Ribosome inactivating proteins from plants inhibiting viruses

Many plants contain ribosome inactivating proteins (RIPs) with N-glycosidase activity, which depurinate large ribosomal RNA and arrest protein synthesis. RIPs so far tested inhibit replication of mRNA as well as DNA viruses and these proteins, isolated from plants, are found to be effective against a broad range of viruses such as human immunodeficiency virus (HIV), hepatitis B virus (HBV) and herpes simplex virus (HSV). Most of the research work related to RIPs has been focused on antiviral activity against HIV; however, the exact mechanism of antiviral activity is still not clear. The mechanism of antiviral activity was thought to follow inactivation of the host cell ribosome, leading to inhibition of viral protein translation and host cell death. Enzymatic activity of RIPs is not limited to depurination of the large rRNA, in addition they can depurinate viral DNA as well as RNA. Recently, Phase I/II clinical trials have demonstrated the potential use of RIPs for treating patients with HIV disease. The aim of this review is to focus on various RIPs from plants associated with anti-HIV activity.

Face recognition using kernel ridge regression

In this paper, we present novel ridge regression (RR) and kernel ridge regression (KRR) techniques for multivariate labels and apply the methods to the problem of face recognition. Motivated by the fact that the regular simplex vertices are separate points with highest degree of symmetry, we choose such vertices as the targets for the distinct individuals in recognition and apply RR or KRR to map the training face images into a face subspace where the training images from each individual will locate near their individual targets. We identify the new face image by mapping it into this face subspace and comparing its distance to all individual targets. An efficient cross-validation algorithm is also provided for selecting the regularization and kernel parameters. Experiments were conducted on two face databases and the results demonstrate that the proposed algorithm significantly outperforms the three popular linear face recognition techniques (Eigenfaces, Fisherfaces and Laplacianfaces) and also performs comparably with the recently developed Orthogonal Laplacianfaces with the advantage of computational speed. Experimental results also demonstrate that KRR outperforms RR as expected since KRR can utilize the nonlinear structure of the face images. Although we concentrate on face recognition in this paper, the proposed method is general and may be applied for general multi-category classification problems.

HIV infection of dendritic cells subverts the IFN induction pathway via IRF-1 and inhibits type 1 IFN production

Many viruses have developed mechanisms to evade the IFN response. Here, HIV-1 was shown to induce a distinct subset of IFN-stimulated genes (ISGs) in monocyte-derived dendritic cells (DCs), without detectable type I or II IFN. These ISGs all contained an IFN regulatory factor 1 (IRF-1) binding site in their promoters, and their expression was shown to be driven by IRF-1, indicating this subset was induced directly by viral infection by IRF-1. IRF-1 and -7 protein expression was enriched in HIV p24 antigen-positive DCs. A HIV deletion mutant with the IRF-1 binding site deleted from the long terminal repeat showed reduced growth kinetics. Early and persistent induction of IRF-1 was coupled with sequential transient up-regulation of its 2 inhibitors, IRF-8, followed by IRF-2, suggesting a mechanism for IFN inhibition. HIV-1 mutants with Vpr deleted induced IFN, showing that Vpr is inhibitory. However, HIV IFN inhibition was mediated by failure of IRF-3 activation rather than by its degradation, as in T cells. In contrast, herpes simplex virus type 2 markedly induced IFNβ and a broader range of ISGs to higher levels, supporting the hypothesis that HIV-1 specifically manipulates the induction of IFN and ISGs to enhance its noncytopathic replication in DCs.

Membrane nanotubes in myeloid cells in the adult mouse cornea represent a novel mode of immune cell interaction

Membrane nanotubes (MNTs) are newly discovered cellular extensions that are either blind-ended or can connect widely separated cells. They have predominantly been investigated in cultured isolated cells, however, previously we were the first group to demonstrate the existence of these structures in vivo in intact mammalian tissues. We previously demonstrated the frequency of both cell–cell or bridging MNTs and blind-ended MNTs was greatest between major histocompatibility complex (MHC) class II+ cells during corneal injury or TLR ligand-mediated inflammation. The present study aimed to further explore the dynamics of MNT formation and their size, presence in another tissue, the dura mater, and response to stress factors and an active local viral infection of the murine cornea. Confocal live cell imaging of myeloid-derived cells in inflamed corneal explants from Cx3cr1GFP and CD11ceYFP transgenic mice revealed that MNTs form de novo at a rate of 15.5 μm/min. This observation contrasts with previous studies that demonstrated that in vitro these structures originate from cell–cell contacts. Conditions that promote formation of MNTs include inflammation in vivo and cell stress due to serum starvation ex vivo. Herpes simplex virus-1 infection did not cause a significant increase in MNT numbers in myeloid cells in the cornea above that observed in injury controls, confirming that corneal epithelium injury alone elicits MNT formation in vivo. These novel observations extend the currently limited understanding of MNTs in live mammalian tissues.

Implications of Kungurian (Early Permian) conodonts from Hatahoko, Japan, for correlation between the Tethyan and international timescales

The correlation between the fusulinid-based Tethyan and the conodont-based international timescales of the Permian System has become one of themost disputed issues among the Permian community during the past two decades. In this paper,we document a conodont fauna consisting of four species including Sweetognathus guizhouensis, Pseudohindeodus augustus, Hindeodus permicus and a new genus Meiognathus pustulus from the lower part of a large exotic limestone block at Hatahoko in the Nyukawa area, Gifu Prefecture, central Japan, which all suggest aKungurian age. The Kungurian age indicated by the conodonts is consistent with the age of the associated brachiopods, but conflicts with the Murgabian age indicated by the associated fusulinids including Cancellina nipponica, Neoschwagerina simplex, Neofusulinella praecursor etc. This co-occurrence of Kungurian conodonts and Murgabian fusulinids in central Japan suggests that previously unrecognized temporal distributions of some key fusulinid or conodont elements need to be clarified and that the intensively-disputed correlation problem between theKungurian containing theMurgabian fusulinids at the Luodian section in Guizhou, South China with the strata containing the ammonoid Waagenoceras in Oman and Sicily was caused by artificial conodont taxonomic discrepancies. The Luodian section in South China could serve as a key reference section for the correlation of the Kungurian Stage (late Early Permian) between the Tethyan and international timescales. 

CD8+ T cells from a novel T cell receptor transgenic mouse induce liver-stage immunity that can be boosted by blood-stage infection in rodent malaria

To follow the fate of CD8+ T cells responsive to Plasmodium berghei ANKA (PbA) infection, we generated an MHC I-restricted TCR transgenic mouse line against this pathogen. T cells from this line, termed PbT-I T cells, were able to respond to blood-stage infection by PbA and two other rodent malaria species, P. yoelii XNL and P. chabaudi AS. These PbT-I T cells were also able to respond to sporozoites and to protect mice from liver-stage infection. Examination of the requirements for priming after intravenous administration of irradiated sporozoites, an effective vaccination approach, showed that the spleen rather than the liver was the main site of priming and that responses depended on CD8α+ dendritic cells. Importantly, sequential exposure to irradiated sporozoites followed two days later by blood-stage infection led to augmented PbT-I T cell expansion. These findings indicate that PbT-I T cells are a highly versatile tool for studying multiple stages and species of rodent malaria and suggest that cross-stage reactive CD8+ T cells may be utilized in liver-stage vaccine design to enable boosting by blood-stage infections.

Estudo sobre a biodegradação do herbicida trufluralina por bactérias isoladas de solo agrícola e proposta metodológica para o ensino de biodegradação

Trifluralina (a, a, a,trifluoro-2,6-dinitro-N, N-dipropil-p-toluidina) (TFL) é um herbicida pré-emergente, incorporado ao solo que tem sido usado na agricultura desde a década de sessenta; ele é moderadamente persistente em vários tipos de solos do Brasil. O objetivo deste estudo foi isolar - de um solo agrícola contaminado por quatro décadas - e caracterizar bactérias resistentes a TFL, determinar suas habilidades em degradar a TFL, investigar a presença de gens degardadores que possam estar envolvidos na degradação da TFL e propor um método de ensino teórico prático sobre a biodegradação, para cursos de graduação. Oito bactérias foram isoladas, pela técnica de subculturas repetidas em meio contendo TFL como única fonte de carbono, e identificadas, pelo método bioquímico e seqüenciamento do rDNA 16S como Klebsiella oxytoca, Herbaspirillum seropedicae, 3 strains of Bacillus simplex, 2 de Pseudomonas montelli e uma outra Pseudomonas sp. Uma terceira bactéria (iaolado #9), não identificada, que crescia ao redor de cristais de TFL em meio sólido, foi isolada; esta é uma técnica nova que poderá ser útil no isolamento de bactérias que são resistentes a outros compostos pouco solúveis em água. Todas as bactérias isoladas foram submetidas ao teste de biodegradação, em um meio contendo sais minerais, 0,1% succinato, 0,1 % de extrato de leveduras e 50 mg. L-1 de TFL Cinco bactérias reduziram a concentração de TFL no meio, após trinta dias de incubação: Klebsiella oxytoca (24,6 %), Herbaspirillum seropedicae (16,4 %), Bacillus simplex 2 (25.0 %), Bacillus simplex 3 (16.0 %) e isolado 9 (21.0 %). Uma bactéria conhecida como degradadora da TFL, Brevundimonas diminuta (NCIMB 10329) degradou a TFL, neste meio, de maneira semelhantes ao das bactérias isoladas. Os DNAs extraídos das quatro bactérias identificadas degradadoras da TFL, foram sondados para os gens catbólicos ndoB, todC, xylX, catA e xylE, os quais codificam as enzimas naftaleno 1,2-dioxigenase, toluene dioxigenase, toluate 1,2-dioxigenase, catecol 1,2-dioxigenase e catecol 2,3-dioxigenase, respectivamente. Técnicas de PCR e hibridização demonstraram que os DNAs de todas estas quatro bactérias foram fortemente hibridizadas para o gen ndoB, contudo, usando a técnica de ¨zonas claras¨, observou-se que nenhuma delas degradou naftaleno. Estes resultados indicam a presença de gens dioxigenases, nestas bactérias degradadoras da TFL, que poderiam estar transformando a TFL como substrato principal, ou como cometabolismo. O conhecimento sobre processos de biodegradação é necessário para os graduados dos cursos de agronomia, química, biologia, tecnologia de alimentos, etc. Neste trabalho, também, propomos o estudo teórico e prático da compostagem o qual estimula o interesse dos estudantes em aprender sobre o metabolismo envolvido neste, e em outros, processos biotecnológicos.

Programação linear e planejamento da empresa agricola

Este trabalho tem por objetivo o estudo do m~todo da Programação Linear e de sua aplicação no planejamento da empresa agricola, tendo em vista a demonstração de sua importancia como instrumento analItico no processo de tomada de decisões. Inicialmente ~ feita uma abordagem sobre a problematica da programação e seus fundamentos, sendo examinados os principais conceitos sobre os quais o assunto se baseia. A discussão encaminhada no sentido de chamar a atenção para o fato de que, em vista das pressões a que esta submetida a agricultura no estagio contemporâneo do desenvolvimento, - que lhe exigem melhores niveis de desempenho traduzidos em Indices mais elevados de produtividade, generalizou-se o consenso sobre a conveniência de se dar maior atenção i programação das atividades do setor agrIcola, tanto a nIvel de Região como no ambito de sua area empresarial. Entre os varias modelos gerados com o objetivo de oferecer respostas aos problemas de organização da produção, otimização de resuItados c racionalização do uso dos fatores, o da Programação Linear considerado o mais geralmente aceito pela comunidade cientIfica, por sua estrutura teórico sofisticada e pela capacidade de dar respostas exatas e diretas a questões de maior complexidade. A parte teórica do metodo da Programação Linear ~ examinada no Capítulo 11. De maneira sumária são abordados os seus fundamentos matemáticos e discutidos os principais teoremas sobre conjuntos convexos, corno elementos do espaço das soluções possiveis. A álgebra do algorÍtmo Simplex ~ abordada em sua forma ardinária, com a demonstração dos passos sucessivos do algoritmo, ato encontro do vertice característico da solução otima. No Capítulo 111 os conceitos teoricos da Programação Linear sao aplicados aos dados de uma empresa agricola, com a finalidade de demonstrar a operacionalização do algoritmo Simplcx ! através de programas de computador.

Análise química e avaliação da atividade antiviral de hypericum connatum lam.

Plantas da família Guttiferae apresentam diversas atividades biológicas sendo Hypericum o gênero mais importante devido às atividades antidepressiva, antibacteriana e antiviral de algumas espécies. Hypericum connatum, utilizado no sul do Brasil para o tratamento de feridas de boca, demonstrou atividade contra o lentivírus, responsável pela imunodeficiência felina. Objetivo: isolar e identificar as substâncias majoritárias de H. connatum e testar a atividade de extratos obtidos das partes aéreas e raízes e das substâncias isoladas frente a duas cepas do herpesvírus simples tipo 1 (HSV-1). Método: foram obtidos frações nhexano, diclorometano e metanol e extratos bruto, aquosos a diferentes temperaturas e hidro-etanólico das partes aéreas e raízes. As frações n-hexano e metanólica das partes aéreas foram submetidas à coluna cromatográfica para o isolamento de substâncias. Os extratos e substâncias isoladas foram testados frente ao HSV-1, cepas KOS e ATCC-VR733. Determinou-se a concentração máxima não tóxica (CMNT) à célula e a concentração que provoca alteração morfológica em 50% das células (CC50) pela técnica da alteração morfológica celular, utilizando-se células VERO, linhagem ATCC CCL-81. A avaliação da atividade antiviral foi realizada em placas de microtitulação e medida pela inibição do efeito citopático (ECP) provocado pelo vírus. Resultados e Conclusões: da fração n-hexano foi isolado hiperbrasilol B, da fração metanólica foram isolados amentoflavona, hiperosídeo e guaijaveriana, além de um flavonol de estrutura ainda não definida (HCN3). A fração n-hexano e o extrato bruto das raízes inibiram o ECP das cepas KOS e ATCC-VR733. Os demais extratos testados não apresentaram atividade antiviral. Dentre as substâncias analisadas, hiperbrasilol B, amentoflavona e HCN3 foram ativos frente às duas cepas. Os flavonóides hiperosídeo e guaijaverina não apresentaram atividade anti-HSV-1.

Prevalência da infecção pelo Papilomavírus humano, Chlamydia Trachomatis e Herpes Simples do tipo 2 em adolescentes atendidas em unidades de saúde pública de Natal

This study assessed the level of knowledge, attitude and practice of Pap smear and human papillomavirus (HPV), in addition to analyzing the prevalence of genital HPV infection, Herpes Simplex Type 2 (HSV-2) and Chlamydia trachomatis in teenagers. The study consisted of two approaches, one based only on interviews conducted with adolescents enrolled in public schools or in public health facilities in the city of Natal. The other approach involved only a group of 132 adolescents enrolled among those admitted to two health units in Natal-RN. This second group of participants two specimens were collected for laboratory analysis: one was directed to prepare the blade for the Pap test, and other processed for DNA extraction for molecular analysis, focusing on the detection of HPV, HSV-2 and C . trachomatis. The presence of DNA of the three pathogens was investigated by the technique of polymerase chain reaction (PCR). The presence of each of the three pathogens was analyzed in terms of socio-demographic characteristics, as well as sexual and reproductive activity to identify risk factors for infection and development of lesions of the uterine cervix. The results show that the adolescents in this study had levels of knowledge and attitude very low, both in relation to cytology to HPV as though they have made a reasonable percentage of adequate practice exam and prevention of HPV infection. The overall prevalence of HPV infection was 54.5% and 48.2% in adolescents with normal cytology and 86.4% in those with abnormal cytology. We observed a higher proportion of cases of infection in the age group of 18 to 21. The prevalence of HPV infection was slightly higher among pregnant teenagers. The overall prevalence of HSV-2 infection was 13.6% and 11.8% in women with normal cytology and 22.7% in those with abnormal cytology. A higher proportion of cases of infection was found in the age group from 14 to 17, with a slightly higher prevalence among pregnant women. The C. trachomatis was found with an overall prevalence of 19.7% and 21.8% in adolescents with normal cytology and 9.1% in those with abnormal cytology. The prevailing rate was highest in the age group 18 to 21 years and in nonpregnant

Prevalência da infecção genital pelo vírus herpes simples tipo 1 e 2 em mulheres grávidas e não grávidas de Natal/RN

Herpes simplex is a virus that can be transmitted sexually and is potentially associated with vertical transmission. This study evaluated the prevalence of genital infection by herpes simplex virus (HSV) types 1 and 2 in pregnant and nonpregnant care in the city of Natal / RN, including a total of 222 women, 92 pregnant and 130 nonpregnant. The participants answered a questionnaire to obtain data and socio-demographic characteristics, as well as potential risk factors for sexually transmitted diseases. After the interview, we collected two cervical specimens, one for the Pap test and the other for DNA extraction and analyzed by polymerase chain reaction (PCR) to detect both virus serotypes. Then the women underwent a clinical examination by colposcopy. For statistical analysis, we used the chi-square and logistic regression by SSPS 17.0 Statistic. Most women were up to 30 years of age, nonwhite ethnicity, married, elementary education, family income below the poverty level; initiated sexual activity with age up to 18 years; had more than one sexual partner lifelong and was not pregnant, but has had at least one child. The HSV-1 showed a prevalence of 26.1% among pregnant women and 30.0% in non-pregnant women. While HSV-2 prevalence was found with 10.9% and 19.2% in pregnant and nonpregnant women, respectively. The largest proportion of morphological changes of the uterine cervix was detected among nonpregnant women, both in cytology and in colposcopy. The women were nonwhite ethnicity, married, became pregnant aged less than or equal to 18 years and who had one to two pregnancies had a lower risk of acquiring genital HSV infection. There was a high prevalence of genital HSV infection, HSV-1 is more prevalent than HSV-2. No association was found between morphological changes of the uterine cervix and the presence of the virus in pregnant and nonpregnant women, nor between genital HSV infection and the classic risk factors for sexually transmitted diseases

Compositional nutrient diagnosis of corn using the Mahalanobis distance as nutrient imbalance index

Parent, L. E., Natale, W. and Ziadi, N. 2009. Compositional nutrient diagnosis of corn using the Mahalanobis distance as nutrient imbalance index. Can. J. Soil Sci. 89: 383-390. Compositional nutrient diagnosis (CND) provides a plant nutrient imbalance index (CND - r(2)) with assumed chi(2) distribution. The Mahalanobis distance D(2), which detects outliers in compositional data sets, also has a chi(2) distribution. The objective of this paper was to compare D(2) and CND - r(2) nutrient imbalance indexes in corn (Zea mays L.). We measured grain yield as well as N, P, K, Ca, Mg, Cu, Fe, Mn, and Zn concentrations in the ear leaf at silk stage for 210 calibration sites in the St. Lawrence Lowlands [2300-2700 corn thermal units (CTU)] as well as 30 phosphorus (2300-2700 CTU; 10 sites) and 10 nitrogen (1900-2100 CTU; one site) replicated fertilizer treatments for validation. We derived CND norms as mean, standard deviation, and the inverse covariance matrix of centred log ratios (clr) for high yielding specimens (>= 9.0 Mg grain ha(-1) at 150 g H(2)O kg(-1) moisture content) in the 2300-2700 CTU zone. Using chi(2) = 17 (P < 0.05) with nine degrees of freedom (i.e., nine nutrients) as a rejection criterion for outliers and a yield threshold of 8.6 Mg ha(-1) after Cate-Nelson partitioning between low- and high-yielders in the P validation data set, D(2) misclassified two specimens compared with nine for CND -r(2). The D(2) classification was not significantly different from a chi(2) classification (P > 0.05), but the CND - r(2) classification differed significantly from chi(2) or D(2) (P < 0.001). A threshold value for nutrient imbalance could thus be derived probabilistically for conducting D(2) diagnosis, while the CND - r(2) nutrient imbalance threshold must be calibrated using fertilizer trials. In the proposed CND - D(2) procedure, D(2) is first computed to classify the specimen as possible outlier. Thereafter, nutrient indices are ranked in their order of limitation. The D(2) norms appeared less effective in the 1900-2100 CTU zone.

Desenvolvimento e teste de um modelo estatístico de identificação das proporções de água produzida em zonas de produção de um campo maduro de petróleo

This study investigates the chemical species produced water from the reservoir areas of oil production in the field of Monte Alegre (onshore production) with a proposal of developing a model applied to the identification of the water produced in different zones or groups of zones.Starting from the concentrations of anions and cátions from water produced as input parameters in Linear Discriminate Analysis, it was possible to estimate and compare the model predictions respecting the particularities of their methods in order to ascertain which one would be most appropriate. The methods Resubstitution, Holdout Method and Lachenbruch were used for adjustment and general evaluation of the built models. Of the estimated models for Wells producing water for a single production area, the most suitable method was the "Holdout Method and had a hit rate of 90%. Discriminant functions (CV1, CV2 and CV3) estimated in this model were used to modeling new functions for samples ofartificial mixtures of produced water (producedin our laboratory) and samples of mixtures actualproduced water (water collected inwellsproducingmore thanonezone).The experiment with these mixtures was carried out according to a schedule experimental mixtures simplex type-centroid also was simulated in which the presence of water from steam injectionin these tanks fora part of amostras. Using graphs of two and three dimensions was possible to estimate the proportion of water in the production area

Flexural strength of acrylic resin repairs processed by different methods: water bath, microwave energy and chemical polymerization

Denture fractures are common in daily practice, causing inconvenience to the patient and to the dentists. Denture repairs should have adequate strength, dimensional stability and color match, and should be easily and quickly performed as well as relatively inexpensive. Objective: The aim of this study was to evaluate the flexural strength of acrylic resin repairs processed by different methods: warm water-bath, microwave energy, and chemical polymerization. Material and methods: Sixty rectangular specimens (31x10x2.5 mm) were made with warm water-bath acrylic resin (Lucitone 550) and grouped (15 specimens per group) according to the resin type used to make repair procedure: 1) specimens of warm water-bath resin (Lucitone 550) without repair (control group); 2) specimens of warm water-bath resin repaired with warm water-bath; 3) specimens of warm water-bath resin repaired with microwave resin (Acron MC); 4) specimens of warm water-bath resin repaired with autopolymerized acrylic resin (Simplex). Flexural strength was measured with the three-point bending in a universal testing machine (MTS 810 Material Test System) with load cell of 100 kgf under constant speed of 5 mm/min. Data were analyzed statistically by Kruskal-Wallis test (p<0.05). Results: The control group showed the best result (156.04 +/- 1.82 MPa). Significant differences were found among repaired specimens and the results were decreasing as follows: group 3 (43.02 +/- 2.25 MPa), group 2 (36.21 +/- 1.20 MPa) and group 4 (6.74 +/- 0.85 MPa). Conclusion: All repaired specimens demonstrated lower flexural strength than the control group. Repairs with autopolymerized acrylic resin showed the lowest flexural strength.

Crystal structure of human purine nucleoside phosphorylase complexed with acyclovir

In human, purine nucleoside phosphorylase (HsPNP) is responsible for degradation of deoxyguanosine and genetic deficiency of this enzyme leads to profound T-cell mediated immunosuppression. PNP is therefore a target for inhibitor development aiming at T-cell immune response modulation and has been submitted to extensive structure-based drug design. This work reports the first crystallographic Study of human PNP complexed with acyclovir (HsPNP:Acy). Acyclovir is a potent clinically useful inhibitor of replicant herpes simplex virus that also inhibits human PNP but with a relatively lower inhibitory activity (K-i=90muM). Analysis of the structural differences among the HsPNP:Acy complex, PNP apoenzyme, and HsPNP:Immucillin-H provides explanation for inhibitor binding, refines the purine-binding site, and can be used for future inhibitor design. (C) 2003 Published by Elsevier B.V.