Identification, content, and distribution of type VI collagen in bovine tendons

Tendon composition changes according to differentiation, mechanical load, and aging. In this study, we attempted to identify, localize, and quantify type VI collagen in bovine tendons. Type VI collagen was identified by the electrophoretic behavior of the alpha chains and Western blotting, and by rotary shadowing. Type VI collagen was extracted from powdered tendon with three sequential 24-h extractions with 4 M guanidine-HCl. The amount of type VI collagen was determined by enzyme-linked immunosorbent assay for purely tensional areas and for the compressive fibrocartilage regions of the deep flexor tendon of the digits, for the corresponding fetal and calf tendons, and for the extensor digital tendon. The distal fibrocartilaginous region of the adult tendon was richer in type VI collagen than the tensional area, reaching as much as 3.3 mg/g (0.33%) of the wet weight. Calf tendons showed an accumulation of type VI at the fibrocartilage site. Immunocytochemistry demonstrated that type VI collagen was evenly distributed in the tensional areas of tendons but was highly concentrated around the fibrochondrocytes in the fibrocartilages. The results demonstrate that tendons are variable with regard to the presence and distribution of type VI collagen. The early accumulation of type VI collagen in the region of calf tendon that will become fibrocartilage in the adult suggests that it is a good marker of fibrocartilage differentiation. Furthermore, the distribution of type VI collagen in tendon fibrocartilage indicates that it organizes the pericellular environment and may represent a survival factor for these cells.

Identification and Analyses of Candidate Genes for Rpp4-Mediated Resistance to Asian Soybean Rust in Soybean

Asian soybean rust is a formidable threat to soybean (Glycine max) production in many areas of the world, including the United States. Only five sources of resistance have been identified (Resistance to Phakopsora pachyrhizi1 [Rpp1], Rpp2, Rpp3, Rpp4, and Rpp5). Rpp4 was previously identified in the resistant genotype PI459025B and mapped within 2 centimorgans of Satt288 on soybean chromosome 18 (linkage group G). Using simple sequence repeat markers, we developed a bacterial artificial chromosome contig for the Rpp4 locus in the susceptible cv Williams82 (Wm82). Sequencing within this region identified three Rpp4 candidate disease resistance genes (Rpp4C1-Rpp4C3 [Wm82]) with greatest similarity to the lettuce (Lactuca sativa) RGC2 family of coiled coil-nucleotide binding site-leucine rich repeat disease resistance genes. Constructs containing regions of the Wm82 Rpp4 candidate genes were used for virus-induced gene silencing experiments to silence resistance in PI459025B, confirming that orthologous genes confer resistance. Using primers developed from conserved sequences in the Wm82 Rpp4 candidate genes, we identified five Rpp4 candidate genes (Rpp4C1-Rpp4C5 [PI459025B]) from the resistant genotype. Additional markers developed from the Wm82 Rpp4 bacterial artificial chromosome contig further defined the region containing Rpp4 and eliminated Rpp4C1 (PI459025B) and Rpp4C3 (PI459025B) as candidate genes. Sequencing of reverse transcription-polymerase chain reaction products revealed that Rpp4C4 (PI459025B) was highly expressed in the resistant genotype, while expression of the other candidate genes was nearly undetectable. These data support Rpp4C4 (PI459025B) as the single candidate gene for Rpp4-mediated resistance to Asian soybean rust.

Influence of HLA-DRB-1 alleles on the production of antibody against CSP, MSP-1, AMA-1, and DBP in Brazilian individuals naturally infected with Plasmodium vivax

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

Variation of genetic expression during development, revealed by esterase patterns in Aedes aegypti (Diptera, Culicidae)

Polyacrylamide gel electrophoresis was used to analyze esterase patterns during development of Aedes aegypti from the cities of Marília and São José do Rio Preto (SJRP), Brazil. The zymograms showed a total of 23 esterase bands, 22 of which were in the specimens from Marília and 19 in those from SJRP. These esterase bands were considered to be the product of 23 alleles distributed tentatively in eight genetic loci. Most of the alleles were developmentally regulated. The larval stage expressed the greatest number of them (19 alleles, from the eight loci, in Marília; and 17 alleles, from seven loci, in SJRP). The pupal stage expressed 10 alleles from seven loci, in both populations, and the adult stage expressed 8 alleles from five and six loci in SJRP and Marília, respectively. Some alleles that were active in every stage were developmentally controlled at the level of expression (amount of product). A single allele was constitutively and highly expressed, in larvae, pupae, and adults, in both populations. Differences in esterase synthesis among stages are probably due to regulatory mechanisms acting in agreement with the requirements of a variable number of processes in which esterases are involved. The larval stage is the most active in developmental processes and shows very intense intake of food and very high mobility. These features may demand increased esterase production at that stage. Comparison of the two populations examined showed (besides the existence of alleles that they do not share) that they exhibit differences in the control of expression of other alleles. Such findings may reflect genetic differences between founders in each population, but the possibility of involvement of the intensive use of insecticides in SJRP is also discussed.

Restoration of a degraded area using soil tillage systems in Campinas, São Paulo, Brazil

In Brazil the intensive agriculture use, mainly pasture, is the main cause of the presence of extensive areas of degraded lands. This study aimed to assess the impact of different soil management practices in a pasture degraded area used as garbage disposal. The experiment was performed at the Faculdade de Engenharia Agrícola, Universidade Estadual de Campinas - UNICAMP, in Campinas, state of São Paulo, Brazil, from 1990 to 1996. This area has undergone a process of recovery through removal of trash deposited on the surface, in 1985, levelling of soil, followed by application of limestone, subsoiling, planting of legumes (Crotalaria juncea) and crop rotation (soybean and maize). Since 1990 only popcorn maize was grown and established plots managed with different soil tillage systems, including harrow, chisel plow, moldboard plow, no tillage, disk plow and revolving hoe. One plot was planted exclusively with guinea grass (Panicum maximum) to serve as a reference for minimum loss of soil and another grown on a downhill direction to correspond to the expected maximum erosion. There were differences in sediment loss, nutrient loss and productivity of the popcorn maize in the period analyzed. The chisel plow and no tillage treatments caused the slightest loss of soil and nutrients, compared to other tillage systems. The results show that the soil management systems influenced the physical and chemical characteristics of soil, allowing an economical and environmental recovery of the area, providing the conditions for grain agricultural production.

Delineation of specific management areas for coffee cultivation based on the soil-relief relationship and numerical classification

Predicting and mapping productivity areas allows crop producers to improve their planning of agricultural activities. The primary aims of this work were the identification and mapping of specific management areas allowing coffee bean quality to be predicted from soil attributes and their relationships to relief. The study area was located in the Southeast of the Minas Gerais state, Brazil. A grid containing a total of 145 uniformly spaced nodes 50 m apart was established over an area of 31. 7 ha from which samples were collected at depths of 0. 00-0. 20 m in order to determine physical and chemical attributes of the soil. These data were analysed in conjunction with plant attributes including production, proportion of beans retained by different sieves and drink quality. The results of principal component analysis (PCA) in combination with geostatistical data showed the attributes clay content and available iron to be the best choices for identifying four crop production environments. Environment A, which exhibited high clay and available iron contents, and low pH and base saturation, was that providing the highest yield (30. 4l ha-1) and best coffee beverage quality (61 sacks ha-1). Based on the results, we believe that multivariate analysis, geostatistics and the soil-relief relationships contained in the digital elevation model (DEM) can be effectively used in combination for the hybrid mapping of areas of varying suitability for coffee production. © 2012 Springer Science+Business Media New York.

Observation of a diffractive contribution to dijet production in proton-proton collisions at √s=7 TeV

The cross section for dijet production in proton-proton collisions at √s=7 TeV is presented as a function of a variable that approximates the fractional momentum loss of the scattered proton in single-diffractive events. The analysis is based on an integrated luminosity of 2.7 nb-1 collected with the CMS detector at the LHC at low instantaneous luminosities, and uses events with jet transverse momentum of at least 20 GeV. The dijet cross section results are compared to the predictions of diffractive and nondiffractive models. The low- data show a significant contribution from diffractive dijet production, observed for the first time at the LHC. The associated rapidity gap survival probability is estimated. © 2013 CERN. Published by the American Physical Society.

Performance of laying hens and economic viability of different climatization systems

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

Reinforcing production cooperation and dialogue spaces: the role of SMEs

Foreword by Alicia Bárcena and Jorge Valdez

Contributions of the old urban homegardens for food production and consumption in Rio Claro, Southeastern Brazil

Urban homegardens are green areas of households within the city limits and they have the potential to provide families with a cheap alternative for diet improvement, and to complement the income of the families who sell cultivated products. This research analyzes the contributions of old urban homegardens on food consumption and household economy. Data related to homegardens composition were collected by interviews and by collecting cultivated plants. Diets were assessed through a retrospective method (last 24 hours food recall) and administered every two months, during a year, to include seasonal variations. The diet of the sampled population was found to be dependent on certain foods, indicating a narrow food niche (Levins index = 25.9; Levins standardized index = 0.23). Variations in interviewees' diet are related to the consumption of fruits and vegetables, which are partly supplied by homegardens. Spices and teas consumed were obtained from homegardens, revealing its importance in food consumption and health. Among the 98 species found in homegardens, only 38% appeared in the interviewees' diet, indicating an under-exploitation of these homegardens. Our study found that the main role of homegardens is to supply variation in the diet, contributing to the consumption of different types of products.

Investigation of the genetic diversity of domestic Capra hircus breeds reared within an early goat domestication area in Iran

Background: Iran is an area of particular interest for investigating goat diversity. Archaeological remains indicate early goat domestication (about 10 000 years ago) in the Iranian Zagros Mountains as well as in the high Euphrates valley and southeastern Anatolia. In addition, mitochondrial DNA data of domestic goats and wild ancestors (C. aegagrusor bezoar) suggest a pre-domestication management of wild populations in southern Zagros and central Iranian Plateau. In this study genetic diversity was assessed in seven Iranian native goat breeds, namely Markhoz, Najdi, Taleshi, Khalkhali, Naini, native Abadeh and Turki-Ghashghaei. A total of 317 animals were characterized using 14 microsatellite loci. Two Pakistani goat populations, Pahari and Teddy, were genotyped for comparison.Results: Iranian goats possess a remarkable genetic diversity (average expected heterozygosity of 0.671 across loci, 10.7 alleles per locus) mainly accounted for by the within-breed component (G(ST) = 5.9%). Positive and highly significant F-IS values in the Naini, Turki-Ghashghaei, Abadeh and Markhoz breeds indicate some level of inbreeding in these populations. Multivariate analyses cluster Iranian goats into northern, central and western groups, with the western breeds relatively distinct from the others. Pakistani breeds show some relationship with Iranian populations, even if their position is not consistent across analyses. Gene flow was higher within regions (west, north, central) compared to between regions but particularly low between the western and the other two regions, probably due to the isolating topography of the Zagros mountain range. The Turki-Ghashghaei, Najdi and Abadeh breeds are reared in geographic areas where mtDNA provided evidence of early domestication. These breeds are highly variable, located on basal short branches in the neighbor-joining tree, close to the origin of the principal component analysis plot and, although highly admixed, they are quite distinct from those reared on the western side of the Zagros mountain range.Conclusions: These observations call for further investigation of the nuclear DNA diversity of these breeds within a much wider geographic context to confirm or re-discuss the current hypothesis (based on maternal lineage data) of an almost exclusive contribution of the eastern Anatolian bezoar to the domestic goat gene pool.

The contribution of multiple use forest management to small farmers' annual incomes in the Eastern Amazon

Small-scale farmers in the Brazilian Amazon collectively hold tenure over more than 12 million ha of permanent forest reserves, as required by the Forest Code. The trade-off between forest conservation and other land uses entails opportunity costs for them and for the country, which have not been sufficiently studied. We assessed the potential income generated by multiple use forest management for farmers and compared it to the income potentially derived from six other agricultural land uses. Income from the forest was from (i) logging, carried out by a logging company in partnership with farmers' associations; and (ii) harvesting the seeds of Carapa guianensis (local name andiroba) for the production of oil. We then compared the income generated by multiple-use forest management with the income from different types of agrarian systems. According to our calculations in this study, the mean annual economic benefits from multiple forest use are the same as the least productive agrarian system, but only 25% of the annual income generated by the most productive system. Although the income generated by logging may be considered low when calculated on an annual basis and compared to incomes generated by agriculture, the one-time payment after logging is significant (US$5,800 to US$33,508) and could be used to implement more intensive and productive cropping systems such as planting black pepper. The income from forest management could also be used to establish permanent fields in deforested areas for highly productive annual crops using conservation agriculture techniques. These techniques are alternatives to the traditional land use based on periodic clearing of the forest. Nevertheless, the shift in current practices towards adoption of more sustainable conservation agriculture techniques will also require the technical and legal support of the State to help small farmers apply these alternatives, which aim to integrate forest management in sustainable agricultural production systems.

Trends in Brazilian academic production in the History of Mathematics Education: directions from EBRAPEM

The number of papers on History of Mathematics Education presented at EBRAPEM (Brazilian Meeting of Graduate Students in Mathematics Education) has increased significantly between 2003 and 2008. This article presents a study with the aim of identifying themes, periods in focus, and sources and theoretical and methodological references used by the authors of the papers on History of Mathematics Education published in the proceedings of VII, VIII, IX, X, XI and XII EBRAPEM. The study indicates that the approach of ongoing research in History of Mathematics Education in Brazil has been similar to the approach of research in History of Education in general. However, the institutional separation between these two areas of investigation is noted as a factor rendering communication between both groups of researchers difficult.

Mechanical strain induces the production of spheroid mineralized microparticles in the aortic valve through a RhoA/ROCK-dependent mechanism

Calcific aortic valve disease (CAVD) is a chronic disorder characterized by an abnormal mineralization of the leaflets, which is accelerated in bicuspid aortic valve (BAV). It is suspected that mechanical strain may promote/enhance mineralization of the aortic valve. However, the effect of mechanical strain and the involved pathways during mineralization of the aortic valve remains largely unknown. Valve interstitial cells (VICs) were isolated and studied under strain conditions. Human bicuspid aortic valves were examined as a model relevant to increase mechanical strain. Cyclic strain increased mineralization of VICs by several-fold. Scanning electron microscope (SEM) and energy dispersive X-ray (EDX) analyses revealed that mechanical strain promoted the formation of mineralized spheroid microparticles, which coalesced into larger structure at the surface of apoptotic VICs. Apoptosis and mineralization were closely associated with expression of ENPP1. Inhibition of ENPP1 greatly reduced mineralization of VIC cultures. Through several lines of evidence we showed that mechanical strain promoted the export of ENPP1-containing vesicles to the plasma membrane through a RhoA/ROCK pathway. Studies conducted in human BAV revealed the presence of spheroid mineralized structures along with the expression of ENPP1 in areas of high mechanical strain. Mechanical strain promotes the production and accumulation of spheroid mineralized microparticles by VICs, which may represent one important underlying mechanism involved in aortic valve mineralization. RhoA/ROCK-mediated export of ENPP1 to the plasma membrane promotes strain-induced mineralization of VICs.

Production of active recombinant eIF5A: reconstitution in E. coli of eukaryotic hypusine modification of eIF5A by its coexpression with modifying enzymes

Eukaryotic translation initiation factor 5A (eIF5A) is the only cellular protein that contains the polyamine-modified lysine, hypusine [Nε-(4-amino-2-hydroxybutyl)lysine]. Hypusine occurs only in eukaryotes and certain archaea, but not in eubacteria. It is formed post-translationally by two consecutive enzymatic reactions catalyzed by deoxyhypusine synthase (DHS) and deoxyhypusine hydroxylase (DOHH). Hypusine modification is essential for the activity of eIF5A and for eukaryotic cell proliferation. eIF5A binds to the ribosome and stimulates translation in a hypusine-dependent manner, but its mode of action in translation is not well understood. Since quantities of highly pure hypusine-modified eIF5A is desired for structural studies as well as for determination of its binding sites on the ribosome, we have used a polycistronic vector, pST39, to express eIF5A alone, or to co-express human eIF5A-1 with DHS or with both DHS and DOHH in Escherichia coli cells, to engineer recombinant proteins, unmodified eIF5A, deoxyhypusine- or hypusine-modified eIF5A. We have accomplished production of three different forms of recombinant eIF5A in high quantity and purity. The recombinant hypusine-modified eIF5A was as active in methionyl-puromycin synthesis as the native, eIF5A (hypusine form) purified from mammalian tissue. The recombinant eIF5A proteins will be useful tools in future structure/function and the mechanism studies in translation.