Ocorrência de animais persistentemente infectados pelo vírus da diarréia viral bovina em rebanhos bovinos nos Estados de Minas Gerais e São Paulo

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

Detecção de anticorpos contra o vírus da diarréia viral bovina no soro sangüíneo, no leite individual e no leite de conjunto em tanque de expansão de rebanhos não vacinados

Pela pesquisa de anticorpos contra o vírus da diarréia viral bovina (BVD), utilizando o teste de ELISA indireto, foi estudada a correlação existente entre a proporção de vacas lactantes e a presença de anticorpos no leite de conjunto do tanque de expansão. Para isso foram analisadas amostras de soro sangüíneo e de leite individual de 376 vacas lactantes não vacinadas, provenientes de 10 propriedades localizadas nas regiões Sul do Estado de Minas Gerais e Nordeste do Estado de São Paulo, assim como uma amostra do leite do tanque de expansão de cada rebanho. em todas as propriedades foram encontradas vacas reagentes no soro sangüíneo, cuja freqüência variou de 12,28 a 100,00%. Já a análise do leite individual não revelou animais reagentes em duas propriedades, e nas demais a freqüência variou de 5,26 a 70,83%. Foram detectados anticorpos no leite do tanque de expansão das propriedades cuja proporção de soros sangüíneos reagentes foi igual a ou maior que 82,86%, e cuja proporção de leites individuais reagentes foi igual a ou maior que 32,14%.

Ocorrência da diarréia viral bovina nas regiões sul do Estado de Minas Gerais e nordeste do Estado de São Paulo

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

Experimental infection of Newcastle disease virus in pigeons (Columba livia): Humoral antibody response, contact transmission and viral genome shedding

The aim of this study was to evaluate the humoral antibody response, the genome viral excretion and the contact transmission of pathogenic chicken origin Newcastle disease virus (NDV) from experimentally infected pigeons (Columba livia) to in-contact pigeon. The antibody response to infection was assessed by the hemagglutination inhibition (HI) test and the genome viral excretion was detected by RT-PCR. Viral strain induced high antibody levels, both in inoculated and in sentinel birds. The pathogenic viral strain for chickens was unable to produce clinical signs of the disease in experimentally infected pigeons, although it induced the Immoral antibody response and produced NDV genome shedding. NDV genome was detected intermittently throughout the experimental period, from 5 days post-infection (dpi) to 24 dpi. Therefore, viral genome shedding occurred for 20 days. The viral genome was detected in all birds, between I I and 13 dpi. Furthermore, the high infectivity of the virus was confirmed, as all non-inoculated sentinel pigeons showed antibody levels as high as those of inoculated birds. (C) 2007 Elsevier B.V. All rights reserved.

Algoritmo memético com infecção viral: uma aplicação ao problema do caixeiro viajante assimétrico

The Combinatorial Optimization is a basic area to companies who look for competitive advantages in the diverse productive sectors and the Assimetric Travelling Salesman Problem, which one classifies as one of the most important problems of this area, for being a problem of the NP-hard class and for possessing diverse practical applications, has increased interest of researchers in the development of metaheuristics each more efficient to assist in its resolution, as it is the case of Memetic Algorithms, which is a evolutionary algorithms that it is used of the genetic operation in combination with a local search procedure. This work explores the technique of Viral Infection in one Memetic Algorithms where the infection substitutes the mutation operator for obtaining a fast evolution or extinguishing of species (KANOH et al, 1996) providing a form of acceleration and improvement of the solution . For this it developed four variants of Viral Infection applied in the Memetic Algorithms for resolution of the Assimetric Travelling Salesman Problem where the agent and the virus pass for a symbiosis process which favored the attainment of a hybrid evolutionary algorithms and computational viable

Molecular mapping of the viral determinants of systemic wilting induced by a Lettuce mosaic virus (LMV) isolate in some lettuce cultivars

The isolate AF199 of Lettuce mosaic virus (LMV, genus Potyvirus) causes local lesions followed by systemic wilting and plant death in the lettuce cultivars Ithaca and Vanguard 75. Analysis of the phenotype of virus chimeras revealed that a region within the PI protein coding region (nucleotides 112-386 in the viral genome) and/or another one within the CI protein coding region (nucleoticles 5496-5855) are sufficient together to cause the lethal wilting in Ithaca, but not in Vanguard 75. This indicates that the determinants of this particular symptom are different in these two lettuce cultivars. The wilting phenotype was not directly correlated with differences in the deduced amino acid sequence of these two regions. Furthermore, transient expression of the LMV-AF 199 proteins, separately or in combination, did not induce local necrosis or any other visible reaction in the plants. Together, these results Suggest that the systemic wilting reaction might be Clue to RNA rather than protein sequences. (c) 2004 Elsevier B.V. All rights reserved.

Avaliação preliminar da viabilidade de produção in vitro de um isolado brasileiro de baculovírus Spodoptera frugiperda MNPV

In vivo production of viral biopesticides is the major source of viral insecticides currently in the marketplace. However, this system presents limitations during production scale-up. For the Spodoptera frugiperda nucleopolyhedrovirus (SfMNPV), the insect used for replication has cannibalistic characteristics, thus production is even more difficult. Insect cells are commonly used for in vitro baculovirus production. Most of these cell lines are derived from Lepidoptera species. The Sf21 cell line is derived from Spodoptera frugiperda caterpillar ovarian tissue, and its clonal isolate Sf9 has been used for biopesticide production due to its ease of growth in suspension cultures. In this work, the in vitro production capabilities of a Brazilian SfMNPV isolate obtained from cornfields was evaluated. Comparison of polyhedra production was carried out using both Sf21 and Sf9 cells, based on volumetric and specific yields. Both cell lines were cultivated in Hyclone medium supplemented with different fetal bovine serum concentrations (2,5 and 5%). The best results were obtained using Sf9 cells supplemented with 5% serum. These results were further confirmed quantitively through kinetic parameter estimation for both cells lines and different serum concentrations. After seven successive passages, this system still presented high specific polyhedra production

Antiviral effect of Guazuma ulmifolia and Stryphnodendron adstringens on poliovirus and bovine herpesvirus

Crude extract (CE) and aqueous (AqF) and ethyl acetate (EtOAcF) fractions of Guazuma ulmifolia LAM., Sterculiaceae and the corresponding AqF, EtOAcF of Stryphnodendron adstringens (MART.) COVILLE, Leguminosae were tested for their antiviral activity against poliovirus 1 (P-1) and bovine herpesvirus 1 (BHV-1) in HEp2 cultured cells. The antiviral activity was monitored by plaque assay and immunolluorescence assay (IFA) under virucidal and therapeutic protocols. The therapeutic protocol demonstrated statistically significant positive results with both plants and for both virus strains. The highest percentages of viral inhibition were found for G. ulmifolia EtOAcF which inhibited BHV-1 and P-1 replication by 100% and 99%, respectively (p < 0.05, Student's t-test). For S. adstringens, AqF was the most efficient, inhibiting BHV-1 and P-1 by 97% and 93%, respectively (p < 0.05). In the virucidal protocol, G. ulmifolia CE inhibited the replication of BHV-1 and P-1 by 60% and 26%, respectively (p < 0.05), while, for S. adstringens, inhibition of 62% (p < 0.05) was demonstrated only with EtOAcF for P-1. IFA demonstrated that the greatest reduction in fluorescent cell number occurred with G. ulmifolia, under the therapeutic protocol for both virus strains. However, AqF and EtOAcF of S. adstringens were most efficient with the virucidal protocol for P-1 In conclusion, we demonstrated that G. ulmifolia and S. adstringens inhibited BHV-1 and P-1 replication, as well as, blocked the synthesis of viral antigens in infected cell cultures.

CCR5 genotype and plasma ß-chemokine concentration of Brazilian HIV-infected individuals

The 32-bp deletion in the HIV-1 co-receptor CCR5 confers a high degree of resistance to HIV-1 infection in homozygous individuals for the deleted allele and partial protection against HIV-1 during disease progression in heterozygotes. Natural ligands for CCR5, MIP-1alpha, MIP-1ß and RANTES, have been shown to inhibit HIV replication in CD4+ T cells. In the present study, we examined the CCR5 genotype by PCR and the plasma levels of RANTES and MIP-1alpha by ELISA among blood donors (N = 26) and among HIV-1-infected individuals (N = 129). The control group consisted of healthy adult volunteers and HIV-1-infected subjects were an asymptomatic and heterogeneous group of individuals with regard to immunologic and virologic markers of HIV-1 disease. The frequency of the CCR5 mutant allele (delta32ccr5) in this population was 0.032; however, no delta32ccr5 homozygote was detected. These results could be related to the intense ethnic admixture of the Brazilian population. There was no correlation between circulating ß-chemokines (MIP-1alpha, RANTES) and viral load in HIV-infected individuals. RANTES concentrations in plasma samples from HIV+ patients carrying the homozygous CCR5 allele (CCR5/CCR5) (28.23 ng/ml) were higher than in the control samples (16.07 ng/ml; P<0.05); however, this HIV+ patient group (mean 26.23 pg/ml) had significantly lower concentrations of MIP-1alpha than those observed in control samples (mean 31.20 pg/ml; P<0.05). Both HIV-1-infected and uninfected individuals heterozygous for the delta32ccr5 allele had significantly lower concentrations of circulating RANTES (mean 16.07 and 6.11 ng/ml, respectively) than CCR5/CCR5 individuals (mean 28.23 and 16.07 ng/ml, respectively; P<0.05). These findings suggest that the CCR5 allele and ß-chemokine production may affect the immunopathogenesis of HIV-1.

A Management Tool for the Replication of Operating Systems in Wireless Communication Networks

To simplify computer management, various administration systems based on wired connections adopt advanced techniques to manage software configuration. Nevertheless, the strong relation between hardware and software makes for an individualism of that management, besides penalizing computational mobility and ubiquity. All these issues lead to degradation of scalability, flexibility and the facility to install and maintain distributed applications. This article presents an environment for centralized wireless communication network management, named WSE-OS (Wireless Sharing Environment - Operating Systems): a model based on Virtual Desktop Infrastructure (VDI) which associates virtualization techniques and safe remote access systems to create a distributed architecture as a base for a managing system. WSE-OS is capable of accomplishing the replication of operating system images using wireless communication network, besides offering abstraction of hardware to its clients, making the management more flexible and independent of wired connections. Results obtained from this work indicate that WSE-OS allows disseminating, through a single software configuration, the execution of data related to operating system images in client computers. WSE-OS can also be used as a management tool for operating systems in a wireless network.

Estratégias de Produção in vitro de Bioinseticida Viral: influências do Isolado, da Cinética e do Modo de operação

Societal concerns about environmental sustainability has lead to the development of ecologically-friendly alternatives to chemical insecticides for crop protection. One such alternative is biological pest control. In particular, baculoviruses are well suited as insect biopesticides due to their narrow host specificity and relative ease of propagation. In Brazil, the baculovirus Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) is the main biological control agent employed for the soybean pest, Anticarsia gemmatalis. This baculovirus biopesticide is currently produced using caterpillars, but increasing market demand for the product has encouraged the development of an in vitro manufacturing process, which can be scaled up to much higher virus productivities. In this study, three wild-type AgMNPV isolates (AgMNPV-2D, AgMNPV-MP2 and AgMNPV-MP5) and a recombinant form (vAgEGT-LacZ) were characterised in terms of occlusion body (OB) production and infection kinetics, to enable future optimisation of the in vitro production process. These viruses were propagated using a Spodoptera frugiperda (IPLB-SF21) insect cell line grown in shaker-flask batch cultures. Among the virus isolates tested, AgMNPV-MP5 was found to be the best producer, yielding (5.3±0.85)x108 OB/mL after 8 days post infection. The characterisation of vAgEGT-LacZ propagation in suspension cell cultures has not been previously reported in the literature; hence it became the main focus for this thesis. In particular, it was carried out a study on the effect of the multiplicity of infection (MOI) on OB production. Five successive batches were performed getting a final production (8.9±1.42)x1014 occlusion bodies, considering that production is related for a bioreactor with final volume of 10m3. A low MOI associated with a fed-batch process for vAgEGT-LacZ production was found to support a 3-fold higher OB yield when compared to the default batch process (1.8x107 and 5.3x107 OB/mL, respectively). This yield is competitive with regards to the production process.

The genetics of hypertension modifies the renal cell replication response induced by experimental diabetes

To investigate whether the genetics of hypertension modifies renal cell responses in experimental diabetes, we studied the renal cell replication and its regulation by two cyclin-dependent kinase (Cdk) inhibitors, p27(Kip1) and p21(Cip1), in prehypertensive spontaneously hypertensive rats (SUR) and their genetically normotensive counterparts, Wistar Kyoto (WKY) rats, with and without streptozotocin-induced diabetes. In diabetic SIIR, the number of proliferating glomerular (0.6 +/- 0.3 positive cells/50 glomeruli) and tubulointerstitial (2.8 +/- 0.6 positive tubulointerstitial cells/50 grid fields) cells assessed by the bromodeoxyuridine technique was significantly (P = 0.0002) lower than in control SIIR (13.2 +/- 1.7 and 48.6 +/- 9.7, respectively) and control (14.0 +/- 1.8 and 63.9 +/- 10.6) and diabetic (14.3 +/- 3.5 and 66.4 +/- 11.5) WKY rats. Proliferating cell nuclear antigen, another marker of cell proliferation, was significantly reduced in replicating glomerular (P = 0.0002) and tubulointerstitial (P < 0.0001) cells in diabetic SHR. In freshly isolated glomeruli, the level of p27(Kip1) detected by Western blotting was significantly higher In diabetic SIIR than in nondiabetic SHR (1.52 +/- 0.14 vs. 1.00 +/- 0.10% of control, P = 0.014). The expression of p21(Cip1) in isolated glomeruli did not differ among the groups of rats. In conclusion, the response of renal cell replication to diabetes differs markedly between prehypertensive SIIR and their WKY control rats. The decreased glomerular cell proliferation in prehypertensive diabetic SIIR is at least partly mediated by a higher expression of the Cdk inhibitor p27(Kip1).

DNA viruses in human cancer: An integrated overview on fundamental mechanisms of viral carcinogenesis

The first experimental data suggesting that neoplasm development in animals might be influenced by infectious agents were published in the early 1900s. However, conclusive evidence that DNA viruses play a role in the pathogenesis of some human cancers only emerged in the 1950s, when Epstein-Barr virus (EBV) was discovered within Burkitt lymphoma cells. Besides EBV, other DNA viruses consistently associated with human cancers are the hepatitis B virus (HBV), human papillomavirus (HPV), and Kaposi sarcoma herpesvirus (KSHV). Although each virus has unique features, it is becoming clearer that all these oncogenic agents target multiple cellular pathways to support malignant transformation and tumor development. (c) 2006 Elsevier B.V.. All rights reserved.

HIV, EBV and KSHV: Viral cooperation in the pathogenesis of human malignancies

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