Effect of catechins and high-temperature-processed green tea extract on scavenging reactive oxygen species and preventing Aß1-42 fibrils' formation in brain microvascular endothelium.

The present study investigated the effect of high-temperature-processed green tea extract (HTP_GTE) and its bioactive components on the reduction of reactive oxygen species (ROS) and amyloid-beta (Aß) protein in human microvascular endothelial cells. Compared to Aß1-42-only treatment, pretreatment of HTP_GTE was revealed to effectively inhibit ROS generation (P<0.05). HTP_GTE and catechins not only inhibit Aß1-42 fibril formation but also destabilize preformed Aß1-42 fibrils. The presence of HTP_GTE, Aß1-42 fibril formation was significantly inhibited in a dose-dependent manner at 12.5-100 µg/ml of HTP_GTE, showing 86-56%, respectively. Treatment of various concentrations of HTP_GTE and catechins steadily destabilized the preformed Aß1-42 fibrils for 24 h in a dose-dependent manner. It was observed that the gallated groups such as epigallocatechin gallate, epicatechin gallate, gallocatechin gallate, and catechin gallate more effectively disturbed Aß1-42 fibril formation and destabilized the preformed Aß1-42 fibrils than the non-gallated group. Taken together, these findings supported that sterilized green tea could be promising natural anti-amyloidogenic agents associated with therapeutic approaches in Alzheimer's disease by scavenging ROS generation and Aß fibril in the brain tissue.

Impact of commercial cigarette smoke condensate on brain tissue co-cultured with astrocytes and blood-brain barrier endothelial cells.

The purpose of the current study was to investigate the effect of two commercial cigarette smoke condensates (CCSC) on oxidative stress and cell cytotoxicity in human brain (T98G) or astrocytes (U-373 MG) in the presence of human brain microvascular endothelial cells (HBMEC). Cell viability of mono-culture of T98G or U-373 MG was markedly decreased in a concentration-dependent manner, and T98G was more susceptible than U-373 MG to CCSC exposure. Cytotoxicity was less prominent when T98G was co-cultured with HBMEC than when T98G was co-cultured with U-373 MG. Significant reduction in trans-epithelial electric resistance (TEER), a biomarker of cellular integrity was noted in HBMEC co-cultured with T98G (HBMEC-T98G co-culture) and U-373 MG co-cultured with T98G (U-373 MG-T98G co-culture) after 24 or 48 hr CCSC exposure, respectively. TEER value of U-373 MG co-cultured with T98G (79-84%) was higher than HBMEC co-cultured with T98G (62-63%) within 120-hr incubation with CCSC. Reactive oxygen species (ROS) generated by CCSC in mono-culture of T98G and U-373 MG reached highest levels at 4 and 16 mg/ml, respectively. ROS production by T98G fell when co-cultured with HBMEC or U-373MG. These findings suggest that adverse consequences of CCSC treatment on brain cells may be protected by blood-brain barrier or astrocytes, but with chronic exposure toxicity may be worsened due to destruction of cellular integrity.

The effect of tobacco smoke exposure on the generation of reactive oxygen species and cellular membrane damage using co-culture model of blood brain barrier with astrocytes.

Brain tissue is known to be vulnerable to the exposure by tobacco smoke. Tobacco smoke can induce generation of reactive oxygen species (ROS), causing inflammatory activity and blood-brain barrier (BBB) impairment. The aim of the present study was to investigate the effect of tobacco smoke on cell cytotoxicity, generation of ROS, and cellular membrane damage in astrocytes and BBB using a co-culture system. Cell viability of U373MG cells was reduced in a dose-dependent manner, ranging from 96.7% to 40.3% by tobacco smoke condensate (TSC). Cell viability of U373MG co-cultured with human brain microvascular endothelial cells (HBMECs) was 104.9% at the IC50 value of TSC. Trans-epithelial electric resistance values drastically decreased 80% following 12-h incubation. The value was maintained until 48 h and then increased at 72-h incubation (85%). It then decreased to 75% at 120 h. Generation of ROS increased in a dose-dependent manner, ranging from 102.7% to 107.9%, when various concentrations of TSC (4-16 mg/mL) were administered to the U373MG monoculture. When TSC was added into U373MG co-cultured with HBMECs, production of ROS ranged from 101.7% to 102.6%, slightly increasing over 12 h. Maximum exposure-generated ROS of 104.8% was reached at 24 h. Cell cytotoxicity and oxidative stress levels in the U373MG co-culture model system with HBMECs were lower than U373MG monoculture. HBMECs effectively acted as a barrier to protect the astrocytes (U373MG) from toxicity of TSC.

Pertussis Toxin Exploits Specific Host Cell Signaling Pathways for Promoting Invasion and Translocation of Escherichia coli K1 RS218 in Human Brain-derived Microvascular Endothelial Cells.

Pertussis toxin (PTx), an AB5 toxin and major virulence factor of the whooping cough-causing pathogen Bordetella pertussis, has been shown to affect the blood-brain barrier. Dysfunction of the blood-brain barrier may facilitate penetration of bacterial pathogens into the brain, such as Escherichia coli K1 (RS218). In this study, we investigated the influence of PTx on blood-brain barrier permissiveness to E. coli infection using human brain-derived endothelial HBMEC and TY10 cells as in vitro models. Our results indicate that PTx acts at several key points of host cell intracellular signaling pathways, which are also affected by E. coli K1 RS218 infection. Application of PTx increased the expression of the pathogen binding receptor gp96. Further, we found an activation of STAT3 and of the small GTPase Rac1, which have been described as being essential for bacterial invasion involving host cell actin cytoskeleton rearrangements at the bacterial entry site. In addition, we showed that PTx induces a remarkable relocation of VE-cadherin and ß-catenin from intercellular junctions. The observed changes in host cell signaling molecules were accompanied by differences in intracellular calcium levels, which might act as a second messenger system for PTx. In summary, PTx not only facilitates invasion of E. coli K1 RS218 by activating essential signaling cascades; it also affects intercellular barriers to increase paracellular translocation.

Growth Performance of Early Finishing Gilts as Affected by Different Net Energy Concentrations in Diets.

The objectives of the current experiment were to study the response of the growth performance of early finishing gilts to different net energy (NE) concentrations in diets, and to compare the NE values of diets between calculated NE values and measured NE values using French and Dutch CVB (Centraal Veevoederbureau; Central Bureau for Livestock Feeding) NE systems. In a metabolism trail, the NE concentrations in five diets used for the growth trial were determined based on digestible nutrient concentrations, digestible energy, and metabolizable energy using a replicated 5×5 Latin square design with 10 barrows (initial body weight [BW], 39.2±2.2 kg). In a growth trial, a total of 60 early finishing gilts (Landrace×Yorkshire; initial BW, 47.7±3.5 kg) were allotted to five dietary treatments of 8.0, 9.0, 10.0, 11.0, and 12.0 MJ NE/kg (calculated, as-is basis) with 12 replicate pens and one pig per pen in a 42-d feeding experiment. The NE and amino acid (AA) concentrations in all diets were calculated based on the values from NRC (2012). Ratios between standardized ileal digestible AA and NE concentrations in all diets were closely maintained. Pigs were allowed ad libitum access to feed and water. Results indicated that calculated NE concentrations in diets (i.e., five dietary treatments) were close to measured NE concentrations using French NE system in diets. The final BW was increased (linear and quadratic, p<0.05) with increasing NE concentrations in diets. Furthermore, average daily gain (ADG) was increased (linear and quadratic, p<0.01) with increasing NE concentrations in diets. There was a quadratic relationship (p<0.01) between average daily feed intake and NE concentrations in diets. Feed efficiency (G:F) was also increased (linear, p<0.01) as NE concentrations in diets were increased. The NE intake per BW gain (kcal NE/kg of BWG) was increased (linear, p<0.01) with increasing NE concentrations in diets that were predicted from both French and Dutch CVB NE systems. Linear regression indicated that predictability of daily NE intake from the BW of pigs was very low for both French (R(2), 0.366) and Dutch CVB (R(2), 0.374) NE systems. In conclusion, increasing NE concentrations in diets increase BW, ADG, G:F, and NE intake per BW gain of early finishing gilts. The BW of early finishing gilts is not a good sole variable for the prediction of daily NE intake.

Oxidative Stress Induced by Cigarette Smoke Extracts in Human Brain Cells (T98G) and Human Brain Microvascular Endothelial Cells (HBMEC) in Mono- and Co-Culture.

The objective of the current study was to examine oxidative stress induced by cigarette smoke extract (CSE) or cigarette smoke condensate (CSC) in human brain cells (T98G) and human brain microvascular endothelial cells (HBMEC) in mono- and co-culture systems. Cell viability of T98G cells exposed to CSC (0.05-4 mg/ml) was significantly decreased compared to CSE (0.025-20%). There were no marked differences between quantities of reactive oxygen species (ROS) generation by either CSE (2, 4, and 10%) or CSC (0.2, 0.4, and 0.8 mg/ml) treatment compared to control. However, a significant effect was noted in ROS generation following CSC incubation at 4mg/ml. Cellular integrity of HBMEC decreased to 74 and 64% within 120 h of exposure at the IC50 value of CSE and CSC, respectively. This study suggests that chronic exposure to cigarette smoking might initiate damage to the blood-brain barrier.

Numerical study on electronic and optical properties of organic light emitting diodes.

In this paper, we present a finite element method (FEM) study of space charge effects in organic light emitting diodes. Our model includes a Gaussian density of states to account for the energetic disorder in organic semiconductors and the Fermi-Dirac statistics to account for the charge hopping process between uncorrelated sites. The physical model cover all the key physical processes in OLEDs, namely charge injection, transport and recombination, exciton diffusion, transfer and decay as well as light coupling, and thin-film-optics. The exciton model includes generation, diffusion, and energy transfer as well as annihilation. We assumed that the light emission originates from oscillating and thus embodied as excitons and embedded in a stack of multilayer. The out-coupled emission spectrum has been numerically calculated as a function of viewing angle, polarization, and dipole orientation. We discuss the accumulation of charges at internal interfaces and their signature in the transient response as well as the electric field distribution.

Breakdown characteristics of high-side lateral double-diffused metal oxide semiconductor devices.

In this paper, we discuss the breakdown characteristics of an LDMOSFET (Lateral Double-diffused Metal Oxide Semiconductor Field Effect Transistor) structure for the optimization of engineering parameters such as the gap between the DEEP N-WELL and the source region and the doping concentration of the N(ADJUST)-layer. The fabricated device exhibits the breakdown voltage (BVdss) over 110 V and the specific on-resistance as low as 2.20 mOmega cm2. The robust breakdown characteristics seem to be due to the migration of the location wherein maximum impact ionization occurs from the gate region to the drain side, which was confirmed by the numerical simulation which verifies that the maximum impact ionization rate of the proposed structure is 2.44 x 10(16) cm(-3) s(-1), while that of the conventional structure being 6.69 x 10(19) cm(-3) s(-1).

Finite element method simulation of the molding process for thermal nano-imprint lithography.

We made a numerical study on the deformation of a viscoelastic polymethyl methacrylene (PMMA) resist when a rigid SiO2 stamp with a rectangular line pattern is imprinted into the PMMA resist for thermal nano-imprint lithography (NIL). The stress distribution in the polymer resist during the molding process is calculated by a finite element method (FEM). Our simulation results reveal that the asymmetric von Mises stress is distributed over the polymer around the external line, which seems to be due to the squeezing flow under the flat space. The stress seems to be concentrated at the sidewall close to the centerline of the whole structure. Our simulation also reveals that a micro gap is formed between the replicated structure and the outer wall of the mold.

Escherichia coli outer membrane protein A adheres to human brain microvascular endothelial cells.

Escherichia coli K1 is the most common gram-negative bacterium causing neonatal meningitis. The outer membrane protein A (OmpA) assembles a beta-barrel structure having four surface-exposed loops in E. coli outer membrane. OmpA of meningitis-causing E. coli K1 is shown to contribute to invasion of the human brain microvascular endothelial cells (HBMEC), the main cellular component of the blood-brain barrier (BBB). However, the direct evidence of OmpA protein interacting with HBMEC is not clear. In this study, we showed that OmpA protein, solubilized from the outer membrane of E. coli, adhered to HBMEC surface. To verify OmpA interaction with the HBMEC, we purified N-terminal membrane-anchoring beta-barrel domain of OmpA and all surface-exposed loops deleted OmpA proteins, and showed that the surface-exposed loops of OmpA were responsible for adherence to HBMEC. These findings indicate that the OmpA is the adhesion molecule with HBMEC and the surface-exposed loops of OmpA are the determinant of this interaction.