Plastic compression of a collagen gel forms a much improved scaffold for ocular surface tissue engineering over conventional collagen gels

We compare the use of plastically compressed collagen gels to conventional collagen gels as scaffolds onto which corneal limbal epithelial cells (LECs) are seeded to construct an artificial corneal epithelium. LECs were isolated from bovine corneas (limbus) and seeded onto either conventional uncompressed or novel compressed collagen gels and grown in culture. Scanning electron microscopy (SEM) results showed that fibers within the uncompressed gel were loose and irregularly ordered, whereas the fibers within the compressed gel were densely packed and more evenly arranged. Quantitative analysis of LECs expansion across the surface of the two gels showed similar growth rates (p > 0.05). Under SEM, the LECs, expanded on uncompressed gels, showed a rough and heterogeneous morphology, whereas on the compressed gel, the cells displayed a smooth and homogeneous morphology. Transmission electron microscopy (TEM) results showed the compressed scaffold to contain collagen fibers of regular diameter and similar orientation resembling collagen fibers within the normal cornea. TEM and light microscopy also showed that cell–cell and cell–matrix attachment, stratification, and cell density were superior in LECs expanded upon compressed collagen gels. This study demonstrated that the compressed collagen gel was an excellent biomaterial scaffold highly suited to the construction of an artificial corneal epithelium and a significant improvement upon conventional collagen gels.

The plastic self organising map

A novel extension to Kohonen's self-organising map, called the plastic self organising map (PSOM), is presented. PSOM is unlike any other network because it only has one phase of operation. The PSOM does not go through a training cycle before testing, like the SOM does and its variants. Each pattern is thus treated identically for all time. The algorithm uses a graph structure to represent data and can add or remove neurons to learn dynamic nonstationary pattern sets. The network is tested on a real world radar application and an artificial nonstationary problem.

A simple device for multiplex ELISA made from melt-extruded plastic microcapillary film

We present a simple device for multiplex quantitative enzyme-linked immunosorbant assays (ELISA) made from a novel melt-extruded microcapillary film (MCF) containing a parallel array of 200µm capillaries along its length. To make ELISA devices different protein antigens or antibodies were immobilised inside individual microcapillaries within long reels of MCF extruded from fluorinated ethylene propylene (FEP). Short pieces of coated film were cut and interfaced with a pipette, allowing sequential uptake of samples and detection solutions into all capillaries from a reagent well. As well as being simple to produce, these FEP MCF devices have excellent light transmittance allowing direct optical interrogation of the capillaries for simple signal quantification. Proof of concept experiments demonstrate both quantitative and multiplex assays in FEP MCF devices using a standard direct ELISA procedure and read using a flatbed scanner. This new multiplex immunoassay platform should find applications ranging from lab detection to point-of-care and field diagnostics.

A novel bogie design made of glass fibre reinforced plastic

This paper presents a completely new design of a bogie-frame made of glass fibre reinforced composites and its performance under various loading conditions predicted by finite element analysis. The bogie consists of two frames, with one placed on top of the other, and two axle ties connecting the axles. Each frame consists of two side arms and a transom between. The top frame is thinner and more compliant and has a higher curvature compared with the bottom frame. Variable vertical stiffness can be achieved before and after the contact between the two frames at the central section of the bogie to cope with different load levels. Finite element analysis played a very important role in the design of this structure. Stiffness and stress levels of the full scale bogie presented in this paper under various loading conditions have been predicted by using Marc provided by MSC Software. In order to verify the finite element analysis (FEA) models, a fifth scale prototype of the bogie has been made and tested under quasi-static loading conditions. Results of testing on the fifth scale bogie have been used to fine tune details like contact and friction in the fifth scale FEA models. These conditions were then applied to the full scale models. Finite element analysis results show that the stress levels in all directions are low compared with material strengths.

Characterization and antimicrobial efficacy against E. coli of a helium/air plasma at atmospheric pressure created in a plastic package

A plasma source, sustained by the application of a floating high voltage (±15 kV) to parallel-plate electrodes at 50 Hz, has been achieved in a helium/air mixture at atmospheric pressure (P = 105 Pa) contained in a zip-locked plastic package placed in the electrode gap. Some of the physical and antimicrobial properties of this apparatus were established with a view to ascertain its performance as a prototype for the disinfection of fresh produce. The current–voltage (I–V) and charge–voltage (Q–V) characteristics of the system were measured as a function of gap distance d, in the range (3 × 103 ≤ Pd ≤ 1.0 × 104 Pa m). The electrical measurements showed this plasma source to exhibit the characteristic behaviour of a dielectric barrier discharge in the filamentary mode and its properties could be accurately interpreted by the two-capacitance in series model. The power consumed by the discharge and the reduced field strength were found to decrease quadratically from 12.0 W to 4.5 W and linearly from 140 Td to 50 Td, respectively, in the range studied. Emission spectra of the discharge were recorded on a relative intensity scale and the dominant spectral features could be assigned to strong vibrational bands in the 2+ and 1− systems of N2 and ${\rm N}_2^+$ , respectively, with other weak signatures from the NO and OH radicals and the N+, He and O atomic species. Absolute spectral intensities were also recorded and interpreted by comparison with the non-equilibrium synthetic spectra generated by the computer code SPECAIR. At an inter-electrode gap of 0.04 m, this comparison yielded typical values for the electron, vibrational and translational (gas) temperatures of (4980 ± 100) K, (2700 ± 200) K and (300 ± 100) K, respectively and an electron density of 1.0 × 1017 m−3. A Boltzmann plot also provided a value of (3200 ± 200 K) for the vibrational temperature. The antimicrobial efficacy was assessed by studying the resistance of both Escherichia coli K12 its isogenic mutants in soxR, soxS, oxyR, rpoS and dnaK selected to identify possible cellular responses and targets related with 5 min exposure to the active gas in proximity of, but not directly in, the path of the discharge filaments. Both the parent strain and mutants populations were significantly reduced by more than 1.5 log cycles in these conditions, showing the potential of the system. Post-treatment storage studies showed that some transcription regulators and specific genes related to oxidative stress play an important role in the E. coli repair mechanism and that plasma exposure affects specific cell regulator systems.

Plastic and evolutionary responses to heat stress in a temperate dung fly: negative correlation between basal and induced heat tolerance?

Extreme weather events such as heat waves are becoming more frequent and intense. Populations can cope with elevated heat stress by evolving higher basal heat tolerance (evolutionary response) and/or stronger induced heat tolerance (plastic response). However, there is ongoing debate about whether basal and induced heat tolerance are negatively correlated and whether adaptive potential in heat tolerance is sufficient under ongoing climate warming. To evaluate the evolutionary potential of basal and induced heat tolerance, we performed experimental evolution on a temperate source 4 population of the dung fly Sepsis punctum. Offspring of flies adapted to three thermal selection regimes (Hot, Cold and Reference) were subjected to acute heat stress after having been exposed to either a hot-acclimation or non-acclimation pretreatment. As different traits may respond differently to temperature stress, several physiological and life history traits were assessed. Condition dependence of the response was evaluated by exposing juveniles to different levels of developmental (food restriction/rearing density) stress. Heat knockdown times were highest, whereas acclimation effects were lowest in the Hot selection regime, indicating a negative association between basal and induced heat tolerance. However, survival, adult longevity, fecundity and fertility did not show such a pattern. Acclimation had positive effects in heat-shocked flies, but in the absence of heat stress hot-acclimated flies had reduced life spans relative to nonacclimated ones, thereby revealing a potential cost of acclimation. Moreover, body size positively affected heat tolerance and unstressed individuals were less prone to heat stress than stressed flies, offering support for energetic costs associated with heat tolerance. Overall, our results indicate that heat tolerance of temperate insects can evolve under rising temperatures, but this response could be limited by a negative relationship between basal and induced thermotolerance, and may involve some but not other fitness-related traits.

Transport of Pirarucu Arapaima gigas juveniles in plastic bag

Este estudo examinou as respostas de estresse de juvenis de pirarucu transportados em sistema fechado. Pirarucu (Arapaima gigas) é um peixe nativo da bacia Amazônica, da família Osteoglossidae que possui respiração aérea obrigatória. Foi realizado um transporte de curta duração (6 h) em sacos de polietileno inflados com ar atmosférico (grupo ar) ou com oxigênio puro (grupo oxi). O oxigênio dissolvido foi o único parâmetro de qualidade da água que apresentou diferença estatística entre os grupos, e como esperado, o oxigênio estava supersaturado para o grupo oxi. Não houve mortalidade após o transporte em ambos os grupos. Os peixes se alimentaram 36 h após o transporte e apresentaram um consumo de ração habitual após 72 h. As respostas fisiológicas foram semelhantes nos dois grupos. O cortisol não apresentou mudança significativa durante o período de amostragem. Ao contrário da maioria das espécies, os valores de cortisol se apresentaram inalterados nos dois grupos durante a amostragem, enquanto a glicose teve um aumento significativo até 12 h após o transporte. Os resultados mostram que o transporte de pirarucu em sacos de polietileno pode ser realizado com ar atmosférico ou oxigênio puro, uma vez que as respostas de estresse, a qualidade da água e o comportamento alimentar após 36 h foram similar entre os grupos.

Soil, water and nutrient losses by interrill erosion from green cane cultivation

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

Availability of soil water under tillage systems, mulch management and citrus rootstocks

The increased availability of soil water is important for the management of non-irrigated orange orchards. The objective of this study was to evaluate the availability of soil water in a Haplorthox (Rhodic Ferralsol) under different tillage systems used for orchard plantation, mulch management and rootstocks in a "Pera" orange orchard in northwest Parana, Brazil. An experiment in a split-split-plot design was established in 2002, in an area cultivated with Brachiaria brizantha grass in which three tillage systems (no tillage, conventional tillage and strip-tillage) were used for orchard plantation. This grass was mowed twice a year between the rows, representing two mulch managements in the split plots (no mulching and mulching in the plant rows). The split-split-plots were represented by two rootstocks ("Rangpur" lime and "Cleopatra" mandarin). The soil water content in the plant rows was evaluated in the 0-20 cm layer in 2007 and at 0-20 and 20-40 cm in 2008-2009. The effect of soil tillage systems prior to implantation of orange orchards on soil water availability was less pronounced than mulching and the rootstocks. The soil water availability was lower when "Pera" orange trees were grafted on "Cleopatra" mandarin than on "Rangpur" lime rootstocks. Mulching had a positive influence on soil water availability in the sandy surface layer (020 cm) and sandy clay loam subsurface (20-40 cm) of the soil in the spring. The production of B. brizantha between the rows and residue disposal in the plant rows as mulch increased water availability to the "Pera" orange trees.

Uso de malhas pigmentadas e mulching em túneis para cultivo de rúcula: efeito no ambiente e nas plantas modelo

O uso de malhas pigmentadas em cultivos de hortaliças folhosas permite a melhor adequação do ambiente às plantas, com destaque para a rúcula. Essa hortaliça vem conquistando maior espaço no mercado consumidor brasileiro desde o final da década de 90. Essa pesquisa teve por objetivo avaliar as condições ambientais proporcionadas pelo uso de telas pigmentadas na cobertura de túneis de cultivo, relacionando com as respostas agronômicas da rúcula, cultivada dentro desses túneis sobre diferentes coberturas de solo. As coberturas de túneis foram: a Chromatinet® azul, Chromatinet® vermelha, tela aluminizada prata, Sombrite® 50% e filme plástico transparente de polietileno de baixa densidade de 100µ. As coberturas de solo, também denominadas mulchings, dentro dos túneis foram: o filme plástico de polietileno de cor preta; de polietileno de dupla-face nas cores preta e branca, com a face branca voltada para cima; casca de arroz e a ausência de mulching. O delineamento utilizado foi o de blocos ao acaso com 24 tratamentos e três repetições. Nas condições do experimento, o emprego de algumas coberturas de túnel e de solo modificou o ambiente e melhorou as respostas agronômicas das plantas de rúcula.

Dinâmica de transposição de herbicida através de palha de aveia-preta utilizando diferentes pontas de pulverização

A transposição da palha por herbicidas aplicados em pré ou pós-emergência durante a aplicação é determinante na sua eficiência, dinâmica e impacto ambiental. O experimento foi conduzido no Núcleo de Pesquisas Avançadas em Matologia - FCA/UNESP, campus de Botucatu-SP, tendo como objetivo avaliar o desempenho de diferentes modelos de pontas de pulverização na transposição em quantidades crescentes de palha de aveia-preta (Avena strigosa). Os tratamentos foram constituídos pelo monitoramento do traçador corante Azul Brilhante (FDC-1) a 3.000 ppm, pulverizado com as pontas de pulverização XR11002-VS, TJ60-11002VS, FL-5VS, DG11002-VS, TXVK-8, TT11002-VP e AI11002-VS, utilizando, respectivamente, as pressões de trabalho de 1,4; 2,0; 1,5; 2,0; 4,9; 3,0 e 3,0 kgf cm² e volume de calda de 200, 200, 428, 200, 213 e 270 L ha-1 sobre quantidades de 0, 1, 2, 4, 6, 8, 10 e 12 t ha-1 de palha de aveia-preta. O delineamento utilizado foi o inteiramente casualizado, com sete tratamentos e cinco repetições, as quais foram constituídas de caixas plásticas com palha acondicionada sobre um fundo falso de área conhecida, sendo este lavado após as aplicações, para posterior quantificação do traçador em espectrofotometria. O modelo de Mitscherlich simplificado (Y = 10 ^ (2 - (C*X))) mostrou ajuste satisfatório para os dados originais de traçador que transpôs a palha, apresentando coeficientes de determinação (R²) elevados, oscilando entre 0,9782 e 0,9971. Todos os modelos de pontas de pulverização mostraram-se similares na transposição da palha pelo traçador. As porcentagens médias de transposição foram de 43,00; 18,77; 3,73; 0,78; 0,17; 0,04 e 0,01% para as quantidades de 1, 2, 4, 6, 8, 10 e 12 t ha-1 de palha, respectivamente.

Effects of Different Periods of Clomazone plus Hexazinona Permanence in Soil and Sugarcane Mulch before the Occurrence of Rain on Weed Control Efficacy

The objective of this work was to evaluate the efficacy of the formulated mixture clomazone and hexazinone applied in soil or combined with sugarcane mulch, after different periods of permanence without the occurrence of rain. The experiment was carried out in vases under greenhouse conditions in Botucatu, São Paulo, Brazil. The weeds Brachiaria decumbens, Ipomoea grandifolia, Ipomoea hederifolia and Euphorbia heterophylla were sown and covered or not with sugarcane mulch. The experiment was arranged in a complete randomized design with four repetitions. The treatments were displaced in a 3x6 factorial scheme, with the factors being six periods without rain (0, 3, 7, 15, 30, and 60 days) and three modes of herbicide application(on the soil without sugarcane mulch, on and under sugarcane mulch). Control evaluations were carried out at 10, 21, 35 and 42 days after the occurrence of rain. The clomazone + hexazinone mixture promoted an excellent weed control for all the species studied when applied on, under, or without sugarcane mulch. However, the control levels tended to reduce for periods over 60 days without rain.

MIGRATION MEASUREMENTS FROM PLASTIC PACKAGING: A SIMULATION STUDY ON INFLUENCE OF INITIAL CONCENTRATION PROFILE

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)