TEMPO-Oxidized Cellulose Nanofibril Films Incorporating Graphene Oxide Nanofillers.

To design a new system of novel TEMPO-oxidized cellulose nanofibrils (TOCNs)/graphene oxide (GO) composite, 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-mediated oxidation was utilized. For the better dispersion of GO into the matrix of nanofibrillated cellulose (NFC), a unique process combining high-intensity homogenization and ultrasonication was adopted with varying degrees of oxidation and GO percent loadings (0.4 to 2.0 wt%). Despite the presence of carboxylate groups and GO, the X-ray diffraction test showed that the crystallinity of the bio-nanocomposite was not altered. In contrast, scanning electron microscopy showed a significant morphological difference in their layers. The thermal stability of the TOCN/GO composite shifted to a lower temperature upon oxidation, and dynamic mechanical analysis signified strong intermolecular interactions with the improvement in Young's storage modulus and tensile strength. Fourier transform infrared spectroscopy was employed to observe the hydrogen bonds between GO and the cellulosic polymer matrix. The oxygen permeability of the TOCN/GO composite decreased, while the water vapor permeability was not significantly affected by the reinforcement with GO. Still, oxidation enhanced the barrier properties. Ultimately, the newly fabricated TOCN/GO composite through high-intensity homogenization and ultrasonification can be utilized in a wide range of life science applications, such as the biomaterial, food, packaging, and medical industries.

Solid State Polymerization of Biodegradable Poly(butylene sebacate-co-terephthalate): A Rapid, Facile Method for Property Enhancement.

Poly(butylene sebacate-co-terephthalate) (PBSeT) has generated attention as a promising biopolymer for preparing bioplastics. However, there are limited studies on the synthesis of PBSeT, impeding its commercialization. Herein, with a view to addressing this challenge, biodegradable PBSeT was modified using solid state polymerization (SSP) with various ranges of time and temperature. The SSP used three different temperatures below the melting temperature of PBSeT. The polymerization degree of SSP was investigated using Fourier-transform infrared spectroscopy. The changes in the rheological properties of PBSeT after SSP were investigated using a rheometer and an Ubbelodhe viscometer. Differential scanning calorimetry and X-ray diffraction showed that the crystallinity of PBSeT was higher after SSP. The investigation revealed that after SSP for 40 min at 90 °C, PBSeT exhibited higher intrinsic viscosity (increased from 0.47 to 0.53 dL/g), crystallinity, and complex viscosity than PBSeT polymerized at other temperatures. However, a high SSP processing time resulted in a decrease in these values. In this experiment, SSP was most effectively performed in the temperature range closest to the melting temperature of PBSeT. This indicates that SSP could be a facile and rapid method for improving the crystallinity and thermal stability of synthesized PBSeT.

Synthesis and Characterization of Poly(Butylene Sebacate-Co-Terephthalate) Copolyesters with Pentaerythritol as Branching Agent.

Poly(butylene sebacate-co-terephthalate) (PBSeT) copolyesters are prepared by melt polymerization via two-step transesterification and polycondensation using pentaerythritol (PE) as a branching agent. The effects of the incorporated PE on its chemical, thermal, mechanical, and degradation properties, along with the rheological properties of its melt, are investigated. The highest molecular weight and intrinsic viscosity along with the lowest melt flow index were achieved at a PE content of 0.2 mol%, with minimal reduction in the tensile strength and the highest tear strength. The addition of PE did not significantly influence the thermal behavior and stability of the PBSeT copolyesters; however, the elongation at break decreased with increasing PE content. The sample with 0.2 mol% PE exhibited a higher storage modulus and loss modulus as well as a lower loss angle tangent than the other samples, indicating improved melt elasticity. The incorporation of more than 0.2 mol% PE enhanced the enzymatic degradation of copolyesters. In summary, including within 0.2 mol%, PE effectively improved both the processability-related characteristics and degradation properties of PBSeT copolyesters, suggesting their potential suitability for use in agricultural and packaging materials.

Maleic Anhydride-Grafted PLA Preparation and Characteristics of Compatibilized PLA/PBSeT Blend Films.

Poly(butylene sebacate-co-terephthalate) (PBSeT) is a biodegradable flexible polymer suitable for melt blending with other biodegradable polymers. Melt blending with a compatibilizer is a common strategy for increasing miscibility between polymers. In this study, PBSeT polyester was synthesized, and poly(lactic acid) (PLA) was blended with 25 wt% PBSeT by melt processing with 3-6 phr PLA-grafted maleic anhydride (PLA-g-MAH) compatibilizers. PLA-g-MAH enhanced the interfacial adhesion of the PLA/PBSeT blend, and their mechanical and morphological properties confirmed that the miscibility also increased. Adding more than 6 phr of PLA-g-MAH significantly improved the mechanical properties and accelerated the cold crystallization of the PLA/PBSeT blends. Furthermore, the thermal stabilities of the blends with PLA-g-MAH were slightly enhanced. PLA/PBSeT blends with and without PLA-g-MAH were not significantly different after 120 h, whereas all blends showed a more facilitated hydrolytic degradation rate than neat PLA. These findings indicate that PLA-g-MAH effectively improves PLA/PBSeT compatibility and can be applied in the packaging industry.

Characterization of PLA/PBSeT Blends Prepared with Various Hexamethylene Diisocyanate Contents.

Poly (lactic acid) (PLA) is the most widely available commercial bioplastic that is used in various medical and packaging applications and three-dimensional filaments. However, because neat PLA is brittle, it conventionally has been blended with ductile polymers and plasticizers. In this study, PLA was blended with the high-ductility biopolymer poly (butylene-sebacate-co-terephthalate) (PBSeT), and hexamethylene diisocyanate (HDI) was applied as a crosslinking compatibilizer to increase the miscibility between the two polymers. PLA (80%) and PBSeT (20%) were combined with various HDI contents in the range 0.1-1.0 parts-per-hundred rubber (phr) to prepare blends, and the resulting physical, thermal, and hydrolysis properties were analyzed. Fourier-transform infrared analysis confirmed that -NH-C=OO- bonds had formed between the HDI and the other polymers and that the chemical bonding had influenced the thermal behavior. All the HDI-treated specimens showed tensile strengths and elongations higher than those of the control. In particular, the 0.3-phr-HDI specimen showed the highest elongation (exceeding 150%) and tensile strength. In addition, all the specimens were hydrolyzed under alkaline conditions, and all the HDI-treated specimens degraded faster than the neat PLA one.

Synthesis, Characterization and Properties of Biodegradable Poly(Butylene Sebacate-Co-terephthalate).

In this study, poly(butylene sebacate-co-terephthalate) (PBSeT) was successfully synthesized using various ratios of sebacic acid (Se) and dimethyl terephthalate (DMT). The synthesized PBSeT showed a high molecular weight (Mw, 88,700-154,900 g/mol) and good elastomeric properties. In particular, the PBSeT64 (6:4 sebacic acid/dimethyl terephthalate mole ratio) sample showed an elongation at break value of over 1600%. However, further increasing the DMT content decreased the elongation properties but increased the tensile strength due to the inherent strength of the aromatic unit. The melting point and crystallization temperature were difficult to observe in PBSeT64, indicating that an amorphous copolyester was formed at this mole ratio. Interestingly, wide angle X-ray diffraction (WAXD) curves was shown in the cases of PBSeT46 and PBSeT64, neither the crystal peaks of PBSe nor those of poly(butylene terephthalate) (PBT) are observed, that is, PBSeT64 showed an amorphous form with low crystallinity. The Fourier-transform infrared (FT-IR) spectrum showed C-H peaks at around 2900 cm-1 that reduced as the DMT ratio was increased. Nuclear magnetic resonance (NMR) showed well-resolved peaks split by coupling with the sebacate and DMT moieties. These results highlight that elastomeric PBSeT with high molecular weight could be synthesized by applying DMT monomer and showed promising mechanical properties.

Cytotoxic effects of ex vivo-expanded natural killer cell-enriched lymphocytes (MYJ1633) against liver cancer.

BACKGROUND: Adoptive transfer of immune cells such as T cells and natural killer (NK) cells has emerged as a targeted method of controlling the immune system against cancer. Despite their significant therapeutic potential, efficient methods to generate adequate numbers of NK cells are lacking and ex vivo-expansion and activation of NK cells is currently under intensive investigation. The primary purpose of this study was to develop an effective method for expansion and activation of the effector cells with high proportion of NK cells and increasing cytotoxicity against liver cancer in a short time period. METHODS: Expanded NK cell-enriched lymphocytes (NKL) designated as "MYJ1633" were prepared by using autologous human plasma, cytokines (IL-2, IL-12 and IL-18) and agonistic antibodies (CD16, CD56 and NKp46) without an NK cell-sorting step. The characteristics of NKL were compared to those of freshly isolated PBMCs. In addition, the cytotoxic effect of the NKL on liver cancer cell was examined in vitro and in vivo. RESULTS: The total cell number after ex vivo-expansion increased about 140-fold compared to that of freshly isolated PBMC within 2 weeks. Approximately 78% of the expanded and activated NKL using the house-developed protocol was NK cell and NKT cells even without a NK cell-sorting step. In addition, the expanded and activated NKL demonstrated potent cytotoxicity against liver cancer in vitro and in vivo. CONCLUSION: The house-developed method can be a new and effective strategy to prepare clinically applicable NKL for autologous NK cell-based anti-tumor immunotherapy.

Halloysite Nanocapsules Containing Thyme Essential Oil: Preparation, Characterization, and Application in Packaging Materials.

Halloysite nanotubes (HNTs), which are natural nanomaterials, have a hollow tubular structure with about 15 nm inner and 50 nm outer diameters. Because of their tubular shape, HNTs loaded with various materials have been investigated as functional nanocapsules. In this study, thyme essential oil (TO) was encapsulated successfully in HNTs using vacuum pulling methods, followed by end-capping or a layer-by-layer surface coating process for complete encapsulation. Nanocapsules loaded with TO were mixed with flexographic ink and coated on a paper for applications as food packaging materials. Scanning electron microscopy and transmission electron microscopy were used to characterize the morphology of the nanocapsules and to confirm the TO loading of the nanocapsules. Fourier transform infrared spectroscopy and thermogravimetric analyses analysis were used to complement the structural information. In addition, the controlled release of TO from the nanocapsules showed sustained release properties over a period of many days. The results reveal that the release properties of TO in these nanocapsules could be controlled by surface modifications such as end-capping and/or surface coating of bare nanocapsules. The packaging paper with TO-loaded HNT capsules was effective in eliminating against Escherichia coli during the first 5 d and showed strong antibacterial activity for about 10 d.

Microwave-assisted step reduced extraction of seaweed (Gelidiella aceroso) cellulose nanocrystals.

In the present work, cellulose nanocrystals were isolated from seaweed by microwave-assisted alkali treatment, bleaching, and an acid hydrolysis process. Microwave-assisted alkali treatment reduces the heating time and eliminates the traditional dewaxing process. This is different from the commonly adopted procedure for cellulose nanocrystal (CNC) synthesis, in which CNC synthesis generally follows the dewaxing process. Further, samples obtained after each stage of treatment were characterized and final samples were freeze-dried for further characterization. TEM results revealed that isolated CNCs had a 32nm average diameter and an average length of 408nm. FTIR and XRD data showed that after each stage of chemical treatment, no cellulosic components were removed. The adopted method is faster than the previous traditional method used for isolation of CNCs from seaweed fibers.

Leakage assessment of flexible pouches using dye penetration test with correlation to modeled bacterial aerosol challenge test.

Package integrity is a primary measure of a package's ability to keep the contained product inside and to keep potential contaminants out. In this study, injecting and vacuum dye penetration methods were applied for the assessment of the package integrity of retortable flexible pouches having various sizes of micro-channels. The purpose of this study is to evaluate the usefulness of dye penetration as a physical test that can be incorporated into a stability protocol and compare the results of the dye penetration test with those from the bacterial aerosol challenge test. The study found a direct correlation between the results of the vacuum dye penetration test and those of the microbial test. The critical leak size that can ensure the flexible package integrity was 15 µm. To detect defective pouches, the dye vacuum testing had a sensitivity similar to that of bioaerosol challenge test.

Application of chitosan-incorporated LDPE film to sliced fresh red meats for shelf life extension.

Chitosan lactate was impregnated as an antimicrobial additive into low density polyethylene (LDPE) with different concentrations. The antimicrobial effectiveness was tested with three pathogenic bacteria, specifically Listeria monocytogenes, Escherichia coli and Salmonella enteritidis. Also, these chitosan incorporated films were applied on red meat surfaces to determine the effectiveness of chitosan on color shelf life extension and microbial growth inhibition. Chitosan was exposed to 0.1% peptone water containing the three pathogens in separate tests and inhibited microbial growth a higher levels with increasing concentration of chitosan in the film matrix. Oxygen permeability was not affected by the incorporation of chitosan, while the water vapor permeability increased with the addition of chitosan. Film elongation decreased with the addition of chitosan. When chitosan incorporated films were applied on fresh red meat, microorganisms on the meat surface were not inhibited but significant extension of red color shelf life were observed in refrigerated, sliced red meats.

Utilization of monolinolein as a substrate for conjugated linoleic acid production by Bifidobacterium breve LMC 520 of human neonatal origin.

This study was designed to isolate bifidobacteria from human intestines that efficiently converts monolinolein, a monoglyceride form of linoleic acid, into conjugated linoleic acid (CLA), as well as to optimize culture conditions for improving CLA production during milk fermentation. Among 150 screened neonatal bifidobacteria, Bifidobacterium breve LMC 520 showed the highest CLA-producing ability and was tested with different types of fat substrates at various concentrations to determine the optimal conditions for CLA production. Monolinolein was tested as a substrate for CLA production. The incubation time optimized for CLA production was 24 h, and CLA production was proportionally increased with monolinolein concentration. The incubation of LMC 520 with commercial starter strains caused minimal reduction in CLA production. Our results demonstrate that the CLA-producing ability of B. breve LMC 520 could offer beneficial effects when utilized as a starter culture for the development of functional dairy products.

Rectal water absorption in seawater-adapted Japanese eel Anguilla japonica.

Marine teleosts drink large amounts of seawater to compensate for continuous osmotic water loss. We investigated a possible significant role of the rectum in water absorption in seawater-adapted eel. In rectal sacs filled with balanced salt solution (BSS) and incubated in isotonic BSS, water absorption was greater in seawater-adapted eel than in freshwater eel. Since rectal fluid osmolality was slightly lower than plasma osmolality in seawater-adapted eel, effects of rectal fluid osmolality on water absorption were examined in rectal sacs filled with artificial rectal fluid with different osmolality. Rectal water absorption was greater at lower rectal fluid osmolality, suggesting that an osmotic gradient between the blood and rectal fluid drives the water movement. Ouabain, a specific inhibitor of Na+/K(+)-ATPase, inhibited water absorption in rectal sacs, indicating that an osmotic gradient favorable to rectal water absorption was created by ion uptake driven by Na+/K(+)-ATPase. Expression levels of aquaporin 1 (AQP1), a water-selective channel, were significantly higher in the rectum than in the anterior and posterior intestines. Immunoreaction for Na+/K(+)-ATPase was detected in the mucosal epithelial cells in the rectum with more intense staining in the basal half than in the apical half, whereas AQP1 was located in the apical membrane of Na+/K(+)-ATPase-immunoreactive epithelial cells. The rectum is spatially separated from the posterior intestine by a valve structure and from the anus by a sphincter. Such structures allow the rectum to swell as intestinal fluid flows into it, and a concomitant increase in hydrostatic pressure may provide an additional force for rectal water absorption. Our findings indicate that the rectum contributes greatly to high efficiency of intestinal water absorption by simultaneous absorption of ions and water.

Human vitamin E requirements assessed with the use of apples fortified with deuterium-labeled alpha-tocopheryl acetate.

BACKGROUND: Little is known about factors that modulate dietary alpha-tocopherol bioavailability. OBJECTIVES: The study aimed to assess the efficacy of vitamin E-fortified apples as a low-fat vitamin E delivery system, the influence of fat on vitamin E absorption, and human vitamin E requirements by using plasma alpha-tocopherol kinetics at a dosage of alpha-tocopherol found in food. DESIGN: Apples fortified with deuterium-labeled alpha-tocopheryl acetate were consumed by 5 participants at a breakfast containing 0%, 6%, or 21% kcal from fat in 3 sequential trials. The trials were separated by a 2-wk washout period. Blood samples were obtained up to 72 h, and plasma was analyzed for labeled and unlabeled alpha-tocopherol. RESULTS: Compared with observations in the 0% fat trial, the maximum observed plasma d6-alpha-tocopherol concentrations (Cmax) and the areas under the curve increased 2- and 3-fold during the 6% and 21% fat trials, respectively. The mean (+/-SD) estimated percentage d6-alpha-tocopherol absorbed increased from 10 +/- 4% during the 0% fat trial to 20 +/- 3% and 33 +/- 5% during the 6% and 21% fat trials, respectively. The mean time to Cmax (9 +/- 2 h), fractional disappearance rates (0.022 +/- 0.003 pools/d), and half-lives (32 +/- 4 h) did not differ significantly between the trials. With the use of fractional disappearance rates and baseline plasma alpha-tocopherol concentrations, the estimated daily plasma alpha-tocopherol efflux was 13-14 mg. The estimated rate of alpha-tocopherol delivery to tissues was 5 mg/d. CONCLUSIONS: Given an estimated 33% absorption, the amount of dietary vitamin E needed daily to replace irreversible losses is

Incorporation of a high concentration of mineral or vitamin into chitosan-based films.

Mineral or vitamin E was incorporated into chitosan-based films: 10-200% (w/w chitosan) Gluconal Cal (GC), a mixture of calcium gluconate and lactate; 5-20% zinc lactate (ZL); and 5-20% alpha-tocopheryl acetate (VE) with acetylated monoglyceride (AM). The functionality of film-forming solutions and dried films was analyzed with standard procedures, and mathematical equations were developed to coordinate selected film functionality with the type and concentration of incorporated mineral or vitamin E. GC incorporation significantly increased pH and decreased viscosity of film-forming solutions, but not the addition of ZL or VE. The water barrier property of the films was improved by increasing the concentration of mineral or vitamin E in the film matrix. The tensile strength of the films was more significantly affected by GC or VE addition than film elongation, puncture strength, and puncture deformation. While a major endothermic peak around 200 degrees C was observed in DSC thermograms of chitosan-based films, only 200% GC incorporation altered this endothermic peak. This study demonstrated the capability of chitosan-based film matrix to carry a high concentration of mineral or vitamin E. Such films may be used for wrapping or coating to enhance the nutritional value of foods.