JNC Corporation

There are few reports of the adoption of continuous processes in bioproduction, particularly the implementation of end‐to‐end continuous or integrated processes, due to difficulties such as feed adjustment and incorporating virus filtration. Here, we propose an end‐to‐end integrated continuous process for a monoclonal antibody (mAb) with three integrated process segments: upstream production processes with pool‐less direct connection, pooled low pH virus inactivation with pH control and a total flow‐through integrated polishing process in which two columns were directly connected with a virus filter. The pooled virus inactivation step defines the batch, and high impurities reduction and mAb recovery were achieved for batches conducted in succession. Viral clearance tests also confirmed robust virus reduction for the flow‐through two‐column chromatography and the virus filtration steps. Additionally, viral clearance tests with two different hollow fiber virus filters operated at flux ranging from 1.5 to 40 LMH (liters per effective surface area of filter in square meters per hour) confirmed robust virus reduction over these ranges. Complete clearance with virus logarithmic reduction value ≥4 was achieved even with a process pause at the lowest flux. The end‐to‐end integrated continuous process proposed in this study is amenable to production processes, and the investigated virus filters have excellent applicability to continuous processes conducted at constant flux.

Objective: ε-Poly-l-lysine (PLL) is a cationic polymer consisting of 25-35 l-lysine residues. Our previous study revealed that fluorescently labelled PLL can stain the stratum corneum (SC) via ionic interactions between PLL and SC constituents. In this study, to further clarify the mechanisms underlying the interaction between PLL and the SC, the staining properties of fluorescent PLL were compared with that of fluorescently labelled anionic dextran (aDex), which has approximately the same molecular weight as PLL. Methods: SC samples were collected by non-invasive tape stripping and stained with fluorescent PLL and/or fluorescent aDex. Fluorescence images were acquired using a fluorescence microscope and then analysed. Results: The SC could be stained with either fluorescent PLL or aDex, both of which were inhibited by the addition of high concentrations of salt solutions. In particular, aDex staining was inhibited at a lower salt concentration than PLL staining. Moreover, PLL staining was inhibited under acidic conditions, while aDex staining was inhibited under neutral to alkaline conditions. Double staining of SC with both fluorescent polymers produced heterogeneous staining patterns: corneocytes stained with both polymers, corneocytes stained with PLL or aDex in a mutually exclusive manner, and unstained corneocytes. Staining of SC samples from the face was more extensive than staining of SC samples from the inside of the upper arm with both polymers. In addition, pretreatment of the SC with ethanol resulted in enhanced staining with both polymers. These results suggest that double staining of SC with both polymers can provide information on the damaged SC. Conclusion: Staining of SC with fluorescent PLL depends on its properties of a cationic and hydrophobic polymer with appropriate molecular size, which can distinguish the damaged SC. Double staining of SC with fluorescent PLL and aDex is a novel approach to obtain information for the analysis of skin conditions.

Background ε‐Poly‐L‐lysine (PLL) is a cationic polymer consisting of 25 to 35 L‐lysine residues that adheres to the surface of skin as well as hair. However, the properties of PLL regarding its adhesion to the skin remain to be elucidated. In this study, we examined the staining of stratum corneum (SC) with fluorescence‐labeled PLL and explored its relationship with skin condition. Materials and methods Alexa Fluor 488‐labeled PLL (AF‐PLL) was reacted with tape‐stripped stratum corneum (SC), and the staining properties were monitored by fluorescence microscopy. Clinical study was performed by measuring the water content of the cheek SC and transepidermal water loss (TEWL), and the tape‐stripped SC was subjected to staining with AF‐PLL. Results AF‐PLL staining of the SC was inhibited at acidic pH or by the addition of high concentration of salt solution, suggesting the involvement of ionic interaction between PLL and the SC, at least in part. The AF‐PLL staining was inhibited by unlabeled PLL or various alkyl amines, but not by L‐lysine monomer. AF‐PLL staining was observed inside the corneocytes as well as surrounding cornified envelope. Clinical study revealed that AF‐PLL staining intensity of the SC was negatively correlated with its water content and positively correlated with its TEWL. Conclusion PLL can efficiently adhere to SC and AF‐PLL staining of SC can be applied to evaluate skin conditions.

In this study, ruthenium‐on‐carbon‐catalyzed hydrogenation of CBDK was developed for the production of cis‐CBDO under solvent‐free conditions. Isomerization using a ruthenium catalyst and hydrogen gas may improve the selective formation of cis‐CBDO. Moreover, the reaction mechanisms for cis‐selectivity was proposed based on density functional theory (DFT) calculations.

Herein, we reported an ultrapure blue multiple‐resonance‐induced thermally activated delayed fluorescence (MR‐TADF) material (ν‐DABNA‐O‐Me) with a high photoluminescence quantum yield and a large rate constant for reverse intersystem crossing. Because of restricted π‐conjugation of the HOMO rather than the LUMO induced by oxygen atom incorporation, ν‐DABNA‐O‐Me shows a hypsochromic shift compared to the parent MR‐TADF material (ν‐DABNA). An organic light‐emitting diode based on this material exhibits an emission at 465 nm, with a small full‐width at half‐maximum of 23 nm and Commission Internationale de l'Eclairage coordinates of (0.13, 0.10), and a high maximum external quantum efficiency of 29.5 %. Moreover, ν‐DABNA‐O‐Me facilitates a drastically improved efficiency roll‐off and a device lifetime compared to ν‐DABNA, which demonstrates significant potential of the oxygen atom incorporation strategy.

Polyethyleneimine (PEI) possesses various molecular weights (MWs), structures, and virus capture capacities. However, whether PEI can capture porcine circovirus (PCV) and animal cell-derived prion protein (PrPC) that may contaminate source materials is unclear. Therefore, we conducted a feasibility study to assess the effectiveness of PEI in removing PCV and PrPC as a model of pathogenic prions. The removal performance of PCV was evaluated by quantitative PCR using PEIs with various MWs, structures, and ion exchange capacities in Tris (pH 7.5) and acetate (pH 5.5) buffers under neutral (pH 7.5) to acidic (pH 5.5) conditions. Removal performances of PrPC were also evaluated by western blotting using PEIs with various MWs and structures. Tris buffer did not affect the ability of PEI-modified resins to remove PCV, whereas acetate buffer affected removal performances, except those of PEI-10K-Br and PEI-70K-Br, which showed high ion-exchange capacities. PrPC was captured by PEIs with high MWs, especially PEI-70K-Br, which was the most effective. The results of this feasibility study suggested that PEI-modified resin could remove PCV and PrPC. PEI-70K-Br with an ion-exchange capacity of at least 0.3 meq/mL appears suitable as a PEI molecule for pathogen capture or removal of PCV or PrPC from biological materials.

Carbazole‐based DABNA analogues (CzDABNAs) were synthesized from triarylamine by regioselective one‐shot single and double borylation. The reaction proceeded selectively at the ortho position of the carbazolyl group, where the highest occupied molecular orbital is mainly localized owing to the difference in the electron‐donating abilities of the diarylamino and carbazolyl groups. The facile and scalable method enabled synthesis of CzDABNAs, exhibiting narrowband thermally activated delayed fluorescence with emission spectra ranging from deep blue to green. The organic light‐emitting diode devices employing these products as emitters exhibited deep‐blue, sky‐blue, and green emission with high external quantum efficiencies of 19.5, 21.8, and 26.7 %, respectively.

Carbazole‐based DABNA analogues (CzDABNAs) were synthesized from triarylamine by regioselective one‐shot single and double borylation. The facile and scalable method enabled synthesis of CzDABNAs exhibiting narrowband thermally activated delayed fluorescence and the preparation of highly efficient organic light‐emitting diode devices showing deep‐blue, sky‐blue, and green emission. Abstract Carbazole‐based DABNA analogues (CzDABNAs) were synthesized from triarylamine by regioselective one‐shot single and double borylation. The reaction proceeded selectively at the ortho position of the carbazolyl group, where the highest occupied molecular orbital is mainly localized owing to the difference in the electron‐donating abilities of the diarylamino and carbazolyl groups. The facile and scalable method enabled synthesis of CzDABNAs, exhibiting narrowband thermally activated delayed fluorescence with emission spectra ranging from deep blue to green. The organic light‐emitting diode devices employing these products as emitters exhibited deep‐blue, sky‐blue, and green emission with high external quantum efficiencies of 19.5, 21.8, and 26.7 %, respectively.

Palladium catalysts immobilized on cellulose particles (Pd/CLP) and on a cellulose‐monolith (Pd/CLM) were developed. These composites were applied as hydrogenation catalysts and their catalyst activities were evaluated. Although both catalysts catalyzed the deprotection of benzyloxycarbonyl‐protected aromatic amines (Ar‐N‐Cbz) and aromatic benzyl esters (Ar‐CO2Bn), only Pd/CLM could accomplish the hydrogenolysis of aliphatic‐N‐Cbz and aliphatic‐CO2Bn protective groups. The difference in the physical structure of the cellulose supports induced unique chemoselectivity. Aliphatic‐N‐Cbz and aliphatic‐CO2Bn groups were tolerated under the Pd/CLP‐catalyzed hydrogenation conditions, while Ar‐N‐Cbz, Ar‐CO2Bn, alkene, alkyne, azido and nitro groups could be smoothly reduced.

The commercially available 3 types of selective media in Japan were compared for the detection of Bacillus cereus. When assessed inclusivity using 25 B. cereus strains, MYP agar, NGKG agar, and chromogenic X-BC agar demonstrated excellent inclusivity. For exclusivity study using 50 non-B. cereus strains, MYP, NGKG, and X-BC allowed to grow 11, 7, and 3 strains, respectively. Of the grown bacteria on each strains tested, only 2 strains of B. thuringiensis formed typical B. cereus colonies on all selective media tested. The NGKG and X-BC were compared with MYP as a reference using artificially contaminated food (fried rice, plain rice, fried noodle, and potato salad ), since MYP is recommended in ISO 7932: 2004. The both correlation coefficients between NGKG and MYP, and X-BC and MYP were 0.999. Therefore, we demonstrated that NGKG and X-BC can be adapted to ISO 7932: 2004 method for selected food as well as MYP.

Luminescent materials that exhibit narrowband emission are vital for full-colour displays. Here, we report a thermally activated delayed-fluorescence material that exhibits ultrapure blue emission with full-width at half-maximum of just 14 nm. The emitter consists of five benzene rings connected by two boron and four nitrogen atoms and two diphenylamino substituents. The multiple resonance effect of the boron and nitrogen atoms induces significant localization of the highest occupied and lowest unoccupied molecular orbitals on different atoms to minimize not only the vibronic coupling between the ground state (S0) and the singlet excited state (S1) but also the energy gap between the S1 state and triplet excited state (T1). Organic light-emitting diode devices employing the emitter emit light at 469 nm with full-width at half-maximum of 18 nm with an external quantum efficiency of 34.4% at the maximum and 26.0% at 1,000 cd m⁻².

In the luminous ostracod Cypridina (presently Vargula) hilgendorfii, Cypridina luciferyl sulfate (3‐enol sulfate of Cypridina luciferin) is converted to Cypridina luciferin by a sulfotransferase with 3′‐phosphoadenosine‐5′‐phosphate (PAP) as a sulfate acceptor. The resultant Cypridina luciferin is used for the luciferase–luciferin reaction of Cypridina to emit blue light. The luminescence stimulation with major organic cofactors was examined using the crude extracts of Cypridina specimens, and we found that the addition of coenzyme A (CoA) to the crude extracts significantly stimulated luminescence intensity. Further, the light‐emitting source in the crude extracts stimulated with CoA was identified as Cypridina luciferyl sulfate, and we demonstrated that CoA could act as a sulfate acceptor from Cypridina luciferyl sulfate. In addition, the sulfate group of Cypridina luciferyl sulfate was also transferred to adenosine 5′‐monophosphate (5′‐AMP) and adenosine 3′‐monophosphate (3′‐AMP) by a sulfotransferase. The sulfated products corresponding to CoA, 5′‐AMP, and 3′‐AMP were identified using mass spectrometry. This is the first report that CoA can act as a sulfate acceptor in a sulfotransferase reaction. This article is protected by copyright. All rights reserved.

MC-Media PadTM EB (MMP-EB) , a novel sheet culture method for the enumeration of Enterobacteriaceae, has been evaluated. When both inclusivity and exclusivity of MMP-EB were assessed using 104 microbes including 51 Enterobacteriaceae strains, all tested Enterobacteriaceae strains grew and formed obvious red-colored colonies and all tested non-Enterobacteriaceae strains were shown different appearance from Enterobacteriaceae strains. For the comparison study of the method, MMP-EB was compared with violet red bile glucose agar (VRBG) according to ISO 21528-2:2017 and PetrifilmTM Enterobacteriaceae Count Plate (Petrifilm EB) method using 100 naturally contaminated food samples. The correlation coefficients between MMP-EB and VRBG, and MMP-EB and Petrifilm EB were 0.940 and 0.972, respectively. Furthermore, there were no statistically significant difference between MMP-EB and both reference methods. Our results demonstrated that MMP-EB was a suitable alternative method for the enumeration of Enterobacteriaceae in food samples.

BD mCCDA Clear-HT (CCHT; Nippon Becton Dickinson Company, Ltd.) , a novel chromogenic selective medium was evaluated for its superior capacity to isolate Campylobacter jejuni/ coli. When CCHT was assessed using 142 microbes including 42 Campylobacter jejuni/ coli strains, all Campylobacter strains were found to form purple-colored colonies on CCHT whereas all the other microbes failed to grow. CCTH was then compared with commercially available selective media using 100 stool samples including 40 Campylobacter positive samples. CCHT detected Campylobacter jejuni/ coli from 39 of 40 (97.5%) stool samples whereas it allowed competitive bacteria to grow as false positive colonies from 1 (1.0%) of 100 stool samples. The values of relative sensitivity (%) and specificity (%) for CCHT were 97.5 and 98.3 in this study. Our results demonstrated that CCHT had the highest detection ratio for Campylobacter jejuni/ coli and the highest inhibition ratio against competitive bacteria among all selective media compared.

Introduction of an amino group at the para position of doubly sterically-hindered aryl azides significantly enhances the reactivity with cyclooctynes. The distortability of the azido group is synergistically enhanced by...

Degranulation refers to the secretion of inflammatory mediators, such as histamine, serotonin, and proteases, that are stored within the granules of mast cells and that trigger allergic reactions. The amount of these released mediators has been measured biochemically using cell mass. To investigate degranulation in living single cells, fluorescence microscopy has traditionally been used to observe the disappearance of granules and the appearance of these discharged granules within the plasma membrane by membrane fusion and the movement of granules inside the cells. Here, we developed a method of video-rate bioluminescence imaging to directly detect degranulation from a single mast cell by measuring luminescence activity derived from the enzymatic reaction between Gaussia luciferase (GLase) and its substrate coelenterazine. The neuropeptide Y (NPY), which was reported to colocalize with serotonin in the secretory granules, fused to GLase (NPY-GLase) was efficiently expressed in rat basophilic leukemia (RBL-2H3) cells, a mast-cell line, using a preferred human codon-optimized gene. Bioluminescence imaging analysis of RBL-2H3 cells expressing NPY-GLase and adhered on a glass-bottomed dish showed that the luminescence signals from the resting cells were negligible, while the luminescence signals of the secreted NPY-GLase were repeatedly detected after the addition of an antigen. In addition, this imaging method was applicable for observing degranulation in RBL-2H3 cells that adhered to the extracellular matrix (ECM). These results indicated that video-rate bioluminescence imaging using GLase will be a useful tool for detecting degranulation in single mast cells adhered to a variety of ECM proteins.

The four types of chromogenic selective media that are commercially available in Japan were compared for establishing a Japanese standard method for detecting Cronobacter spp. based on ISO/TS 22964:2006. When assessed using 9 standard Cronobacter spp. strains and 29 non-Cronobacter strains, Enterobacter sakazakii isolation agar, ChromocultTMEnterobacter sakazakii agar, CHROMagarTME. sakazakii, and XM-sakazakii agar demonstrated excellent inclusivity and exclusivity. Using the ISO/TS 22964:2006 method, the recovered numbers of 38 Cronobacter spp. strains, including 29 C. sakazakii isolates obtained from each medium, were equivalent, indicating that there was no significant difference (p > 0.05) among the four types of chromogenic selective media. Thus, we demonstrated that these four chromogenic selective media are suitable alternatives when using the standard method for detecting Cronobacter spp. in Japan, based on the ISO/TS 22964:2006.

A calamitic-liquid-crystal compound with 1,3-dioxane and fluorinated aryl units exhibits a highly polar and fluid mesophase (MP phase) in which anomalously large dielectric permittivity (>10 000), ferroelectric-like polarization switching, and second-harmonic generation are observed by Hiroya Nishikawa, Hirotsugu Kikuchi, and co-workers in article number 1702354. The experimental data clearly shows that a unidirectional ferroelectric-like polar order is generated along the director, although there is no layer structure.

A novel azobenzene compound for isomerization type photoalignment film has been developed. It has 4-aminophenethyl groups at both sides of azobenzene core. Photoalignment film (PAF) containing it shows high transmittance and almost the same alignment-ability compared with our conventional isomerization type PAF.