Influence of Protein Adsorption on Aggregation in Prefilled Syringes.

Protein aggregate formation in prefilled syringes (PFSs) can be influenced by protein adsorption and desorption at the solid-liquid interface. Although inhibition of protein adsorption on the PFS surface can lead to a decrease in the amount of aggregation, the mechanism underlying protein adsorption-mediated aggregation in PFSs is unclear. This study investigated protein aggregation caused by protein adsorption on silicone oil-free PFS surfaces [borosilicate glass (GLS) and cycloolefin polymer (COP)] and the factors affecting the protein adsorption on the PFS surfaces. The adsorbed proteins formed multilayered structures that consisted of two distinct types of layers: proteins adsorbed on the surface of the material and proteins adsorbed on top of the proteins on the surface. A pH-dependent electrostatic interaction was the dominant force for protein adsorption on the GLS surface, while hydrophobic effects were dominant for protein adsorption on the COP surface. When the repulsion force between proteins was weak, protein adsorption on the adsorbed protein layer was increased for both materials and as a result, protein aggregation increased. Therefore, a formulation with high colloidal stability can minimize protein adsorption on the COP surface, leading to reduced protein aggregation.

Culturing Chinese hamster ovary cells on cyclo olefin polymer triggers epithelial-mesenchymal transition and spheroid formation, which increases the foreign gene expression driven by the Moloney murine leukemia virus long terminal repeat promoter.

Chinese hamster ovary (CHO) cells are frequently used for recombinant protein production (RPP) as a host. While the RPP has been proven successful, there is still a compelling need for further improvement. Cyclo olefin polymer (COP) is a plastic material widely utilized due to its properties including its low protein absorption. We applied this as a raw material for RPP cell culture to see if the COP is suitable. A recombinant CHO cell line expressing the human erythropoietin (hEPO) gene under the control of the Moloney murine leukemia virus-long terminal repeat (MMLV-LTR) was established. When the cells were cultured in a dish made from COP, the cells attached to the bottom, and then started to float and form spheroids. RNASeq data analysis suggested the epithelial-mesenchymal transition (EMT) was triggered with receptor tyrosine kinase activation shortly after cultivation. It coincided with the hEPO transcription increase. After the cell floating, though EMT marker gene expression subsided, a hEPO expression increase sustained. When fibronectin was applied to COP dish surface, the cell floating was suppressed and hEPO expression decreased. We then treated cells with MßCD, a drug that destroys the lipid raft, eliminating molecules in the raft. This facilitated cell floating and spheroid formation coincided with hEPO expression enhancement. These results suggest interactions between a cell and COP surface might trigger the EMT and the subsequent event, both of which activated the MMLV-LTR promoter. Thus, employing COP for culturing cells, a potent RPP system could be established with its advantage for efficient protein purification.

Association between coffee and green tea intake and pneumonia among the Japanese elderly: a case-control study.

A large prospective cohort study in the United States examined the association between coffee intake and overall and cause-specific mortality and showed a inverse association between pneumonia and influenza deaths and coffee intake. In Japan, the mortality rate of pneumonia in elderly people is high, and its prevention is an important issue. The present study investigated the association between coffee and green tea intake and pneumonia among the elderly. The design was a hospital-based case control study. The cases were patients over 65 years old newly diagnosed as pneumonia. As a control, patients with the same sex and age (range of 5 years) who visited the same medical institution around the same time (within 2 months after examination of the case) for a disease other than pneumonia were selected. There were two controls per case. Odds ratio (OR) and 95% confidence interval (CI) for pneumonia of coffee and green tea intake during the past month were calculated using a conditional logistic regression model. A total of 199 cases and 374 controls were enrolled. When compared to those who do not drink coffee, the OR for pneumonia of those who drink less than one cup of coffee per day was 0.69 (95% CI 0.39-1.21), OR of those who drink one cup was 0.67 (0.38-1.18), and OR of those who drink two or more cups was 0.50 (0.28-0.88) (Trend p = 0.024). No association was found between pneumonia and green tea consumption. This study suggested a preventive association between coffee intake over 2 cups per day and pneumonia in the elderly.

Identification of black microplastics using long-wavelength infrared hyperspectral imaging with imaging-type two-dimensional Fourier spectroscopy.

Despite recent progress in focal plane array Fourier transform infrared spectroscopy (FPA-FT-IR) for automatic microplastic (MP) discrimination, the analysis time is still too long (e.g., 9 h for a sample with a diameter of 47 mm) and the equipment is expensive. As a solution, a hyperspectral camera restricted to the near-infrared or short-wavelength infrared band could be applied. However, with these bands, the minimum discriminable size is only about 100 µm, and discrimination among darkly colored plastics is difficult. The long-wavelength infrared (LWIR) band is reportedly effective for discrimination among darkly colored plastics. In this study, we constructed a palm-sized LWIR hyperspectral camera (105 mm × 90 mm × 50 mm, 1.25 kg) for imaging-type two-dimensional Fourier spectroscopy. Our system used a general-purpose, inexpensive, and compact microbolometer for the LWIR band. This system could record the absorbance of black MPs (polystyrene, polyethylene, and polypropylene) in a 3.8 mm × 3.0 mm area in 36 s, which was less than 1/6th of the time required for FPA-FT-IR. Additionally, our system could obtain spectra for a 12 µm × 12 µm area. Because our device is cheaper and more compact than a FPA-FT-IR, it will be easier to take out in the field or on a research vessel.

Optical coherent transients in 167Er3+ at telecom-band wavelength.

We demonstrate optical coherent transients in a Λ-like hyperfine energy-level system of Er1673+ in yttrium orthosilicate (Y2SiO5) with telecom-band photons at a zero magnetic field. Spectral hole burning was used to study the temperature dependence of the induced spectral antihole. We find that temperatures below 3.0 K suppress population dissipation induced by electron-phonon interactions sufficiently to enable population initialization in the Λ-like system. Further, the pulse area dependence of photoluminescence (PL) from the Λ-like system was measured at 2.2 K. An optical pump power dependence of PL intensity shows Rabi oscillations that contain two full Rabi cycles at the frequency of 2π×810 kHz. A two-pulse photon echo measurement reveals an optical coherence time of 12 µs. To date, this measured optical coherence time is the longest observed for Er3+ in solids at zero magnetic field. These findings will facilitate optical coherent manipulation of Λ-like Er1673+ electronic states as a quantum memories operating at telecom-band wavelengths.

Practical determination of LODP (limit of detection for microarray platform) for the evaluation of microarray platforms.

The performance indicator called limit of detection for microarray platform (LODP) was defined in ISO 16578:2013. The methods to determine practical LODP were explored. In general, + 3 SD of the background is used as the signal strength of limit of detection and criteria for dividing positive and negative results. Since the negative signal had been defined differently for each microarray platform, signals obtained from Non-Probe Spots (NPS) installed on the microarrays were defined as the "background" of microarrays. LODP was determined as the lowest concentration of which the average signal exceeded Avg. + 3 SD of the background (NPS) and the signal was significantly different from those of the lower and higher adjacent concentration points measured with a diluted series of reference materials. For reliable qualitative analysis, the positive results can be defined as signals higher than those corresponding to LODP and negative results as lower signals, without determining limit of detection for all target probes. The use of LODP also enables comparisons of platform performances without checking sequence dependencies, and assists to select reliable and fitting platforms for experimental purposes.

Control of Protein Adsorption to Cyclo Olefin Polymer by the Hofmeister Effect.

Cyclo olefin polymer (COP) is an attractive plastic because it has low protein adsorption despite its hydrophobic chemical structure. Here, the adsorption of model proteins to the COP was evaluated in comparison with a representative plastic, polystyrene (PSt), using reflectometry interference spectroscopy (RIfS) technology. The effects of different salts on adsorption were then examined. The adsorption of bovine serum albumin onto COP increased in the presence of kosmotropic salts, whereas adsorption of IgG increased in the presence of chaotropic salts. By contrast, the adsorption of these 2 proteins to PSt was unaffected by these Hofmeister salts. Langmuir-Freundlich model of COP adsorption suggested that the COP surface is more homogeneous for protein binding than the PSt surface. Furthermore, RIfS and sum frequency generation analyses indicated that water molecules bind more weakly to COP than to PSt. Our data propose a novel viewpoint of the way protein binds to COP surface that is different from the way it binds to PSt.

Sweeping of Adsorbed Therapeutic Protein on Prefillable Syringes Promotes Micron Aggregate Generation.

This study evaluated how differences in the surface properties of prefillable syringe barrels and in-solution sampling methods affect micron aggregates and protein adsorption levels. Three syringe types (glass barrel with silicone oil coating [GLS/SO+], glass barrel without silicone oil coating [GLS/SO-], and cyclo-olefin polymer [COP] barrel syringes) were tested with 3 therapeutic proteins (adalimumab, etanercept, and infliximab) using 2 sampling methods (aspiration or ejection). After quiescent incubation, solutions sampled by aspiration exhibited no significant change in micron aggregate concentration in any syringes, whereas those sampled by ejection exhibited increased micron aggregates in both GLS syringe types. Micron aggregate concentration in ejected solutions generally increased with increasing density of adsorbed proteins. Notably, COP syringes contained the lowest micron aggregate concentrations, which were independent of the sampling method. Correspondingly, the adsorbed protein density on COP syringes was the lowest at 1-2 mg/m2, which was much less compared with that on GLS syringes and was calculated to be equivalent to only 1-2 protein layers, as visually confirmed by high-speed atomic force microscopy. These data indicate that low-adsorption prefillable syringes should be used for therapeutic proteins because protein aggregate concentration in the ejected solution is elevated by increased protein adsorption to the syringe surface.

Signatures of Γ1-Γ5 mixed-mode polaritons in polarized reflectance spectra of ZnO.

Theoretical and experimental studies were carried out on exciton-polaritons excited in ZnO. Polaritons in which both Γ(1) and Γ(5) excitons couple to electromagnetic waves simultaneously are shown to exist, and their signatures are observed in polarized reflectance spectra measured under k is perpendicular to a and E is parallel to c configurations for an m-plane sample. Theoretical calculations reveal that the mixed-mode polaritons consist of one Γ(1) transverse mode and two Γ(5) longitudinal modes. It is also shown that the signatures are sensitive to the valence band ordering.

Development of a compact magnetic resonance imaging system for a cold room.

A compact magnetic resonance imaging (MRI) system for a cold (-5 degrees C) room has been developed to acquire MR images below the freezing point of water. The MRI system consists of a 1.0 T permanent magnet, a higher-order shim coil set, and a gradient coil probe, installed in the cold room, and a compact MRI console installed in a room at normal temperature (20-25 degrees C). The most difficult problem for the installation of the MRI system in the cold room was the degradation of the field homogeneity of the permanent magnet shimmed at 25 degrees C. To overcome this problem, higher-order shim coils were developed and the temperature variation of the magnetic field distribution was measured using a standard phantom with and without shim coil currents. As a result, it was confirmed that the homogeneity (the difference between the minimum and maximum values) of the magnetic field in the 17x17x19 mm(3) rectangular parallelepiped region was improved from 117 to 59 ppm using an appropriate combination of shim coil currents. A snowpack immersed in dodecane (C(12)H(26)) was imaged using a driven-equilibrium three-dimensional (3D) spin-echo sequence at -5 degrees C. The visualized 3D structure of the snowpack demonstrated the effectiveness of our approach.

[The prophylaxis of wound infection in pull-percutaneous endoscopic gastrostomy: Correlation with methicillin resistant staphylococcus aureus positive patient in the throat].

In this study, we elucidated the efficacy of our prophylactic method for wound infection in pull-percutaneous endoscopic gastrostomy (PEG). The total 29 patients received the pull-PEG. The first 8 patients received the oral sterilization with povidone iodine and antibiotics at the time of pull-PEG (Group-I). The frequency of wound infection in this group was 50.0% (4/8). It was revealed that all infections were induced by methicillin resistant staphylococcus aureus (MRSA). 3 patients were MRSA positive in the throat. In Group-II, we eradicated MRSA in the throat before the pull-PEG by combination mupirocin calcium hydrate with the Group-I treatment. In contrast in Group-I, the frequency of wound infection was significantly reduced in Group-II (4.8%: 1/21). The results showed that our eradication method was very useful for prevention of the wound infection in pull-PEG treatment.