Effect of aluminium on migratory and invasive properties of MCF-7 human breast cancer cells in culture

Aluminium (Al) has been measured in human breast tissue, nipple aspirate fluid and breast cyst fluid, and recent studies have shown that at tissue concentrations, aluminium can induce DNA damage and suspension growth in human breast epithelial cells. This paper demonstrates for the first time that exposure to aluminium can also increase migratory and invasive properties of MCF-7 human breast cancer cells. Long-term (32 weeks) but not short-term (1 week) exposure of MCF-7 cells to 10-4M aluminium chloride or 10-4M aluminium chlorohydrate increased motility of the cells as measured by live cell imaging (cumulative length moved by individual cells), by a wound healing assay and by migration in real time through 8m pores of a membrane using xCELLigence technology. Long-term exposure (37weeks) to 10-4M aluminium chloride or 10-4M aluminium chlorohydrate also increased the ability of MCF-7 cells to invade through a matrigel layer as measured in real time using the xCELLigence system. Although molecular mechanisms remain to be characterized, the ability of aluminium salts to increase migratory and invasive properties of MCF-7 cells suggests that the presence of aluminium in the human breast could influence metastatic processes. This is important because mortality from breast cancer arises mainly from tumour spread rather than from the presence of a primary tumour in the breast.

Implication of methodological uncertainties for mid-Holocene sea surface temperature reconstructions

We present and examine a multi-sensor global compilation of mid-Holocene (MH) sea surface temperatures (SST), based on Mg/Ca and alkenone palaeothermometry and reconstructions obtained using planktonic foraminifera and organic-walled dinoflagellate cyst census counts. We assess the uncertainties originating from using different methodologies and evaluate the potential of MH SST reconstructions as a benchmark for climate-model simulations. The comparison between different analytical approaches (time frame, baseline climate) shows the choice of time window for the MH has a negligible effect on the reconstructed SST pattern, but the choice of baseline climate affects both the magnitude and spatial pattern of the reconstructed SSTs. Comparison of the SST reconstructions made using different sensors shows significant discrepancies at a regional scale, with uncertainties often exceeding the reconstructed SST anomaly. Apparent patterns in SST may largely be a reflection of the use of different sensors in different regions. Overall, the uncertainties associated with the SST reconstructions are generally larger than the MH anomalies. Thus, the SST data currently available cannot serve as a target for benchmarking model simulations. Further evaluations of potential subsurface and/or seasonal artifacts that may contribute to obscure the MH SST reconstructions are urgently needed to provide reliable benchmarks for model evaluations.

Orbital and millenial-scale environmental changes between 64 and 25 ka BP recorded in Black Sea sediments

High-resolution pollen and dinoflagellate cyst records from sediment core M72/5-25-GC1 were used to reconstruct vegetation dynamics in northern Anatolia and surface conditions of the Black Sea between 64 and 20 ka BP. During this period, the dominance of Artemisia in the pollen record indicates a steppe landscape and arid climate conditions. However, the concomitant presence of temperate arboreal pollen suggests the existence of glacial refugia in northern Anatolia. Long-term glacial vegetation dynamics reveal two major arid phases ~64–55 and 40–32 ka BP, and two major humid phases ~54–45 and 28–20 ka BP, correlating with higher and lower summer insolation, respectively. Dansgaard–Oeschger (D–O) cycles are clearly indicated by the 25-GC1 pollen record. Greenland interstadials are characterized by a marked increase in temperate tree pollen, indicating a spread of forests due to warm/wet conditions in northern Anatolia, whereas Greenland stadials reveal cold and arid conditions as indicated by spread of xerophytic biomes. There is evidence for a phase lag of ~500 to 1500 yr between initial warming and forest expansion, possibly due to successive changes in atmospheric circulation in the North Atlantic sector. The dominance of Pyxidinopsis psilata and Spiniferites cruciformis in the dinocyst record indicates brackish Black Sea conditions during the entire glacial period. The decrease of marine indicators (marine dinocysts, acritarchs) at ~54 ka BP and increase of freshwater algae (Pediastrum, Botryococcus) from 32 to 25 ka BP reveals freshening of the Black Sea surface water. This freshening is possibly related to humid phases in the region, to connection between Caspian Sea and Black Sea, to seasonal freshening by floating ice, and/or to closer position of river mouths due to low sea level. In the southern Black Sea, Greenland interstadials are clearly indicated by high dinocyst concentrations and calcium carbonate content, as a result of an increase in primary productivity. Heinrich events show a similar impact on the environment in the northern Anatolia/Black Sea region as Greenland stadials.

High Matrix Metalloproteinase Activity Is a Hallmark of Periapical Granulomas

Introduction: The inability to distinguish periapical cysts from granulomas before performing root canal treatment leads to uncertainty in treatment outcomes because cysts have lower healing rates. Searching for differential expression of molecules within cysts or granulomas could provide information with regard to the identity of the lesion or suggest mechanistic differences that may form the basis for future therapeutic intervention. Thus, we investigated whether granulomas and cysts exhibit differential expression of extracellular matrix (ECM) molecules. Methods: Human periapical granulomas, periapical cysts, and healthy periodontal ligament tissues were used to investigate the differential expression of ECM molecules by microarray analysis. Because matrix metalloproteinases (MMP) showed the highest differential expression in the microarray analysis, MMPs were further examined by in situ zymography and immunohistochemistry. Data were analyzed by using one-way analysis of variance followed by the Tu-key test. Results: We observed that cysts and granulomas differentially expressed several ECM molecules, especially those from the MMP family. Compared with cysts, granulomas exhibited higher MMP enzymatic activity in areas stained for MMP-9. These areas were composed of polymorphonuclear cells (PMNs) in contrast to cysts. Similarly, MMP-13 was expressed by a greater number of cells in granulomas compared with cysts. Conclusion: Our findings indicate that high enzymatic MIMP activity in PMNs together with MMP-9 and MMP-13 stained cells could be a molecular signature of granulomas unlike periapical cysts. (J Endod 2009;35:1234-1242)

Growth, encystment and survival of Acanthamoeba castellanii grazing on different bacteria

Free-living amoebae of the genus Acanthamoeba are widely distributed in soil and water collections, where trophozoites (vegetative, multiplicative stages) feed mainly by phagocytosis and thus control bacterial populations in the environment. Here, we examined the growth, encystment and survival of Acanthamoeba castellanii receiving different bacteria (Escherichia coli, Pseudomonas aeruginosa, Enterobacter cloacae, Bacillus subtilis, Bacillus megaterium, Micrococcus luteus, and Staphylococcus aureus) in nonnutrient saline. All bacteria assayed induced a dose-dependent proliferative response, in most cases maximized with a bacterial dose of 1 x 10(9) mL(-1); except for M. luteus, trophozoites grew better with viable than with heat-killed bacteria. In addition, Acanthamoeba growth was improved by adding bacteria on alternate days. Single-dose experiments indicated a temporal association between the growth of trophozoite and bacterial consumption, and higher consumption of M. luteus, E. coli and P. aeruginosa, bacterial species that allowed the highest trophozoite yields. Long-term Acanthamoeba-bacteria incubation revealed that encystment was significantly delayed by almost all the bacteria assayed (including S. aureus, which elicited a poor growth response) and that the presence of bacteria markedly increased cyst yield; final cyst recovery clearly depended on both the dose and the type of the bacterium given, being much higher with E. coli, M. luteus and P. aeruginosa.

Estudo in vitro da habilidade predatória de monacrosporium robustum sobre heterodera glycines

O fungo nematófago Monacrosporium robustum foi detectado, isolado e identificado pela primeira vez de solos do Brasil, em maio de 1999, no Laboratório de Nematologia do Departamento de Fitossanidade da Universidade Estadual Paulista (UNESP), Campus de Jaboticabal, São Paulo, tendo sido o potencial como agente de biocontrole do nematóide de cisto da soja, Heterodera glycines raça 3 observado ao microscópio eletrônico de varredura (MEV) (Maia & Santos, 1999). Na presente pesquisa, detalhes das estruturas de captura, tamanho, forma e septação dos conídios, bem como nematóides capturados pelo fungo foram documentados. Monacrosporium robustum produz micélio hialino, e as estruturas de captura são constituídas por ramificações adesivas, na forma de protuberâncias verticais que surgem das hifas, medindo, em média, 10 µm de comprimento e 5 µm de diâmetro. Uma substância gelatinosa desprende- se dessas estruturas, ao contato com o nematóide, aprisionando-o. Os conídios do fungo são hialinos, fusóides com dois ou quatro septos, às vezes, cinco. Conídios jovens são asseptados e piriformes. Sob condições de laboratório, esse fungo predou 100% dos ovos e dos juvenis de segundo estádio de H. glycines e formas ativas de Panagrellus sp., no período de 72 h da exposição desses nematóides ao fungo.

Detecção de Heterodera glycines em plantio de soja mediante espectrorradiometria no visível e infravermelho próximo

O ataque do nematóide de cisto da soja, Heterodera glycines, limita o potencial de expansão e maior produtividade de áreas plantadas com soja (Glycine Max). O conhecimento da distribuição espacial desse patógeno na lavoura é fundamental, para elaboração de estratégias de manejo. A área em estudo estava localizada em lavoura de soja, variedade BRS133, localizada no Município de Florínea, SP, com solos naturalmente infestados por H. glycines. Foram obtidas medidas de espectrorradiometria de campo, 112 dias após o plantio, nas regiões do visível e do infravermelho próximo do espectro eletromagnético, a fim de se conhecer o padrão da resposta espectral de plantas atacadas pelo fitonematóide. Paralelamente, foram retiradas amostras de solo e encaminhadas ao Laboratório de Nematologia, Departamento de Fitossanidade da Universidade Estadual Paulista Júlio de Mesquita Filho, Campus de Jaboticabal, onde foram processadas para determinação da densidade populacional do nematóide. As medidas do espectrorradiômetro foram transformadas em índice vegetativo, com diferença normalizada (NDVI), que foi relacionado com a densidade populacional do nematóide, peso da matéria fresca e número de vagens por planta. Observou-se que diferentes densidades de população estão diretamente relacionados com a resposta espectral das plantas expressa, através dos valores do NDVI.

Partial protection against tissue cysts formation in pigs vaccinated with crude rhoptry proteins of Toxoplasma gondii

A vaccine containing crude Toxoplasma gondii rhoptry proteins incorporated in the immunostimulating complexes (ISCOM) adjuvant was tested in pigs for protecting against tissue cyst formation. For this, 38 mixed breed pigs were divided into four groups, G1 (vaccinated challenged, n = 10) received two doses (100 mu g/dose) of the rhoptry vaccine at days 0 and 21, G2 (vaccinated challenged, n = 10) received viable tachyzoites (7 x 10(7)) of the RH strain at day 0, G3 (unvaccinated challenged, n = 10) and G4 (unvaccinated unchallenged, n = 8). Pigs were challenged with 4 x 10(4) VEG strain oocysts 57 days later. The G1 pigs produced high IgG antibody levels in the indirect enzyme-linked immunosorbent assay (ELISA) after the second dose of rhoptry vaccine, but were not clinically protected against a high dose oocyst challenge. Partial protection was observed in G1 at the chronic phase of infection, when compared with G3. Pigs in group 2 developed high antibody levels and were protected against clinic signs. T gondii was not detected in two (G1) and three (G2) pigs by mouse bioassay. The results indicate partial protection in pigs vaccinated with a rhoptry vaccine. (c) 2005 Elsevier B.V. All rights reserved.

Taenia saginata: Production and characterization of monoclonal antibodies against Taenia saginata metacestode antigens

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

Ultrastructural Aspects of Spermatogenesis in Phalloceros caudimaculatus (Teleostei, Poeciliidae)

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

Cisto dentígero: estudo epidemiológico, correlação clinicopatológica e caracterização de uma possível variante inflamatória

The aim of this study was to evaluate the clinical and pathological features of cases diagnosed as dentigerous cyst by the Department of Oral Pathology, School of Dentristy at the Federal University of Rio grande do Norte, attempting the possible correlation between histomorphological findings and epidemiological data contained at the files of the patients, in order to define a suggested variation of lesion named inflammatory dentigerous cyst. It was verified that dentigerous cyst are more frequently present in the earfy three decades of life, with the majority of cases occuring in the second decade (40,740/0 ), and also male (57,41%) and white patients (68,52%) were most affected. In relation to anatomic site, the dentigerous cyst was more prevalent at anterior maxila and posterior mandible, showing, usually, a slow growth pattem. The majority of lesions were asymptomatic and the radiographic observed was frequently na unilocular radiolucency. In regard to the histomorphological analysis, it was noticed that the lesions showed commonly a thin epithelium, with a capsule of fibrous connective tissue, richly vascularized and collagenized with an intense mononuclear inflammatory infiltrate. Finally, clinicopathological was performed and it was find out that cysts that showed a thick epithelium, with a high degree of vascularization and collagenization, intense inflammatory infiltrate in the cystic capsule, were located in the pre-molares region, in patients under 12 years old and the majority showing painfull sintomatology, properly, compatible with inflammatory dentigerous cyst. The findings of the present study indicate that, probably, there is a variant of the dentigerous cyst, and therefore, we suggest the denomination inflammatory follicular cyst for this entity

Expressão imuno-histoquímica das proteínas IFN-γ e TGF-β1 em cistos radiculares e cistos dentígeros

Odontogenic cysts are pathologic cavities covered by odontogenic epithelium and filled by liquid, desquamated cells or other materials. The intraosseous lesions, such as radicular cyst and dentigerous cyst, present a potential of expansion capable of promoting the destruction of the surrounding osseous tissue. The mechanisms related to this process of expansion are the proliferation of cystic epithelium, the increase of the osmolarity of the cystic fluid and the synthesis of reabsorption factors such as IFN-γ and TGF-β1. The aim of this study was to evaluate and compare the immunohistochemical expression of IFN-γ and TGF-β1 between radicular cysts and dentigerous cysts in order to understand the role and behavior of these proteins in the expansion of these cysts. We selected 20 cases of radicular cyst and 20 cases of dentigerous cyst chosen from the files of UFRN s Laboratory of Oral Pathology. After analyzing the clinical data, the cases underwent the routine staining technique (HE) and immunohistochemistry for the appearance of IFN-γ and TGF-β1 in the epithelium and capsule of these cysts. The statistical analysis using the Mann-Whitney test revealed no statistically significant difference in immunoexpression of IFN-γ between the epithelium (p = 0.565) and capsules (p = 0.414) of radicular cysts and dentigerous cysts. Moreover, there was no statistically significant difference of immunoexpression of TGF-β1 between the epithelium (p = 0.620) and capsules (p = 0.056) of radicular cysts and dentigerous cysts. The Wilcoxon test revealed no statistically significant difference between IFN-γ and TGF-β1 imunoexpressions in the epithelium (p = 0.225) and capsules (p = 0.370) of radicular cysts. There was no statistically significant difference between IFN-γ and TGF-β1 imunoexpressions in the epithelium (p = 0.361) of dentigerous cysts. However, there was a statistically significant difference between IFN-γ and TGF-β1 immunoexpressions in the capsule (p = 0.001) of dentigerous cysts, being TGF-β1 the factor which presented the most significant immunoexpression. Given these results, we conclude that there was no difference in immunohistochemical expression of IFN-γ and TGF-β1 between radicular and dentigerous cysts and that TGF-β1 was more significant than the IFN-γ in the capsule of dentigerous cysts

Imunoexpressão do EGFR e da podoplanina em cistos radiculares e dentígeros

The radicular cysts (RCs) and dentigerous (DCs), despite having different etiologies, form a pathological cavity lined by epithelium, which grows due to the buildup of fluid inside, as the surrounding bone is reabsorbed and the epithelium will being induced to proliferate. The epithelial proliferation, which has been identified as one of the key processes in the growth of odontogenic cystic lesions, is influenced by growth factors such as EGFR (epidermal growth receptor factor) and podoplanin (PDPN), many of which may have its production stimulated mainly during inflammatory processes. The objective of this research was to evaluate and compare the immunohistochemical expression of EGFR and PDPN in 30 cases of RCs and 30 cases of DCs, semiquantitatively, in light microscopy, associating it with the degree of inflammation, cellular localization of immunostaining and with the immunostained epithelial layers. Data were statistically analyzed by Chi-square test and Fisher exact test, considering a significance level of 5 %. The results showed high immunoreactivity of both proteins in the lesions studied, only statistically significant difference was observed in immunostaining of PDPN (p=0.033), which proved higher in RCs. The other analyzed parameters showed no relevant significant differences. We conclude that, as EGFR and PDPN showed high immunoreactivity in cystic lesions analyzed, these proteins participate the pathogenesis of these lesions through the epithelial stimulation process, despite having different etiologies. Furthermore, it can infer that the higher immunostaining of PDNP in RCs that DCs showed no distinction indicator between the two lesions, regarding their etiologies, once this protein also showed a considerable expression in DCs, independent of the intensity of the inflammatory infiltrate

Cistos subependimários diagnosticados pelo ultra-som transfontanelar

Objetivos: avaliar a evolução ultra-sonográfica de cistos subependimários (CSE) do ângulo externo dos ventrículos laterais, e a evolução clínica dos pacientes. Comparar os pacientes com CSE isolados e os com CSE associados a outras lesões. Métodos: foram diagnosticados um a vários cistos no ângulo externo de um ou de ambos os ventrículos laterais, por meio de ultra-som transfontanelar (US) de rotina, realizado nos primeiros dias de vida, caracterizando os CSE. Durante o período de 1981-2000, 66 recém-nascidos tiveram CSE evidenciados na UTI neonatal do Hospital de Port-Royal. Foram constituídos dois grupos: G-I, com CSE isolados (n=21), e G-II, com CSE associados a outras lesões (n=45). Resultados: os recém-nascidos do GI apresentaram maior maturidade, melhores condições de nascimento e menor morbidade respiratória em relação a GII. A incidência de malformações congênitas foi elevada em ambos os grupos. Houve baixa taxa de infecção bacteriana e ausência de infecção congênita. Os CSE foram uni ou bilaterais, únicos ou múltiplos (colar de pérolas), sem diferença entre os grupos estudados, e predominaram à esquerda. US seriados foram realizados em 49/66 pacientes (74%), mostrando aumento no tamanho do cisto em 21/49 (45%), no primeiro mês de vida, enquanto 12 CSE (24%) desapareceram. O óbito ocorreu em dez recém-nascidos com lesões neurológica graves (quatro leucomalácias periventriculares, cinco hemorragias peri e intraventriculares), e somente um com hérnia diafragmática não apresentava outras lesões ao US transfontanelar. Conclusões: as características dos CSE não diferiram quando esses estavam associados a outras lesões. O nítido predomínio no lado esquerdo sugere uma etiologia vascular. Foi encontrada uma alta taxa de malformações associadas, alertando para a possibilidade de uma etiologia malformativa. Ambas hipóteses sugerem um desvio de desenvolvimento, e não de uma fetopatia viral.

Aneurisma cerebral e cisto de aracnóide: a propósito de caso com hemorragia intracística

Relatamos um caso de aneurisma da bifurcação da artéria carótida interna, cuja ruptura se deu para dentro de cisto de aracnóide da fissura silviana. em revisão da literatura apenas 3 casos foram descritos. Discutimos ainda os aspectos clínicos atípicos do caso, as características dos achados cirúrgicos e uma correlação etiopatogênica entre as duas patologias.