Добірка наукової літератури з теми "Materiale solubile e cellulato"

Deoiled coconut cake powder (DCCP) was hydrolyzed to reduce the ratio of insoluble/soluble dietary fiber (RIS) by partially converting insoluble dietary fiber to soluble using Celluclast 1.5 L, a commercial cellulase preparation in citrate buffer medium. Firstly, the influence of citrate buffer amount, enzyme concentration, pH, and retention time on the enzymatic hydrolysis efficiency was investigated. Then, response surface methodology (RSM) was employed to optimize the process in which the insoluble and soluble dietary fiber contents were the responses. The results revealed that 10.3 g buffer/g of materials, 3.7 U/g of the materials, and 60 min of retention time were the optimal conditions for the enzymatic hydrolysis to obtain the insoluble and soluble contents of 68.21%db and 8.18%db, respectively. Finally, DCCP or hydrolyzed DCCP (HDCCP) was partially substituted for wheat flour at different replacement ratios in a cookie recipe at 0, 10, 20, 30, and 40%. The cookies with a 10% replacement ratio of hydrolyzed deoiled coconut cake powders had a lower RIS by more than two folds those of DCCP and had the same sensorial score as the control sample. This study proposed that Celluclast 1.5 L effectively reduced RIS by partially converting insoluble to soluble dietary fiber, improving the soluble dietary fiber content in fiber-enriched cookies.

Objective: To establish a set of standardized parameters this will assist in identification of raw material of Caralluma species. Materials: The present study reports, detailed set of parameters from four species, Caralluma lasiantha, Caralluma umbellata, Caralluma attenuata and Caralluma diffusa, for powder characteristics, physicochemical evaluation, HPTLC fingerprint profile, and quantitative estimation of phytoconstituents that would contribute in the correct identity of the raw materials. Results: Various prominent cellular components were observed under the microscopic study, extractive values showed the presence of more water soluble compounds and phytochemical analysis revealed the presence nearly eight classes of phytoconstituents. HPTLC analysis showed the marked differences among species. Conclusion: The results of the present study serve as valuable information for correct identification of plant and determine its genuinity.

Azolla filiculoides is an aquatic fern with the potential to become a source of raw materials in a biorefinery system, e.g., a source of soluble and insoluble carbohydrates, proteins, carotenoids, and polyphenols. The fiber chemical content was determined as cellulose (19.2% dry basis) and hemicellulose (7.6% dry basis) content. Azolla has no lignin as a cell wall structure material. Cellulase treatment showed no effect in ethanolic extraction, but polyphenols were found in the enzyme solution at the end of the reaction. The phenolic acids and flavonoids contents of those with health promoting activity were determined, with gallic, syringic, rosmarinic, and p-coumaric the most abundant acids; kaempferol, apigenin, and quercetin were the most abundant flavonoids. The results show that A. filiculoides is a valuable source of bioactive components and cellulosic materials.

Low-temperature (cryo) electron microscopy has been established as a promising approach for alleviating technical problems encountered in electron microscopy of life science materials. Cryofixation freezes in the native situation of biological material, with the rapid freezing process preventing redistribution of water-soluble elements in cellular and non-cellular compartments. Thus it becomes possible to correlate tissue morphology with chemical and physical properties.The use of cryo techniques puts stringent demands on the electron microscope technology: an ultra-high vacuum, including special measures to tackle the water vapour released by the specimen; stable, low-drift cryo-temperature specimen holders; special functions providing low-dose imaging conditions; electron dose indicators; and a TV system for observation under low-level illumination. The new CM-CRYO combines these features with the ease of use of the CM microscope concept and the quality of the TWIN objective lens system.

Fruit ripening and softening are highly complex processes, and there is an interplay and coordination between the metabolic pathways that are involved in the biological processes. In this study, we aimed to elucidate the variation in the characters and possible causes of cell wall materials and morphological structure during apple fruits development. We studied the cell wall material (CWM), structure, cellular morphology, hydrolase activity, and the transcriptional levels of the related genes in four apple varieties ‘Ruixue’ and ‘Ruixianghong’ and their parents (‘Pink Lady’ and ‘Fuji’) during fruit development. The decrease in the contents of CWMs, sodium carbonate soluble pectin, hemicellulose, and cellulose were positively correlated with the decline in the hardness during the fruit development. In general, the activities of polygalacturonase, β-galactosidase, and cellulase enzymes increased during the late developmental period. As the fruit grew, the fruit cells of all of the cultivars gradually became larger, and the cell arrangement became more relaxed, the fruit cell walls became thinner, and the intercellular space became larger. In conclusion, the correlation analysis indicated that the up-regulation of the relative expression levels of ethylene synthesis and cell wall hydrolase genes enhanced the activity of the cell wall hydrolase, resulting in the degradation of the CWMs and the depolymerization of the cell wall structure, which affected the final firmness of the apple cultivars in the mature period.

Green brackish algae Chaetomorpha sp. are easily found in shrimp ponds in Mekong Delta, Vietnam. They can also be co-cultured with shrimps in brackish water shrimp ponds to increase shrimp health and yield. Chaetomorpha sp. algae contain high amount of protein from 10 to 20% w/w db, including water soluble protein and alkaline-soluble protein with over 88% total protein. Dried material were used for protein extraction by using cellulase enzyme (Crestone Conc., Genecor) and NaOH solution. In this research, we optimize the extraction condition of protein from green algae Chaetomorpha sp. by using response surface methodology (RSM). At optimal extraction conditions, dried material was used for protein extraction by using cellulase enzyme (Crestone Conc., Genecor) with the enzyme dosage of 121 UI/g db at 400C during 90 mins. After extraction, the slurry was centrifuged to separate the algae biomass residue to extract the alkaline-soluble protein. The protein extraction yield by using cellulase enzyme was 38.921 mg/g db. After that the, algae biomass residue was extracted by a 1.2% NaOH solution for 78 mins at 500C. The protein extraction yield was 68.651 mg/g db. The total protein extraction yield was 105.755 mg/g db. The extraction yield was increased 10.33% when using the response surface methodology. Concentrated algae protein can be used as a good protein source for food and feed products.

The present study reports on a biological model based on fibroblast proliferation applied to 3 different types of flat-plate dialysis membrane, in order to ascertain whether the artificial materials currently used in hemodialysis cause in vitro cellular proliferation. The study plan we followed involved plate membrane isolation from non-used dialyzers and used dialyzers, observed through scanning electron microscopy (SEM) both before and after testing with human fibroblasts by means of cell culture. Fibroblast growth was assessed by phase contrast light microscopy examination and cytometric DNA content evaluation. Our investigations proved that the artificial materials we considered interact with fibroblast cultures. Noticeable proliferative response was observed both after contact with unused material and on mediation by the protein layer absorbed on the membrane surface at the end of dialysis sessions. In this last case fibroblast proliferative activity appeared higher than that observed with unused membranes, showing that the soluble molecules entrapped in the protein layer appeared able to exert a biological activity even in in vitro tests

Despite significant efforts have been directed toward reducing waste generation and encouraging alternative waste management strategies, landfills still remain the main option for Municipal Solid Waste (MSW) disposal in many countries. Hence, landfills and related impacts on the surroundings are still current issues throughout the world. Actually, the major concerns are related to the potential emissions of leachate and landfill gas into the environment, that pose a threat to public health, surface and groundwater pollution, soil contamination and global warming effects. To ensure environmental protection and enhance landfill sustainability, modern sanitary landfills are equipped with several engineered systems with different functions. For instance, the installation of containment systems, such as bottom liner and multi-layers capping systems, is aimed at reducing leachate seepage and water infiltration into the landfill body as well as gas migration, while eventually mitigating methane emissions through the placement of active oxidation layers (biocovers). Leachate collection and removal systems are designed to minimize water head forming on the bottom section of the landfill and consequent seepages through the liner system. Finally, gas extraction and utilization systems, allow to recover energy from landfill gas while reducing explosion and fire risks associated with methane accumulation, even though much depends on gas collection efficiency achieved in the field (range: 60-90% Spokas et al., 2006; Huitric and Kong, 2006). Hence, impacts on the surrounding environment caused by the polluting substances released from the deposited waste through liquid and gas emissions can be potentially mitigated by a proper design of technical barriers and collection/extraction systems at the landfill site. Nevertheless, the long-term performance of containment systems to limit the landfill emissions is highly uncertain and is strongly dependent on site-specific conditions such as climate, vegetative covers, containment systems, leachate quality and applied stress. Furthermore, the design and operation of leachate collection and treatment systems, of landfill gas extraction and utilization projects, as well as the assessment of appropriate methane reduction strategies (biocovers), require reliable emission forecasts for the assessment of system feasibility and to ensure environmental compliance. To this end, landfill simulation models can represent an useful supporting tool for a better design of leachate/gas collection and treatment systems and can provide valuable information for the evaluation of best options for containment systems depending on their performances under the site-specific conditions. The capability in predicting future emissions levels at a landfill site can also be improved by combining simulation models with field observations at full-scale landfills and/or with experimental studies resembling landfill conditions. Indeed, this kind of data may allow to identify the main parameters and processes governing leachate and gas generation and can provide useful information for model refinement. In view of such need, the present research study was initially addressed to develop a new landfill screening model that, based on simplified mathematical and empirical equations, provides quantitative estimation of leachate and gas production over time, taking into account for site-specific conditions, waste properties and main landfill characteristics and processes. In order to evaluate the applicability of the developed model and the accuracy of emissions forecast, several simulations on four full-scale landfills, currently in operative management stage, were carried out. The results of these case studies showed a good correspondence of leachate estimations with monthly trend observed in the field and revealed that the reliability of model predictions is strongly influenced by the quality of input data. In particular, the initial waste moisture content and the waste compression index, which are usually data not available from a standard characterisation, were identified as the key unknown parameters affecting leachate production. Furthermore, the applicability of the model to closed landfills was evaluated by simulating different alternative capping systems and by comparing the results with those returned by the Hydrological Evaluation of Landfill Performance (HELP), which is the most worldwide used model for comparative analysis of composite liner systems. Despite the simplified approach of the developed model, simulated values of infiltration and leakage rates through the analysed cover systems were in line with those of HELP. However, it should be highlighted that the developed model provides an assessment of leachate and biogas production only from a quantitative point of view. The leachate and biogas composition was indeed not included in the forecast model, as strongly linked to the type of waste that makes the prediction in a screening phase poorly representative of what could be expected in the field. Hence, for a qualitative analysis of leachate and gas emissions over time, a laboratory methodology including different type of lab-scale tests was applied to a particular waste material. Specifically, the research was focused on mechanically biologically treated (MBT) wastes which, after the introduction of the European Landfill Directive 1999/31/EC (European Commission, 1999) that imposes member states to dispose of in landfills only wastes that have been preliminary subjected to treatment, are becoming the main flow waste landfilled in new Italian facilities. However, due to the relatively recent introduction of the MBT plants within the waste management system, very few data on leachate and gas emissions from MBT waste in landfills are available and, hence, the current knowledge mainly results from laboratory studies. Nevertheless, the assessment of the leaching characteristics of MBT materials and the evaluation of how the environmental conditions may affect the heavy metals mobility are still poorly investigated in literature. To gain deeper insight on the fundamental mechanisms governing the constituents release from MBT wastes, several leaching experiments were performed on MBT samples collected from an Italian MBT plant and the experimental results were modelled to obtain information on the long-term leachate emissions. Namely, a combination of experimental leaching tests were performed on fully-characterized MBT waste samples and the effect of different parameters, mainly pH and liquid to solid ratio (L/S,) on the compounds release was investigated by combining pH static-batch test, pH dependent tests and dynamic up-flow column percolation experiments. The obtained results showed that, even though MBT wastes were characterized by relatively high heavy metals content, only a limited amount was actually soluble and thus bioavailable. Furthermore, the information provided by the different tests highlighted the existence of a strong linear correlation between the release pattern of dissolved organic carbon (DOC) and several metals (Co, Cr, Cu, Ni, V, Zn), suggesting that complexation to DOC is the leaching controlling mechanism of these elements. Thus, combining the results of batch and up-flow column percolation tests, partition coefficients between DOC and metals concentration were derived. These data, coupled with a simplified screening model for DOC release, allowed to get a very good prediction of metal release during the experiments and may provide useful indications for the evaluation of long-term emissions from this type of waste in a landfill disposal scenario. In order to complete the study on the MBT waste environmental behaviour, gas emissions from MBT waste were examined by performing different anaerobic tests. The main purpose of this study was to evaluate the potential gas generation capacity of wastes and to assess possible implications on gas generation resulting from the different environmental conditions expected in the field. To this end, anaerobic batch tests were performed at a wide range of water contents (26-43 %w/w up to 75 %w/w on wet weight) and temperatures (from 20-25 °C up to 55 °C) in order to simulate different landfill management options (dry tomb or bioreactor landfills). In nearly all test conditions, a quite long lag-phase was observed (several months) due to the inhibition effects resulting from high concentrations of volatile fatty acids (VFAs) and ammonia that highlighted a poor stability degree of the analysed material. Furthermore, experimental results showed that the initial waste water content is the key factor limiting the anaerobic biological process. Indeed, when the waste moisture was lower than 32 %w/w the methanogenic microbial activity was completely inhibited. Overall, the obtained results indicated that the operative conditions drastically affect the gas generation from MBT waste, in terms of both gas yield and generation rate. This suggests that particular caution should be paid when using the results of lab-scale tests for the evaluation of long-term behaviour expected in the field, where the boundary conditions change continuously and vary significantly depending on the climate, the landfill operative management strategies in place (e.g. leachate recirculation, waste disposal methods), the hydraulic characteristics of buried waste, the presence and type of temporary and final cover systems.

Abstract This work draws inspiration from totipotent cellular systems to design smart materials whose compositions and properties can be learned or evolved. Totipotency refers to the inherent genetic potential of a single cell to adapt and produce all types of differentiated cells within an organism. To study this principal and apply it synthetically, tissue-like compartmentalized assemblies are constructed via lipid membrane-separated aqueous droplets in a hydrophobic medium through the droplet interface bilayer (DIB) method. Within our droplets, we explore synthetic totipotency via cell-free reactions including actin polymerization and cell free protein synthesis (CFPS). The transcription and translation of our CFPS reactions are controlled by stimuli-responsive riboswitches (RS). Via this scheme, adaptable material properties and functions are achieved in vitro via protein production from cell-free machinery administered through RS governance. Here, we present thermally or chemically-triggered riboswitches for orthogonal production of representative fluorescent protein products, as well functional proteins. To characterize the material properties of target proteins, we study the formation of polymerized actin shells to stabilize organically-encased droplets and span DIBs. We present a modified protocol for chemically-triggered actin polymerization as well as a thermally triggered actin RS. We characterize theophylline (TP)-triggered production of alpha hemolysin (α-HL) through CFPS and synthesized an organic-soluble trigger that can be sensed from the oil phase by a RS in an aqueous bioreactor droplet. We also demonstrate increased droplet conductivity when CFPS α-HL products are incorporated in DIBs. This interdisciplinary work involves cell culture, gene expression, organic synthesis, vesicle formation, protein quantification, tensiometry, droplet aspiration, microplate fluorescence/absorption experiments, fluorescent microscopy, and electrophysiology. This project is an essential design analysis for creating smart, soft materials using synthetic biology and provides motivation for artificial tissues capable of adapting in response to external stimuli.

External forces, cell adhesion and soluble signaling molecules influence fundamental functions of cells like shape, migration, proliferation or differentiation. Thus, investigating how mechanical forces affect 3D cell structure and function is of crucial significance in order to gain a better understanding healthy and malignant cell behavior during embryogenesis, regeneration or malignancy [1]. Micromanipulation of cells in a controlled environment is a widely used approach for understanding cellular responses with respect to external mechanical forces. While experimental data provide optical information about the overall cell shape, the 3D deformation state of intracellular structures is not accessible by direct observations and measurements. However, the continuous description of the intracellular deformation state can be calculated as a numerical solution of the boundary value problem given by the partial differential equations of structural mechanics, including a set of canonic material constants (stiffness, compressibility), and the boundary conditions derived from time series of images, e.g. change of visible cell contours. The main idea of our approach is to reformulate the problem of finding optimal modeling parameters as an image registration problem. That is the optimal set of modeling parameters corresponds to the minimum of a suitable similarity measure between computationally predicted and experimentally observed deformations. In this article, we focus on the numerical analysis of uniaxial stretching of a rat embryonic fibroblast 52 (REF 52) based on a series of 2D images reflecting the successive alteration of cell contours during deformation. The goal of this study consists in finding an optimal set of material constants within a non-linear hyperelastic material law, which is able to reproduce results of experimental observations.

Many preservation methods have utilized sugars such as trehalose as protectants against injury during cell preservation processing, especially during drying (1–5). As mammalian cells do not synthesize trehalose, research in the mammalian cell desiccation field has focused on the development of strategies to enable trehalose delivery into the intracellular milieu. Numerous techniques have been explored ranging from microinjection (2) to the creation or utilization of membrane pores (1,3). Fluid phase endocytosis has shown great promise as an effective strategy for non-invasively delivering water-soluble materials into the intracellular space (4, 5). In this technique trehalose is transported across the cell membrane in membrane-bound cellular compartments called endosomes. Cells incubated in cell culture medium containing trehalose have been shown to take up considerable amounts of trehalose by this technique (4, 5). How much of this trehalose actually become available for protection of biomolecules during the dehydration process has yet to be determined.

The uptake, internalization and intracellular degradation of 125I-labeled rt-PA (125I-rt-PA) by isolated rat hepatocytes was investigated. Incubation at 37°C resulted in internalization of 125I-rt-PA, followed by the appearance of labeled trichloroacetic acid-soluble (TCA) material in the inclubation media due to degradation of rt-PA. Degradation of rt-PA was inhibited by the presence of NH4Cl (10mM) or chloroquine (ImM) (lysosoma tropic agents) in the incubation media. This suggests that rt-PA degradation occurs intracellularly, perhaps within the lysosomes. 125I-rt-PA was taken up by rat hepatocytes through a specific, high affinity mechanism. Scatchard analysis of the data indicated that 106 molecules of rt-PA were taken up per cell/hour and the calculated dissociation constant was lOnM. Uptake of 125I-rt-PA was not inhibited by glycopeptides isolated from rt-PA nor by several other glycoproteins known to be cleared by identified hepatic receptors. These results suggest that the uptake of rt-PA by rat hepatocytes involves a receptor specific for t-PA and is not mediated by a carbohydrate specific receptor.