Induced albumin secretion from HepG2 spheroids prepared using poly(ethylene glycol) derivative with oleyl groups

We developed a poly(ethylene glycol) (PEG) derivative with oleyl groups, so-called “cell adhesive”, for the promotion of human hepatocellular carcinoma HepG2 cell spheroids. Our approach was based on crosslinking of the cell membrane with a cell adhesive via a hydrophobic interaction. A cell adhesive, PEG derivative with hydrophobic oleyl groups at both ends was synthesized and characterized. HepG2 spheroids formed when the adhesive was added to cell suspensions. The size of the spheroids increased with time in culture. In addition, Ammonia elimination of HepG2 spheroid with cell adhesive was 3.4 times higher than that without cell adhesive. Furthermore, albumin secretion from HeG2 spheroids grown with the cell adhesive for 7 days was 3.3 times that from HepG2 spheroids grown without cell adhesive. Fluorescence microscopy showed greater albumin staining in spheroids grown with cell adhesive compared with spheroids grown without adhesive. This cell adhesive may be useful not only for single type of cells but also for multi types of cells to form artificial organs. This cell adhesive will be a key material for liver tissue engineering when it will apply to primary hepatocytes.     

Microfluidic device with chemical gradient for single-cell cytotoxicity assays

Here, we report the fabrication of a chemical gradient microfluidic device for single-cell cytotoxicity assays. This device consists of a microfluidic chemical gradient generator and a microcavity array that enables entrapment of cells with high efficiency at 88 ± 6% of the loaded cells. A 2-fold logarithmic chemical gradient generator that is capable of generating a serial 2-fold gradient was designed and then integrated with the microcavity array. High density single-cell entrapment was demonstrated in the device without cell damage, which was performed in 30 s. Finally, we validated the feasibility of this device to perform cytotoxicity assays by exposing cells to potassium cyanide (0-100 μM KCN). The device captured images of 4000 single cells affected by 6 concentrations of KCN and determined cell viability by counting the effected cells. Image scanning of the microcavity array was completed within 10 min using a 10× objective lens and a motorized stage. Aligning cells on the microcavity array eases cell counting, observation, imaging, and evaluation of singular cells. Thus, this platform was able to determine the cytotoxicity of chemicals at a single-cell level, as well as trace the cytotoxicity over time. This device and method will be useful for cytotoxicity analysis and basic biomedical research.     

Recovery of Phenol through the Decomposition of Tar under Hydrothermal Alkaline Conditions

Decomposition of the tar residue from oil distillation was carried out under hydrothermal conditions using a batch reactor at 623–673 K and 25–40 MPa, with and without K₂CO₃ as a catalyst. The reaction scheme for tar decomposition was determined as follows: the liquefaction and dissolution process of tar occur first and then intermediate chemical compounds are transformed into lighter molecular weight species. The presence of K₂CO₃ activates the dissociation of molecular hydrogen to facilitate hydrogenation reactions. The main products from the decomposition of tar were phenol, biphenyl, diphenylether (DPE), and diphenylmethane (DPM). These results indicate that hydrolysis was important in the cleavage of the macromolecular structure of tar under both catalytic and non‐catalytic hydrothermal conditions. This method can be developed for efficient tar liquefaction to generate high yields of valuable chemicals in an environmentally friendly way.     

The Ets-1 and Ets-2 transcription factors activate the promoters for invasion-associated urokinase and collagenase genes in response to epidermal growth factor

Transgenic 3.L2 T cells are stimulated by Hb(64-76)/I-Ek and are positively selected on I-Ek plus self-peptides. To this pool of self-peptides we have added a single, well-defined 3.L2 TCR antagonist (A72) in vivo. We find that mice expressing both the 3.L2 TCR and A72 have a minimal loss of T cells expressing the clonotypic TCR in the thymus and spleen. Importantly, the proliferative response of 3.L2 x A72 splenocytes is significantly reduced compared with splenocytes from 3.L2 mice. This reduced response can be attributed to peripheral antagonism. Thus we have identified a new class of self-ligands whose predominant effect is constitutive peripheral antagonism rather than negative selection. The net effect of these ligands is to avoid potential self-reactivity while maintaining as large a repertoire as possible.     

Aberrant gastric venous drainage into the medial segment: demonstration by color Doppler sonography

BACKGROUND: Aberrant gastric venous drainage (AGVD) into the posterior edge of the medial segment of the liver (segment IV) is the main cause of pseudolesion on computed tomography (CT) during arterial portography. We estimated the prevalence of AGVD into the medial segment of the liver with color and power Doppler ultrasound (US). METHODS: Screening gray-scale and color Doppler and power Doppler US were performed in 100 consecutive patients. AGVD was defined as a venous structure that ascended parallel to the main portal vein and drained independently into segment IV. RESULTS: AGVDs were observed in eight of 100 patients (8%) with color and power Doppler US. Power Doppler US depicted these veins more clearly than did color Doppler US. Gray-scale US did not show any AGVDs. Two of eight patients with AGVDs detected by color Doppler US underwent celiac arteriography and CT during arterial portography (CTAP). In these two patients, celiac arteriography directly demonstrated AGVDs draining into segment IV, which revealed nontumorous perfusion defects (pseudolesions) on CTAP. CONCLUSION: Color and power Doppler US are useful imaging methods for demonstrating AGVDs.     

Refined crystal structure of the seryl-tRNA synthetase from Thermus thermophilus at 2.5 A resolution

The three-dimensional structure of the seryl-tRNA synthetase from Thermus thermophilus has been determined and refined at 2.5 A resolution. The final model consists of a dimer of 421 residues each and 190 water molecules. The R-factor is 18.4% for all the data between 10 and 2.5 A resolution. The structure is very similar to that of the homologous enzyme from Escherichia coli, with an r.m.s. difference of 1.5 A for the 357 alpha-carbon atoms considered equivalent. The comparison of the two structures indicates increased hydrophobicity, reduced conformational entropy and reduced torsional strain as possible mechanisms by which thermostability is obtained in the enzyme from the thermophile.     

Multifunctional Hydrophobized Microparticles for Accelerated Wound Healing after Endoscopic Submucosal Dissection

Endoscopic submucosal dissection (ESD) provides strong therapeutic benefits for early gastrointestinal cancer as a minimally invasive treatment. However, there is currently no reliable treatment to prevent scar contracture resulting from ESD which may lead to cicatricial stricture. Herein, a multifunctional colloidal wound dressing to promote tissue regeneration after ESD is demonstrated. This sprayable wound dressing, composed of hydrophobized microparticles, exhibits the multifunctionality necessary for wound healing including tissue adhesiveness, blood coagulation, re‐epithelialization, angiogenesis, and controlled inflammation based on hydrophobic interaction with biological systems. An in vivo feasibility study using swine gastric ESD models reveals that this colloidal wound dressing suppresses fibrosis and accelerates wound healing. Multifunctional colloidal and sprayable wound dressings have an enormous therapeutic potential for use in a wide range of biomedical applications including accelerated wound healing after ESD, prevention of perforation, and the treatment of inflammatory diseases.     

Mechanical Reinforcement of Low‐Concentration Alginate Fibers by Microfluidic Embedding of Multiple Cores

This paper presents mechanically reinforced low‐concentration alginate fibers by embedding inner cores of high‐concentration alginate. 3D structures by stacking multiple polydimethylsiloxane (PDMS) layers allow the microfluidic formation and control of the isolated cores in the continuous flow. The alginate hydrogel fibers are simply spun, and the compartments, central core, surrounding cores, and outer shell layer are successfully verified. The results demonstrate the great potential for the development of complex fibrous materials, particularly for biological applications, which require specific morphology and composition of the fibers.