Surface plasmon resonance-based inhibition assay for real-time detection of Cryptosporidium parvum oocyst

A surface plasmon resonance (SPR)-based inhibition assay method using a polyclonal anti-mouse IgM arrayed Cryptosporidium sensor chip was developed for the real-time detection of Cryptosporidium parvum oocysts. The Cryptosporidium sensor chip was fabricated by subsequent immobilization of streptavidin and polyclonal anti-mouse IgM (secondary antibody) onto heterogeneous self-assembled monolayers (SAMs). The assay consisted of the immunoreaction step between monoclonal anti-C. parvum oocyst (primary antibody) and oocysts, followed by the binding step of the unbound primary antibody onto the secondary antibody surface. It enhanced not only the immunoreaction yield of the oocysts by batch reaction but also the accessibility of analytes to the chip surface by antibody–antibody interaction. Furthermore, the use of optimum concentration of the primary antibody maximized its binding response on the chip. An inversely linear calibration curve for the oocyst concentration versus SPR signal was obtained in the range of 1×106–1×102 oocysts ml-1. The oocyst detection was also successfully achieved in natural water systems. These results indicate that the SPR-based inhibition assay using the Cryptosporidium sensor chip has high application potential for the real-time analysis of C. parvum oocyst in laboratory and field water monitoring.

Architecture for real-time coordination of multiple autonomous mobile units

Interest on using teams of mobile robots has been growing, due to their potential to cooperate for diverse purposes, such as rescue, de-mining, surveillance or even games such as robotic soccer. These applications require a real-time middleware and wireless communication protocol that can support an efficient and timely fusion of the perception data from different robots as well as the development of coordinated behaviours. Coordinating several autonomous robots towards achieving a common goal is currently a topic of high interest, which can be found in many application domains. Despite these different application domains, the technical problem of building an infrastructure to support the integration of the distributed perception and subsequent coordinated action is similar. This problem becomes tougher with stronger system dynamics, e.g., when the robots move faster or interact with fast objects, leading to tighter real-time constraints. This thesis work addressed computing architectures and wireless communication protocols to support efficient information sharing and coordination strategies taking into account the real-time nature of robot activities. The thesis makes two main claims. Firstly, we claim that despite the use of a wireless communication protocol that includes arbitration mechanisms, the self-organization of the team communications in a dynamic round that also accounts for variable team membership, effectively reduces collisions within the team, independently of its current composition, significantly improving the quality of the communications. We will validate this claim in terms of packet losses and communication latency. We show how such self-organization of the communications can be achieved in an efficient way with the Reconfigurable and Adaptive TDMA protocol. Secondly, we claim that the development of distributed perception, cooperation and coordinated action for teams of mobile robots can be simplified by using a shared memory middleware that replicates in each cooperating robot all necessary remote data, the Real-Time Database (RTDB) middleware. These remote data copies, which are updated in the background by the selforganizing communications protocol, are extended with age information automatically computed by the middleware and are locally accessible through fast primitives. We validate our claim showing a parsimonious use of the communication medium, improved timing information with respect to the shared data and the simplicity of use and effectiveness of the proposed middleware shown in several use cases, reinforced with a reasonable impact in the Middle Size League of RoboCup.

The cloning and reconstitution of a bovine adenovirus type 2 E3 deletion mutant and the sequencing and analysis of the early 4 region

Recombinant Adenoviruses (Ads) have been shown to have potential applications in three areas: gene therapy, high level protein expression and recombinant vaccines.' At least three different locations within the Ad genome can be deleted and subsequently used for the insertion of foreign sequences. These include the Early 3 (E3), Early 1 (E1) and Early 4 (E4) regions. Viral vectors of this type have been well studied in Human Ads 2 and 5, however one has not yet been constructed for Bovine Adenovirus Type 2 (BAV2). The E3 region is located between 76.6 and 86 m.u. on the r-strand and is transcribed in a rightward direction. The gene products of the Early 3 region (E3) have been shown to be non-essential for viral replication, in vitro, but are required for host immunosurveillance. This study represents the cloning and reconstitution of a BAV2 E3 deletion mutant. A deletion of 1800bp was made within the E3 region of BAV2 and the thymidine kinase gene was subsequently inserted in the deleted area . . The plasmid pdlE3-4tk1 (23.4Kbp) was constructed and used to to facilitate homologous recombination with the wild type BAV2 to produce a mutant. Southern Blotting and Hybridization results suggest the presence of a BAV2 E3 deletion mutant with thymidine kinase sequences present. The E4 region of Human Adenovirus types 2 and 5 is located at the extreme right end of the genome (91.3 map units - 99.1 map units) and is transcribed in a leftward direction giving rise to a complicated set of differentially spliced mRNAs. Essentially there are 7 open reading frames (ORFs) encoding for at least 7 polypeptides. The gene products encoded by the E4 region have been shown to be essential for the expression of late viral genes, host cell shutoff and normal viral growth. We have cloned and sequenced the right end segment between 90.5 map units and 100 map units of the BAV2 genome. The results show several open reading frames which encode polypeptides exhibiting homology to three polypeptides encoded by the E4 region of human adenovirus type 2. These include the 14kDa protein encoded by ORF1, the 34kDa protein encoded by ORF6 and the 13kDa protein encoded by ORF3. The nucleotide sequence, restriction enzyme map, and ORF map of the E4 region could be very useful in future molecular manipulation of this region and could possibly explain the slow growth rate of BAV2 in MDBK cells.

Bt cotton, pesticides, labour and health - A case study of smallholder farmers in the Makhathini Flats, Republic of South Africa

This paper describes some of the results of a detailed farm-level survey of 32 small-scale cotton farmers in the Makhathini Flats region of South Africa. The aim was to assess and measure some of the impacts (especially in terms of savings in pesticide and labour as well as benefits to human health) attributable to the use of insect-tolerant Bt cotton. The study reveals a direct cost benefit for Bt growers of SAR416 ($51) per hectare per season due to a reduction in the number of insecticide applications. Cost savings emerged in the form of lower requirements for pesticide, but also important were reduced requirements for water and labour. The reduction in the number of sprays was particularly beneficial to women who do some spraying and children who collect water and assist in spraying. The increasing adoption rate of Bt cotton appears to have a health benefit measured in terms of reported rates of accidental insecticide poisoning. These appear to be declining as the uptake of Bt cotton increases. However, the understanding of refugia and their management by local farmers are deficient and need improving. Finally, Bt cotton growers emerge as more resilient in absorbing price fluctuations.

Bt cotton, pesticides, labour and health - A case study of smallholder farmers in the Makhathini Flats, Republic of South Africa

This paper describes some of the results of a detailed farm-level survey of 32 small-scale cotton farmers in the Makhathini Flats region of South Africa. The aim was to assess and measure some of the impacts (especially in terms of savings in pesticide and labour as well as benefits to human health) attributable to the use of insect-tolerant Bt cotton. The study reveals a direct cost benefit for Bt growers of SAR416 ($51) per hectare per season due to a reduction in the number of insecticide applications. Cost savings emerged in the form of lower requirements for pesticide, but also important were reduced requirements for water and labour. The reduction in the number of sprays was particularly beneficial to women who do some spraying and children who collect water and assist in spraying. The increasing adoption rate of Bt cotton appears to have a health benefit measured in terms of reported rates of accidental insecticide poisoning. These appear to be declining as the uptake of Bt cotton increases. However, the understanding of refugia and their management by local farmers are deficient and need improving. Finally, Bt cotton growers emerge as more resilient in absorbing price fluctuations.

An approach to chemotaxonomy to the fatty acid content of some Malvaceae species

Malvales is an order of flowering plants with a controversial circumscription. The relationships between taxa, particularly Malvaceae, Bombacaceae, Sterculiaceae, and Tiliaceae, are not well delineated. Several studies have reported the fatty acid compositions of Malvaceae plants but not for taxonomic purposes. In the present study, the fatty acid composition of oilseeds from seven species belonging to the Malvaceae family was determined by capillary gas chromatography/mass spectrometry (GC/MS), and the quantitative distribution of fatty acids was analyzed by a cluster analysis With Euclidean Distance and UPGMA. The oil content in the seeds was very low (8.3-11.8%). The profile of fatty acids showed that there were two distinct groups: species rich in palmitic acid (Herissantia tiubae, Sidastrum paniculatum and Sida rhombifolia) and species rich in linoleic acid (other Sida species). The fatty acid profiles found for Sida species are consistent with other reported data. Although our data support a distinction between Sida and Sidastrum, more species should be analyzed to evaluate the real taxonomic value of differences in fatty acid content for distinguishing Malvaceae. (C) 2010 Elsevier Ltd. All rights reserved.

Separating the wheat from the chaff: unbiased filtering of background tandem mass spectra improves protein identification

Only a small fraction of spectra acquired in LC-MS/MS runs matches peptides from target proteins upon database searches. The remaining, operationally termed background, spectra originate from a variety of poorly controlled sources and affect the throughput and confidence of database searches. Here, we report an algorithm and its software implementation that rapidly removes background spectra, regardless of their precise origin. The method estimates the dissimilarity distance between screened MS/MS spectra and unannotated spectra from a partially redundant background library compiled from several control and blank runs. Filtering MS/MS queries enhanced the protein identification capacity when searches lacked spectrum to sequence matching specificity. In sequence-similarity searches it reduced by, on average, 30-fold the number of orphan hits, which were not explicitly related to background protein contaminants and required manual validation. Removing high quality background MS/MS spectra, while preserving in the data set the genuine spectra from target proteins, decreased the false positive rate of stringent database searches and improved the identification of low-abundance proteins.

Bayesian coalescent analysis reveals a high rate of molecular evolution in GB virus C

GB virus C/hepatitis G (GBV-C) is an RNA virus of the family Flaviviridae. Despite replicating with an RNA-dependent RNA polymerase, some previous estimates of rates of evolutionary change in GBV-C suggest that it fixes mutations at the anomalously low rate of similar to 100(-7) nucleotide substitution per site, per year. However, these estimates were largely based on the assumption that GBV-C and its close relative GBV-A (New World monkey GB viruses) codiverged with their primate hosts over millions of years. Herein, we estimated the substitution rate of GBV-C using the largest set of dated GBV-C isolates compiled to date and a Bayesian coalescent approach that utilizes the year of sampling and so is independent of the assumption of codivergence. This revealed a rate of evolutionary change approximately four orders of magnitude higher than that estimated previously, in the range of 10(-2) to 10(-3) sub/site/year, and hence in line with those previously determined for RNA viruses in general and the Flaviviridae in particular. In addition, we tested the assumption of host-virus codivergence in GBV-A by performing a reconciliation analysis of host and virus phylogenies. Strikingly, we found no statistical evidence for host-virus codivergence in GBV-A, indicating that substitution rates in the GB viruses should not be estimated from host divergence times.

Consumption smoothing in Latin America: an empirical assessment of present value models

The study aims to assess the empirical adherence of the permanent income theory and the consumption smoothing view in Latin America. Two present value models are considered, one describing household behavior and the other open economy macroeconomics. Following the methodology developed in Campbell and Schiller (1987), Bivariate Vector Autoregressions are estimated for the saving ratio and the real growth rate of income concerning the household behavior model and for the current account and the change in national cash ‡ow regarding the open economy model. The countries in the sample are considered separately in the estimation process (individual system estimation) as well as jointly (joint system estimation). Ordinary Least Squares (OLS) and Seemingly Unrelated Regressions (SURE) estimates of the coe¢cients are generated. Wald Tests are then conducted to verify if the VAR coe¢cient estimates are in conformity with those predicted by the theory. While the empirical results are sensitive to the estimation method and discount factors used, there is only weak evidence in favor of the permanent income theory and consumption smoothing view in the group of countries analyzed.

PZT FOR MEASURING ENERGY FLUENCE RATE of X-RAY USED IN SUPERFICIAL CANCER THERAPY

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

Effect of microwave disinfection on the hardness and adhesion of two resilient liners

Statement of problem. Microwave irradiation has been suggested for denture disinfection. However, the effect of this procedure on the hardness and bond strength between resilient liners and denture base acrylic resin is not known.Purpose. This study evaluated the effect of water storage time and microwave disinfection on the hardness and peel bond strength of 2 silicone resilient lining materials to a heat-polymerized acrylic resin.Material and methods. Acrylic resin (Lucitone 199) specimens (75 X 10 X 3 mm) were stored in water at 37 degrees C (2 or 30 days) before bonding (n = 160). The resilient lining materials (GC Reline Extra Soft and Dentusil) were bonded to the denture base and divided into the following 4 groups (n = 10): Tests performed immediately after bonding (control); specimens immersed in water (200 mL) and irradiated twice, with 650 W for 6 minutes; specimens irradiated daily for 7 total cycles of disinfection; specimens immersed in water (37 degrees C) for 7 days. Specimens were submitted to a 180-degree peel test (at a crosshead speed of 10 mm/min) and the failure values (MPa) and mode of failure were recorded. Pretreatment and posttreatment hardness measurements (Shore A) of the resilient materials were also performed. Three-way analysis of variance, followed by the Tukey HSD test, was performed (alpha=.05).Results. The analysis revealed that, for all conditions, the mean failure strengths of GC Reline Extra Soft (0.95-1.19 MPa) were significantly higher (P<.001) than those of Dentusil (0.45-0.50 MPa). The adhesion of the liners was not adversely affected by water storage time of Lucitone 199 or microwave disinfection. All peel test failures were cohesive. There was a small but significant difference (P<.001) between the pretreatment (34.33 Shore A) and posttreatment (38.69 Shore A) hardness measurements.Conclusion. Microwave disinfection did not compromise the hardness of either resilient liners or their adhesion to the denture base resin Lucitone 199.

Dissociated time course recovery between rate of force development and peak torque after eccentric exercise

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

Inferences of body energy reserves on conception rate of suckled Zebu beef cows subjected to timed artificial insemination followed by natural mating

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

Survival rate of osseointegrated implants in atrophic maxillae grafted with calvarial bone: a retrospective study

The aim of this study was to evaluate the clinical survival rate of osseointegrated implants placed in the atrophic maxilla that has been reconstructed by means of autogenous bone grafts harvested from a cranial calvarial site. Further, we sought to analyse the level of pen-implant bone after prosthetic rehabilitation and to determine subjective patient satisfaction with the treatment performed. This study conformed to the STROBE guidelines regarding retrospective studies. Twenty-five patients who had received osseointegrated implants with late loading in the reconstructed atrophic maxilla were included in the study. The survival rate and level of pen-implant bone loss were evaluated. A questionnaire related to the surgical and prosthetic procedures was completed. The observed implant survival rate was 92.35%. The mean bone loss recorded was 1.76 mm in the maxilla and 1.54 mm in the mandible. The results of the questionnaire indicated a high level of patient satisfaction, little surgical discomfort, and that the patients would recommend the procedure and would undergo the treatment again. From the results obtained, it is concluded that the cranial calvarial site is an excellent donor area; calvarial grafts provided stability and maintenance of bone volume over the course of up to 11 years.

Characterization of geothermal reservoirs’ parameters by inverse problem resolution and geostatistical simulations

BTES (borehole thermal energy storage)systems exchange thermal energy by conduction with the surrounding ground through borehole materials. The spatial variability of the geological properties and the space-time variability of hydrogeological conditions affect the real power rate of heat exchangers and, consequently, the amount of energy extracted from / injected into the ground. For this reason, it is not an easy task to identify the underground thermal properties to use when designing. At the current state of technology, Thermal Response Test (TRT) is the in situ test for the characterization of ground thermal properties with the higher degree of accuracy, but it doesn’t fully solve the problem of characterizing the thermal properties of a shallow geothermal reservoir, simply because it characterizes only the neighborhood of the heat exchanger at hand and only for the test duration. Different analytical and numerical models exist for the characterization of shallow geothermal reservoir, but they are still inadequate and not exhaustive: more sophisticated models must be taken into account and a geostatistical approach is needed to tackle natural variability and estimates uncertainty. The approach adopted for reservoir characterization is the “inverse problem”, typical of oil&gas field analysis. Similarly, we create different realizations of thermal properties by direct sequential simulation and we find the best one fitting real production data (fluid temperature along time). The software used to develop heat production simulation is FEFLOW 5.4 (Finite Element subsurface FLOW system). A geostatistical reservoir model has been set up based on literature thermal properties data and spatial variability hypotheses, and a real TRT has been tested. Then we analyzed and used as well two other codes (SA-Geotherm and FV-Geotherm) which are two implementation of the same numerical model of FEFLOW (Al-Khoury model).