Production of bioactive cellulose films reconstituted from ionic liquids

A new method for introducing enzymes into cellulosic matrixes which can be formed into membranes, films, or beads has been developed using a cellulose-in-ionic-liquid dissolution and regeneration process. Initial results on the formation of thin cellulose films incorporating dispersed laccase indicate that active enzyme-encapsulated films can be prepared using this methodology and that precoating the enzyme with a second, hydrophobic ionic liquid prior to dispersion in the cellulose/ionic liquid solution can provide an increase in enzyme activity relative to that of untreated films, presumably by providing a stabilizing microenvironment for the enzyme.     

Radiation dose verification of an X-ray based blood irradiator using EBT3 radiochromic films calibrated using Gamma Knife machine

AimBlood irradiators (BI) initial acceptance testing and routine annual dosimetry checks require radiation dose measurements in order to comply with regulatory requirements.BackgroundTraditionally thermo-luminescence dosimeters (TLD) have been used to measure the dose. The EBT3 film is reported to be a better dosimeter for low energy X-rays than its predecessors EBT2 and EBT. To the best of our knowledge, the use of EBT3 films to perform dosimetry on X-ray based BI has not been reported yet.Materials and methodsWe performed routine radiation dosimetry checks using EBT3 films on a new X-ray based BI and compared the results with TLD dosimetry. Calibration films were irradiated with radiation beam from a Co-60 Gamma Knife (GK) radiosurgery machine and, alternatively, using an Ir-192 high dose rate (HDR) brachytherapy device. The films were calibrated to cover a wide dose range from 1 to 40 Gy. Such a wide dose range has not been reported yet in BI film dosimetry.ResultsWe obtained a relative difference of about 6.6% between doses measured using TLD and those measured using EBT3 films. Both irradiation methods using GK or HDR were found to be adequate for the calibration of the EBT3 Gafchromic films.ConclusionsWe recommend the use of EBT3 films in routine X-ray based BI dosimetry checks. The presented method takes advantage of available radiotherapy equipment that can be efficiently used for EBT3 films calibration. The method is fast, reproducible and saves valuable medical physicist's time.     

The physical action of cellulases revealed by a quartz crystal microbalance study using ultrathin cellulose films and pure cellulases

The effects of fungal cellulases on model cellulose films were studied using a high-resolution quartz crystal microbalance (QCM) sensitive to minute changes of the nanometer thick model cellulose films. It was found that endoglucanases not only produce new end groups but also cause a swelling of the cellulose film. The cellobiohydrolases degraded the films quickly, which was detected as a rapid decrease in the remaining amount of cellulose on the QCM crystal. However, changing viscoelastic properties of the films also indicated a softening of the film during the degradation. A defined mixture of selected cellulases caused a significantly higher rate of degradation than only cellobiohydrolases. Cellulase synergism is discussed with the endoglucanase swelling effects and film softening added.     

Enhancement of the wet properties of transparent chitosan-acetic-acid-salt films using microfibrillated cellulose

This report presents a new route to enhance the wet properties of chitosan-acetic-acid-salt films using microfibrillated cellulose (MFC). The enhancement makes it easier to form chitosan-acetic-acid-salt films into various shapes at room temperature in the wet state. Chitosan with MFC was compared with the well-known buffer treatment. It was observed that films containing 5 wt % MFC were visually identical to the buffered/unbuffered films without MFC. Field-emission scanning electron microscopy indicated that MFC formed a network with uniformly distributed fibrils and fibril bundles in the chitosan matrix. The addition of MFC reduced the risk of creases and deformation in the wet state because of a greater wet stiffness. The wet films containing MFC were also extensible. Although the stiffness, strength and extensibility were highest for the buffered films, the wet strength of the MFC-containing unbuffered films was sufficient for wet forming operations. The effects of MFC on the mechanical properties of the dry chitosan films were small or absent. It was concluded that the addition of MFC is an acceptable alternative to buffering for shaping chitosan films/products in the wet state. The advantages are that the "extra" processing step associated with buffering is unnecessary and that the film matrix remains more water-soluble.     

Antibacterial copper–nickel bilayers and multilayer coatings by pulsed laser deposition on titanium

Biofouling, especially microfouling, is a major concern with the use of titanium (Ti) in the marine environment as a condenser material in cooling water systems. Earlier, copper–nickel (Cu/Ni) alloys were extensively used in marine environments due to their high corrosion and biofouling resistance. However, the choice of condenser material for the new fast breeder reactor in Kalpakkam is Ti to avoid steam side corrosion problems, which may pose a threat to steam generator parts having sodium as the secondary coolant. This study evaluates the surface modification of Ti using nano films of copper (Cu) and nickel (Ni) to utilize the antibacterial property of copper ions in reducing microfouling. The surface modification of Ti was carried out by the deposition of a Cu/Ni bilayer and (Cu/Ni)₁₀ multilayer films using a pulsed laser deposition technique. Various surface characterization studies revealed that the deposited Cu/Ni films were thin and nanocrystalline in nature. The antibacterial properties were evaluated using total viable count and epifluorescence microscopic techniques. The results showed an apparent decrease in bacterial attachment on multilayered and bilayered Cu/Ni thin films on Ti surfaces. Comparative studies between the two types of films showed a bigger reduction in numbers of microorganisms on the multilayers.     

Tuning the mechanical and physicochemical properties of cross-linked protein films

Films derived from natural sources such as proteins provide an advantage over synthetic films due to their noncytotoxicity, biodegradability, and vast functionality. A new protein source gained from the cataractous eye protein isolate (CEPI) obtained after surgery has been investigated for this purpose. Glycerol has been employed as the plasticizer and glutaraldehyde (GD) as a cross-linker. Fourier transform infrared spectroscopy was employed to characterize the films. Nanoindentation and thermogravimetric analyses reveal improved mechanical and thermal properties of the cross-linked films. The films with 20% (w/w) GD exhibited properties such as the highest modulus and low water solubility. It is possible to tune the properties based on the extent of cross-linking. All the films were completely degraded by the enzyme trypsin. The similarity of these films was checked by using the prepared films as a delivery vehicle for a model compound, ampicillin sodium. The encapsulation efficiency was found to be 74%, and in vitro release studies showed significant amounts of drug release at physiological pH. This study will help us understand how the properties of protein films can be tuned to obtain the desired physicochemical properties. These biodegradable protein films could find use in pharmaceutical industries as delivery carriers.     

An electrochemical investigation of hemoglobin and catalase incorporated in collagen films

Collagen, an electrochemically inert protein, formed films on pyrolytic graphite (PG) electrodes, which provided a suitable microenvironment for heme proteins to transfer electron directly with the underlying electrodes. Hemoglobin (Hb) and catalase (Cat) incorporated in collagen films exhibited a pair of well-defined and quasi-reversible cyclic voltammetric peaks at around -0.35 V and -0.47 V (vs. SCE) in pH 7.0 buffers, respectively, characteristic of the protein heme Fe(III)/Fe(II) redox couples. UV-vis spectra showed that the heme proteins in collagen films retained their near-native conformations in the medium pH range. The results of scanning electron microscopy (SEM) demonstrated that the interaction between heme proteins and collagen made the morphology of dry protein-collagen films different from the collagen films alone. The electrochemical parameters such as apparent heterogeneous electron transfer rate constant (k(s)) and formal potential (E degrees ') of the films were estimated by using square wave voltammograms (SWV) and nonlinear regression analysis. The heme protein-collagen film electrodes were also used to catalyze the reduction of nitrite, oxygen and hydrogen peroxide, indicating potential applications of the films for the fabrication of a new type of biosensor that does not use mediators.     

An effective calibration technique for radiochromic films using a single-shot dose distribution in Gamma Knife®

PurposeA method of calibrating radiochromic films for Gamma Knife^® (GK) dosimetry was developed. The applicability and accuracy of the new method were examined.MethodsThe dose distribution for a sixteen millimeter single-shot from a GK was built using a reference film that was calibrated using the conventional multi-film calibration (MFC) method. Another film, the test film, from a different set of films was irradiated under the same conditions as the reference film. The calibration curve for the second set of films was obtained by assigning the dose distribution of the reference film to the optical density of the test film, point by point. To assess the accuracy of this single-film calibration (SFC) method, differences between gamma index pass rates (GIPRs) were calculated.ResultsThe SFC curves were successfully obtained with estimated errors of 1.46%. GIPRs obtained with the SFC method for films irradiated using a single-shot showed differences less than one percentage point when dose difference criterion (ΔD) was 2% and the distance to agreement criterion (Δd) was 1 mm. The GIPRs of the SFC method when the films were irradiated following a virtual target treatment plan were consistent with the GIPRs of the MFC method, with differences of less than 0.2 percentage points for ΔD = 1% and Δd = 1 mm.ConclusionThe accuracy of the SFC method is comparable to that of conventional multi-film calibration method for GK film dosimetry.     

Touring the Dream Factory: The Dream-Film Connection in The Wizard of Oz and A Nightmare on Elm Street

This essay explores the complex interplay of dreams and film, using an analysis of The Wizard of Oz (1939) and A Nightmare on Elm Street (1984) to illustrate how an interdisciplinary approach can provide fruitful insights into how dreams influence films, and films influence dreams. The essay suggests that reflecting on the dreams-film connection can deepen our understanding of the cultural dimensions of human development, particularly in the context of modern American culture. By using the methodological resources of psychology, sociology, history, film criticism, and theology, the essay argues more generally that the interdisciplinary analysis of films offers significant new possibilities for the development of dream studies. Kelly Bulkeley, Ph.D.     

Development of in situ zymography to localize active matrix metalloproteinase-7 (matrilysin-1).

Matrix metalloproteinase-7 (MMP-7) is upregulated during carcinogenesis and its expression correlates with metastasis of human endometrial and gastrointestinal carcinomas. In the present study, we have developed a new method to localize the activity of MMP-7 within tissues. Polyethylene terephthalate films were uniformly coated with crosslinked carboxymethylated transferrin (CCm-Tf) as a substrate and incubated with frozen tissue sections mounted on the films. CCm-Tf on the films was degraded selectively by MMP-7, but showed little or no susceptibility to MMP-1, -2, -3, -9, or -13; MT1-MMP; MT3-MMP; or ADAMTS4. Although some serine proteinases such as elastase also digested CCm-Tf, CCm-Tf films impregnated with serine proteinase inhibitors prevented the digestion. When frozen sections of human endometrial carcinoma and lung carcinoma tissues were incubated on CCm-Tf films or those treated with proteinase inhibitors, the activity was detected in the carcinoma cell nests, where MMP-7 was immunolocalized. The present in situ zymography using CCm-Tf may be a useful method to analyze the functions of MMP-7 in pathophysiological conditions.     

Preparation of polysaccharide supramolecular films by vine-twining polymerization approach

In this study, we investigated the preparation of polysaccharide supramolecular films through the formation of inclusion complexes by amylose in vine-twining polymerization using carboxymethyl cellulose-graft-poly(?-caprolactone) (CMC-g-PCL) as a new guest polymer. First, hydrogels were prepared by phosphorylase-catalyzed enzymatic polymerization in the presence of CMC-g-PCL according to the vine-twining polymerization manner. The XRD result of a powdered sample obtained by lyophilization of the resulting hydrogel indicated the presence of inclusion complexes of amylose with the PCL graft-chains between intermolecular (CMC-g-PCL)s, which acted as supramolecular cross-linking points for the hydrogelation. Then, the supramolecular films were obtained by adding water to the powdered samples, followed by drying. The mechanical properties of the selected films examined by tensile testing were superior to those of a CMC film. The effect of the supramolecular cross-linking structures on the mechanical properties of the films was evaluated further by several investigations.     

Identification of a fungus able to secrete enzymes that degrade regenerated cellulose films and analyses of its extracellular hydrolases

The LW03 strain was isolated from Chinese farmland soil and found to be able to secrete certain enzymes degrading regenerated cellulose films at low temperature. The LW03 strain was systematically identified as Rhizopus arrhizus var. arrhizus by morphological, physiological, and molecular methods. Incubation of regenerated cellulose films with the extracted crude enzyme of LW03 was done to measure morphological changes by using scanning electron microscopy. Microscopic observations showed that the morphology of the regenerated cellulose films changed drastically due to enzymatic hydrolysis. The extracellular hydrolases of LW03 strain incubated on bran medium were also assessed. The predominant activity in the crude enzyme was glucoamylase activity, followed by acid proteinase, phytase and pectinase activity. Interestingly, activities of β-glucosidase, endoglucanase, exoglucanase, and cellulase were also observed, but at a much lower extent. Based on initial evidence, the crude enzyme is most likely to contain some new constituents capable of degrading regenerated cellulose films.     

Preparation and characterization of novel nanocomposite films formed from silk fibroin and nano-TiO2

This paper describes the synthesis and characterization of new regenerated silk fibroin (SF)/nano-TiO(2) composite films. The preparation method, based on the sol-gel technique using butyl titanate as oxide precursor, could avoid reagglomeration of the prepared nanoparticles. Samples were characterized mainly by X-ray diffraction (XRD), ultra-violet (UV) spectroscopy, atomic force microscopy (AFM), Fourier transform infrared (FT-IR) spectroscopy, and thermogravimetric analysis (TGA). The UV and AFM results indicated that TiO(2) nanoparticles could be well dispersed inside the SF film, and the size of TiO(2) was about 80nm. The XRD and FT-IR analysis implied that the formation of nano-TiO(2) particles may induce the conformational transition of silk fibroin to a typical Silk II structure partly with the increasing of crystallinity in the composite films. Compared to the pure SF films, the mechanical and thermal properties of composite films were improved, and the solubility in water was decreased due to the conformational transition of silk fibroin to Silk II structure.     

Effects of UV-blocking films on the dispersal behavior of Encarsia formosa (Hymenoptera: Aphelinidae)

The parasitoid Encarsia formosa Gahan (Hymenoptera: Aphelinidae) has been used successfully for the control of Trialeurodes vaporariorum (Westwood) (Homoptera: Aleyrodidae). The development of UV-blocking plastic films has added a new component to future integrated pest management systems by disrupting insect pest infestation when UV light is excluded. Because both T. vaporariorum and E. formosa are reported to have similar spectral efficiency, there was a need to identify the impact of UV-blocking films on the dispersal behavior of both the pest and the natural enemy. In field studies, using choice-chamber experiments, E. formosa showed some preference to disperse into compartments where less UV light was blocked. However, further studies indicated that the effect was primarily attributable to the different light diffusion properties of the films tested. Thus, unlike its whitefly host, when the UV-absorbing properties of the films were similar, but the light diffusion properties differed, E. formosa adults preferred to disperse into compartments clad with films that had high light diffusion properties. When the plastic films differed most in their UV-absorbing capacity and had no light-diffusion capability, the initial dispersal of E. formosa between treatments was similar, although a small preference toward the environment with UV light was observed over time. When parasitoid dispersal was measured 3 h after release, more parasitoids were found on plants, suggesting that the parasitoids would search plants for whitefly hosts, even in a UV-blocked light environment. The potential for the integration of UV-blocking films with E. formosa in an advanced whitefly management system is discussed.     

Polarized dosimetry method for Gafchromic EBT3

PurposeTo analyze the changes in the polarization state of the flatbed scanner light caused by the EBT3 films and to propose a new method for correcting the lateral effects.Methods and materialsThe polarization changes induced by radiochromic films are analyzed using linear polarizing film. Based on the results, the linear polarizing films are used in the scanning process of the EBT3 films. This method is tested against the conventional EBT3 dosimetry using a series of simple regular beams and 21 cases of IMRT.ResultsThe mean results are statically different from the conventional dosimetry with EBT3. Depending on the transmission axis of the polarizing sheet, the results are better or worse compared to conventional dosimetry EBT3 film. When the transmission axis of the polarizing sheet is parallel to the coating direction, the dosimetry results are better and its variability is smaller. However, when the polarizer transmission axis is perpendicular to the coating direction, results are worse as well as its variability.ConclusionUsing a polarized film with the polarization axis parallel to the coating direction of the radiochromic film, and preferably above it, significantly improves the dosimetry results and is an easy and inexpensive way to correct the lateral artifacts of the conventional EBT3 dosimetry.     

Material properties of plasticized hardwood xylans for potential application as oxygen barrier films

Free films based on glucuronoxylan isolated from aspen wood were prepared by casting from aqueous solutions and drying in a controlled environment. Addition of xylitol or sorbitol facilitated film formation and thus examination of the material properties of these films. The mechanical properties of the films were evaluated using tensile testing and dynamic mechanical analysis in a controlled ambient relative humidity. The strain at break increased, and the stress at break and Young's modulus of the films decreased with increasing amounts of xylitol and sorbitol due to plasticization. At high amount of plasticizer, it was found that films with xylitol gave lower extensibility. Wide-angle X-ray scattering analysis showed that xylitol crystallized in a distinct phase, which we believe contributes to the more brittle behavior of these films. The effect of the plasticizers on the glass transition temperature was determined using dynamic mechanical analysis and differential scanning calorimetry. An increased amount of plasticizer shifted the glass transition to lower temperatures. The effect of moisture on the properties of plasticized films was investigated using water vapor sorption isotherms and by humidity scans in dynamic mechanical analysis. Sorption isotherms showed a transition from type II to type III when adding plasticizer. The films showed low oxygen permeability and thus have a potential application in food packaging.     

Development of polythene films for food packaging activated with an antilisterial bacteriocin from Lactobacillus curvatus 32Y

AIMS: The aims of this work were to (i) use a bacteriocin produced by Lactobacillus curvatus 32Y active against Listeria monocytogenes to activate polythene films by different methods, (ii) implement a large-scale process for antilisterial polythene films production and (iii) verify the efficacy of the developed films in inhibiting the growth of L. monocytogenes during the storage of meat products. METHODS AND RESULTS: The film was made active by using the antilisterial bacteriocin 32Y by Lact. curvatus with three different procedures: soaking, spraying and coating. The antimicrobial activity of the activated films was tested in plate assays against the indicator strain L. monocytogenes V7. All the used procedures yielded active polythene films although the quality of the inhibition was different. The coating was therefore employed to develop active polythene films in an industrial plant. The antimicrobial activity of the industrially produced films was tested in experiments of food packaging involving pork steak and ground beef contaminated by L. monocytogenes V7 at roughly 10(3) CFU cm(-2) and gram respectively. The results of the challenge tests showed the highest antimicrobial activity after 24 h at 4 degrees C, with a decrease of about 1 log of the L. monocytogenes population. CONCLUSIONS: Antimicrobial packaging can play an important role in reducing the risk of pathogen development, as well as extending the shelf life of foods. SIGNIFICANCE AND IMPACT OF THE STUDY: Studies of new food-grade bacteriocins as preservatives and development of suitable systems of bacteriocin treatment of plastic films for food packaging are important issues in applied microbiology and biotechnology, both for implementing and improving effective hurdle technologies for a better preservation of food products.     

Under-response correction for EBT3 films in the presence of proton spread out Bragg peaks

We present a study of the under-response of the new Gafchromic EBT3 films and a procedure to accurately perform 2D and 3D proton dosimetry measurements for both pristine and spread out Bragg peaks (SOBP) of any energy. These new films differ from the previous EBT2 generation by a slightly different active layer composition, which we show has not effected appreciably their response. The procedure and the beam quality correction factor curve have been benchmarked using 29 MeV modulated proton beams. In order to show the correction to apply when EBT3 films are used as treatment verification tools in anthropomorphic phantoms, two simulation studies involving clinical energies are presented: a SOBP for eye treatments and a SOBP to treat 20 cm deep and 5 cm thick tumours. We find maximum under-responses of 37%, 30% and 7.7% for the modulated 29 MeV beam, eye and deep tumour treatment, respectively, which were attained close to the end of the peak tails, due to a higher proportion of very low energy protons. The maximum deviations between corrected and uncorrected doses were for the three cases, respectively, 20.7%, 8.3% and 2.1% of the average dose across flat region of the SOBP. These values were obtained close to the distal edge of the SOBPs, where the proportion of low energy protons was not as high as on the tail, but there still was a number of protons high enough to deposit a reasonable amount of dose in the films.     

Assessment of Size of Small Volume Foeto-maternal Bleeds. A New Method of Quantification of the Kleihauer Technique

The risk of Rh-isoimmunization is probably related to the volume of foeto-maternal bleeding. With the Kleihauer technique foetal cells may be detected in maternal blood, but estimation of the size of foetal bleed the cells represent is at present difficult and open to serious error.A new method of quantifying the technique by using a standard volume of maternal blood, and a simple machine for the preparation of comparable blood films, is described. The new method eliminates many of the errors of previous techniques, and should be capable of automation for large-scale screening programmes.     

Polymer manipulation and nanofabrication in real time using transmission electron microscopy

Here we present time-resolved in situ transmission electron microscopy (TEM) observations and real-time manipulation of nematic ordered cellulose and ultradrawn polyethylene films. Drawn films of these two polymers exhibited a unique response to the low-dose electron beam. Electron beam damage was minimal based on retention of an organized electron diffraction pattern. Increased electron dosage appeared to melt the polymer with subsequent movement and attraction toward preferred electron concentrations within the beam. This discovery allowed the preferential, directed manipulation of polymer chain aggregates in two dimensions. These findings provide a basis for a new technique to manipulate and simultaneously observe dynamic assembly at the molecular level of structures using TEM.