Rpe65-Gene Transfer Using an Integration-Deficient Lentiviral Vector

Purpose: We previously demonstrated efficient retinal rescue of RPE65 mouse models (Rpe65-/- (Bemelmans et al, 2006) and Rpe65R91W/R91W mice) using a HIV1-derived lentiviral vector encoding for the mouse RPE65 cDNA. In order to optimize a lentiviral vector as an alternative tool for RPE65-derived Leber Congenital Amaurosis clinical trials, we evaluated the efficiency of an integration-deficient lentiviral vector (IDLV) encoding the human RPE65 cDNA to restore retinal function in the Rpe65R91W/R91W mice. Methods: An HIV-1-derived lentiviral vector expressing either the hrGFPII or the human Rpe65 cDNA under the control of a 0.8 kb fragment of the human Rpe65 promoter (R0.8) was produced by transient transfection of 293T cells. A LQ-integrase mutant was used to generate the IDLV vectors. IDLV-R0.8-hRPE65 or hrGFPII were injected subretinally into 1 month-old Rpe65R91W/R91W mice. Functional rescue was assessed by ERG (1 and 3 months post-injection) and cone survival by immunohistology. Results: An increased light sensitivity was detected by scotopic ERG in animals injected with IDLV-R0.8-hRPE65 compared to hrGFPII-treated animals or untreated mice. However the improvement was delayed compared to integration-proficient LV and observed at 3 months but not 1 month post-injection. Immunolabelling of cone markers showed an increased number of cones in the transduced area compared to control groups. Conclusions: The IDLV-R0.8-hRPE65 vectors allow retinal improvement in the Rpe65R91W/R91W mice. Both rod function and cone survival were demonstrated even if there is a delay in the rescue as assessed by scotopic ERG. Integration-deficient vectors minimize insertional mutagenesis and thus are safer candidates for human application. Further experiments using large animals are now needed to validate correct gene transfer and expression of the RPE65 gene as well as tolerance of the vector after subretinal injection before envisaging a clinical trial application.

Soil biological attributes in pastures of different ages in a crop-livestock integrated system

The objective of this work was to evaluate the effect of the pasture (Urochloa brizantha) component age on soil biological properties, in a crop-livestock integrated system. The experiment was carried out in a Brazilian savannah (Cerrado) area with 92 ha, divided into six pens of approximately 15 ha. Each pen represented a different stage of the pasture component: formation, P0; one year, P1; two years, P2; three years, P3; and final with 3.5 years, Pf. Samples were taken in the 0-10 cm soil depth. The soil biological parameters - microbial biomass carbon (MBC), microbial biomass respiration (C-CO2), metabolic quotient (qCO2), microbial quotient (q mic), and total organic carbon (TOC) - were evaluated and compared among different stages of the pasture, and between an adjacent area under native Cerrado and another area under degraded pasture (PCD). The MBC, q mic and TOC increased and qCO2 reduced under the different pasture stages. Compared to PCD, the pasture stages had higher MBC, q mic and TOC, and lower qCO2. The crop-livestock integrated system improved soil microbiological parameters and immobilized carbon in the soil in comparison to the degraded pasture.

Correlation of physical and chemical attributes of soil with sugarcane yield

The objective of this work was to evaluate the correlation between sugarcane yield and some physical and chemical attributes of soil. For this, a 42‑ha test area in Araras, SP, Brazil, was used. Soil properties were determined from samples collected at the beginning of the 2003/2004 harvest season, using a regular 100x100 m grid. Yield assessment was done with a yield monitor (Simprocana). Correlation analyses were performed between sugarcane yield and the following soil properties: pH, pH CaCl2, N, C, cone index, clay content, soil organic matter, P, K, Ca, Mg, H+AL, cation exchange capacity, and base saturation. Correlation coefficients were respectively ‑0.05, ‑0.29, 0.33, 0.41, ‑0.27, 0.22, 0.44, ‑0.24, trace, ‑0.06, 0.01, 0.32, 0.14, and 0.04. Correlations of chemical and physical attributes of soil with sugarcane yield are weak, and, per se, they are not able to explain sugarcane yield variation, which suggests that other variables, besides soil attributes, should be analysed.

Chemical attributes of a Hapludox soil after nine years of pig slurry application

The objective of this study was to evaluate the pig slurry application effects on chemical attributes of a Hapludox soil managed under no-tillage system. Treatments consisted of 50, 100 and 200 m³ ha-1 per year of pig slurry application, and a control with replacement of P and K exported through harvested grains. Attributes related to soil chemical reaction, exchange complex, and nutrient contents were determined in soil samples collected in the ninth year of experimentation from 0 - 0.025, 0.025 - 0.05, 0.05 - 0.10, 0.10 - 0.20, 0.20 - 0.40 and 0.40 - 0.60 m soil depths. The continuous application of high doses of pig slurry on the Oxisol surface under no-tillage acidifies the soil and increases Al, P, Cu, and Zn contents down to 0.2-m depth, and K levels down to 0.6-m depth.

Metabolic attributes, yield and stability of milk in Jersey cows fed diets containing sodium citrate and sodium bicarbonate

The objective of this work was to evaluate the inclusion of sodium citrate and sodium bicarbonate in the diet of lactating Jersey cows, and its effects on the metabolic attributes, productivity and stability of milk. We evaluated urinary pH, levels of glucose and urea in blood, body weight, body condition score, milk yield, milk stability (ethanol test), and milk physicochemical properties of 17 cows fed diets containing sodium citrate (100 g per cow per day), sodium bicarbonate (40 g per cow per day) or no additives. Assessments were made at the 28th and 44th days. Supply of sodium citrate or bicarbonate has no influence on the metabolic attributes, productivity, body weight, and body condition score of the cows, neither on the composition and stability of milk.

Microbiological and chemical attributes of a Hapludalf soil with swine manure fertilization

The objective of this work was to evaluate the microbiological and chemical attributes of a soil with a seven‑year history of urea and swine manure application. In the period from October 2008 to October 2009, soil samples were collected in the 0-10 cm layer and were subjected to the treatments: control, without application of urea or manure; and with the application of urea, pig slurry, and deep pig litter in two doses, in order to supply one or two times the recommended N doses for the maize (Zea mays)/black oat (Avena strigosa) crop succession. The carbon of the microbial biomass (MB‑C) and the basal respiration (C‑CO2) were analyzed, and the metabolic (qCO2) and microbial quotient (qmic) were calculated with the obtained data. Organic matter, pH in water, available P and K, and exchangeable Ca and Mg were also determined. The application of twice the dose of deep pig litter increases the MB‑C and C‑CO2 values. The qmic and qCO2 are little affected by the application of swine manure. The application of twice the dose of deep pig litter increases the values of pH in water and the contents of available P and of exchangeable Ca and Mg in the soil.

Chemical and biological attributes of a lowland soil affected by land leveling

The objective of this work was to evaluate the relationship between soil chemical and biological attributes and the magnitude of cuts and fills after the land leveling process of a lowland soil. Soil samples were collected from the 0 - 0.20 m layer, before and after leveling, on a 100 point grid established in the experimental area, to evaluate chemical attributes and soil microbial biomass carbon (MBC). Leveling operations altered the magnitude of soil chemical and biological attributes. Values of Ca, Mg, S, cation exchange capacity, Mn, P, Zn, and soil organic matter (SOM) decreased in the soil profile, whereas Al, K, and MBC increased after leveling. Land leveling decreased in 20% SOM average content in the 0 - 0.20 m layer. The great majority of the chemical attributes did not show relations between their values and the magnitude of cuts and fills. The relation was quadratic for SOM, P, and total N, and was linear for K, showing a positive slope and indicating increase in the magnitude of these attributes in cut areas and stability in fill areas. The relationships between these chemical attributes and the magnitude of cuts and fills indicate that the land leveling map may be a useful tool for degraded soil recuperation through amendments and organic fertilizers.

A model of the wind direction signature in the Stokes emission vector from the ocean surface at microwave frequencies

This paper presents a model of the Stokes emission vector from the ocean surface. The ocean surface is described as an ensemble of facets with Cox and Munk's (1954) Gram-Charlier slope distribution. The study discusses the impact of different up-wind and cross-wind rms slopes, skewness, peakedness, foam cover models and atmospheric effects on the azimuthal variation of the Stokes vector, as well as the limitations of the model. Simulation results compare favorably, both in mean value and azimuthal dependence, with SSM/I data at 53° incidence angle and with JPL's WINDRAD measurements at incidence angles from 30° to 65°, and at wind speeds from 2.5 to 11 m/s.

Personalized drug administrations using Support Vector Machine

The decision-making process regarding drug dose, regularly used in everyday medical practice, is critical to patients' health and recovery. It is a challenging process, especially for a drug with narrow therapeutic ranges, in which a medical doctor decides the quantity (dose amount) and frequency (dose interval) on the basis of a set of available patient features and doctor's clinical experience (a priori adaptation). Computer support in drug dose administration makes the prescription procedure faster, more accurate, objective, and less expensive, with a tendency to reduce the number of invasive procedures. This paper presents an advanced integrated Drug Administration Decision Support System (DADSS) to help clinicians/patients with the dose computing. Based on a support vector machine (SVM) algorithm, enhanced with the random sample consensus technique, this system is able to predict the drug concentration values and computes the ideal dose amount and dose interval for a new patient. With an extension to combine the SVM method and the explicit analytical model, the advanced integrated DADSS system is able to compute drug concentration-to-time curves for a patient under different conditions. A feedback loop is enabled to update the curve with a new measured concentration value to make it more personalized (a posteriori adaptation).

A new disruption vector (pDHO) to obtain heterothallic strains from both Saccharomyces cerevisiae and Saccharomyces pastorianus

Yeasts are responsible for several traits in fermented beverages, including wine and beer, and their genetic manipulation is often necessary to improve the quality of the fermentation product. Improvement of wild-type strains of Saccharomyces cerevisiae and Saccharomyces pastorianus is difficult due to their homothallic character and variable ploidy level. Homothallism is determined by the HO gene in S. cerevisiae and the Sc-HO gene in S. pastorianus. In this work, we describe the construction of an HO disruption vector (pDHO) containing an HO disruption cassette and discuss its use in generating heterothallic yeast strains from homothallic Saccharomyces species.

How a haemosporidian parasite of bats gets around: the genetic structure of a parasite, vector and host compared.

Parasite population structure is often thought to be largely shaped by that of its host. In the case of a parasite with a complex life cycle, two host species, each with their own patterns of demography and migration, spread the parasite. However, the population structure of the parasite is predicted to resemble only that of the most vagile host species. In this study, we tested this prediction in the context of a vector-transmitted parasite. We sampled the haemosporidian parasite Polychromophilus melanipherus across its European range, together with its bat fly vector Nycteribia schmidlii and its host, the bent-winged bat Miniopterus schreibersii. Based on microsatellite analyses, the wingless vector, and not the bat host, was identified as the least structured population and should therefore be considered the most vagile host. Genetic distance matrices were compared for all three species based on a mitochondrial DNA fragment. Both host and vector populations followed an isolation-by-distance pattern across the Mediterranean, but not the parasite. Mantel tests found no correlation between the parasite and either the host or vector populations. We therefore found no support for our hypothesis; the parasite population structure matched neither vector nor host. Instead, we propose a model where the parasite's gene flow is represented by the added effects of host and vector dispersal patterns.

Definition of linear color models in the RGB vector color space to detect red peaches in orchard images taken under natural illumination

This work proposes the detection of red peaches in orchard images based on the definition of different linear color models in the RGB vector color space. The classification and segmentation of the pixels of the image is then performed by comparing the color distance from each pixel to the different previously defined linear color models. The methodology proposed has been tested with images obtained in a real orchard under natural light. The peach variety in the orchard was the paraguayo (Prunus persica var. platycarpa) peach with red skin. The segmentation results showed that the area of the red peaches in the images was detected with an average error of 11.6%; 19.7% in the case of bright illumination; 8.2% in the case of low illumination; 8.6% for occlusion up to 33%; 12.2% in the case of occlusion between 34 and 66%; and 23% for occlusion above 66%. Finally, a methodology was proposed to estimate the diameter of the fruits based on an ellipsoidal fitting. A first diameter was obtained by using all the contour pixels and a second diameter was obtained by rejecting some pixels of the contour. This approach enables a rough estimate of the fruit occlusion percentage range by comparing the two diameter estimates.

SOIL ATTRIBUTES THREE YEARS AFTER MAKING RIDGES AND OR DRAINS FOR PLANTING GRAPEVINE ROOTSTOCK

RESUMO Os atributos do solo estão relacionados com o desenvolvimento e a produção das culturas, tanto anuais como perenes. O presente trabalho teve como objetivo determinar os atributos químicos e físicos do solo três anos após a aplicação de sistemas de manejo para implantação de porta-enxertos de videira, em um Nitossolo Vermelho com textura muito argilosa. Os sistemas de manejo consistiram na confecção ou não de camalhões e/ou de drenos antes da implantação das mudas de porta-enxertos Dog Ridge. Os atributos químicos (pH em água e teores de matéria orgânica e de macronutrientes) foram determinados em amostras coletadas na linha da cultura, nas camadas de 0-0,1; 0,1-0,2; 0,2-0,3 e 0,3-0,4 m de profundidade. Os atributos físicos do solo (teor de argila, densidade, porosidade, resistência à penetração e estabilidade dos agregados) foram determinados em amostras coletadas com estrutura preservada nas camadas de 0,025-0,075; 0,125-0,175; 0,225-0,275 e 0,325-0,375 m de profundidade. Também foram determinadas a massa seca de raízes do portaenxerto nas camadas de 0-0,2 e 0,2-0,4 m de profundidade e suas correlações com os atributos do solo. A confecção de camalhões alterou significativamente a maioria dos atributos químicos do solo e proporcionou maior uniformidade nos valores em profundidade, criando condições mais favoráveis ao crescimento radicular nas camadas mais profundas. Os atributos físicos foram menos afetados do que os químicos pela confecção de camalhões ou drenos, e não foram observados valores restritivos ao crescimento radicular nas camadas amostradas após quatro anos da aplicação dos sistemas de manejo do solo. A confecção de drenos alterou menos os atributos químicos e físicos do solo do que a confecção de camalhões.