Detection of three Allexivirus species infecting garlic in Brazil

Garlic viruses often occur in mixed infections under field conditions. In this study, garlic samples collected in three geographical areas of Brazil were tested by Dot-ELISA for the detection of allexiviruses using monoclonal specific antibodies to detect Garlic virus A (GarV-A), Garlic virus B (GarV-B), Garlic virus C (GarV-C) and a polyclonal antiserum able to detect the three virus species mentioned plus Garlic virus D (GarV-D). The detected viruses were biologically isolated by successive passages through Chenopodium quinoa. Reverse Transcriptase Polimerase Chain Reaction (RT-PCR) was performed using primers designed from specific regions of the coat protein genes of Japanese allexiviruses available in the Genetic Bank of National Center of Biotechnology Information (NCBI). By these procedures, individual garlic virus genomes were isolated and sequenced. The nucleotide and amino acid sequence analysis and the one with serological data revealed the presence of three distinct allexiviruses GarV-C, GarV-D and a recently described allexivirus, named Garlic mite-borne filamentous virus (GarMbFV), in Brazil.

Sequence similarity between the viral cp gene and the transgene in transgenic papayas

The Papaya ringspot virus (PRSV) coat protein transgene present in 'Rainbow' and 'SunUp' papayas disclose high sequence similarity (>89%) to the cp gene from PRSV BR and TH. Despite this, both isolates are able to break down the resistance in 'Rainbow', while only the latter is able to do so in 'SunUp'. The objective of this work was to evaluate the degree of sequence similarity between the cp gene in the challenge isolate and the cp transgene in transgenic papayas resistant to PRSV. The production of a hybrid virus containing the genome backbone of PRSV HA up to the Apa I site in the NIb gene, and downstream from there, the sequence of PRSV TH was undertaken. This hybrid virus, PRSV HA/TH, was obtained and used to challenge 'Rainbow', 'SunUp', and an R2 population derived from line 63-1, all resistant to PRSV HA. PRSV HA/TH broke down the resistance in both papaya varieties and in the 63-1 population, demonstrating that sequence similarity is a major factor in the mechanism of resistance used by transgenic papayas expressing the cp gene. A comparative analysis of the cp gene present in line 55-1 and 63-1-derived transgenic plants and in PRSV HA, BR, and TH was also performed.

Heritability of blood pressure in the swiss population: the family-based skipogh study

Objective: Blood pressure is known to aggregate in families. Yet, heritability estimates are population-specific and no Swiss data have been published so far. Moreover, little is known on the heritability of the white-coat effect. We investigated the heritability of various blood pressure (BP) traits in a Swiss population-based sample. Methods: SKIPOGH (Swiss Kidney Project on Genes in Hypertension) is a family-based multi-centre (Lausanne, Bern, Geneva) cross-sectional study that examines the role of genes in determining BP levels. Office and 24-hour ambulatory BP were measured using validated devices (A&D UM-101 and Diasys Integra). We estimated the heritability of systolic BP (SBP), diastolic BP (DBP), heart rate (HR), pulse pressure (PP), proportional white-coat effect (i.e. [office BP-mean ambulatory daytime BP]/mean ambulatory daytime BP), and nocturnal BP dipping (difference between mean ambulatory daytime and night-time BP) using a maximum likelihood method implemented in the SAGE software. Analyses were adjusted for age, sex, body mass index (BMI), and study centre. Analyses involving PP were additionally adjusted for DBP. Results: The 517 men and 579 women included in this analysis had a mean (}SD) age of 46.8 (17.8) and 47.8 (17.1) years and a mean BMI of 26.0 (4.2) and 24.2 (4.6) kg/m2, respectively. Heritability estimates (}SE) for office SBP, DBP, HR, and PP were 0.20}0.07, 0.20}0.07, 0.39}0.08, and 0.16}0.07 (all P<0.01). Heritability estimates for 24-hour ambulatory SBP, DBP, HR, and PP were, respectively, 0.39}0.07, 0.30}.08, 0.19}0.09, and 0.25}0.08 (all P<0.05). The heritability of the white-coat effect was 0.29}0.07 for SBP and 0.31}0.07 for DBP (both P<0.001). The heritability of nocturnal BP dipping was 0.15}0.08 for SBP and 0.22}0.07 for DBP (both P<0.05). Conclusions: We found that the white-coat effect is significantly heritable. Our findings show that BP traits are moderately heritable in a multi-centric study in Switzerland, in line with previous population-based studies, justifying the ongoing search for genetic determinants in this field.

Analysis of preharvest sprouting in three Brazilian wheat populations

The objective of this work was to evaluate the possibility of obtaining recombinant inbred wheat lines more resistant to preharvest sprouting, independently of colour genes, in three red-grained Brazilian wheat populations. The results showed statistical significance among lines within all populations, which presented a normal distribution and transgressive segregation for preharvest sprouting. The normal distribution of the lines from all red-grained populations suggests that sprouting, excluding the genes expressing seed coat pigmentation, is, probably, controlled by many genes. These findings also indicate that it may be possible to improve resistance to preharvest sprouting, independently of the colour genes.

New species emergence via recombination among isolates of the Brazilian tomato infecting Begomovirus complex

Partial nucleotide sequences of five tomato infecting Begomovirus isolates were determined from DNA-A fragments, corresponding to the 5' region of the replication associated protein gene, the intergenic region and the 5' region of the coat protein gene. Isolate DFM shared 95% identity with Tomato mottle leaf curl virus (TMoLCV), isolates 34, PA-05, and Ta4 were 88% identical to Tomato yellow vein streak virus and isolate DF-BR3 shared 77% identity with TMoLCV. Recombination analysis indicated that isolate DF-BR3 was a chimaera, and it provided evidence that there is a complex and actively recombining population of tomato infecting begomoviruses in Brazil.

Prediction of cottonseed longevity

The objective of this work was to determine the viability equation constants for cottonseed and to detect the occurrence and depletion of hardseededness. Three seedlots of Brazilian cultivars IAC-19 and IAC-20 were tested, using 12 moisture content levels, ranging from 2.2 to 21.7% and three storage temperatures, 40, 50 and 65ºC. Seed moisture content level was reached from the initial value (around 8.8%) either by rehydration, in a closed container, or by drying in desiccators containing silica gel, both at 20ºC. Twelve seed subsamples for each moisture content/temperature treatment were sealed in laminated aluminium-foil packets and stored in incubators at those temperatures, until complete survival curves were obtained. Seed equilibrium relative humidity was recorded. Hardseededness was detected at moisture content levels below 6% and its releasing was achieved either naturally, during storage period, or artificially through seed coat removal. The viability equation quantified the response of seed longevity to storage environment well with K E = 9.240, C W = 5.190, C H = 0.03965 and C Q = 0.000426. The lower limit estimated for application of this equation at 65ºC was 3.6% moisture content.

The role of the c-myc gene in pigment cell development and homeostasis

Résumé La dérégulation de c-Myc est un événement fréquent de la transformation cellulaire. Une régulation positive de cette oncoprotéine a été démontrée dans divers mélanomes cutanés primaires et métastatiques et est associée à un pronostic défavorable (Grover et al., 1996; Zhuang et al., 2008). c-Myc est considéré comme une molécule centrale impliquée dans plusieurs processus de l'homéostasie cellulaire. En raison de sa contribution importante dans la progression tumorale, la fonction de c-Myc a été étudiée intensément. Cependant nous connaissons peu le rôle de ce facteur de transcription dans l'embryogenèse et dans la spécification tissulaire. Un déficit total de c-Myc pendant l'embryogenèse conduit à la mort embryonnaire avant 10.5 jours de gestation. Cette mort est causée par de multiples imperfections du développement touchant la taille de l'embryon, le coeur, le péricarde, le tube neural et les cellules sanguines (Davis et al., 1993; Trumpp et al., 2001). Récemment, il a été montré que la plupart de ces anomalies sont secondaires et résultent d'une insuffisance du placenta dans les embryons c-myc-/- (Dubois et al., 2008). Sachant que c-Myc est important dans la maintenance des lignées de la crête neurale (Wei et al., 2007), nous nous sommes intéressés au rôle de c-Myc dans le développement des cellules pigmentaires et à leur homéostasie après la naissance. Un allèle floxé de c-myc (Trumpp et al., 2001) a été utilisé pour supprimer ce gène spécifiquement dans la lignée mélanocytaire à l'aide d'une souris transgénique Tyr::Cre (Delmas et al., 2003). L'ablation des deux allèles de c-myc dans les mélanocytes des souris c-myccKO conduit au phénotype de grisonnement des poils, observé directement après la naissance et associé à une diminution du nombre de mélanocytes dans le bulbe des follicules pileux. Les cellules pigmentaires restantes expriment les marqueurs mélanogéniques (Tyr, TRP-1, Dct and MITF) et semblent être fonctionnelles puisqu'elles peuvent produire et transférer la mélanine. De plus, la capacité de prolifération des mélanocytes déficients en c-Myc dans le bulbe des follicules pileux ne semble pas être affectée chez les nouveaux-nés. Les cellules souches mélanocytaires sont présentes, mais en nombre réduit, dans le bulge des follicules pileux à la fin de la morphogenèse chez les souris c-myccKO âgées de huit jours. Ces cellules sont maintenues sans changement durant le premier cycle pileux (vérifié à l'âge de trente jours), ce qui sous-entend que la fonction de c-Myc n'est pas nécessaire pour ce processus. Ceci explique pourquoi, en supposant que des cellules souches mélanocytaires fonctionnelles sont présentes dans la peau, nous n'observons pas de dilution de couleur de la robe liée à l'âge. Cependant, la présence de ces cellules souches mélanocytaires dans la peau c-myccKO ne suffit pas à assurer une quantité normale de mélanocytes différenciés dans le bulbe des follicules pileux. Cette population de cellules pigmentaires matures est sévèrement affectée par la suppression de c-Myc, ce qui contribue amplement au phénotype de grisonnement des poils. De plus, c-Myc paraît être important pour le développement des mélanocytes. Ainsi, le nombre de mélanoblastes diminue dans les embryons c-myccKO à partir du douzième jour de gestation. A treize jours de gestation, au stade où les mélanoblastes pénètrent dans l'épiderme et prolifèrent, les mélanoblastes déficients en c-Myc ne s'adaptent pas aux signaux de prolifération et se retrouvent en nombre réduit dans l'épiderme. Finalement, nous nous sommes intéressés, au rôle de N-Myc, un homologue proche de c-Myc, dans la lignée mélanocytaire. Nos expériences ont montré que. N-Myc était superflu pour le développement et l'homéostasie des mélanocytes, une seule copie du gène c-myc étant suffisante pour maintenir une pigmentation normale de la robe des souris c-mycc-myccKO/+~N_ myccKO/KO. Cependant, le rôle essentiel de N-Myc dans la maintenance des cellules mélanocytaires précurseurs apparaît lorsque c-Myc est absent, puisque la suppression simultanée des deux Myc résulte en une perte complète de la coloration de la robe. Ceci implique la présence d'un mécanisme compensatoire entre c- et N-Myc dans la lignée mélanocytaire, avec un rôle prédominant de c-Myc. Summary Deregulation of c-Myc is known to be a common event in cellular transformation. Upregulation of this oncoprotein was shown in a variety of primary and metastatic cutaneous melanomas and has been associated with a poor prognosis (Grover et al., 1996; Zhuang et al., 2008). c-myc is seen as a central molecule involved in many aspects of cellular homeostasis. c-Myc function has been intensively studied mostly because of its significant contribution to tumour progression. However little is known on the role of this transcription factor in embryogenesis and tissue specification. Complete loss of c-Myc during embryogenesis results in embryonic death before E10.5 due to multiple developmental defects including embryonic size, heart, pericardium, neural tube and blood cells (Davis et al., 1993; Trumpp et al., 2001). Recently it was discovered that most of these abnormalities are secondary and results of placental insufficiency in c-Myc-/- embryos (Dubois et al., 2008). Here, we focused on the role of c-Myc in pigment cell development and homeostasis after birth, knowing that c-Myc is important in the maintenance of neural crest lineages (Wei et al., 2007). A floxed allele of c-Myc (Trumpp et al., 2001) was used to specifically delete this gene in the melanocyte lineage using Tyr::Cre transgenic mice (Delmas et al., 2003). Removal of both c-Myc alleles in melanocytes of c-MyccKO mouse led to the grey hair phenotype which is seen directly after birth and was associated with a decrease in the melanocyte number in the bulb of the hair follicle. The remaining population of pigment cells express melanogenic markers (Tyr, TRP-1, Dct and MITF) and seem functionally normal since they can produce and transfer melanin. Furthermore proliferation capacity of c-Myc deficient melanocytes in the bulb of hair follicle seems not to be affected in newborn animals. Melanocyte stem cells (MSCs) are present but reduced in numbers in the bulge of the hair follicle at the end of morphogenesis in 8 days old c-MyccKO mice. These cells are maintained through the first hair cycle (as verified at P30) without any further changes, suggesting that c-Myc function is not required for this process. This explains why we did not detect any agerelated coat color dilution, assuming a presence of functional MSCs in the skin. Importantly, presence of MSCs in c-MyccKO skin was not sufficient for assuring a normal number of differentiated melanocytes in the bulb of the hair follicle. This population of mature pigmented cells is severely affected upon c-myc deletion thus largely contributing to the grey hair phenotype. Moreover, c-Myc appears to be important for melanocyte development. Thus, melanoblast number is affected in c-MyccKO embryos day 12 of gestation onwards. At E13.5, when melanoblasts enter the epidermis and proliferate, c-myc deficient melanoblasts failed to adapt to proliferation signals and are therefore reduced in number in the epidermis. Finally, we addressed the role of N-Myc, a closest homologue of c-Myc, in the melanocyte lineage. In these experiments, N-Myc was dispensable for melanocyte development and homeostasis, and even one copy of the c-myc gene was sufficient to maintain normal coat color pigmentation in c-mycc-mycCKO/+ ,N-myccKO/KO mice. However the crucial role of N-Myc in maintenance of melanocyte precursor cells became apparent when c-myc is eliminated since simultaneous deletion of both Myc results in complete loss of coat color pigmentation. This suggests compensatory mechanisms between c- and N-Myc with a predominant role of c-Myc in melanocyte lineage.

Portland Cement Concrete Curing Compound Performance Phase I and Phase II, MLR-02-03, 2003

Testing the efficiency of Portland Cement Concrete (PCC) curing compounds is currently done following Test Method Iowa 901-D, May 2002. Concrete test specimens are prepared from mortar materials and are wet cured 5 hours before the curing compound is applied. All brands of curing compound submitted to the Iowa Department of Transportation are laboratory tested for comparative performance under the same test conditions. These conditions are different than field PCC paving conditions. Phase I tests followed Test Method Iowa 901-D, but modified the application amounts of the curing compound. Test results showed that the application of two coats of one-half thickness each increased efficiency compared to one full thickness coat. Phase II tests also used the modified application amounts, used a concrete mix (instead of a mortar mix) and applied curing compound a few minutes after molding. Measurements of losses, during spraying of the curing compound, were noted and were found to be significant. Test results showed that application amounts, testing techniques, concrete specimen mix design and spray losses do influence the curing compound efficiency. The significance of the spray losses indicates that the conventional test method being used (Iowa 901 D) should be revised.

Penetration Characteristics of Asphalt in a Recycled Mixture, MLR-79-03, 1979

Samples of both recycled and nonrecycled asphaltic concrete were extracted in increments by the Abson Recovery Method and the penetration of the asphalt from each increment determined. The recycled projects were plantsite operations containing various amounts of virgin gravel. Cored samples were taken from the pavements on Kossuth County roads that were constructed as recycled projects in 1975, 1976, and 1977. Cored samples were also taken from a Kossuth County paving project done in 1975, that was not recycled. Comparison mix samples from 1978 construction projects in Marshall and Woodbury Counties of non - recycled projects are included. The test data from the penetrations of the recovered asphalt indicates that mixing of the old and new asphalt occurs very extensively in the hot recycling process. In laboratory controlled conditions it is difficult to coat aggregates with different penetration asphalts and prevent them from mixing.

Thin Maintenance Surfaces for Municipalities, TR-507, 2007

As streets age, officials must deal with rehabilitating and reconstructing these pavements to maintain a safe and comfortable ride. In light of nationwide budget shortfalls, cost-effective methods of extending pavement service life must be developed or the overall condition of street systems will continue to fall. Thin maintenance surfaces (TMSs) are a set of cost-effective preventive maintenance surfacing techniques that can be used to extend the life of bituminous pavement—pavement built with hot mix asphalt, hot mix asphalt overlays of portland cement concrete pavements, built-up seal coat (chip seal), stabilized materials, or a combination of these. While previous phases of TMS research have provided information about the uses of thin maintenance surfaces in rural settings, urban areas have different road maintenance challenges that should be considered separately. This research provides city street officials with suggestions for TMS techniques that street departments can easily test and include into their current programs. This research project facilitated the construction of TMS test sections in Cedar Rapids, Council Bluffs, and West Des Moines (all urban settings in Iowa). Test section sites and surfaces were selected to suit the needs of municipalities and were applied to roads with an array of various distresses and maintenance needs. Condition surveys of each test section were performed before construction, after construction, and after the first winter to record the amount and severity of existing distress and calculate the pavement condition index. Because conditions of the test sections varied greatly, determining which surface was most successful by comparing case studies was not feasible. However, some general conclusions can be made from this research. TMSs are suitable preventive maintenance techniques for a municipal street department’s program for preserving existing pavements. Careful attention should be paid to proper planning, quality control during construction, aggregate and binder selection, and aggregate embedment in order to support successful TMS application.

Hydro-Surface Preparation and Coating for Painted Structural Steel, TR-407, 1999

Cities and counties in Iowa have more than 8,890 steel bridges, most of which are painted with red lead paint. The Iowa Department of Transportation (Iowa DOT) maintains less than 35 bridges coated with red lead paint, including seven of the large border bridges over the Mississippi and Missouri Rivers. Because of the federal and state regulations for bridge painting, many governmental agencies have opted not to repaint, or otherwise maintain, lead paint coatings. Consequently, the paint condition on many of these bridges is poor, and some bridges are experiencing severe rusting of structural members. This research project was developed with two objectives: 1) to evaluate the effectiveness of preparing the structural steel surface of a bridge with high pressure water jetting instead of abrasive blasting and 2) to coat the structural steel surface with a moisture-cured polyurethane paint under different surface preparation conditions.

Resistance of genetically modified potatoes to Potato virus Y under field conditions

The objective of this work was to evaluate the resistance of genetically modified clones of potato to Potato virus Y (PVY) under field conditions. Genetically modified plants were compared with nontransformed plants of the same cultivar. The plots were flanked with potato plants infected with both PVYº and PVY N strains (spread lines), in order to provide the experimental area with the source of virus, which was naturally spread by the native aphid population. The experiment was weekly monitored by visual inspections and by DAS-Elisa in the plants produced from the harvested tubers, in order to evaluate the resistance of transgenic plants throughout the plant growth cycle. By the end of the third year, no infection symptoms were observed in the 1P clone; clone 63P showed 1% of infection, in contrast to about 90% of nontransformed plants infected. The stable expression of resistance to PVY provided by the coat protein gene was obtained in genetically modified clones of potato plants cultivar Achat under field conditions, during three consecutive years.

Stability of Citrus tristeza virus protective isolates in field conditions

The objective of this work was to monitor the maintenance of Citrus tristeza virus (CTV) protective isolates stability in selected clones of 'Pêra' sweet orange (Citrus sinensis), preimmunized or naturally infected by the virus, after successive clonal propagations. The work was carried out in field conditions in the north of Paraná State, Brazil. Coat protein gene (CPG) analysis of 33 isolates collected from 16 clones of 'Pêra' sweet orange was performed using single strand conformational polymorphism (SSCP). Initially, the isolates were characterized by symptoms of stem pitting observed in clones. Then viral genome was extracted and used as template for the amplification of CPG by reverse transcription polimerase chain reaction (RTPCR). RTPCR products electrophoretic profiles were analyzed using the Jaccard coefficient and the UPGMA method. The majority of the clones had weak to moderate stem pitting symptoms and its CTV isolates showed alterations in the SSCP profiles. However, the stability of the protective complex has been maintained, except for isolates from two analised clones. Low genetic variability was observed within the isolates during the studied years.

Bond Contribution to Whitetopping Performance on Low Volume Roads, Construction Report, HR-341, 1993

This research was initiated in 1991 as a part of a whitetopping project to study the effectiveness of various techniques to enhance bond strength between a new Portland cement concrete (PCC) overlay and an existing asphalt cement concrete (ACC) pavement surface. A 1,676 m (5,500 ft) section of county road R16 in Dallas County, Iowa was divided into 12 test sections. The various techniques used to enhance bond were power brooming, power brooming with air blast, milling, cement and water grout, and emulsion tack coat. As a part of these bonding techniques, two pavement thicknesses were placed; two different concrete proportions were used; and two sections were planed to a uniform cross-slope.

Screening of papaya accessions resistant to Papaya lethal yellowing virus and capacity of Tetranychus urticae to transmit the virus

The objective of this work was to produce a polyclonal antiserum against the coat protein (CP) of Papaya lethal yellowing virus (PLYV) and to determine its specificity and sensibility in the diagnosis of the virus, as well as to evaluate the genetic resistance to PLYV in papaya (Carica papaya) accessions and to investigate the capacity of the two-spotted spider mite Tetranychus urticae to acquire and transmit PLYV to the plants. Sixty-five papaya accessions were evaluated. For each accession, ten plants were mechanically inoculated using PLYV-infected plant extracts, and three plants were mock inoculated with phosphate buffer alone and used as negative controls. Ninety days after inoculation, newly-emerging systemic leaves were collected from the inoculated plants, and viral infection was diagnosed by indirect Elisa, using polyclonal antiserum sensible to the in vitro-expressed PLYV CP. Viral transmission by T. urticae was evaluated in greenhouse. The experiments were repeated twice. Polyclonal antiserum recognized the recombinant PLYV CP specifically and discriminated PLYV infection from infections caused by other plant viruses. Out of the 65 papaya accessions evaluated, 15 were considered resistant, 18 moderately resistant, and 32 susceptible. The two-spotted spider mite T. urticae was capable of acquiring PLYV, but not of transmitting it to papaya.