Polymer gels as artificial soft tissue

Soft tissue consists a main body of biological systems. It is made of hydrogels mainly composed of extra cellular matrix (ECM), which is the three-dimensional highly organized and inter-woven network of proteins and polysaccharides. ECM has three symbolic roles. They are pertinent and excellent mechanical performance, extremely low friction, and effective and selective transportation of solute molecules for cell viability. One of the important goals for gel science would be to design and create hydrogels with critical parameters, which natural tissues possess and eventually to replace them with artificial one.     

Optimization of biohydrogen production by Clostridium butyricum EB6 from palm oil mill effluent using response surface methodology

Clostridium butyricum EB6 successfully produced hydrogen gas from palm oil mill effluent (POME). In this study, central composite design and response surface methodology were applied to determine the optimum conditions for hydrogen production (P_c) and maximum hydrogen production rate (R_max) from POME. Experimental results showed that the pH, temperature and chemical oxygen demand (COD) of POME affected both the hydrogen production and production rate, both individually and interactively. The optimum conditions for hydrogen production (P_c) were pH 5.69, 36 °C, and 92 g COD/l; with an estimated P_c value of 306 ml H₂/g carbohydrate. The optimum conditions for maximum hydrogen production rate (R_max) were pH 6.52, 41 °C and 60 g COD/l; with an estimated R_max value of 914 ml H₂/h. An overlay study was performed to obtain an overall model optimization. The optimized conditions for the overall model were pH 6.05, 36 °C and 94 g COD/l. The hydrogen content in the biogas produced ranged from 60% to 75%.     

Mutation Analysis of Radioresistant Early-Stage Cervical Cancer

Radiotherapy is a definitive treatment for early-stage cervical cancer; however, a subset of this disease recurs locally, necessitating establishment of predictive biomarkers and treatment strategies. To address this issue, we performed gene panel-based sequencing of 18 stage IB cervical cancers treated with definitive radiotherapy, including two cases of local recurrence, followed by in vitro and in silico analyses. Simultaneous mutations in KRAS and SMAD4 (KRAS^mt/SMAD4^mt) were detected only in a local recurrence case, indicating potential association of this mutation signature with radioresistance. In isogenic cell-based experiments, a combination of activating KRAS mutation and SMAD4 deficiency led to X-ray resistance, whereas either of these factors alone did not. Analysis of genomic data from 55,308 cancers showed a significant trend toward co-occurrence of mutations in KRAS and SMAD4. Gene Set Enrichment Analysis of the Cancer Cell Line Encyclopedia dataset suggested upregulation of the pathways involved in epithelial mesenchymal transition and inflammatory responses in KRAS^mt/SMAD4^mt cancer cells. Notably, irradiation with therapeutic carbon ions led to robust killing of X-ray-resistant KRAS^mt/SMAD4^mt cancer cells. These data indicate that the KRAS^mt/SMAD4^mt signature is a potential predictor of radioresistance, and that carbon ion radiotherapy is a potential option to treat early-stage cervical cancers with the KRAS^mt/SMAD4^mt signature.     

Polyunsaturated fatty glyceride syntheses by microbial lipases

The degree of glyceride syntheses by lipase TOYO (Chromobacterium viscosum) and lipase OF (Candida cylindracea) using individual free fatty acids C_18∶1, C_18∶2, C_18∶3, C_18∶4, C_20∶4, C_20∶5 and C_22∶6 were compared. Lipase TOYO incorporated each of the fatty acids into glycerol at levels of greater than 89%. Lipase OF incorporated most of the fatty acids at levels above 70% (docosahexaenoic acid incorporation was 63%). It was concluded that these two lipases are feasible for producing glycerides from unsaturated fatty acids.     

Fluorescent in situ hybridization analysis of open lactic acid fermentation of kitchen refuse using rRNA-targeted oligonucleotide probes

Reproducible amounts of lactic acid accumulate in minced kitchen refuse under open conditions with intermittent pH neutralization [Sakai et al., Food Sci. Technol. Res., 6, 140 (2000)]. Here, we showed that such pH-controlled open fermentation of kitchen refuse reproducibly resulted a selective proliferation of a major lactic acid bacterial (LAB) species. In one experiment, the predominant microorganisms isolated during the early phase (6 h) were Gammaproteobacteria. In contrast, those that predominated during the late phase (48 h) were always Lactobacillus plantarum in three independent experiments. To further quantify the microbial community within open lactic acid fermentation, we performed fluorescent in situ hybridization (FISH) analysis targeting 16S (23S) rRNA. We designed two new group-specific DNA probes: LAC722(L) was active for most LAB including the genera Lactobacillus, Pediococcus, Leuconostoc and Weisella, whereas Lplan477 was specific for L. plantarum and its related species. We then optimized sample preparation using lysozyme and hybridization conditions including temperature, as well as the formamide concentration and the salt concentration in the washing buffer. We succeeded in quantification of microorganisms in semi-solid, complex biological materials such as minced kitchen refuse by taking color microphotographs in modified RGB balance on pre-coated slides. FISH analysis of the fermentation of kitchen refuse indicated that control of the pH swing leads to domination by the LAB population in minced kitchen refuse under open conditions. We also confirmed that L. plantarum, which generates lactic acid in high quantities but with low optical activity, became the dominant microorganism in kitchen refuse during the late phase of open fermentation.     

Terephthalic acid synthesis at higher concentrations in high‐temperature liquid water. 2. Eliminating undesired byproducts

We synthesized terephthalic acid (TPA) from p‐xylene at an initial concentration above its solubility limit in high‐temperature liquid water (HTW). The nominal p‐xylene loading at the reaction conditions was 0.4 mol L^?1, which is the highest reported to date for generation of high TPA yields (>70 mol %) in HTW. The presence of two liquid phases during the reaction did not appear to accelerate the rate, unlike behavior reported for some other organic reactions done “on water” at lower temperatures. Adding oxygen gas in a large increment during synthesis produced a black liquid and a black solid byproduct, which is a previously undocumented problem. Adding oxygen in smaller increments prevented formation of the liquid and solid byproducts and also provided high selectivities (90 mol %) and yields (>70 mol %) of TPA. These results demonstrate the feasibility of HTW as a medium for TPA synthesis at p‐xylene concentrations even higher than its solubility limit. © 2009 American Institute of Chemical Engineers AIChE J, 2009     

Quantification of cell adhesion force with AFM: distribution of vitronectin receptors on a living MC3T3-E1 cell

Distribution of vitronectin (VN) receptors on a living murine osteoblastic cell was successfully measured by atomic force microscopy (AFM). First, the distribution of the integrin beta(5) subunit which constitutes a part of the VN receptor on the cell was confirmed by conventional immunohistochemistry after fixing the cell. To visualize the distribution of the receptor on a living cell by an independent and potentially a more quantitative method, the AFM was used with a microbead attached to the cantilever tip to increase the area of contact and VN was immobilized on the microbead. Force measurements were then performed over a large area of a living murine osteoblastic cell using the microbead covered with VN.     

Photoluminescence enhancement by isolating β-FeSi2 layers from defective layers

We have investigated an optimal annealing process in order to enhance 1.55 μm light emission from semiconducting β-FeSi₂ and found that two steps annealing at 600 °C and 800 °C is effective to its enhancement. Rutherford backscattering spectroscopy and SEM observations revealed that pronounced surface segregation of Fe atoms during annealing at 600 °C caused surface precipitate of β-FeSi₂. The enhancement of light emission is attributed spatial isolation of the surface β-FeSi₂ (light emitting layer) from damaged and defective layers with nonradiative recombination centers.     

Intra-molecular mobility of holes along rod-like helical Si backbones in optically active polysilanes

Intra-molecular mobility of positive charge carriers in poly(n-hexyl-[S]-2-methylbutylsilane) (PHMBS) films doped with N,N′-bis(2,5-di-tert-buthylphenyl)-3,4,9,10-perylenedicarboximide (PDI) is studied by time-resolved microwave conductivity (TRMC) measurements. PHMBS with optically active side chains has a rod-like tightly locked Si catenation with unlikely long persistence length compared with conventional dialkyl-substituted polysilanes. It was found that PDI is a suitable electron acceptor for PHMBS as it provides the high product of photo carrier generation yield ? as ～0.08% under an excitation at 355 nm. The efficient electron transfer reaction from PHMBS to PDI gives clear conductivity transients ascribed to the positive charges on the Si catenation of PHMBS by TRMC measurements. The estimated value of isotropic mobility along the tightly locked Si catenation is 0.36 cm² V^?1 s^?1, suggesting the presence of mobile holes on the backbones of PHMBS.