Severity of tegumentary leishmaniasis is not exclusively associated with Leishmania RNA virus 1 infection in Brazil

Leishmania RNA virus (LRV) has been shown to be a symbiotic component of Leishmania parasites in South America. Nested retro-transcription polymerase chain reaction was employed to investigate LRV1 presence in leishmaniasis lesions from Brazil. In endemic areas of Rio de Janeiro (RJ), no LRV1 infection was observed even with mucosal involvement. LRV1 was only detected in Leishmania (V.) guyanensis cutaneous lesions from the northern region, which were obtained from patients presenting with disease reactivation after clinical cure of their primary lesions. Our results indicated that the severity of leishmaniasis in some areas of RJ, where Leishmania (V.) brazi-liensis is the primary etiological agent, was not associated with Leishmania LRV1 infection.

Quantification of C4d deposition and hepatitis C virus RNA in tissue in cases of graft rejection and hepatitis C recurrence after liver transplantation

Histology is the gold standard for diagnosing acute rejection and hepatitis C recurrence after liver transplantation. However, differential diagnosis between the two can be difficult. We evaluated the role of C4d staining and quantification of hepatitis C virus (HCV) RNA levels in liver tissue. This was a retrospective study of 98 liver biopsy samples divided into four groups by histological diagnosis: acute rejection in patients undergoing liver transplant for hepatitis C (RejHCV+), HCV recurrence in patients undergoing liver transplant for hepatitis C (HCVTx+), acute rejection in patients undergoing liver transplant for reasons other than hepatitis C and chronic hepatitis C not transplanted (HCVTx-). All samples were submitted for immunohistochemical staining for C4d and HCV RNA quantification. Immunoexpression of C4d was observed in the portal vessels and was highest in the HCVTx- group. There was no difference in C4d expression between the RejHCV+ and HCVTx+ groups. However, tissue HCV RNA levels were higher in the HCVTx+ group samples than in the RejHCV+ group samples. Additionally, there was a significant correlation between tissue and serum levels of HCV RNA. The quantification of HCV RNA in liver tissue might prove to be an efficient diagnostic test for the recurrence of HCV infection.

Cholesterol oxidase interference on the emergence and viability of cotton boll weevil larvae

The aim of this work was to evaluate the influence of the enzyme cholesterol oxidase (Coase) on emergence and viability of larvae of the cotton boll weevil (Anthonomus grandis Boheman, 1843). A series of bioassays was performed with eggs and neonate larvae exposed to different enzyme concentrations in artificial diet. Larval survival was affected at all enzyme concentrations tested, and the six-day LD50 was 53 mug/mL (CI 95%: 43-59). Coase also interfered with hatching of larvae after eggs were floated for 15 min in Coase solution at different concentrations. Observations at the light and electronic microscopic level of midguts from larvae fed on artificial diet containing 53 mug/mL of Coase and collected at six days revealed highly vacuolated regions in the epithelial cells as well as partial degradation of the basal membrane and microvilli.

Interference of genotypes x environments interaction in the genetic control of resistance to Asian rust soybean

The objectives of this work were to identify parents resistant to Asian soybean rust using diallel crosses, obtain information on the genetic control of soybean resistance to the pathogen and verify whether the combining ability estimates interact with the environment (year or time of assessment). The F1 generation was obtained in a greenhouse from crosses between five contrasting parents for the trait resistance to soybean rust, in a complete diallel without reciprocals. Two rust-severity assessments were carried out on individual soybean plants of 25 treatments (parents and F2 and F3 populations) in 2006/2007 and 2007/2008, in an experimental field at Embrapa Soja, Londrina, PR, Brazil. Additive effects predominated in the genetic control of soybean resistance to Asian rust, and the interaction of the segregant populations with the environment, although significant, did not alter the genetic parameter's general combining ability (GCA) and specific combining ability estimates, indicating that estimates obtained in one year and one assessment can be extrapolated to others. BR01-18437 inbred line is resistant to Asian rust and showed high GCA effects. This line should be used as parent if the objective is the resistance to Phakopsora pachyrhizi.

COMPARISON OF RNA EXTRACTION METHODS FOR Passiflora edulis SIMS LEAVES

ABSTRACT Functional genomic analyses require intact RNA; however, Passiflora edulis leaves are rich in secondary metabolites that interfere with RNA extraction primarily by promoting oxidative processes and by precipitating with nucleic acids. This study aimed to analyse three RNA extraction methods, Concert™ Plant RNA Reagent (Invitrogen, Carlsbad, CA, USA), TRIzol® Reagent (Invitrogen) and TRIzol® Reagent (Invitrogen)/ice -commercial products specifically designed to extract RNA, and to determine which method is the most effective for extracting RNA from the leaves of passion fruit plants. In contrast to the RNA extracted using the other 2 methods, the RNA extracted using TRIzol® Reagent (Invitrogen) did not have acceptable A260/A280 and A260/A230 ratios and did not have ideal concentrations. Agarose gel electrophoresis showed a strong DNA band for all of the Concert™ method extractions but not for the TRIzol® and TRIzol®/ice methods. The TRIzol® method resulted in smears during electrophoresis. Due to its low levels of DNA contamination, ideal A260/A280 and A260/A230 ratios and superior sample integrity, RNA from the TRIzol®/ice method was used for reverse transcription-polymerase chain reaction (RT-PCR), and the resulting amplicons were highly similar. We conclude that TRIzol®/ice is the preferred method for RNA extraction for P. edulis leaves.

Transgenic passionfruit expressing RNA derived from Cowpea aphid-borne mosaic virus is resistant to passionfruit woodiness disease

Sixteen transgenic yellow passionfruit (Passiflora spp.) plants (R0) were obtained which express a non-translatable transgenic RNA corresponding to the 3' region of the NIb gene and the 5' region of the CP gene, derived from the genome of a Brazilian isolate of Cowpea aphid-borne mosaic virus (CABMV). The transgenic plants were propagated by stem cuttings and challenged by sap inoculation with isolates CABMV-MG1 and CABMV-PE1. One transgenic plant (TE5-10) was resistant to the isolate CABMV-MG1, but susceptible to CABMV-PE1. The remaining transgenic plants developed systemic symptoms, equal to non-transformed plants, when inoculated with either isolate. The absence of virus in TE5-10 plants was confirmed by indirect ELISA. Transcription analysis of the transgene demonstrated that the TE5-10 plant did not accumulate transgenic mRNA, even before inoculation. After inoculation, viral RNA was only detected in plants inoculated with CABMV-PE1. These results confirm that the transgenic plant TE5-10 is resistant to isolate CABMV-MG1, and suggest that the resistance mechanism is post-transcriptional gene silencing, which is already activated in the transgenic plants before virus inoculation.

Viróides e virusóides: relíquias do mundo de RNA

Até meados do século XX, os vírus eram considerados os representantes mais simples da escala biológica. A descoberta dos RNAs satélites e dos viróides por volta de 1970 foi surpreendente, pois comprovou-se a existência de uma nova classe de moléculas auto-replicativas ainda mais simples, denominada agentes sub-virais. Há indícios de que os viróides e virusóides (que formam uma classe de RNAs satélites), teriam feito parte do "Mundo de RNA" (que precedeu o mundo atual baseado no DNA e proteínas), podendo ser considerados fósseis moleculares dessa era antiga. A simplicidade desses agentes sub-virais e o fato de que a molécula de RNA deve interagir diretamente com fatores do hospedeiro para o desenvolvimento do seu ciclo infeccioso colocam esses patógenos como um modelo para o estudo de processos metabólicos celulares. Nos últimos anos, tem-se observado um volume grande de publicações visando elucidar aspectos da interação viróide/hospedeiro, como os mecanismos da patogênese, movimento dos viróides nas plantas hospedeiras, silenciamento gênico e atividades das ribozimas. Mudanças recentes ocorridas na taxonomia desses patógenos com a criação de famílias, gêneros e espécies, além da descoberta de novos viróides, também têm sido verificadas. A presente revisão visa atualizar o leitor quanto aos recentes avanços nas pesquisas com viróides, principalmente na taxonomia, filogenia e em vários aspectos moleculares da interação viróide/hospedeiro. Estão incluídas também algumas características dos virusóides e sua relação evolutiva com os viróides.

Interference of age and repetition of the same noxious stimulus on hyperalgesia

Pain in animals has been recognized for less than one century. Several authors confirm that animals are capable to process, register and modulate nociceptive stimuli in a very similar way to human kind and there are several evidences registering the impact of pain sensation over vital systems interfering on disease outcome. Nevertheless, despite some evidences that animals, as human beings, can store information from past painful experiences less is known about how this so called pain memory works. The aims of this study were: to evaluate if the response to a painful stimuli differs during different stages of life and if repetition of a same acute stimuli in the same animal interferes with expression of hyperalgesia. Thus, 60 rats were selected and arranged in 3 equal groups: 3 months, 6 months, and 9 months of age. All animals were injected 5% formalin solution in the plantar face of hind paw under volatile general anesthesia. Von Frey filaments were applied at 1h, 24h and 48h after sensitization. Injection was repeated twice with a 30-day interval, each time in a different hind paw. Results showed that younger rats express lower hyperalgesia thresholds in the first stimulation compared to elder animals and that repetition of same stimulus diminishes hyperalgesia thresholds when it begins during infant period and augments hyperalgesia thresholds when it begins during elder ages.

Detection of rabies virus nucleoprotein-RNA in several organs outside the Central Nervous System in naturally-infected vampire bats

Rabies is a neurological disease, but the rabies virus spread to several organs outside the central nervous system (CNS). The rabies virus antigen or RNA has been identified from the salivary glands, the lungs, the kidneys, the heart and the liver. This work aimed to identify the presence of the rabies virus in non-neuronal organs from naturally-infected vampire bats and to study the rabies virus in the salivary glands of healthy vampire bats. Out of the five bats that were positive for rabies in the CNS, by fluorescent antibody test (FAT), viral isolation in N2A cells and reverse transcription - polymerase chain reaction (RT-PCR), 100% (5/5) were positive for rabies in samples of the tongue and the heart, 80% (4/5) in the kidneys, 40% (2/5) in samples of the salivary glands and the lungs, and 20% (1/5) in the liver by RT-PCR test. All the nine bats that were negative for rabies in the CNS, by FAT, viral isolation and RT-PCR were negative for rabies in the salivary glands by RT-PCR test. Possible consequences for rabies epidemiology and pathogenesis are discussed in this work.

Initialism as a mechanism of weed interference: can a crop plant be blinded?

Initialism is a new word proposed to indicate the "shade-avoidance syndrome". Plants detect the presence of neighbor plants very early in the growing season through changes in light quality. They modify the allocation of photosynthesis products privileging shoot growth over the roots. One of the hypotheses of the authors is that, when weed management is timely scheduled, a "blind" crop could be more productive because it would avoid an imbalance on the shoot:root ratio (S:R). Two strategies were developed to test this hypothesis: a) to use the classical Yoda's Law to screen several crops for insensitivity to S:R imbalance; b) to evaluate several growth regulators to control the plant responses to crowding. Experimental results confirm that both strategies can yield insensitive plants. The possibilities of the use of this knowledge are discussed.

Modeling of weeds interference periods in bean

The research objective was to determine the effects of spacing and seeding density of common bean to the period prior to weed interference (PPI) and weed period prior to economic loss (WEEPPEL). The treatments consisted of periods of coexistence between culture and the weeds, with 0 to 10, 0 to 20, 0 to 30, 0 to 40, 0 to 50, 0 to 60, 0 to 70, and 0 to 80 days and a control maintained without weeds. In addition to the periods of coexistence, there were still studies with an inter-row of 0.45 and 0.60 m, 10 and 15 plants m-1. The experimental delineation used was randomized blocks with four repetitions per treatment. The grain productivity of the culture had a reduction of 63, 50, 42 and 57% when the coexistence with the weed plants was during the entire cycle of the culture for a row spacing of 0.45 m and a seeding density of 10 and 15 plants per meter; and a row spacing of 0.60m and a seeding density of 10 and 15 plants per meter, respectively. The PPI occurred in 23, 27, 13, and 19 days after crop emergence and WEEPPEL in 10, 9, 8, and 8 days, respectively.

Effect of weed interference on soybean cultivars at two sowing times

This study aimed to evaluate the effects of weed interference on soybean cultivars at two sowing times in Urutaí, GO. The treatments were arranged in split-plots, and the sowing times (November 16 and December 16, 2009) were allocated in the plots; the soybean cultivars [BRSGO Amaralina, P98C81 (semi-late cycle) BRSGO Raissa, BRSGO Indiara, P98Y11 (median cycle) and BRSGO 7560, BRSGO Caiapônia, Emgopa 302RR (early cycle)] were allocated in the split-plots; and the coexistence or non coexistence of soybean cultivars with weeds, throughout their cycle, was allocated in the split-plots. Non coexistence was established by manual weeding. The experiment was arranged in randomized blocks with four replications. It was verified that the optimal time for sowing soybeans was the month of November, and that under these conditions, the cultivars had higher competitive ability against weeds. Late sowing affected the cycle, development, and yield of the soybean cultivars; this effect was greater under the influence of the weed community.

Weed interference periods in the 'Fécula Branca' cassava

This study aimed to determine the periods of weed interference in the first cycle of 'Fécula Branca' cassava. The experiment was arranged in a randomized block design, with four repetitions. The treatments consisted of eight periods of weed control (25, 50, 75, 100, 125, 150, and 175 days after planting - DAP) and eight periods of coexistence between the weed community and the crop (25, 50, 75, 100, 125, 150, and 175), besides control without weeds and control with weeds until harvest (322 DAP). The predominant weed species with higher relative importance were: Avena sativa, Sorghum halepense, Conyza Canadensis, Euphorbia heterophylla, Raphanus raphanistrum, and Commelina benghalensis. It was concluded that, accepting losses of 1% for root and starch production, the period before interference (PBI) was 66 and 70 DAP; the total period of interference prevention (TPIP) was 88 and 91 DAP and the critical period of interference (CPI) was between 66-88 and 70-91 DAP, respectively. For losses of 5% for root and starch production, the PBI was 87 and 88 DAP, and the TPIP 80 and 81 DAP, respectively; in this case, there was no CPI. Considering the average prices of R$ 218.90 t-1 and R$ 1,191.84 t-1, paid in 2012 for root and starch production, respectively, and control cost of R$ 300.00 ha-1 , the root and starch production for the period prior to economic loss (WEEPPEL) could be estimated to be 20 and 24 DAP, respectively.

Interference of Sorghum sudanense and Eleucine indica in the soybean and corn cultivation

The natural infestations are composed of numerous species that compete for environmental resources such as water, light, nutrients and space. The objective of this study was to evaluate the interference of mixed infestations Sorghum sudanense (sudangrass) and Eleusine indica (goosegrass) in the presence of soybean and corn. The experimental design was completely randomized with four replications and the experimental units consisted of plastic pots with a volume capacity of 8 L. The treatments were associations of plants S. sudanense and E. indica in the proportions 8:0, 6:2, 4:4,2:6 and 0:8, respectively, corresponding to 100, 75, 50, 25 and 0% S. sudanense and the reverse for E. indica. In all treatments remained constant four soybean or corn plants per experimental unit. The variables analyzed in the weeds were shoot dry weight, root, total and height of plants. The competitive analysis was accomplished through diagrams applied to replacement series experiment and indexes of competiveness. The results indicated that E. indica was more competitive than S. sudanense in mixed infestations with corn. Rather, S. sudanense was more competitive than E. indica, in mixed infestations with soybean, demonstrating differences in competitiveness among the weeds.

Weed interference on growth and yield of transgenic maize

Plants kept under competition tend to modify their morphology to optimize the use of production factors. This study aimed to evaluate the effects of competition between transgenic maize and five weed species on the growth and yield of transgenic maize hybrid. The experiment used a randomized block design with four replicates in a factorial 5 x 2 + 6 scheme consisting of a combination of maize under competition with five weed species (Bidens pilosa, Commelina benghalensis, Brachiaria brizantha, Sorghum arundinaceum and Ipomoea triloba) in two weed densities (15 or 30 plants m-2) plus six treatments corresponding to maize and weed species without competition. All the means for dry matter accumulated by maize plants in the stem and leaf in the density of 15 plants m ² were higher than the means for plants in coexistence with 30 plants m-². Number of kernels, diameter and length of cob were not affected by competition with weeds. The weeds that most interfered with maize biomass production were S.arundinaceum and B.brizantha. Leaf dry mass accumulation was more sensitive than the production of stem. It was observed that maize was usually very competitive with weeds, and there was a strong decrease in dry matter accumulation of all the weeds in the study when in coexistence with the crop.