Effect of a hepatocyte growth factor/heparin-immobilized collagen system on albumin synthesis and spheroid formation by hepatocytes

A hepatocyte growth factor (HGF)/heparin-immobilized collagen system was used as a synthetic extracellular matrix for hepatocyte culture. The albumin synthesis, nucleus numbers and morphology of the hepatocytes were determined separately to evaluate the hepatocyte number and hepatocyte-specific function under this system. The benefits of the HGF/heparin-immobilized collagen system for hepatocyte culture were confirmed by three types of culture methods in vitro, namely 2D film cultures, 2D gel cultures and 3D gel cultures. In 2D collagen film cultures, hepatocytes exhibited the highest albumin synthesis (1.42 μg/well/day) in HGF/heparin-immobilized collagen films at 7 days of culture. Heparin inhibited hepatocyte adhesion while HGF promoted hepatocyte migration, and spheroid formation was easily detected in HGF/heparin-immobilized collagen films. In 2D collagen gel cultures, albumin synthesis of around 15 μg/well/day was detected and maintained for more than 18 days on HGF/heparin-immobilized collagen gels. Similar findings were obtained in 3D HGF/heparin-immobilized collagen gel cultures, which exhibited albumin synthesis of up to 30 μg/well/day. The albumin synthesis by hepatocytes was two-fold higher in 3D gel cultures compared with 2D gel cultures, and was maintained for over 2 weeks compared with 2D film cultures using the HGF/heparin-immobilized collagen system. Taken together, the HGF/heparin-immobilized collagen system was effective for albumin synthesis by hepatocytes in both 2D film cultures and 3D gel cultures, and therefore shows good potential for tissue engineering use.     

Measurement of particle concentration in powder coating process using capacitance computed tomography and wavelet analysis

Capacitance computed tomography techniques were used to visualize particles movement in the draft tube of a spouted fluidized bed for the coating process of drug production. A total of 512 frames images of the particle concentration distribution were obtained at 10-millisecond intervals over a coating time of 5 min using a capacitance computed tomography system. The three-dimensional capacitance CT images (time and two-dimensional space images) were decomposed to wavelet time and space levels to extract the dominant particle distribution feature using three-dimensional discrete wavelet multiresolution at different coating times. As a result, the time and space dominant particle distribution with a specific frequency level can be visualized.     

The Chemistry of PCR Primers: Concept and Application

DNA is a typical organic compound with marked differences from other chemicals and biopolymers because DNA can be amplified by the enzyme polymerase. DNA can be, in principle, amplified from a single copy by the polymerase chain reaction (PCR). In this review, we focus our attention on the chemistry of PCR primers. Because PCR is basic technology in biology research fields, we sometimes use chemically labeled primers without any awareness of the chemistry they leave behind. We would like to emphasize that chemically labeled primers contain a lot of potential for different chemistry ideas and much study is still necessary to advance PCR for single-nucleotide polymorphism (SNP) typing, genetic diagnosis, and other fields. Two categories of primers, affinity-capture primers and signaling primers, are discussed from the viewpoints of their chemical concepts and applications. Affinity-capture primers are used for purification, isolation, and manipulation of PCR products by high specificity and affinity to the cognate molecules by molecule molecule interactions, whereas signaling primers report the hybridization and/or progress of PCR amplification by a signal change, in most cases by a fluorescence change. The content of this review may be useful for a better understanding of the chemistry of PCR primers and, more importantly, for the invention of novel PCR chemistry.     

Severity of delayed (“secondary”) cerebral energy failure after acute hypoxia-ischemia is related to the time integral of acute ATP depletion

The aim of this study was to reproduce the delayed (secondary) cerebral energy failure previously described in birth-asphyxiated newborn infants and to investigate relationships between primary insult severity and the extent of the delayed energy failure. Phosphorus (³¹P) magnetic resonance spectroscopy (MRS) at 7 T was used to study the brains of 12 newborn piglets during an acute, reversible, cerebral hypoxic-ischemic episode which continued until nucleotide triphosphates (NTP) were depleted. After reperfusion and reoxygenation, spectroscopy was continued for 48 h. High-energy metabolite concentrations returned to near normal levels after the insult, but later they fell as delayed energy failure developed. The time integral of NTP depletion in the primary insult correlated strongly with the minimum [phosphocreatine (PCr)]/[inorganic orthophosphate (P_i)] observed 24–48 h after the insult. (Linear regression analysis gave slope –8.04 h^–1; ordinate intercept=1.23;r=0.92;P<0.0001.) This model is currently being used to investigate the therapeutic potential of various cerebroprotective strategies including hypothermia.     

Profiling and Imaging of Phospholipids in Brains of Abcd1‐Deficient Mice

ABCD1 is a gene responsible for X‐linked adrenoleukodystrophy (X‐ALD), and is critical for the transport of very long‐chain fatty acids (VLCFA) into peroxisomes and subsequent β‐oxidation. VLCFA‐containing lipids accumulate in X‐ALD patients, although the effect of ABCD1‐deficiency on each lipid species in the central nervous system has not been fully characterized. In this study, each phospholipid and lysophospholipid species in Abcd1‐deficient mice brains were profiled by liquid chromatography‐mass spectrometry. Among the phospholipid and lysophospholipid species that are significantly more enriched in Abcd1‐deficient mice brains, VLCFA were present in 75, 15, 5, 4, and 1 species of phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, lysophosphatidylcholine, and lysophosphatidylethanolamine, respectively. Most VLCFA were incorporated at the sn‐1 position of phosphatidylcholine and phosphatidylethanolamine. Among the phospholipid species that are significantly less enriched in Abcd1‐deficient mice brains, odd‐numbered saturated or mono‐unsaturated fatty acyl moieties are contained in all phosphatidylcholine species. In addition, a number of phosphatidylglycerol, phosphatidylinositol, and phosphatidylserine species contained highly unsaturated fatty acyl moieties. Intriguingly, 44:1 phosphatidylcholine with VLCFA was mainly distributed in the gray matter, such as the cortex, but not in the white matter in the cerebrum and cerebellum. These results show that ABCD1‐deficiency causes metabolic alternation of long‐chain fatty acids and VLCFA. Moreover, our results imply a molecular mechanism for the incorporation of saturated or monounsaturated VLCFA into the sn‐1 position of phospholipids, and also indicate that the distribution of phospholipids with VLCFA may correlate with the development of X‐ALD.     

Simulation of the vapor explosion of tin and aluminum oxide fragments in water

The vapor explosion process involves an explosive phase change from a thin liquid film into a vapor with a dramatic change in material properties across a high-pressure shock wave. The energy released during the explosion process can reach a level of chemical explosions associated with detonating high explosives. Because no fuel-air chemical reaction is needed for initiating a vapor explosion, the process can be considered as a green reaction that generates zero pollutants. However, the phase change that accompanies a thousand-fold increase in the vapor volume during the rapid film evaporation is a natural consequence of a sudden change in density across the phase front; this feature closely resembles the process of blast wave generation. The high-pressure vapor produced without a chemical reaction expands as a spherical wave at a uniform velocity predicted through the Rankine-Hugoniot shock conditions. In this paper, a new methodology based on a simple shock wave and a phase field theory is proposed to formulate the vapor explosion process.     

Multifunctional Initiator with Platinum-Acetylide Dendritic Core for Living Polymerization of Isocyanides

A novel multifunctional initiator has been developed by peripheral modification of a Pt-acetylide dendrimer. Pd–Pt μ-ethynediyl units were introduced to the first-generation dendrimer by a divergent method. The dendritic initiator was applicable to the living polymerization of aryl isocyanides, producing high molecular weight polymers with a narrow polydispersity index in quantitative yields. The ³¹P NMR spectrum of the resulting polymer suggested that all Pd–Pt μ-ethynediyl units in the dendritic molecule actually functioned as an initiator.     

Preparation of acetamiprid-loaded polymeric microcapsules: Influence of preparation parameter in emulsion system on microcapsule characteristics

Microencapsulation of pesticides is a promising technique for avoiding high initial doses and multiple applications of the chemicals to agricultural land which cause environmental pollution. It is because the formulation is possible to improve the stability of the chemicals against environmental degradation and control the release rate. In the present study, polymeric microcapsules prepared by the solvent evaporation method via water-in-oil-in-water (W/O/W) emulsion were used as immobilization supports of acetamiprid, a water-soluble pesticide. The pesticide was loaded in the microcapsules by impregnating the polymeric supports with acetamiprid dissolved in an organic solvent. Increased volume ratio (ϕ) of inner aqueous phase to oil phase in the emulsion system contributed to more pores in the microcapsules and increase in the content of acetamiprid in the polymeric supports. Release rate of acetamiprid from the microcapsules could be controlled by changing ϕ.     

Pore Structure and Thermal Stability of Mesoporous Mullite Fibers Prepared by Crystallization and Selective Leaching of Al2O3-SiO2 Glass Fibers

Porous mullite fibers were prepared by crystallization and selective leaching of Al2O3-SiO2 glass fibers using buffered HF-NH4F(BHF) aqueous solutions. The optimum concentration of BHF solution for selective leaching of the fibers was 0.9 mass% HF and 17.0 mass% NH4F. By firing at 1000–1300°C, the glass fibers changed into composite texture of mullite and glassy phase. Since the pores in the fibers were formed by selective leaching of glassy phase among mullite grains, they were tunable by changing the firing conditions of fibers. Pore size of the samples changed from around 4 nm in the 1100°C fired sample to 16 and 40 nm in the 1200 and 1300°C fired samples, respectively. The highest specific surface area obtained was around 30 m2/g, when the fibers were heat treated at 1200°C for 24 h and leached for 20 h in 0.9 mass% HF-17.0 mass% NH4F solution. From the thermal stability tests of the porous mullite fibers, its specific surface area was found to be maintained up to 1200–1300°C.