when the spray mixture was amended with COC, MSO, NIS or OSB at the manufacturer-recommended concentrations, respectively. Among the four surfaces, the largest wetted area was on the abaxial surface, followed by the adaxial surface, the petiole and then the basal stem. Droplet evaporation times were inversely proportional to the wetted areas. The evaporation time of 300-μm diameter droplets ranged from 36 to 142. s on the four surfaces when the spray mixture was amended with an adjuvant, whereas the water-only droplets ranged from 161 to 190. s. The results demonstrated that use of adjuvants offers great potential to improve the homogeneity of sprayed pesticides, to increase spray coverage and to reduce pesticide application rates on soybean plants. These effects could benefit farmers economically and reduce environmental contamination by pesticides. © 2012.

Glycaspis brimblecombei Moore, 1964 (Hemiptera: Psyllidae) on eucalyptus: oviposition non-preference and antibiosis

Forest plantations with Eucalyptus spp. (L'H,r) in Brazil are highly yielding. However, this activity is more and more threatened due to interactions with phytophagous insects, especially by exotic species, such as the red gum lerp psyllid Glycaspis brimblecombei Moore (Hemiptera: Psyllidae). This study aimed to evaluate the resistance in eucalyptus against the psyllid in oviposition and biological development assays, attempting to identify potential genotypes resistant to the pest for forest plantations. In addition, we tested the hypothesis of that concentration of total phenolics and lignin, and amount of epicuticular wax were associated with the expression of resistance. Results showed that there was variation in the levels of resistance among the genotypes assessed. Oviposition non-preference was observed on Eucalyptus citriodora (Hook) in free-choice and no-choice tests, and Clone FP10 was least preferred in the no-choice test. The genotypes E. citriodora and Clone FP6 provided 100 % nymphal mortality, and Clones FP7 and FP9 also affected negatively the G. brimblecombei development by lengthening the duration of the nymphal stage and reducing adult emergence. Clone FP6 had higher concentration of total phenolics and larger contact angle formed between the water droplet and leaf surface, which may be associated with thicker layer of epicuticular wax on the leaves, and one of the causes of high nymphal mortality. Thus, the use of the resistant genotypes of eucalyptus screened against G. brimblecombei is a promising and viable alternative for forest plantations infested with this pest.

MicroRNA-499 expression distinctively correlates to target genes sox6 and rod1 profiles to resolve the skeletal muscle phenotype in nile tilapia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estimating the development of the fennel aphid, Hyadaphis foeniculi (Passerini) (Hemiptera: Aphiididae), using non-linear models

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

OPENING REMARKS - THIRD VERTEBRATE PEST CONFERENCE

To open this Third Vertebrate Pest Conference is a real privilege. It is a pleasure to welcome all of you in attendance, and I know there are others who would like to be meeting with us, but, for one reason or another cannot be. However, we can serve them by taking back the results of discussion and by making available the printed transactions of what is said here. It has been the interest and demand for the proceedings of the two previous conferen- ces which, along with personal contacts many of you have with the sponsoring committee, have gauged the need for continuing these meetings. The National Pest Control Association officers who printed the 1962 proceedings still are supplying copies of that conference. Two reprintings of the 1964 conference have been necessary and repeat orders from several universities indicate that those proceedings have become textbooks for special classes. When Dr. Howard mentioned in opening the first Conference in 1962 that publication of those papers would make a valuable handbook of animal control, he was prophetic, indeed. We are pleased that this has happened, but not surprised, since to many of us in this specialized field, the conferences have provided a unique opportunity to meet colleagues with similar interests, to exchange information on control techniques and to be informed by research workers of problem solving investigations as well as to hear of promising basic research. The development of research is a two-way street and we think these conferences also identify areas of inadequate knowledge, thereby stimulating needed research. We have represented here a number of types of specialists—animal ecologists, public health and transmissible disease experts, control methods specialists, public agency administration and enforcement staffs, agricultural extension people, manufacturing and sale industry representatives, commercial pest control operators, and others—and in addition to improving communications among these professional groups an equally important purpose of these conferences is to improve understanding between them and the general public. Within the term general public are many individuals and also organizations dedicated to appreciation and protection of certain animal forms or animal life in general. Proper concepts of vertebrate pest control do not conflict with such views. It is worth repeating for the record the definition of "vertebrate pest" which has been stated at our previous conferences. "A vertebrate pest is any native or introduced, wild or feral, non-human spe- cies of vertebrate animal that is currently troublesome locally or over a wide area to one or more persons either by being a general nuisance, a health hazard or by destroying food or natural resources. In other words, vertebrate pest status is not an inherent quality or fixed classification but is a circumstantial relationship to man's interests." I believe progress has been made in reducing the misunderstanding and emotion with which vertebrate pest control was formerly treated whenever a necessity for control was stated. If this is true, I likewise believe it is deserved, because control methods and programs have progressed. Control no longer refers only to population reductions by lethal means. We have learned something of alternate control approaches and the necessity for studying the total environment; where reduction of pest animal numbers is the required solution to a problem situation we have a wider choice of more selective, safe and efficient materials. Although increased attention has been given to control methods, research when we take a close look at the severity of animal damage to so many facets of our economy, particularly to agricultural production and public health, we realize it still is pitifully small and slow. The tremendous acceleration of the world's food and health requirements seems to demand expediting vertebrate pest control to effectively neutralize the enormous impact of animal damage to vital resources. The efforts we are making here at problem delineation, idea communication and exchange of methodology could well serve as both nucleus and rough model for a broader application elsewhere. I know we all hope this Third Conference will advance these general objectives, and I think there is no doubt of its value in increasing our own scope of information.