Miracle-dz/UniFAR-FTUs huggingface dataset:
IDENTIFICAÇÃO, PELA TÉCNICA DE PCR-RFLP, DE Aspergillus spp. ISOLADOS DE GRÃOS DE SOJA E MILHO.

A agricultura e um dos fatores mais rentaveis no Brasil, alem de ser uma das maneiras de sustento das familias rurais e urbanas. As culturas mais cultivadas no Brasil sao soja, milho e arroz. Fungos do genero Aspergillus podem contaminar graos de soja e milho durante seu armazenamento e pode produzir micotoxinas, como as aflatoxinas, prejudiciais a saude de quem os consome. O objetivo desse trabalho foi identificar, atraves de tecnicas moleculares, fungos da especie Aspergillus flavus e estabelecer uma correlacao do tempo de armazenamento dos graos com a contaminacao fungica. Foram coletadas sementes de milho e soja em diferentes silos da regiao de Maringa-PR, e posteriormente realizada cultura para fungos em meio de cultivo especifico. Apos o isolamento e caracterizacao dos fungos em meio de cultura, foi realizada a extracao do DNA genomico e amplificacao, pela Reacao em Cadeia da Polimerase (PCR), das regioes IGS e β-tubulina. Os produtos de amplificacao da regiao IGS foram submetidos a clivagem com a enzima de restricao BgI II, para a confirmacao da especie, pela tecnica de PCR-RFLP. Das amostras coletadas 68,75% estavam contaminadas por Aspergillus spp. , seguindo analises microbiologicas. Destas, 31,25% resultou positivo para a especie Aspergillus flavus. Os outros 37,5% foram dadas como negativas, por varios fatores, um deles a sensibilidade da tecnica de PCR. Foi possivel concluir que um pouco menos da metade das amostras encontrava-se contaminada por Aspergillus spp .
Detection of Genetically Modified Food Products in a Commercial Laboratory

Since genetically engineered agricultural plants entered the market, new challenges have been imposed on grain producers, food processors, and food testing laboratories. Service laboratories, especially in the United States, face a lack of standardization of testing methods for bioengineered crops. Food companies need a basis on which to choose a laboratory that can meet their specific analytical needs withe regard to applicable regulations or commercial contracts by providing scientifically valid and reproducible results. This article outlines the prime technical aspects for laboratories providing analysis of food and agricultural products by PCR and ELISA. La detection et la quantification des OGM necessite de nouvelles techniques de laboratoire, comme la PCR. Les normes et accreditations ne sont pas clairement etablies aux Etats-Unis. Les industries qui utilisent les services de ces laboratoires d'analyse doivent examiner les procedures et capacites des laboratoires offrant ces nouveaux services. Cet article presentent les aspects techniques des analyses par PCR et ELISA des produits agricoles modifies genetiquement.
Isolation and characterization of simple sequence repeat loci in Rubus hochstetterorum and their use in other species from the Rosaceae family

The identification of microsatellite loci in Rubus hochstetterorum provides an important tool for the characterization and conservation of wild populations of this species. Crossspecies amplification of markers may be of particular interest for the study of other Rubus species. In this study, 41 simple sequence repeat markers were identified in a genomic library of R. hochstetterorum . Fifteen of the identified microsatellite loci were characterized in a set of 30 samples and revealed to be polymorphic with three to 19 alleles per locus. All the identified markers allowed cross-species amplification in at least one of the other three tested species from the Rosaceae family.
Powdery Mildew Resistance Genes in Spring Barley Cultivars Registered in Slovakia from 2000 to 2010

Spring barley was the second largest crop in Slovakia. Resistance genes in cultivars registered in Slovakia from 2000 to 2010 were identified using the postulation method. Forty-three cultivars including 30 individually harvested plants, two parents ‘Meltan’ and ‘Pax’ and a standard line SK5968 were tested at the seedling stage for their reaction to 40 selected isolates of Blumeria graminis f. sp. hordei. In total, 19 resistance spectra were detected and the following 13 resistance genes were postulated, namely mlo, Mla1, Mla6, Mla12, Mla13, MlaRu4, Mlat, Mlg, MlLa, Ml(Ab), Ml(Hu4), Ml(Ro) and Ml(St). Two unknown resistances were detected in ‘Calcule’ and ‘Spilka’. Two cultivars (‘Madonna’ and ‘Nadir’) exhibited heterogeneity for mildew resistance. The most frequent gene was mlo, which was present in 25 of the tested cultivars. Some new results to previously published data were found.
Macroinvertebrate variation in endorheic depression wetlands in North West and Mpumalanga provinces, South Africa

Aquatic macroinvertebrates are rarely used in wetland assessments due to their variation. However, in terms of biodiversity, these invertebrates form an important component of wetland fauna. Spatial and temporal variation of macroinvertebrate assemblages in endorheic depressions (locally referred to as pans ) in Mpumalanga and North West provinces were compared in wet and dry seasons in 2012 and 2013. A total of 29 taxa were identified from both provinces, with similarities in the structure of communities, with the exception of one or two species, in perennial endorheic depressions in both provinces. Macroinvertebrates sampled in Mpumalanga endorheic depressions were similar to those reported in previous studies completed in the area, and most macroinvertebrates sampled in Mpumalanga and North West were known to be commonly found in temporary habitats. Long-term studies are required to understand better the ecological functioning of the pans in the North West province.
Using morphometric measurements to estimate age of nestling Red-shouldered Hawks in two eastern populations

Abstract Nestling growth may be used to estimate age of nestling raptors, which is valuable for investigating hatch order dynamics and nestling behavior, as well as assessing reproductive rate and back-calculating hatching date. To estimate nestling age, the most valuable parameter to measure growth is one that does not vary greatly with environmental factors, and ideally is applicable over a wide range of populations. We measured growth of nestling Red-shouldered Hawks (Buteo lineatus) in Quebec, Canada, from ages 3 days to near fledging (38 days old), and compared growth of several parameters in different size broods. As a validation study, we measured similar parameters one time in known-age nestling Red-shouldered Hawks in southwestern Ohio. Growth rates for tarsus length, bill length, and tail length differed between nestlings in broods of one and three young, respectively, in Quebec. However, mass gain and growth of secondary feathers (mean length of first and second secondaries) did not differ between brood sizes, although mass gain was more variable than secondary growth. These results suggested that secondary feather length was the most valuable parameter for estimating nestling age in Red-shouldered Hawks. Comparing Ohio nestlings to Quebec nestlings, we found that growth of secondary feathers differed significantly, with Ohio nestlings having smaller secondary length, relative to age estimate. Application of the equation generated with the Quebec data to estimate the age of the Ohio nestlings based on secondary length resulted in estimates that were 2.3 ± 0.3 days (range 0.25–4.5 days; n = 22) younger than the Ohio nestlings' actual ages. Based on this validation study, we suggest that the use of the Quebec age-secondary length equation to estimate age for nestling Red-shouldered Hawks in other parts of eastern North America is acceptable, though with the caveat that such estimates are associated with potential small errors.
Rootstock Influence on the Biochemical Composition and Polyphenol Oxidase Activity of ‘Thompson Seedless’ Grapes and Raisins

‘Thompson Seedless’ grapes grafted on five rootstocks and own-rooted vines were evaluated for yield, fruit composition, and raisin recovery percentage. Biochemical constituents were analyzed in fresh grapes before drying into raisins and also in raisins. Among different rootstocks, raisin recovery was highest in ‘Thompson Seedless’ grapes grafted on 110 R followed by those on ‘Dogridge’, 1103 P and 99 R. Significant differences were observed in total proteins, reducing sugars, phenol contents, and polyphenol oxidase (PPO) activity in both fresh grapes and raisins made from all rootstock scion combinations. Increase in the content of proteins and reducing sugars was observed in raisins compared to fresh grapes. However, there was a reduction in phenolic concentrations in raisins compared to fresh grapes. PPO activity was highest in ‘Thompson Seedless’ grapes grafted on ‘Dogridge’, while it was least in 110 R.
Computational and experimental methods in functional genomics : the good, the bad, and the ugly of systems biology

Dedication To everyone who has ever quietly dreamt of changing his life, but has been hesitant to take the plunge: it's never too late. program and became a best friend and so much more. I hope you're as proud of me as I am of you. Where do these stairs go? v Acknowledgements First, I would like to thank the many members of the Marcotte, Ellington, and Iyer labs, past and present, for showing me how to be both a bench and a computational molecular biologist. An abbreviated list is: Matt Levy, for my first introduction to benchwork; Jonghwan Kim, for teaching basic techniques like PCR (fourteen thousand times) and demonstrating what scientific rigor really means; Ram Swamy, for the immersion in the yeast cell chip; Zhihua Li, for sharing his encyclopedic knowledge of biology and bench techniques; Arun Ramani, for a guided tour of the world of protein interaction networks. Also, the collaborators, without whose time, thought, and energy, none of these projects would have seen the light: everyone above, plus Andy Ellington's unlimited supply of undergraduates and ad hoc resources; the Microarray Core for the tools for cell chip development; and especially Alice Zhao, without whose help the cell chip project would still be dragging on. Finally, I would like to express my profound gratitude to my adviser, Edward Marcotte, for taking a chance on a guy with some curiosity but no skills, training, or knowledge in the life sciences. If you're reading this, it seems to have worked out. Seven years into the postgenomic era, we sit atop a mountain of data whose generation was enabled by gene sequencing. The creation, integration, and analysis of these large scale data sets allow us to move forward toward the complementary goals of determining the individual roles of the thousands of uncharacterized mammalian genes and understanding how they work together to produce a healthy human being – or, perhaps more importantly, how their malfunction results in disease. Collapsing the results of large-scale assays into gene networks provides a useful framework from which we can glean information that advances both of these goals. However, the utility of networks is limited by the quality of the data that goes into them. This study offers seeks to shed some light on the quality and breadth of protein interaction networks, describes a new experimental technique for functional genetic assays in mammalian cell lines, and …
Regulatory networks controlling ANAC092 expression during different stress responses in Arabidopsis thaliana

Senescence is the final developmental stage of a plant leaf, which occurs as the organ reaches the end of its natural lifecycle. In this process, photosynthetic machinery is broken down and recycled as nutrients for use in the rest of the plant. This allows the recovery of the resources invested in the plant leaves as the plant transitions to a reproductive state. While a developmentally controlled process that occurs naturally as the organ ages, leaf senescence can also be induced prematurely by environmental stresses. The onset of senescence facilitates survival of the plant as a whole at the cost of leaf tissue. As such, senescence is controlled by regulatory systems that cross link developmental and stress response signals. One such signalling mechanism is the action of transcription factors. The NAC family transcription factor ANAC092 has been shown to promote the onset and progression of senescence during developmental and stress conditions. Timeseries gene expression data and network inference indicated ANAC092 as being a central hub in the stress response to Botrytis cinerea infection. ANAC092 was shown to be a functional gene during Botrytis cinerea infection, promoting the spread of lesions through the onset of a senescence like process. As such, ANAC092 represents a major cross-link between developmental and pathogenic stress response signals. A number of transcription factors bound to the ANAC092 promoter region in a yeast 1-hybrid experiment, including members of the NAC, MYB, ERF and AtHB transcription factor families. Interestingly, members of the same transcription factor family co-localised to regions of the ANAC092 promoter, suggesting they recognised similar sequences. Gene expression profiles and network inference were used to generate a model of regulation by these transcription factors. It was predicted certain transcription factors would regulate ANAC092 expression during specific conditions. In particular, different members of the NAC family of transcription factors appeared to regulate ANAC092 expression during Botrytis cinerea infection or developmental senescence. These predictions were tested using transgenic Arabidopsis. The NAC transcription factor recognition sequence is known for a limited subset of NAC transcription factors. In an attempt to define the binding motif for a higher proportion of the NACs, a yeast 1-hybrid library of 94 NAC transcription factors was constructed and screened against a range of Arabidopsis promoter regions. Binding motifs were reverse engineered and tested for NAC protein binding.
Analysis of Mycelial Growth Rates and RAPD-PCR Profiles in a Population of Gaeumannomyces graminis var. tritici Originating from Wheat Plants Grown from Fungicide-treated Seed

Linear mycelial growth rates of 70 isolates of Gaeumannomyces graminis var. tritici on agar medium amended or unamended with the fungicide silthiofam were not correlated. Mycelial growth rate was not influenced by the fungicide applied to the seed of the plants from, which the isolates originated. DNA polymorphism determined by randomly amplified polymorphic DNA (RAPD) polymerase chain reaction was used to assess genetic variation among isolates. Thirty RAPD markers generated with five arbitrary 10-mer primers revealed DNA polymorphism suitable for assessing variability in this fungal population. Cluster analysis of RAPD data identified two groups at the 54% similarity level. There was a significant relationship between the presence of 11 markers and sensitivity to silthiofam.
Associations of Plants and Insects in Deciduous Forest

Phytophagous insect larvae (Lepidoptera and Tenthredinoidea) in the leaf-chewing guild were sampled from accessible parts of all the 18 species of common woody plants in a mature upland forest in New York State, in early and in late June. Among the procedures used to describe patterns of associations between plants and insects are principal components analysis and the clus- tering techniques of numerical taxonomy, whereby the faunistic similarity among plant species is described. We introduce a weighting factor for cluster analysis that weights a character (in this case the density of an insect species on a plant) in proportion to its deviation from the norm. During June, both the abundance and variety of insects declined on most plant species, which vary greatly in both these aspects of their fauna. Insect species that are either quite highly specialized or generalized in diet appeared to be prevalent; those with intermediate diet breadth were less so. Clustering plant species by faunal similarity revealed some clusters of taxonomically related species, but the correspondence between faunal similarity and taxonomic affinity is not strong. Few plant species have a highly distinct, specialized fauna. The complexity of distribution of insects over plants indicates that insects are responding to many factors that differentiate plant species.
Type III TGF‐β receptor‐independent signalling of TGF‐β2 via TβRII‐B, an alternatively spliced TGF‐β type II receptor

Transforming growth factor‐β (TGF‐β) signals through membrane‐bound serine/threonine kinase receptors, which upon stimulation phosphorylate Smad proteins and thereby trigger their nuclear translocation and transcriptional activity. Although the three mammalian isoforms of TGF‐β are highly homologous at the level of sequence, analysis of their in vivo function by gene knockouts revealed striking differences, suggesting no significant functional redundancy between TGF‐β1, ‐2 and ‐3. While signal transduction by TGF‐β1 has been well characterized, receptor binding and activation by the TGF‐β2 isoform is less well understood. Here, we show that TβRII‐B, an alternatively spliced variant of the TGF‐β type II receptor, is a TGF‐β2 binding receptor, which mediates signalling via the Smad pathway in the absence of any TGF‐β type III receptor (TβRIII). L6 cells lacking endogenous TβRIII as well as TβRII‐B do not respond to TGF‐β2. Transfection of these cells with TβRII‐B restores TGF‐β2 sensitivity. The expression of TβRII‐B is restricted to cells originating from tissues such as bone where the isoform TGF‐β2 has a predominant role. This reflects the importance of this receptor in TGF‐β isoform‐specific signalling.
Comparative molecular physiology of novel P2X receptors : identification, cloning and functional characterisation

P2X receptors are ATP-gated ion channels with myriad roles in humans and other higher vertebrates. Research over the past decade has described the cloning, pharmacology, and physiological role of this receptor family in a number of invertebrate organisms, as well as in unicellular amoeba and algae. However, questions remain regarding the extant of P2X receptor phylogeny and function in many invertebrates. A greater understanding of invertebrate P2X receptor pharmacology and function may provide insights into structure-function relationships in “higher” homologues, as well as novel roles for this ligand-gated ion channel family. This thesis investigated P2X receptor expression and function within eukaryotic phylogeny, with particular emphasis on invertebrate animals. Homology searching of transcriptomic and genomic datasets identified a number of candidate P2X receptor sequences across a range of phyla. Notably, homologues were bioinformatically identified within orders of the class Insecta, where they had been previously thought to be absent. Homologous sequences were also identified in a multicellular alga of the Charophycea, a class of the Viridiplantae division Charophyta, considered to represent the closest extant taxon to terrestrial plants. Following the identification of a P2X receptor homologue in the cnidarian Hydra vulgaris (AEP) (‘aepP2X’) by bioinformatics approaches, total RNA preparations were used for cDNA synthesis to generate templates for PCR to yield aepP2X clones. These clones encoded proteins that exhibited concentration-dependent ATP-evoked inward currents when expressed heterologously in HEK293 cells (EC50 ca. 120 μM; holding potential of -60 mV). Co-application of the classic P2 receptor antagonists pyridoxalphosphate-6-azophenyl-2', 5'-disulphonic acid (PPADS) and suramin was ineffective, as were phenol red and brilliant blue G (BBG). The synthetic ATP analogues α,β-methylene ATP and β,γ-methylene ATP (1 mM) did not evoke currents at aepP2X. Consistent with most mammalian homologues, aepP2X was found to be a non-selective cation channel with negligible chloride ion permeability. Immunohistochemistry using a custom polyclonal antiserum raised against a C- terminal epitope of aepP2X suggested the expression of this receptor occurs in developing and late-stage nematocysts in whole Hydra polyps. A P2X receptor from the microcrustacean Daphnia pulex (DpuP2XB) was also expressed heterologously in HEK293 cells, and was found to be largely insensitive to extracellular ATP (EC50 > 1 mM), although reduction of total divalent cation concentration of perfusing extracellular solution partially restored ATP sensitivity in DpuP2XB. Whilst a P2X receptor homologue from the placazoan, Trichoplax adhaerens (TadP2XA) did not respond to extracellular ATP in HEK293 cells, a P2X receptor from the lancelet Branchiostoma floridae (BfloP2X) does respond in a concentration-dependent manner to A TP . Currents evoked through BfloP2X displayed rundown with repeated 1 mM ATP applications.
An Immunochemical Study of the Relationship of Human Blood Group Isoantibodies to γ1- and γ2-Globulins

Rabbit antisera were prepared to human γ2-globulin and to specific precipitates containing human isoanti-A and isoanti-B. The difference between the amount of antibody removed by A-anti-A and by dextranantidextran specific precipitate from anti-γ2-globulin sera was found to represent a small amount of additional antibody not detected by agar diffusion nor by the quantitative precipitin method. Antisera to A-anti-A and B-anti-B specific precipitates contain at least 3 antibodies, one of which is to γ1-globulin of S = 20 and another to γ2-globulin of S = 7. The capacity of these sera to give a Coombs test with anti-D and to enhance the saline agglutinating titer of anti-A and anti-B in the Coombs test depends on their content of anti-γ2-globulin.
Effects of tropical rainforest roads on small mammals: inhibition of crossing movements

Along a narrow, unsealed road through rainforest in north-eastern Queensland, movements of small mammals were examined to determine whether the road would inhibit road crossings, thereby causing linear barrier effects. Crossings of a 12- or 20-m-wide road clearing by Melomys cervinipes were severely inhibited, crossing inhibition of Rattus sp. was less severe, while crossings by Uromys caudimaculatus were unaffected. This differential effect was attributed to species differences in size, mobility and behaviour. Baiting on only one side of the road increased crossing rates for all species. During the breeding season, crossings of 20-m clearings by Rattus sp. were almost completely inhibited and were significantly fewer than crossings of 12-m clearings. Clearing width had little effect on crossing rate outside the breeding season. Seasonal dispersal of juvenile and breeding animals appeared to explain this discrepancy in clearing-width effects. Rattus sp. were significantly less likely to cross a road where there was no vegetative cover at the entrance to a road culvert than where there was cover at both culvert entrances. Linear barrier effects for small mammals may be mitigated by narrower road-clearing widths, by replanting of grassy road verges resulting in increased cover at culvert entrances and canopy closure above the road, and by providing more faunal underpasses.
Immunoblotting Analysis of nsLTP1 in Cereal Grains with Antiserum Raised against Recombinant Rice nsLTP1

Nonspecific lipid transfer protein 1 (nsLTP1) is widely distributed in plants and has been recognized as a widely cross-reacting plant pan-allergen. In the present study, a recombinant protein of rice nsLTP1 was prepared with an Escherichia coli expression system, and polyclonal antiserum was raised by immunizing rabbits with this recombinant nsLTP1. The antiserum specifically immunoreacted with rice nsLTP1 and homologous nsLTP1 proteins of other kinds of cereal grains. Moreover, the antiserum was applied to compare nsLTP1 content among rice cultivars and among rice fractions separated by grain-milling. Variation of nsLTP1 contents were observed among rice cultivars, and rice nsLTP1 was proved to be mainly located in the outer layer of rice grains, providing evidence for decreasing nsLTP1 content by grain-milling.