Microvascular density and tumor budding in oral squamous cell carcinoma.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is the most prevalent malignant head and neck tumor, excluding the nonmelanoma skin cancer. Despite recent advances in the diagnosis and treatment, the disease's mortality rate is nonetheless high. The presence of isolated neoplastic cells or small clusters of up to four cells at the tumor's invasive front, named tumor budding, is associated with a worse prognosis in OSCC. Angiogenesis has also been recognized as a determining factor in the progression of malignancies and in the development of metastases. Several studies have investigated the assessment of microvascular density (MVD) as a potential prognostic factor in OSCC. This study aimed to evaluate, in OSCC, differences in MVD between tumors with high-intensity tumor budding and tumors with low-intensity or no tumor budding. In samples with high-intensity tumor budding, differences in MVD between the budding area and the area outside the budding were also evaluated. Moreover, the study assessed differences in MVD concerning clinicopathological characteristics such as sex, age, tobacco smoking, tumor location and tumor size. MATERIAL AND METHODS: One hundred and fifty [150] samples of OSCC were subjected to immunohistochemistry to assess the intensity of tumor budding (by immunostaining for multi-cytokeratin) and MVD (by immunostaining for CD34 and CD105, independently). The data were treated using descriptive and analytical statistics. RESULTS: There were no differences in MVD, assessed by immunostaining for CD34 or CD105, concerning clinicopathological characteristics such as sex, age, tobacco smoking, tumor location and tumor size (p > 0.05). Tumors with high-intensity tumor budding did not show differences in MVD, assessed by immunostaining for CD34 or CD105, when compared to tumors with low-intensity or no tumor budding (p > 0.05). However, in samples with high-intensity tumor budding, the MVD assessed by immunostaining for CD34 was higher in the budding area than in the area outside the budding (p < 0.05). This difference was not observed when MVD was assessed by immunostaining for CD105 (p > 0.05). CONCLUSIONS: The higher MVD in the budding area may be an additional indication that this is a peculiar region of the tumor, associated with biological phenomena related to tumor progression.

SERS Investigation of Cancer Cells Treated with PDT: Quantification of Cell Survival and Follow-up.

In this study Surface Enhanced Raman Spectroscopy (SERS) data recorded from mouse mammary glands cancer cells (4T1 cell line) was used to assess information regarding differences between control, death and viable cells after Photodynamic Therapy (PDT) treatment. The treatment used nanoemulsions (NE/PS) loaded with different chloroaluminumphthalocyanine (ClAlP) photosensitizer (PS) contents (5 and 10 µmol × L-1) and illumination (660 nm wavelength) at 10 J × cm-2 (10 minutes). The SERS data revealed significant molecular alterations in proteins and lipids due to the PDT treatment. Principal Component Analysis (PCA) was applied to analyze the data recorded. Three-dimensional and well reproductive PCA scatter plots were obtained, revealing that two clusters of dead cells were well separated from one another and from control cluster. Overlap between two clusters of viable cells was observed, though well separated from control cluster. Moreover, the data analysis also pointed out necrosis as the main cell death mechanism induced by the PDT, in agreement with the literature. Finally, Raman modes peaking at 608 cm-1 (proteins) and 1231 cm-1 (lipids) can be selected for follow up of survival rate of neoplastic cells after PDT. We envisage that this finding is key to contribute to a quick development of quantitative infrared thermography imaging.

Chemical characterization of the Allium sativum and Origanum vulgare essential oils and their inhibition effect on the growth of some food pathogens / Caracterização química e efeito inibitório dos óleos essenciais de Allium sativum e Origanum vulgare frente ao crescimento de alguns patógenos de alimentos

This study sought to evaluate the chemical composition of the Allium sativum and Origanum vulgare essential oils and their effect on the growth inhibition of microorganisms, such as P. aeruginosa, S. Choleraesuis, A. flavus, A. niger and P. simplicissimum, important food contaminants. The main constituents of the oregano essential oil were 4-terpineol (27.03%), γ-terpinene (20.04%), and β-cymene (6.34%), and the main constituents of the garlic essential oil were diallyl trisulfide (38, 81%), diallyl disulfide (25.23%), and methyl allyl trisulfide (12.52%). Inhibition zones were formed in in vitro tests on the bacteria S. Choleraesuis and P. aeruginosa, except for A. sativum against P. aeruginosa. The inhibition of mycelial growth caused by the oregano essential oil occurred with the concentrations of 0.10, 0.03 and 0.05 mg mL-1 for the A. flavus, A. niger and P. simplicissimum fungi, respectively. The CMI for the garlic oil began at the 0.03 mg mL-1 concentration for all species of fungi. The oils presented an inhibitory effect against the microorganisms studied and constitute an alternative for microbiological control in food.

Objetivou-se avaliar a composição química e o efeito inibitório dos óleos essenciais de Allium sativum e Origanum vulgare frente ao crescimento dos micro-organismos Pseudomonas aeruginosa, Salmonella Choleraesuis, Aspergillus flavus, Aspergillus niger e Penicillium simplicissimum, importantes patógenos causadores de contaminações em alimentos. Para quantificação e identificação dos constituintes químicos dos óleos, utilizou-se cromatógrafo gasoso acoplado a espectrômetro de massas. Os principais constituintes do óleo essencial de orégano foram o 4-terpineol (27,03%), γ-terpineno (20,04%), β-cimeno (6,34%), e do alho, o dialil trissulfeto (38,81%), dialil dissulfeto (25,23%), metil alil trissulfeto (12,52%). Os resultados dos testes in vitro sobre as bactérias S. Choleraesuis e P. aeruginosa indicaram a formação de halo de inibição e revelaram o efeito inibitório para os referidos óleos, exceto para o óleo de A. sativum frente a P. aeruginosa. Para os fungos A. flavus, A. niger e P. simplicissimum a inibição do crescimento micelial provocada pelo óleo essencial de orégano ocorreu a partir das concentrações de 0,10, 0,03 e 0,05 µg mL-1, respectivamente, sendo que a CMI para o óleo de alho iniciou-se a partir da concentração 0,03 µg mL-1 para todas as espécies de fungos. Foi possível verificar que os óleos possuem efeito inibitório sobre os microrganismos estudados, sendo, portanto, uma alternativa no controle microbiológico de alimentos.

Antifungal activity using medicinal plant extracts against pathogens of coffee tree / Atividade antifúngica utilizando extratos de plantas medicinais contra fitopatógenos do cafeeiro

Generally, the medicinal plants have antifungal substances that can be used for the plant protection against phytopathogens. The objective of this study was to know the efficiency of aqueous extracts from medicinal plants against the major etiological agents of coffee tree. The aqueous extracts used were extracted from bulbs of Allium sativum, leaves of Vernonia polysphaera, Cymbopogon citratus, Cymbopogon nardus, Cordia verbenacea, Eucalyptus citriodora, Ricinus communis, Azadirachta indica, Piper hispidinervum and flower buds of Syzygium aromaticum. The etiological agents considered for this study were Cercospora coffeicola, Colletotrichum gloeosporioides, Fusarium oxysporum, Phoma tarda, Rhizoctonia solani and Hemileia vastatrix. The screening for harmful extracts was done based on mycelial growth and conidial germination inhibition. All experiments performed were in vitro conditions. The inhibition of mycelial growth was performed mixing the extracts with the PDA. This mixture was poured in Petri dishes. On the center of the dishes was added one PDA disc with mycelium. It was incubated in a chamber set to 25ºC. The evaluation was done daily by measuring the mycelial growth. The germination assessment was also performed with Petri dishes containing agar-water medium at 2%. These were incubated at 25ºC for 24 hours. After this period the interruption of germination was performed using lactoglycerol. The experiments were conducted in a completely randomized design. The most effective plant extracts against the micelial growth and conidial germination were V. polysphaera, S. aromaticum and A. sativum.

Geralmente, as plantas medicinais têm substâncias antifúngicas que podem ser utilizadas para a proteção das plantas contra fitopatógenos. O objetivo deste estudo foi conhecer a eficiência de extratos aquosos de plantas medicinais contra os principais agentes etiológicos do cafeeiro. Os extratos aquosos utilizados foram extraídos de bulbos de Allium sativum, folhas de Vernonia polysphaera, Cymbopogon citratus, Cymbopogon nardus, Cordia verbenacea, Eucalyptus citriodora, Ricinus communis, Azadirachta indica, Piper hispidinervum e botões florais de Syzygium aromaticum. Os agentes etiológicos considerados neste estudo foram Cercospora coffeicola, Colletotrichum gloeosporioides, Fusarium oxysporum, Phoma tarda, Rhizoctonia solani e Hemileia vastatrix. A triagem dos extratos foi realizada com base no crescimento micelial e na inibição da germinação de conídios. Todos os experimentos foram realizados em condições in vitro. A inibição do crescimento micelial foi realizada misturando-se os extratos com PDA. Esta mistura foi vertida em placas de Petri. No centro das placas foi adicionado um disco de PDA com micélio. Incubou-se em câmara programada para 25°C. A avaliação foi feita diariamente através da medição do crescimento micelial. O experimento sobre a germinação também foi realizado com placas com meio ágar-água a 2%. Estas foram incubadas durante 24 horas. Após este período, a interrupção da germinação foi realizada utilizando lactoglicerol. Os experimentos foram conduzidos em delineamento inteiramente casualizado. Os extratos de plantas mais eficazes contra o crescimento micelial e germinação de conídios foram V. polysphaera, S. aromaticum e A. sativum.

Association of CD28 and CTLA-4 gene polymorphisms with aggressive periodontitis in Brazilians.

OBJECTIVE: Susceptibility to and severity of periodontal disease is influenced by gene polymorphisms related to the immune response. Co-stimulatory molecules, such as CD28 and CTLA-4, are critical in the development of such responses. Our hypothesis is that polymorphisms in genes that code for these molecules may be associated with periodontitis. The aim of the study was to investigate the association between +17 (T/C) CD28 and +49 (A/G) CTLA-4 gene polymorphisms and periodontitis in Brazilians. MATERIALS AND METHODS: Genomic DNA was obtained from oral swabs of 424 individuals categorized into three groups (control group, aggressive, and chronic periodontitis) considering clinical parameters such as probing depth and clinical attachment loss. The genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: There was an association between the T(-) genotype of the CD28 polymorphism and aggressive periodontitis (P = 0.04). Moreover, the A(+) genotype for CTLA-4 was associated with greater clinical attachment loss in non-smokers with aggressive periodontitis (P = 0.006, OR = 16.25, CI = 2.25-117.11). CONCLUSIONS: These findings show that T(-) in CD28 + 17 (T/C) and the A(+) in CTLA-4 +49 (A/G) genotypes are associated with susceptibility to aggressive periodontal disease. Thus, our study highlights these polymorphisms as potential genetic susceptibility markers of periodontitis in Brazilians.

Odontogenic tumours in children and adolescents: a collaborative study of 431 cases.

This study describes the oral and maxillofacial pathological characteristics of a series of odontogenic tumours in children and adolescents from three Brazilian reference centres. The records were reviewed for all odontogenic tumours in patients up to 18 years old based on criteria proposed by the World Health Organization (WHO) in 2005. Data concerning sex, age, skin colour and tumour location were collected and plotted. Four hundred and thirty one odontogenic tumours in children and adolescents were found, accounting for 37.5% of the total number of odontogenic tumours diagnosed. Benign tumours were predominant (99.8% of the cases), and odontoma was the most frequent type (41.4%), followed by keratocystic odontogenic tumours (25.5%) and ameloblastoma (14.6%). Odontogenic tumours were rarely detected in early childhood, and their prevalence increased with age. An almost equal distribution was observed with respect to sex and the site of the lesions. This study is the largest reported retrospective analysis describing odontogenic tumours in children and adolescents to date. The authors detected some variation in the relative frequency of odontogenic tumours compared with similar reports. Additional studies should be conducted based on the new WHO classification and predetermined age parameters to enable comparative analysis among different worldwide populations.

Extratos de plantas de cobertura no desenvolvimento de Sclerotinia sclerotiorum / Plant extracts of cover crops in the development of Sclerotinia sclerotiorum

Extratos vegetais podem interferir no ciclo de vida dos fitopatógenos por promover ou inibir o desenvolvimento. Com base nessa propriedade, este trabalho foi realizado com o objetivo de estudar o efeito dos extratos aquosos das plantas de cobertura crotalária (Crotalaria juncea), braquiária (Brachiaria ruziziensis), capim-mombaça (Panicum maximum cv. mombaça), milheto (Pennisetum glaucum), feijão-guandu-anão (Cajanus cajan) e estilosantes (Stylosanthes capitata x Stylosanthes macrocephala) sobre o crescimento micelial, germinação carpogênica e micelial dos escleródios e germinação dos ascósporos do fungo Sclerotinia sclerotiorum. As etapas do experimento foram desenvolvidas em casa de vegetação, câmaras de crescimento e laboratório. As concentrações dos extratos utilizadas foram de 1%, 5%, 10% e 25%. Para verificar a influência dos extratos sobre a germinação micelial e carpogênica dos escleródios, foi utilizada somente a concentração de 25%. Os resultados sobre o crescimento micelial mostraram que o extrato da planta de Stylosanthes sp. inibiu o desenvolvimento do patógeno apenas na concentração de 25%. Para a germinação micelial dos escleródios, verificou-se que os extratos induziram a germinação 12 horas após a instalação do experimento, exceto o Stylosanthes sp. Na germinação dos ascósporos, apenas o extrato de Stylosanthes sp. impediu a germinação. (AU)

PLANT EXTRACTS OF COVER CROPS IN THE DEVELOPMENT OF SCLEROTINIA SCLEROTIORUM. Plant extracts may interfere with the life cycle of pathogens by promoting or inhibiting development. Based on this property, the present study aimed to evaluate the effect of aqueous extracts of the cover plants crotalaria (Crotalaria juncea), brachiaria (Brachiaria ruziziensis), panicum maximum grass (Panicum maximum cv. mombaça), millet (Pennisetum glaucum), pigeon pea dwarf bean (Cajanus cajan.) and stylosanthes (Stylosanthes macrocephala x Stylosanthes capitata) on the mycelial growth, mycelial and carpogenic germination of sclerotia, and ascospore germination of Sclerotinia sclerotiorum. The stages of the experiment were carried out in a greenhouse, growth chambers and laboratory. The concentrations of the extracts used were 1%, 5%, 10% and 25%. To assess the influence of extracts on the mycelial and carpogenic germination of the sclerotia only the concentration of 25% was used. The results on the mycelial growth showed that the Stylosanthessp. plant extract was effective for inhibiting the pathogen only at concentration of 25%. For mycelial germination of sclerotia, it was found that with the exception of Stylosanthes sp. the extracts induced germination 12 hours after the experiment. In regard to the germination of ascospores, only the extract from Stylosanthes sp. successfully prevented germination. (AU)

Extratos de plantas de cobertura no desenvolvimento de Sclerotinia sclerotiorum / Plant extracts of cover crops in the development of Sclerotinia sclerotiorum

Extratos vegetais podem interferir no ciclo de vida dos fitopatógenos por promover ou inibir o desenvolvimento. Com base nessa propriedade, este trabalho foi realizado com o objetivo de estudar o efeito dos extratos aquosos das plantas de cobertura crotalária (Crotalaria juncea), braquiária (Brachiaria ruziziensis), capim-mombaça (Panicum maximum cv. mombaça), milheto (Pennisetum glaucum), feijão-guandu-anão (Cajanus cajan) e estilosantes (Stylosanthes capitata x Stylosanthes macrocephala) sobre o crescimento micelial, germinação carpogênica e micelial dos escleródios e germinação dos ascósporos do fungo Sclerotinia sclerotiorum. As etapas do experimento foram desenvolvidas em casa de vegetação, câmaras de crescimento e laboratório. As concentrações dos extratos utilizadas foram de 1%, 5%, 10% e 25%. Para verificar a influência dos extratos sobre a germinação micelial e carpogênica dos escleródios, foi utilizada somente a concentração de 25%. Os resultados sobre o crescimento micelial mostraram que o extrato da planta de Stylosanthes sp. inibiu o desenvolvimento do patógeno apenas na concentração de 25%. Para a germinação micelial dos escleródios, verificou-se que os extratos induziram a germinação 12 horas após a instalação do experimento, exceto o Stylosanthes sp. Na germinação dos ascósporos, apenas o extrato de Stylosanthes sp. impediu a germinação.

PLANT EXTRACTS OF COVER CROPS IN THE DEVELOPMENT OF SCLEROTINIA SCLEROTIORUM. Plant extracts may interfere with the life cycle of pathogens by promoting or inhibiting development. Based on this property, the present study aimed to evaluate the effect of aqueous extracts of the cover plants crotalaria (Crotalaria juncea), brachiaria (Brachiaria ruziziensis), panicum maximum grass (Panicum maximum cv. mombaça), millet (Pennisetum glaucum), pigeon pea dwarf bean (Cajanus cajan.) and stylosanthes (Stylosanthes macrocephala x Stylosanthes capitata) on the mycelial growth, mycelial and carpogenic germination of sclerotia, and ascospore germination of Sclerotinia sclerotiorum. The stages of the experiment were carried out in a greenhouse, growth chambers and laboratory. The concentrations of the extracts used were 1%, 5%, 10% and 25%. To assess the influence of extracts on the mycelial and carpogenic germination of the sclerotia only the concentration of 25% was used. The results on the mycelial growth showed that the Stylosanthessp. plant extract was effective for inhibiting the pathogen only at concentration of 25%. For mycelial germination of sclerotia, it was found that with the exception of Stylosanthes sp. the extracts induced germination 12 hours after the experiment. In regard to the germination of ascospores, only the extract from Stylosanthes sp. successfully prevented germination.

Extratos de plantas de cobertura no desenvolvimento de Sclerotinia sclerotiorum

Plant extracts may interfere with the life cycle of pathogens by promoting or inhibiting development. Based on this property, the present study aimed to evaluate the effect of aqueous extracts of the COVER plants crotalaria (Crotalaria juncea), brachiaria (Brachiaria ruziziensis), panicum maximum grass (Panicum maximum cv. mombaça), millet (Pennisetum glaucum), pigeon pea dwarf bean (Cajanus cajan.) and stylosanthes (Stylosanthes macrocephala x Stylosanthes capitata) on the mycelial growth, mycelial and carpogenic germination of sclerotia, and ascospore germination of Sclerotinia sclerotiorum. The stages of the experiment were carried out in a greenhouse, growth chambers and laboratory. The concentrations of the extracts used were 1%, 5%, 10% and 25%. To assess the influence of extracts on the mycelial and carpogenic germination of the sclerotia only the concentration of 25% was used. The results on the mycelial growth showed that the Stylosanthes sp. plant extract was effective for inhibiting the pathogen only at concentration of 25%. For mycelial germination of sclerotia, it was found that with the exception of Stylosanthes sp. the extracts induced germination 12 hours after the experiment. In regard to the germination of ascospores, only the extract from Stylosanthes sp. successfully prevented germination.

Extratos vegetais podem interferir no ciclo de vida dos fitopatógenos por promover ou inibir o desenvolvimento. Com base nessa propriedade, este trabalho foi realizado com o objetivo de estudar o efeito dos extratos aquosos das plantas de cobertura crotalária (Crotalaria juncea), braquiária (Brachiaria ruziziensis), capim-mombaça (Panicum maximum cv. mombaça), milheto (Pennisetum glaucum), feijão-guandu-anão (Cajanus cajan) e estilosantes (Stylosanthes capitata x Stylosanthes macrocephala) sobre o crescimento micelial, germinação carpogênica e micelial dos escleródios e germinação dos ascósporos do fungo Sclerotinia sclerotiorum. As etapas do experimento foram desenvolvidas em casa de vegetação, câmaras de crescimento e laboratório. As concentrações dos extratos utilizadas foram de 1%, 5%, 10% e 25%. Para verificar a influência dos extratos sobre a germinação micelial e carpogênica dos escleródios, foi utilizada somente a concentração de 25%. Os resultados sobre o crescimento micelial mostraram que o extrato da planta de Stylosanthes sp. inibiu o desenvolvimento do patógeno apenas na concentração de 25%. Para a germinação micelial dos escleródios, verificou-se que os extratos induziram a germinação 12 horas após a instalação do experimento, exceto o Stylosanthes sp. Na germinação dos ascósporos, apenas o extrato de Stylosanthes sp. impediu a germinação.

Efeito do silício na intensidade da cercosporiose e na nutrição mineral de mudas de cafeeiro / Effect of silicon on the intensity of brown eye spot and on the mineral nutrition of coffee seedlings

O presente trabalho objetivou estudar o efeito do silício na intensidade da cercosporiose e na nutrição mineral de mudas de cafeeiro. No experimento 1, testou-se seis doses de ácido silícico (0, 0,5; 1; 2; 4 e 6 g kg­1 de solo) em mudas da cultivar Catuaí Vermelho IAC 99 inoculadas com o fungo Cercospora coffeicola. No experimento 2, foram realizadas microanálises de raios-X para a avaliação de nutrientes presentes nas folhas das mudas de cafeeiro das cultivares Topázio MG1190 e Icatu Precoce IAC 3282, inoculadas e não inoculadas com C. coffeicola, com e sem silicato de cálcio (1 g kg-1 de solo). Com o aumento das doses de ácido silícico observou-se redução na área abaixo da curva de progresso do número de lesões por folha (AACPNLF), redução linear nos teores foliares de magnésio e fósforo e aumento nos teores de enxofre e cobre. Os teores foliares de boro apresentaram comportamento quadrático, diminuindo com o aumento das doses de ácido silícico e aumentando a partir da dose de 4 g kg-1 de solo. Em microanálise de raio X, mudas de cafeeiro com cercosporiose apresentam menores picos de potássio e cálcio, independente da cultivar utilizada.

Our objective was to verify the effect of silicon on the intensity of brown eye spot and on the mineral nutrition of coffee seedlings. In the first experiment, 6 doses of silicic acid (0, 0.5, 1, 2, 4 and 6 g kg-1 soil) were tested using a complete randomized block design with 4 replicates with 6 coffee seedlings cultivar Catuaí Vermelho IAC 99 inoculated with the fungus Cercospora coffeicola. In the second experiment, X-ray microanalysis in a scanning electron microscope was performed on 2 coffee cultivars (Topazio MG1190 and Icatu Precoce IAC 3282), inoculated and non-inoculated with C. coffeicola, treated and untreated with calcium silicate (1 g kg-1 of soil). With the increase of the silicic acid doses, there was observed a reduction in the area under the disease progress curve of the number of lesions per leaf (AUPCNLL), coupled with a linear reduction in the foliar contents of magnesium and phosphorus as well as an increase in the contents of sulfur and copper. The foliar contents of boron presented a quadratic behavior, decreasing with the increase of silicic acid and increasing with the dose of 4 g kg-1 of soil. In X-ray microanalysis, coffee seedlings with brown eye spot presented lower peaks of potassium and calcium, regardless of the cultivar used.

Óleo essencial de Baccharis tridentata Vahl: composição química, atividade antioxidante e fungitóxica, e caracterização morfológica das estruturas secretoras por microscopia eletrônica de varredura / Baccharis tridentata Vahl essential oil: chemical composition, and antioxidant and fungitoxic activities and morphological characterization of secretory structures by scanning electron microscopy

Este trabalho teve como objetivos avaliar a composição química do óleo essencial de Baccharis tridentata Vahl, as atividades antioxidante e fungitóxica, e estudar a morfologia das estruturas secretoras do óleo essencial presentes na superfície foliar por meio de microscopia eletrônica de varredura (MEV). A extração do óleo essencial foi realizada por hidrodestilação, as análises quantitativas e qualitativas foram executadas por meio de cromatografia em fase gasosa com detector de ionização de chamas (FID) e acoplada à espectrometria de massas, respectivamente. A atividade antioxidante foi realizada empregando-se os métodos de redução do radical estável DPPH e o ensaio de oxidação do sistema β-caroteno/ácido linoleico. As atividades fungitóxicas foram avaliadas utilizando o teste bioanalítico in vitro, sobre a inibição do crescimento micelial dos fitopatógenos Fusarium oxysporum, Colletotrichum gloeosporioides e Rhizoctonia solani. A composição química revelou a presença de 28 compostos, sendo o α-tujeno (22,93 por cento) o constituinte majoritário; não foi observada atividade antioxidante por meio dos ensaios utilizados, no entanto, observou-se atividade fungitóxica sobre o crescimento micelial dos fitopatógenos estudados. Já os estudos da superfície foliar por MEV revelaram a presença de tricomas glandulares em ambas as superfícies abaxial e adaxial.

This study aimed to evaluate the chemical composition and the antioxidant and fungitoxic activities of Baccharis tridentata essential oil, as well as to study the morphology of its secretory structures present on the leaf surface by scanning electron microscopy (SEM). The essential oil was extracted by hydrodistillation; the quantitative and qualitative analyses were performed on a gas chromatograph equipped with a flame ionization detector (FID) and coupled to a mass spectrometer, respectively. The antioxidant activity was determined by the methods of reduction of the DPPH stable radical and oxidation of the β-carotene/linoleic acid system. Fungitoxic activities were assessed by the in vitro bioanalytical test on the inhibition of the mycelial growth of the plant pathogens Fusarium oxysporum, Colletotrichum gloeosporioides and Rhizoctonia solani. The chemical composition revealed the presence of 28 compounds, with α-thujene (22.93 percent) as the major constituent. No antioxidant activity was observed in the tests used; however, there was fungitoxic activity against the mycelial growth of plant pathogens. Leaf surface studies by SEM revealed the presence of glandular trichomes on both abaxial and adaxial surfaces.

Bioatividade de óleos essenciais no controle de Botrytis cinerea isolado de morangueiro / Essential oils bioactivity in strawberry grey mould control

Objetivou-se avaliar o uso de óleos essenciais sobre isolados de Botrytis cinerea, causador do mofo cinzento em morangueiro. Foram testados óleos essenciais de capim-limão, palmarosa, citronela, cravo, canela, menta, lavanda, tangerina, eucalipto, melaleuca, alecrim e laranja, todos estes analisados em cromatógrafo a gás acoplado a detector de massas, para identificação dos principais componentes dos óleos. Foram avaliados o crescimento micelial, produção e germinação de conídios de B. cinerea, com a incorporação do óleo no meio de cultura. Realizou-se ainda uma avaliação de voláteis e a eficiência de óleos em isolado resistente a fungicida. Para cada teste, diferentes óleos apresentaram eficiência, contudo capim limão, palmarosa, canela e menta demonstraram os melhores efeitos em todos os testes realizados. Todos os tratamentos a base de óleos demonstraram efeito semelhante a um fungicida recomendado para a cultura, a base de tiofanato metílico. Dois tratamentos mostraram-se efetivos no caso de isolado resistente (óleo de capim limão e de canela). Óleos essenciais mostram-se como opção promissora para o desenvolvimento de possíveis produtos fitossanitários para o manejo de doenças em plantas.

This study aimed evaluates essential oils in Botrytis cinerea isolates growth, which causes gray mould on strawberry. Were tested essential oils of lemon grass, palmrose, citronella, clove, cinnamon, mint, lavender, tangerine, eucalyptus, tea tree, rosemary and orange. The oils were analyzed in gas chromatograph attached to mass detector for identifying the mainly oils components. Were evaluated mycelial growth, conidia production and conidia germination of B. cinerea, with oil incorporation in culture medium. Were conducted an evaluation of oils volatile components and the efficiency of oils in fungicide-resistant isolate. For each test, different oils showed efficiency, however lemongrass, palmarosa, cinnamon and mint showed the best effects in all tests. All treatments demonstrated similar effects to recommended fungicide for the culture, which had methyl thiophanate. Two treatments (lemon grass and cinnamon oils), were effective against resistant isolate. Essential oils are shown as promising option for development of possible product for plant disease management.

EFEITO DO SILÍCIO NA INTENSIDADE DA CERCOSPORIOSE E NA NUTRIÇÃO MINERAL DE MUDAS DE CAFEEIRO

ABSTRACT Our objective was to verify the effect of silicon on the intensity of brown eye spot and on the mineral nutrition of coffee seedlings. In the first experiment, 6 doses of silicic acid (0, 0.5, 1, 2, 4 and 6 g kg-1 soil) were tested using a complete randomized block design with 4 replicates with 6 coffee seedlings cultivar Catuaí Vermelho IAC 99 inoculated with the fungus Cercospora coffeicola. In the second experiment, X-ray microanalysis in a scanning electron microscope was performed on 2 coffee cultivars (Topazio MG1190 and Icatu Precoce IAC 3282), inoculated and non-inoculated with C. coffeicola, treated and untreated with calcium silicate (1 g kg-1 of soil). With the increase of the silicic acid doses, there was observed a reduction in the area under the disease progress curve of the number of lesions per leaf (AUPCNLL), coupled with a linear reduction in the foliar contents of magnesium and phosphorus as well as an increase in the contents of sulfur and copper. The foliar contents of boron presented a quadratic behavior, decreasing with the increase of silicic acid and increasing with the dose of 4 g kg-1 of soil. In X-ray microanalysis, coffee seedlings with brown eye spot presented lower peaks of potassium and calcium, regardless of the cultivar used.

RESUMO O presente trabalho objetivou estudar o efeito do silício na intensidade da cercosporiose e na nutrição mineral de mudas de cafeeiro. No experimento 1, testou-se seis doses de ácido silícico (0, 0,5; 1; 2; 4 e 6 g kg1 de solo) em mudas da cultivar Catuaí Vermelho IAC 99 inoculadas com o fungo Cercospora coffeicola. No experimento 2, foram realizadas microanálises de raios-X para a avaliação de nutrientes presentes nas folhas das mudas de cafeeiro das cultivares Topázio MG1190 e Icatu Precoce IAC 3282, inoculadas e não inoculadas com C. coffeicola, com e sem silicato de cálcio (1 g kg-1 de solo). Com o aumento das doses de ácido silícico observou-se redução na área abaixo da curva de progresso do número de lesões por folha (AACPNLF), redução linear nos teores foliares de magnésio e fósforo e aumento nos teores de enxofre e cobre. Os teores foliares de boro apresentaram comportamento quadrático, diminuindo com o aumento das doses de ácido silícico e aumentando a partir da dose de 4 g kg-1 de solo. Em microanálise de raio X, mudas de cafeeiro com cercosporiose apresentam menores picos de potássio e cálcio, independente da cultivar utilizada.

Recruitment of CD8(+) T cells expressing granzyme A is associated with lesion progression in human cutaneous leishmaniasis.

Human infection with Leishmania braziliensis leads to the establishment of cutaneous leishmaniasis (CL), characterized by the appearance of skin lesions that progress from nonulcerated to ulcerated forms. Our goal was to characterize the immunological kinetics associated with this progression, comparing the cellular composition, cytokines and granzyme expression between lesions of patients with early (E-CL) and late stages (L-CL) of CL. Histopathological analysis showed that lesions from L-CL had more exuberant inflammatory infiltrate as compared to E-CL. Although E-CL and L-CL lesions were predominantly mononuclear, lesions from E-CL patients presented higher neutrophil and eosinophil counts than L-CL. While percentages of CD4(+) and of CD68(+) cells were slightly higher in L-CL, a fivefold increase of CD8(+) cells was observed in L-CL, as compared to E-CL. Moreover, CD8(+) T-cells from L-CL expressed significantly higher levels of granzyme A than E-CL. Interestingly, granzyme A expression was positively correlated with intensity of the inflammatory infiltrate in L-CL but not E-CL. Lastly, percentages of IFN-gamma(+) and IL-10(+) cells were higher in L-CL as compared to E-CL, with CD4(+) T-cells and CD68(+) monocytes as the main sources of these cytokines, respectively. These results suggest that recruitment of CD8(+) granzyme A(+) T cells is involved in lesion progression in human CL.

Novel mutations in the SH3BP2 gene associated with sporadic central giant cell lesions and cherubism.

Central giant cell lesion (CGCL) is a reactive bone lesion that occurs mainly in the mandible, characterized by the multinucleated osteoclast-like giant cells in a background of oval to spindle-shaped mononuclear cells. The etiology is unknown and occurs more commonly in young adults. Cherubism, a rare disease found predominantly in females has histologic characteristics indistinguishable from those of CGCL and is caused by mutations mostly present in exon 9 of the SH3BP2 gene. In this study, we investigated four cases of CGCL and one case of cherubism. DNA was extracted from peripheral blood and tumor tissue and all coding and flanking regions of the SH3BP2 amplified by PCR and directly sequenced to identify underlying mutations. Two novel mutations were found; a heterozygous missense mutation c.1442A>T (Q481L) in exon 11 in one sporadic case of CGCL and a heterozygous germline and tumor tissue missense mutation c.320C>T (T107M) in exon 4 in one patient with cherubism. These findings open a new window to investigate the possible relationship between the pathogenesis of the cherubism and CGCL.

Phenotypic and functional characteristics of CD28+ and CD28- cells from chagasic patients: distinct repertoire and cytokine expression.

Chronic human Chagas' disease ranges from an asymptomatic to a severe cardiac clinical form. The involvement of the host's immune response in the development and maintenance of chagasic pathology has been demonstrated by several groups. We have shown that activated T-cells lacking CD28 expression are increased in the peripheral blood of chagasic patients (CP), suggesting a relationship between these cells and disease. In order to better characterize this cell population, determining their possible role in immunoregulation of human Chagas' disease, we evaluated the expression of TCR-Vbeta regions 2, 3.1, 5, 8 and 17, as well as the expression of IFN-gamma, TNF-alpha, IL-4 and IL-10 by CD28+ and CD28- cells from polarized indeterminate and cardiac CP. Flow cytometric analysis demonstrated equivalent TCR-Vbeta usage between CD4+CD28+ and CD4+CD28- cells from all groups (chagasic and healthy controls). However, there was a predominance of Vbeta5 expression in the CD28+ and CD28- populations in the CP groups (indeterminate and cardiac). Interestingly, CD8+CD28- cells from CP, but not from nonchagasic individuals, displayed a reduced frequency of most analysed Vbetas when compared with the CD8+CD28+ subpopulation. Comparison of V-beta expression in CD28+ or CD28- cell populations among individuals from different groups also showed several interesting differences. Functionally, cardiac CP displayed a higher frequency of IFN-gamma, TNF-alpha and IL-4 producing lymphocytes than indeterminate CP. Correlation analysis between the frequency of cytokine expressing cells, and the frequency of CD4+ T-cells with differential expression of CD28 demonstrated that CD4+CD28- T-cells were positively correlated with TNF-alpha in cardiac and with IL-10 in indeterminate CP, suggesting that these cells might have an important regulatory role in human Chagas' disease.

An extreme case of cherubism.

We describe an 8-year-old boy who presented with severe facial swelling. This progressed rapidly and 17 months later he died of gastrointestinal and pulmonary infections. The diagnosis was initially brown tumour associated with hyperparathyroidism, but this was revised in the light of laboratory investigations that were within the reference ranges, and normal appearance of the parathyroids on exploration to that of an extreme case of cherubism that behaved in a locally aggressive manner.

Cysticercosis of the oral cavity: report of seven cases.

Cysticercosis is a condition in which a human acts as the intermediate host of the pork tapeworm Taenia solium. Although cysticercosis is a common disease in some regions of the world and can occur in any body site, oral lesions are rare. This paper reviews the natural history of the disease and reports on seven cases of oral cysticercosis.

Evaluation of p53, PCNA, Ki-67, MDM2 and AgNOR in oral peripheral and central giant cell lesions.

OBJECTIVES: Peripheral giant cell lesion (PGCL) and central giant cell lesion (CGCL) of the jaws have a distinct clinical behaviour. Whether such biological differences are supported by a distinct pattern of proliferation markers or cell cycle associated proteins expression is not known. Therefore the purpose of the present study was to compare the immunohistochemical expression of p53, MDM2, Ki-67, PCNA and the histochemical expression of argyrophilic nuclear organiser region (AgNOR) on PGCL and CGCL of the jaws. MATERIALS AND METHODS: Paraffin wax blocks of 14 cases of PGCL and 12 cases of CGCL were retrieved. A biotin-streptavidin amplified system was used for identification of the antigens. The AgNOR number was also evaluated. RESULTS: Ki-67 immunoreactivity was greater in the mononuclear cells of PGCL compared to CGCL. PCNA and AgNOR staining were similar in PGCL and CGCL. Prominent MDM2 immunoreactivity was observed in all tissues investigated. By contrast, there was no p53 immunoreactivity. CONCLUSIONS: Although CGCL present a more aggressive clinical behaviour, it has a decreased proliferative activity compared to PGCL. Finally, p53, MDM2, PCNA, Ki-67 immunohistochemical expression and AgNOR histochemical expression do not reflect their distinct biological behaviour.

Cell cycle-associated proteins in melanotic neuroectodermal tumor of infancy.

OBJECTIVE: The purpose of the present study was to compare the immunohistochemical expression of cell cycle-associated proteins in neuroblastic and melanocytic cell populations of melanotic neuroectodermal tumor of infancy. STUDY DESIGN: Three cases of melanotic neuroectodermal tumor of infancy were selected. The immunohistochemical expression of MDM-2, p53, proliferating cell nuclear antigen, cyclin D1, and cyclin A was assessed through use of the streptavidin-biotin-peroxidase complex technique. RESULTS: Positive immunostaining for MDM-2, proliferating cell nuclear antigen, cyclin D1, and cyclin A was occasionally observed in the large melanin-containing epithelioid cells. CONCLUSIONS: These data suggest that MDM-2 expression may be important for the development of melanotic neuroectodermal tumor of infancy and that the melanocytic cell population, not the neuroblastic one, is the proliferative component of the tumor.