Hexagonal boron nitride tunnel barriers grown on graphite by high temperature molecular beam epitaxy

We demonstrate direct epitaxial growth of high-quality hexagonal boron nitride (hBN) layers on graphite using high-temperature plasma-assisted molecular beam epitaxy. Atomic force microscopy reveals mono- and few-layer island growth, while conducting atomic force microscopy shows that the grown hBN has a resistance which increases exponentially with the number of layers, and has electrical properties comparable to exfoliated hBN. X-ray photoelectron spectroscopy, Raman microscopy and spectroscopic ellipsometry measurements on hBN confirm the formation of sp2-bonded hBN and a band gap of 5.9 ± 0.1 eV with no chemical intermixing with graphite. We also observe hexagonal moiré patterns with a period of 15 nm, consistent with the alignment of the hBN lattice and the graphite substrate

Retention of progenitor cell phenotype in otospheres from guinea pig and mouse cochlea

Abstract\ud \ud Background\ud Culturing otospheres from dissociated organ of Corti is an appropriate starting point aiming at the development of cell therapy for hair cell loss. Although guinea pigs have been widely used as an excellent experimental model for studying the biology of the inner ear, the mouse cochlea has been more suitable for yielding otospheres in vitro. The aim of this study was to compare conditions and outcomes of otosphere suspension cultures from dissociated organ of Corti of either mouse or guinea pig at postnatal day three (P3), and to evaluate the guinea pig as a potential cochlea donor for preclinical cell therapy.\ud \ud \ud Methods\ud Organs of Corti were surgically isolated from P3 guinea pig or mouse cochlea, dissociated and cultivated under non-adherent conditions. Cultures were maintained in serum-free DMEM:F12 medium, supplemented with epidermal growth factor (EGF) plus either basic fibroblast growth factor (bFGF) or transforming growth factor alpha (TGFα). Immunofluorescence assays were conducted for phenotype characterization.\ud \ud \ud Results\ud The TGFα group presented a number of spheres significantly higher than the bFGF group. Although mouse cultures yielded more cells per sphere than guinea pig cultures, sox2 and nestin distributed similarly in otosphere cells from both organisms. We present evidence that otospheres retain properties of inner ear progenitor cells such as self-renewal, proliferation, and differentiation into hair cells or supporting cells.\ud \ud \ud Conclusions\ud Dissociated guinea pig cochlea produced otospheres in vitro, expressing sox2 and nestin similarly to mouse otospheres. Our data is supporting evidence for the presence of inner ear progenitor cells in the postnatal guinea pig. However, there is limited viability for these cells in neonatal guinea pig cochlea when compared to the differentiation potential observed for the mouse organ of Corti at the same developmental stage

IL-4 Amplifies the Pro-Inflammatory Effect of Adenosine in Human Mast Cells by Changing Expression Levels of Adenosine Receptors

Adenosine inhalation produces immediate bronchoconstriction in asthmatics but not in normal subjects. The bronchospastic effect of adenosine is largely mediated through adenosine-induced mast cell activation, the mechanism of which is poorly understood due to limitations in culturing human primary mast cells. Here, we show that human umbilical cord blood -derived mast cells incubated with the Th2 cytokine IL-4 develop increased sensitivity to adenosine. Potentiation of anti-IgE- induced and calcium ionophore/PMA-induced degranulation was augmented in mast cells cultured with IL-4, and this effect was reduced or abolished by pre-treatment with A2BsiRNA and selective A2B receptor antagonists, respectively. IL-4 incubation resulted in the increased expression of A2B and reduced expression of A2A adenosine receptors on human mast cells. These results suggest that Th2 cytokines in the asthmatic lung may alter adenosine receptor expression on airway mast cells to promote increased responsiveness to adenosine

Estabelecimento de alecrim-pimenta in vitro In vitro establishment of Lippia sidoides Cham

O alecrim-pimenta (Lippia sidoides Cham.) é um arbusto nativo da região do semi-árido do nordeste brasileiro, cujo óleo essencial possui elevado valor comercial devido aos seus constituintes majoritários, o timol e o carvacrol, de potente propriedade antimicrobiana e anti-séptica. Avaliou-se os efeitos de concentrações e tempos de imersão em hipoclorito de sódio, de meios de cultivo e da utilização de antibiótico e antioxidantes no estabelecimento in vitro de alecrim-pimenta. Os experimentos foram conduzidos em delineamento inteiramente casualizado. Foram avaliadas as concentrações 0,2; 0,4; 0,6 e 0,8% de hipoclorito de sódio e 8; 12; 16 e 20 minutos de imersão, em esquema fatorial 4 x 4; as concentrações 0; 50; 100; 150 e 200 mg L-1 do antibiótico cefatoxima sódica; os meios-de-cultura MS, B5 e WPM; e o efeito de antioxidantes (PVP: 0,5 e 2 g L-1; e carvão ativado: 3 e 12 g L-1). A concentração de 0,8% de hipoclorito de sódio proporcionou um número significativamente maior (p<0,01) de folhas por broto: 1,88. Para as demais características, não houve efeito significativo da concentração de hipoclorito de sódio: a contaminação variou de 33,7 a 50,6%; o número de brotos formados, de 1,17 a 1,65 e, o número de folhas por explante, de 1,77 a 3,07. Apesar de não ter havido diferença significativa para os tempos de imersão, os tempos de 12 e 16 minutos tendem a proporcionar menor contaminação, enquanto o aumento de 16 para 20 minutos tende a induzir uma redução do número de brotos formados (de 1,52 para 1,22), número de folhas por explante (de 2,62 para 1,81) e número de folhas por brotos (de 1,70 para 1,24). A utilização de cefotaxima sódica reduziu significativamente a contaminação bacteriana (55,23%, no tratamento testemunha; 9,99%, na concentração de 200 mg L-1), elevando a sobrevivência dos explantes de 0 (testemunha) para 37,32% (200 mg L-1). Os meios-de-cultura proporcionaram resultados estatisticamente iguais. Todos os antioxidantes avaliados, mesmo nas concentrações mais baixas, reduziram a oxidação de 50% na testemunha para menos de 10%. Os resultados indicam o uso de 0,8% de hipoclorito de sódio, com imersão de 16 minutos, 200 mg L-1 de cefotaxima sódica, os meios WPM, MS ou B5 e 3,0 g L-1 de carvão ativado ou 0,5 g L-1 de PVP para o estabelecimento in vitro de segmentos nodais de alecrim-pimenta.<br>Lippia sidoides Cham. is a native shrub from the semi-arid region of Northeast Brazil. Its essential oil has high commercial value, due to the major compounds thymol and carvacrol, which have strong antimicrobial and antiseptic properties. The effect of concentrations and immersion time in sodium hypochlorite, culture media, the use of antibiotic and antioxidants on in vitro establishment of L. sidoides were evaluated. The assays were conducted in a completely randomized design. We evaluated the concentrations 0.2; 0.4; 0.6 and 0.8% of sodium hypochlorite and 8; 12; 16 and 20 minutes of immersion, in a 4 x 4 factorial scheme; the concentrations 0; 50; 100; 150 and 200 mg L-1 of cefotaxime sodium; the medium cultures MS, B5 and WPM; and the effect of antioxidants (PVP: 0.5 and 2 g L-1; and activated charcoal: 3 and 12 g L-1). The concentration of 0.8% of sodium hypochlorite resulted in a significantly higher (p<0.01) number of leaves per shoot: 1.88. For the other characteristics we did not observe any significant effect of sodium hypochlorite concentrations: the contamination varied from 33.7 to 50.6%; the number of new shoots varied from 1.17 to 1.65, and the number of leaves per explant varied from 1.77 to 3.07. Although we did not observe significant difference for immersion times, 12 and 16 minutes of immersion tend to result in minor contamination. Increasing the immersion time from 16 to 20 minutes tends to induce a reduction of new shoots (form 1.52 to 1.22), number of leaves per explant (from 2.62 to 1.81) and number of leaves per shoot (from 1.70 to 1.24). The use of cefotaxime sodium reduced significatively the bacterial contamination (55.23% at the control treatment; 9.99% at the 200 mg L-1 concentration), increasing the survival of explants from 0 (control) to 37.32% (200 mg L-1). The medium cultures offered statistically identical results. All the evaluated antioxidants, even at the lowest concentrations, reduced the oxidation from 50% (control) to as little as 10%. For in vitro establishment of L. sidoides nodal segments, the results indicate immersion of explants for 16 minutes in a 0.8% sodium hypochlorite solution, 200 mg L-1 of cefotaxime sodium, WPM, MS or B5 culture medium, and 3.0 g L-1 of activated charcoal or 0.5 g L-1 of PVP