CaPrx, a Coffea arabica gene encoding a putative class III peroxidase induced by root-knot nematode infection

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

Action mechanisms of petroleum hydrocarbons present in waters impacted by an oil spill on the genetic material of Allium cepa root cells

Chromosomal aberration (CA) assays have been widely used, not only to assess the genotoxic effects of chemical agents, but also to evaluate their action mechanisms on the genetic material of exposed organisms. This is of particular interest, since such analyses provide a better knowledge related to the action of these agents on DNA. Among test organisms, Allium cepa is an outstanding species due to its sensitivity and suitable chromosomal features, which are essential for studies on chromosomal damage or disturbances in cell cycle. The goal of the present study was to analyze the action mechanisms of chemical agents present in petroleum polluted waters. Therefore, CA assay was carried out in A. cepa meristematic cells exposed to the Guaeca river waters, located in the city of Sao Sebastiao, SP, Brazil, which had its waters impacted by an oil pipeline leak. Analyses of the aberration types showed clastogenic and aneugenic effects for the roots exposed to the polluted waters from Guaeca river, besides the induction of cell death. Probably all the observed effects were induced by the petroleum hydrocarbons derived from the oil leakage. (C) 2008 Elsevier B.V. All rights reserved.

Aluminum in corn plants: influence on growth and morpho-anatomy of root and leaf

A toxicidade do alumínio (Al) é um dos fatores mais limitantes para a produtividade. Esta pesquisa foi realizada para avaliar a influência do Al, em solução nutritiva, na altura de plantas, no peso da matéria seca e nas alterações morfoanatômicas de raízes e folhas de milho (Zea mays L.). O experimento foi conduzido em casa de vegetação com tratamentos constituídos de cinco doses de Al (0; 25; 75; 150; e 300 µmol L-1) e seis repetições. As soluções foram constantemente aeradas e o pH foi ajustado a 4,3, inicialmente. A matéria seca da parte aérea e das raízes e a altura das plantas diminuíram significativamente com o aumento da concentração de Al. As raízes de plantas de milho cultivadas em soluções com Al tiveram seu crescimento inibido e apresentaram menos raízes laterais e desenvolvimento do sistema radicular inferior, em comparação com as das plantas-controle. As folhas das plantas crescidas em soluções que continham 75 e 300 µmol L-1 de Al não apresentaram muita diferença anatômica em relação às das plantas-controle. A bainha da folha das plantas exposta ao Al apresentou epiderme uniestratificada revestida por uma fina camada de cutícula e as células da epiderme e do córtex foram as que menos se desenvolveram. No feixe vascular, o metaxilema e protoxilema não tinham paredes secundárias, e o diâmetro de ambos foi muito menor quando comparado com os das plantas-controle.

Root, shoot and leaf traits of the congeneric Styrax species may explain their distribution patterns in the cerrado sensu lato areas in Brazil

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

Effect of enzymatic hydrolysis on some physicochemical properties of root and tuber granular starches

A hidrólise enzimática do amido pode fornecer informações importantes sobre sua estrutura granular. Amidos de mandioca, batata-doce, mandioquinha-salsa e batata foram hidrolisados por α-amilase bacteriana a 37 °C durante 48 horas, e algumas propriedades físico-químicas dos resíduos da hidrólise foram determinadas. O amido de mandioca foi o mais suscetível à enzima com 20,9% de hidrólise, enquanto o amido de batata foi o mais resistente com 5,9%. O tamanho médio dos grânulos variou de 10,8 a 23,4 μm para os amidos de mandioquinha-salsa e batata, respectivamente. Amidos de mandioca e batata-doce apresentaram um padrão de difração de raio-X tipo A, enquanto os amidos de mandioquinha-salsa e batata mostraram padrão tipo B. Todos os amidos nativos mostraram superfície granular lisa e, após hidrólise, os amidos de mandioca, batata-doce e mandioquinha-salsa mostraram alguns grânulos bastante degradados, enquanto o amido de batata apresentou sutil sinal de degradação. O teor de amilose dos amidos diminuiu com a hidrólise para os amidos de mandioca, batata-doce e mandioquinha-salsa, permanecendo inalterado para o amido de batata. Como esperado, a viscosidade intrínseca e as propriedades de pasta diminuíram para todos os amidos hidrolisados. Não houve diferença significativa entre as propriedades térmicas dos amidos nativos e hidrolisados. Estes resultados sugeriram que a hidrólise ocorreu nas áreas cristalinas e amorfas dos grânulos. O padrão de difração do tipo B e o grande tamanho dos grânulos do amido de batata podem ter contribuído para a maior resistência deste amido à hidrólise.

Caracterização do gene da proteína capsidial do Grapevine virus A em videiras afetadas pela acanaladura do lenho de Kober no Estado de São Paulo

O presente trabalho caracteriza o gene codificador da proteína capsidial do isolado do Grapevine virus A (GVA) encontrado no Estado de São Paulo (GVA-SP). RNA total foi extraído de folhas e pecíolos de plantas de videira (Vitis spp.) da variedade 'Kober 5BB' e submetido a RT-PCR usando oligonucleotídeos desenhados para amplificar um fragmento entre as posições 6409 e 7175 do RNA do GVA (GenBank, acesso X75433). Foi obtido um fragmento de tamanho esperado (767 nt) que inclui o gene da proteína capsidial, codificando 198 aminoácidos. A seqüência do GVA-SP apresentou similaridade de nucleotídeos e aminoácidos de, respectivamente, 86-92,3% e 94,5-98% com isolados do GVA da Europa, África e Japão (Acessos X75433, AF441234, AF007415, AB039841) e da região Sul do Brasil (Acesso AF494187), sendo, entretanto, mais similar aos isolados africano e italiano.

Detecção do Grapevine virus A e Grapevine virus B por hibridização dot-blot com sondas moleculares não radioativas

O vírus A da videira (Grapevine virus A, GVA) e o vírus B da videira (Grapevirus virus B, GVB) estão associados à acanaladura do lenho de Kober (Kober stem grooving) e ao fendilhamento cortical da videira (grapevine corky bark), respectivamente. Este trabalho descreve o uso de sondas moleculares de cDNA na detecção de isolados do GVA (GVA-SP) e do GVB (GVB-C-SP e GVB-I-SP) em videiras (Vitis spp.) e fumo (Nicotiana occidentalis). As sondas marcadas com digoxigenina foram produzidas por RT-PCR utilizando oligonucleotídeos específicos para os genes da proteína capsidial. Os RNA totais foram extraídos de 45 plantas de diversas variedades de videira e de 13 plantas de fumo inoculadas mecanicamente com o GVB. Os RNA extraídos das plantas infetadas, indexadas biologicamente, hibridizaram com as sondas, não se verificando reação com plantas sadias. Para confirmar os resultados de hibridização, foram também feitos testes de RT-PCR. A utilização de hibridização dot-blot com sondas de cDNA mostrou-se eficaz na detecção dos vírus com especificidade e sensibilidade, ressaltando-se que, preferencialmente, folhas maduras e ramos dormentes devem ser utilizados nos testes diagnósticos para o GVB e GVA, respectivamente.

Detecção do Grapevine leafroll-associated virus 5 no Estado de São Paulo

O enrolamento da folha da videira (grapevine leafroll) é uma doença atribuída a pelo menos nove vírus sorologicamente distintos, Grapevine leafroll-associated viruses 1 a 9 e designados GLRaV-1 a GLRaV-9. No Brasil, já é conhecida a existência do GLRaV-1, GLRaV-2, GLRaV-3 e GLRaV-6. Neste trabalho, foi demonstrada a ocorrência do GLRaV-5 em amostras de videiras cultivadas no Estado de São Paulo, mediante teste de Biotina-ELISA. O vírus foi detectado com baixa incidência nas cultivares avaliadas, exceto na 'Cardinal', que apresentou 100% de infecção. Este é o primeiro relato da ocorrência do GLRaV-5 no Brasil.

Caracterização do gene da proteína capsidial de dois isolados, patologicamente distintos e sorologicamente semelhantes, do Grapevine virus B em videiras no Estado de São Paulo

No presente trabalho, descreve-se a caracterização do gene codificador da proteína capsidial de dois isolados sintomatologicamente distintos do Grapevine virus B (GVB). Para isto, RNA totais foram extraídos de folhas e pecíolos de videiras (Vitis spp.) infetadas, cultivares Rubi (GVB-C SP) e Itália (GVB-I SP) e utilizados para amplificar, por RT/PCR, um fragmento entre as posições 6425 e 7118 (694 nucleotídeos, nt) do RNA do GVB (GenBank, acesso X75448). O fragmento obtido inclui o gene da proteína capsidial (594 nt) codificando 197 aminoácidos com massa molecular estimada em aproximadamente 21.600 Da. A seqüência do GVB-C SP apresentou maior similaridade de nucleotídeos e aminoácidos deduzidos com o isolado italiano (acesso X75448), enquanto que o GVB-I SP foi mais similar a um outro isolado brasileiro do GVB descrito no Rio Grande do Sul (GVB BR1, acesso AF438410). Os dois isolados paulistas do GVB podem ser diferenciados por digestão com a enzima de restrição EcoRI, uma vez que há um sítio interno no GVB-C SP que está ausente no isolado GVB-I SP.

Structural resistance in immature teeth using root reinforcements in vitro

The purpose of this study was to evaluate in vitro the efficacy of root reinforcements by light-cured composite resin or zirconium fiber post in simulated immature non-vital teeth. Fifty-six bovine incisors teeth were used for this study. The crown of each tooth was removed in the medium third to obtain a standard length of 30 mm. The specimens were divided into four groups (n = 14): G1) the root canals were instrumented and enlarged to simulate immature non-vital teeth and were reinforced with a light-cured composite resin using a translucent curing post (Luminex system); (G2) the specimens were instrumented, enlarged and they received root reinforcement with zirconium fiber post; G3 (positive control): they received similar treatment to the G1 and G2 groups, but did not receive root reinforcement; G4 (negative control): the roots were not weakened and did not receive reinforcement. Every tooth was submitted to compressive force using an Instron testing machine with an angle of 45&DEG; at a speed of 1 mm min(-1) until the fracture. The results showed a markedly increased resistance to fracture in the G1 and G2 (122.38 and 122.08 kgf, respectively). Among the results of G1 and G2 there was not any significant difference (P > 0.05) but they were significantly different from the control groups (P < 0.05). The conclusion is that the use of root reinforcements with zirconiun fiber post or composite resin can increase significantly the structural resistance of the weakened teeth, decreasing the risk of the fracture.

Effects of chlorhexidine and sodium hypochlorite on the microhardness of root canal dentin

Objective. The purpose of this study was to evaluate the effects of endodontic irrigants on the microhardness of root canal dentin.Study design. Thirty extracted single-rooted human teeth were used. The crowns were sectioned at the cementoenamel junction. Each root was transversely sectioned into cervical, middle, and apical segments, resulting in 90 specimens. The 3 sections of each root were separately mounted in an individual silicon device with acrylic resin. The specimens were randomly divided into the following 3 groups (n = 30), according to the irrigant solution used: (1) group 1, control (saline solution); (2) group 2, 2% chlorhexidine gluconate solution; and (3) group 3, 1% sodium hypochlorite (NaOCl). After 15 minutes of irrigation, dentin microhardness was measured on each section at 500 mu m and 1000 mu m from the pulp-dentin interface with a Vickers diamond microhardness tester in Vickers hardness number (VHN).Results. Data obtained were analyzed using analysis of variance and the Tukey test (5%). Specimens irrigated with 2% chlorhexidine (group 2) or 1% NaOCl (group 3) presented lower values of dentin microhardness, with significant difference in relation to the control group (P < .05).Conclusion. It could be concluded that chlorhexidine and NaOCl solutions significantly reduced the microhardness of root canal dentin at 500 mu m and 1000 mu m from the pulp-dentin interface.

In vitro effects of calcium hydroxide and polymyxin B on endotoxins in root canals

Objectives. To evaluate the effects of intracanal medicaments on endotoxins in root canals.Methods. Seventy-five freshly extracted maxillary incisors were used in this study. The crowns of teeth were sectioned near the CEJ in order to standardize the root length to 14 mm. The root canals were instrumented to an apical size #50 file and irrigated with 1% sodium hypochlorite solution and sterilized with 60 Co gamma irradiation. Standardized suspension containing Escherichia coli endotoxin was inoculated into the 60 root canals. The specimens were randomly assigned to 5 groups (n=15), according to the intracanal medicament used: (G1) calcium hydroxide; (G2) polymyxin B; (0) combination neomycin-potymyxin B-hydrocortisone; (G4) positive control (no intracanal medicament); (G5) negative control (no endotoxin and no intracanal medicament). After 7 days, the detoxification of endotoxin was evaluated by Limulus lysate assay and antibody production in B-tymphocytes culture.Results. Groups 1, 2 and 5 presented the best results by Limulus lysate and were significantly different to groups 3 and 4 (p<0.05). Stimulation of antibodies production in cell culture by groups 1 and 6 was smaller and statistically different than groups 2, 3, 4 and 5 (p<0.05). Groups 2 and 5 induced a small increase in the antibodies production in relation to the groups 1 and 6. Groups 3 and 4 induced a significant increase of antibodies production (p<0.05).Conclusions. The calcium hydroxide and polymyxin B intracanal medicaments detoxified endotoxin in root canals and altered the properties of LPS to stimulate the antibody production by B-Lymphocytes. The combination neomycin-polymyxin B-hydrocortisone did not detoxified endotoxin. (C) 2004 Elsevier Ltd. All rights reserved.

Efficacy of Endodontic Treatment for Endotoxin Reduction in Primarily Infected Root Canals and Evaluation of Cytotoxic Effects

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

Evaluation of simulated external root resorptions with digital radiography and digital subtraction radiography

Introduction: Root resorption can cause damage in orthodontic patients. Digital subtraction radiography (DSR) is a useful resource for the detection of mineral losses. The purpose of this study was to compare the efficacy of digital radiography (DR) and DSR in detecting simulated external root resorption. Examiner agreement between the 2 techniques was also evaluated. Methods: Root resorptions of various sizes were simulated on the apical and lingual aspects of 49 teeth from 9 dry human mandibles. The teeth were radiographed in standardized conditions. The radiographs were registered with Regeemy Image Registration and Mosaicking (version 0.2.43-RCB, DPI-INPE, Sao Jose dos Campos, São Paulo, Brazil) and subtracted with Image Tool (University of Texas Health Science Center at San Antonio). The subtracted images and the digital radiographs were evaluated by 3 oral radiologists. Results: No statistically significant differences were found for the methods in the detection of apical root resorptions, independently from lesion size, and of lingual resorptions of 1.2 mm or greater. DSR was significantly better than DR for detection of lingual resorptions up to 1 mm. Resorptions less than 0.5 mm were not precisely detected by either method. DSR provided better intraexaminer and interexaminer agreement than did DR. Conclusions: Both methods are precise for detection of apical root resorptions as small as 0.5 mm and lingual resorptions of 1 mm or more. However, DSR frequently performed better than did DR. (Am J Orthod Dentofacial Orthop 2011;139:324-33)

SEALING ABILITY of CASTOR OIL POLYMER AS A ROOT-END FILLING MATERIAL

Objective: The purpose of this study was to evaluate the sealing ability of castor oil polymer (COP), mineral trioxide aggregate (MTA) and glass ionomer cement (GIC) as root-end filling materials. Forty-five single-rooted human teeth were cleaned and prepared using a step-back technique. The apical third of each root was resected perpendicularly to the long axis direction. All teeth were obturated with gutta-percha and an endodontic sealer. After, a root-end cavity with 1.25-mm depth was prepared using a diamond bur. The specimens were randomly divided into three experimental groups (n = 15), according to the root-end filling material used: G1) COP; G2) MTA; G3) GIC. The external surfaces of the specimens were covered with epoxy adhesive, except the root-end filling. The teeth were immersed in rhodamine B dye for 24 hours. Then, the roots were sectioned longitudinally and the linear dye penetration at the dentin/material interface was determined using a stereomicroscope. ANOVA and Tukey's tests were used to compare the three groups. The G1 group (COP) presented smaller dye penetration, statistically different than the G2 (MTA) and G3 (GIC) groups (p < 0.05). No statistically significant difference in microleakage was observed between G2 and G3 groups (p > 0.05). The results of this study indicate that the COP presented efficient sealing ability when used as a root-end filling material showing results significantly better than MTA and GIC.