Microneedle Arrays Allow Lower Microbial Penetration Than Hypodermic Needles In Vitro

Methods: In this study we determined, for the first time, the ability of microorganisms to traverse microneedle-induced holes using two different in vitro models.

Results: When employing Silescol® membranes, the numbers of Candida albicans, Pseudomonas aeruginosa and Staphylococcus epidermidis crossing the membranes were an order of magnitude lower when the membranes were punctured by microneedles rather than a 21G hypodermic needle. Apart from the movement of C. albicans across hypodermic needle-punctured membranes, where 40.2% of the microbial load on control membranes permeated the barrier over 24 h, the numbers of permeating microorganisms was less than 5% of the original microbial load on control membranes. Experiments employing excised porcine skin and radiolabelled microorganisms showed that the numbers of microorganisms penetrating skin beyond the stratum corneum were approximately an order of magnitude greater than the numbers crossing Silescol® membranes in the corresponding experiments. Approximately 103?cfu of each microorganism adhered to hypodermic needles during insertion. The numbers of microorganisms adhering to MN arrays were an order of magnitude higher in each case.

Conclusion: We have shown here that microneedle puncture resulted in significantly less microbial penetration than did hypodermic needle puncture and that no microorganisms crossed the viable epidermis in microneedle—punctured skin, in contrast to needle-punctured skin. Given the antimicrobial properties of skin, it is, therefore, likely that application of microneedle arrays to skin in an appropriate manner would not cause either local or systemic infection in normal circumstances in immune-competent patients. In supporting widespread clinical use of microneedle-based delivery systems, appropriate animal studies are now needed to conclusively demonstrate this in vivo. Safety in patients will be enhanced by aseptic or sterile manufacture and by fabricating microneedles from self-disabling materials (e.g. dissolving or biodegradable polymers) to prevent inappropriate or accidental reuse.

Re-organisation of the cytoskeleton during developmental programmed cell death in Picea abies embryos

Cell and tissue patterning in plant embryo development is well documented. Moreover, it has recently been shown that successful embryogenesis is reliant on programmed cell death (PCD). The cytoskeleton governs cell morphogenesis. However, surprisingly little is known about the role of the cytoskeleton in plant embryogenesis and associated PCD. We have used the gymnosperm, Picea abies , somatic embryogenesis model system to address this question. Formation of the apical-basal embryonic pattern in P. abies proceeds through the establishment of three major cell types: the meristematic cells of the embryonal mass on one pole and the terminally differentiated suspensor cells on the other, separated by the embryonal tube cells. The organisation of microtubules and F-actin changes successively from the embryonal mass towards the distal end of the embryo suspensor. The microtubule arrays appear normal in the embryonal mass cells, but the microtubule network is partially disorganised in the embryonal tube cells and the microtubules disrupted in the suspensor cells. In the same embryos, the microtubule-associated protein, MAP-65, is bound only to organised microtubules. In contrast, in a developmentally arrested cell line, which is incapable of normal embryonic pattern formation, MAP-65 does not bind the cortical microtubules and we suggest that this is a criterion for proembryogenic masses (PEMs) to passage into early embryogeny. In embryos, the organisation of F-actin gradually changes from a fine network in the embryonal mass cells to thick cables in the suspensor cells in which the microtubule network is completely degraded. F-actin de-polymerisation drugs abolish normal embryonic pattern formation and associated PCD in the suspensor, strongly suggesting that the actin network is vital in this PCD pathway.

Consulta en ABIES.

Se presenta en formato pdf una pequeña guía didáctica sobre cómo realizar la consulta del catálogo bibliográfico con el programa de gestión bibliotecaria Abies. La guía está diseñada para la formación de los usuarios de las bibliotecas escolares.

La informatización de la biblioteca escolar : el programa Abies 2.0.

Se trata de un manual para la instalación y manejo del programa de gestión de bibliotecas ABIES 2.0 y tiene como objetivo ayudar al profesorado encargado de la biblioteca escolar en las cuestiones técnicas y organizativas.

Specific leaf area and leaf area index in developing stands of Fagus sylvatica L. and Picea abies Karst

European beech (Fagus sylvatica L.) and Norway spruce (Picea abies Karst.) are two of the most ecologically and economically important forest tree species in Europe. These two species co-occur in many locations in Europe, leading to direct competition for canopy space. Foliage characteristics of two naturally regenerated pure stands of beech and spruce with fully closed canopies were contrasted to assess the dynamic relationship between foliage adaptability to shading, stand LAI and tree growth. We found that individual leaf size is far more conservative in spruce than in beech. Individual leaf and needle area was larger at the top than at the bottom of the canopy in both species. Inverse relationship was found for specific leaf area (SLA), highest SLA values were found at lowest light availability under the canopy. There was no difference in leaf area index (LAI) between the two stands, however LAI increased from 10.8 to 14.6 m2m-2 between 2009 and 2011. Dominant trees of both species were more efficient in converting foliage mass or area to produce stem biomass, although this relationship changed with age and was species-specific. Overall, we found larger foliage plasticity in beech than in spruce in relation to light conditions, indicating larger capacity to exploit niche openings.

Protection of spruce seedlings against pine weevil attacks by treatment of seeds or seedlings with nicotinamide, nicotinic acid and jasmonic acid

Sustainable methods are required to protect newly planted tree seedlings from insect herbivore attack. To this end, here Norway spruce (Picea abies (L.) Karst.) seeds were treated with 2.5 mM nicotinamide (NIC), 2.5 mM nicotinic acid (NIA), 3 mM jasmonic acid (JA) or 0.2 mM 5-azacytidine (5-Aza), and 6-month-old seedlings grown from these seeds were planted at a reforestation area in central Sweden. Attack by pine weevils (Hylobius abietis) was reduced by 50 per cent by NIC treatment, 62.5 per cent by JA treatment and 25 per cent by 5-Aza treatment, when compared with seedlings grown from untreated seeds. Watering 18-month-old spruce seedlings with 2 mM NIC or 2 mM NIA did reduce attack during the first season in the field by 40 and 53 per cent, respectively, compared with untreated plants. Girdling was also reduced by the different treatments. Analysis of conifer seedlings treated with 5-Aza points at a possible involvement of epigenetic mechanisms in this defensive capacity. This is supported by a reduced level of DNA methylation in the needles of young spruce seedlings grown in a greenhouse from NIC-treated seeds. Seed treatment for seedling defense potentiation is simple, inexpensive and also a new approach for forestry with many potential applications.

Development of a molecular test to determine the vitality status of Norway spruce (Picea abies) seedlings during frozen storage

In boreal forest regions, a great portion of forest tree seedlings are stored indoors in late autumn to prevent seedlings from outdoor winter damage. For seedlings to be able to survive in storage it is crucial that they store well and can cope with the dark and cold storage environment. The aim of this study was to search for genes that can determine the vitality status of Norway spruce (Picea abies (L.) Karst.) seedlings during frozen storage. Furthermore, the sensitivity of the ColdNSure (TM) test, a gene activity test that predicts storability was assessed. The storability of seedlings was tested biweekly by evaluating damage with the gene activity test and the electrolyte leakage test after freezing seedlings to -25 A degrees C (the SELdiff-25 method). In parallel, seedlings were frozen stored at -3 A degrees C. According to both methods, seedlings were considered storable from week 41. This also corresponded to the post storage results determined at the end of the storage period. In order to identify vitality indicators, Next Generation Sequencing (NGS) was performed on bud samples collected during storage. Comparing physiological post storage data to gene analysis data revealed numerous vitality related genes. To validate the results, a second trial was performed. In this trial, gene activity was better in predicting seedling storability than the conventional freezing test; this indicates a high sensitivity level of this molecular assay. For multiple indicators a clear switch between damaged and vital seedlings was observed. A collection of indicators will be used in the future development of a commercial vitality test.

PHENYLBUTANOID AND TAXANE-LIKE METABOLITES FROM NEEDLES OF TAXUS-BREVIFOLIA

The structures for brevifoliol and three baccatin VI derivatives were revised to possess the novel 11(15 --> 1)-abeotaxane tricylic skeleton based on X-ray crystallographic studies. Four more new rearranged taxane derivatives related to brevifoliol and the unusual phenylbutanoid, (-)-rhododendrol, were isolated from the needles of Taxus breuifolia. Their structures were established as 10 beta-benzoxy-5 alpha-(3'-dimethylamino-3'-phenyl)-propanoxy-1 beta-hydroxy- 7 beta,9 alpha,13 alpha-triacetoxy-11(15 --> 1)-abeotaxa-4(20),11-diene; 10 beta-benzoxy-1 beta-hydroxy-5 alpha-(3'-methylamino-3'-phenyl)- propanoxy-7 beta,9 alpha,l3 alpha-triacetoxy-11(15 --> 1)-abeotaxa-4(20),11-diene; 10 beta-benzoxy-5 alpha-cinnamoxy-1 beta-hydroxy-7 beta,9 alpha,13 alpha-triacetoxy-1 1(15 --> 1)-abeotaxa-4(20),11-diene; 10 beta-benzoxy-1 beta,5 alpha-dihydroxy-7 beta,9 alpha,13 alpha-triacetoxy-11(15 --> 1)-abeotaxa-4(20),11-diene and 2(R)-hydroxy-4-(4'-hydroxyphenyl)-butane.

Zirconia needles synthesized inside hexagonal swollen liquid crystals

We report the synthesis of zirconia microneedles by the direct nucleation of particles inside a hexagonal swollen liquid crystal (SLC) (cell parameter a = 27 nm) prepared by mixing with the proper ratio, an aqueous solution of sulfated zirconium colloids, a cationic surfactant (cetylpyridinium chloride), cychlohexane as swelling agent with an oil over water ratio of 2.5 (vol.), and 1-pentanol as cosurfactant. After a slow crystallogenesis that can be enhanced by an initial induction step under moderate temperature, particles in the centimeter range can be obtained, with a very high shape ratio (over 100). These particles are made of crystalline octahydrate zirconium oxychloride containing pores of 20 nm diameter, aligned along the main axis of the liquid crystal, as the fingerprint of the oil cylinders present in the hexagonal phase. The morphology of these particles confirms that the shaping mechanism is based on true liquid crystal templating (TLCT). Further thermal treatment of these particles, after extraction from the SLC, leads to the crystallization of zirconia with the same needlelike morphology as the zirconium oxychloride.

Controlling the growth of zirconia needles precursor from a liquid crystal template

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efficacy of four irrigation needles in cleaning the apical third of root canals

This study aimed to evaluate the influence of irrigation needle gauge and design, and the final root canal diameter on the apical cleaning efficacy. Twelve human mandibular incisors were used. At different stages of root canal widening (sizes 20, 30 and 40 K-files), root canals were filled with radiopaque contrast medium. Four different needles were evaluated: 23G with side opening, 22G with apical opening, 30G with side opening and 30G with apical opening. Irrigation was carried out with 2 mL distilled water. The same tooth was radiographed with a digital system several times to assess the four types of needle in those three stages of canal widening. Pre-irrigation (canals filled with contrast) and post-irrigation (canals with remaining contrast) images were submitted to digital subtraction using the Adobe Photoshop CS4 program. Pre-irrigation (filled with contrast) and subtracted (cleaned by irrigation) areas were outlined by a trained and blinded operator using the Image Tool 3.0 software. Their ratio was calculated to express the percentage of apical cleaning in each stage of canal widening (sizes 20, 30 and 40 K-files) with each of the four needles. Data obtained were subjected to one-way ANOVA and Tukey's tests. The 30G needles with side and apical opening promoted better apical cleaning at all stages of root canal widening (p<0.05). In conclusion, smaller diameter needles were more efficacious in cleaning the apical third of the root canals, regardless of their design.